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Wang QS, Shen SQ, Sun HW, Xing ZX, Yang HL. Interferon-gamma induces autophagy-associated apoptosis through induction of cPLA2-dependent mitochondrial ROS generation in colorectal cancer cells. Biochem Biophys Res Commun 2018; 498:1058-1065. [PMID: 29551681 DOI: 10.1016/j.bbrc.2018.03.118] [Citation(s) in RCA: 26] [Impact Index Per Article: 3.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/09/2018] [Accepted: 03/14/2018] [Indexed: 12/12/2022]
Abstract
Colorectal cancer (CRC) is the second most commonly diagnosed cancer in females and the third in males. In this work, we aim to investigate the possible anti-cancer effects of interferon-gamma (IFN-γ) in CRC cells. We observed that IFN-γ induced mitochondria-derived reactive oxygen species (ROS) production in a time-dependent manner in SW480 and HCT116 cell lines. The IFN-γ-induced mitochondrial ROS generation was dependent on the activation of cytosolic phospholipase A2 (cPLA2). In addition, a mitochondria-targeted antioxidant SS31 and/or cPLA2 inhibitor AACOCF3 abolished the IFN-γ-induced ROS production and subsequent autophagy and apoptosis. Moreover, suppression of autophagy by CQ was able to reduce IFN-γ-induced cell apoptosis. Beclin-1 gene silencing resulted in caspase-3 inactivation, decreased Bax/Bcl-2 ratio and less population of apoptotic cells. Collectively, our results suggested that IFN-γ induces autophagy-associated apoptosis in CRC cells via inducing cPLA2-dependent mitochondrial ROS production.
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Affiliation(s)
- Qiu-Shuang Wang
- Department of General Surgery, Renmin Hospital of Wuhan University, 99 Zhangzhidong Road, Wuhan, Hubei, 430060, PR China
| | - Shi-Qiang Shen
- Department of General Surgery, Renmin Hospital of Wuhan University, 99 Zhangzhidong Road, Wuhan, Hubei, 430060, PR China.
| | - Hua-Wen Sun
- Department of General Surgery, Renmin Hospital of Wuhan University, 99 Zhangzhidong Road, Wuhan, Hubei, 430060, PR China
| | - Zhi-Xiang Xing
- Department of General Surgery, Renmin Hospital of Wuhan University, 99 Zhangzhidong Road, Wuhan, Hubei, 430060, PR China
| | - Hou-Lai Yang
- Department of General Surgery, Renmin Hospital of Wuhan University, 99 Zhangzhidong Road, Wuhan, Hubei, 430060, PR China
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Chethan SG, Singh SK, Nongsiej J, Rakesh HB, Singh RP, Kumar N, Agarwal SK. IFN-τ acts in a dose-dependent manner on prostaglandin production by buffalo endometrial stromal cells cultured in vitro. Reprod Domest Anim 2014; 49:403-8. [PMID: 24612212 DOI: 10.1111/rda.12287] [Citation(s) in RCA: 12] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/08/2013] [Accepted: 01/07/2014] [Indexed: 11/30/2022]
Abstract
Interferon-τ (IFN-τ) has been recognized as the primary embryonic signal responsible for maternal recognition of pregnancy. Uterine endometrium produces both prostaglandin F2α (PGF2α ) and prostaglandin E2 (PGE2 ). PGF2α is responsible for the luteolysis; however, PGE2 favours establishment of pregnancy by its luteoprotective action. In this study, the dose-response effect of recombinant bovine IFN-τ (rbIFN-τ) on prostaglandin (PG) production by buffalo endometrial stromal cells cultured in vitro was studied. Buffalo endometrial stromal cells were isolated by double enzymatic digestion, initially with trypsin III followed by a cocktail of trypsin III, collagenase type II and DNase I and subsequently cultured till confluence. Further, cells were treated with different doses of rbIFN-τ (0.001, 0.01, 0.1, 1.0 and 10 μg/ml) and keeping a separate set of control. Culture supernatant was collected after 6, 12 and 24 h of treatment. PG levels in the culture supernatant were measured by enzyme immune assay (EIA) and total cellular protein estimated by Bradford method. Results indicated that buffalo endometrial stromal cells following rbIFN-τ treatment enhanced the secretion of both PGE2 and PGF2α , and also its ratio in a strict dose-dependent manner with a significant increase (p < 0.01) in PGE2 production at 1 μg/ml dose of rbIFN-τ and maximal stimulation for both PG was observed at 10 μg/ml. Further, both PG production and its ratio were increased significantly (p < 0.01) in a time-dependent fashion in all the groups at 6, 12 and 24 h post-treatment with highest level achieved at 24 h as compared with control. Absolute levels of PGE2 remained higher than PGF2α indicating PGE2 as the major PG produced by endometrial stromal cells. The dose-dependent response of rbIFN-τ signifies the importance of optimum concentration of IFN-τ for the embryonic development especially during the critical period to establish successful pregnancy.
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Affiliation(s)
- S G Chethan
- Division of Animal Reproduction, Indian Veterinary Research Institute, Izatnagar, India
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3
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The pharmacology and activity of non-steroidal anti-inflammatory drugs (NSAIDs): a review of their use as an adjuvant treatment in patients with HBV and HCV chronic hepatitis. ITALIAN JOURNAL OF MEDICINE 2011. [DOI: 10.1016/j.itjm.2011.02.004] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/24/2022] Open
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Bach JH, Jung KM, Choi JS, Jung SY, Chin MR, Ahn KH, Kim SK, Kim DK. Identification of a 42-kDa Group IV cPLA2-activating protein, cPLAPγ, as a GTP-binding protein in the bovine brain. J Biochem 2011; 150:385-94. [PMID: 21613292 DOI: 10.1093/jb/mvr064] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/13/2022] Open
Abstract
Brain tissue contains multiple forms of Phospholipase A(2) (PLA(2)) whose activities are involved in intracellular and intercellular signalling related to normal functions such as long-term potentiation, neurotransmitter release, cell growth and differentiation. Among them, we focused on regulatory mechanism of cPLA(2)α (Group IVA cytosolic PLA(2)) in brain tissue. In the present study, we report the identification of a cPLA(2)-activating protein (cPLAP) in the bovine brain. cPLAP activity appeared as two major peaks with molecular masses of 200 and 42 kDa in a Superose 12 gel filtration FPLC column. The 42-kDa form of cPLAP, designated cPLAPγ, was further purified using a Mono S FPLC column to near homogeneity and characterized to as a GTP-binding protein (G protein). Metabolic labelling and immunoprecipitation studies revealed that cPLAPγ associates with cPLA(2) in vitro and co-immunoprecipitates with [(35)S]-cPLA(2). Notably, cPLAPγ rendered cPLA(2) fully activated at submicromolar concentrations of Ca(2+). These results suggest that cPLAPγ may act as a G protein, activating cPLA(2)α prior to reaching full intracellular Ca(2+) concentrations.
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Affiliation(s)
- Jae Hyung Bach
- Department of Environmental & Health Chemistry, College of Pharmacy, Chung-Ang University, 221 Huksuk-dong, Dongjak-ku, Seoul 156-756, South Korea
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Otranto M, Do Nascimento AP, Monte-Alto-Costa A. Effects of supplementation with different edible oils on cutaneous wound healing. Wound Repair Regen 2010; 18:629-36. [DOI: 10.1111/j.1524-475x.2010.00617.x] [Citation(s) in RCA: 32] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/22/2022]
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Su KP, Huang SY, Peng CY, Lai HC, Huang CL, Chen YC, Aitchison KJ, Pariante CM. Phospholipase A2 and cyclooxygenase 2 genes influence the risk of interferon-alpha-induced depression by regulating polyunsaturated fatty acids levels. Biol Psychiatry 2010; 67:550-7. [PMID: 20034614 PMCID: PMC2982743 DOI: 10.1016/j.biopsych.2009.11.005] [Citation(s) in RCA: 132] [Impact Index Per Article: 8.8] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 02/19/2009] [Revised: 11/04/2009] [Accepted: 11/05/2009] [Indexed: 11/17/2022]
Abstract
BACKGROUND Phospholipase A2 (PLA2) and cyclooxygenase 2 (COX2) are the two key enzymes in the metabolism of polyunsaturated fatty acids, which in turn play an important role in cytokine-induced depression and sickness behavior. METHODS Patients with chronic hepatitis C viral infection (n = 132) were assessed to examine the effects of seven single nucleotide polymorphisms in COX2 and PLA2 genes on the development of depression during interferon (IFN)-alpha treatment; a subsample (n = 63) was assessed for the erythrocyte levels of the three main polyunsaturated fatty acids, docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA), and arachidonic acid. An independent "replication" sample of patients with major depression unrelated to cytokine treatment (n = 82) was also examined. RESULTS Twenty-eight percent of participants developed INF-alpha-induced depression. Participants with the PLA2 BanI GG or the COX2 rs4648308 AG genotypes had a higher risk of IFN-alpha-induced depression (odds ratio = 3.1 and 3.5, respectively). The "at risk" PLA2 genotype was associated with lower EPA levels, and the "at risk" COX2 genotype was associated with lower DHA levels, during IFN-alpha treatment. The PLA2 BanI GG polymorphism was also associated with more somatic symptoms of depression, both in patients with INF-alpha-induced depression and in the replication sample of patients with major depression. CONCLUSIONS Genetic variations in the COX2 and PLA2 genes increase the risk of IFN-alpha-induced depression, possibly by affecting the levels of EPA and DHA. Moreover, PLA2 genotype is associated with somatic symptoms in depression. Our study confirms the role of inflammatory mechanisms in major depression.
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Affiliation(s)
- Kuan-Pin Su
- Department of Psychiatry and Mind-Body Interface Laboratory, China Medical University, Hospital, Taichung, Taiwan
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Gramenzi A, Cursaro C, Margotti M, Balsano C, Spaziani A, Anticoli S, Loggi E, Salerno M, Galli S, Furlini G, Bernardi M, Andreone P. Ketoprofen, peginterferon 2a and ribavirin for genotype 1 chronic hepatitis C: A phase II study. World J Gastroenterol 2009; 15:5946-52. [PMID: 20014458 PMCID: PMC2795181 DOI: 10.3748/wjg.15.5946] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 02/06/2023] Open
Abstract
AIM: To evaluate the safety of adding ketoprofen to pegylated-interferon (PEG-IFN) with or without ribavirin and the effect on viral kinetics, STAT1 activity and expression of 2’-5’-oligoadenylate synthetase (2’-5’OAS) in genotype 1 chronic hepatitis C in a phase II study.
METHODS: Forty-five patients were studied: fifteen were randomized to PEG-IFN plus ribavirin (PR), 16 to PEG-IFN plus ketoprofen and 14 to PR and ketoprofen. The molecular study of IFN-dependent signal transduction was conducted in 9 patients from each group.
RESULTS: The combination of ketoprofen and PEG-IFN with or without ribavirin was safe and well tolerated. An early activation of STAT1 was observed in ketoprofen-treated patients, but this activation was less sustained over time. Conversely, ketoprofen plus PEG-IFN and ribavirin induced an early and sustained increase of 2’-5’OAS transcription starting 24 h after the first dose until the 36th wk. These data are consistent with the clinical results, showing a better sustained virological response and a lower relapse rate in patients receiving ketoprofen plus PEG-IFN and ribavirin.
CONCLUSION: The addition of ketoprofen to the standard therapy of chronic hepatitis C should be explored in larger randomized clinical studies.
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Montero D, Grasso V, Izquierdo MS, Ganga R, Real F, Tort L, Caballero MJ, Acosta F. Total substitution of fish oil by vegetable oils in gilthead sea bream (Sparus aurata) diets: effects on hepatic Mx expression and some immune parameters. FISH & SHELLFISH IMMUNOLOGY 2008; 24:147-155. [PMID: 18158252 DOI: 10.1016/j.fsi.2007.08.002] [Citation(s) in RCA: 38] [Impact Index Per Article: 2.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 06/15/2007] [Revised: 08/01/2007] [Accepted: 08/03/2007] [Indexed: 05/25/2023]
Abstract
The use of vegetable oils in fish nutrition has been extensively studied; and recent work has focused attention on replacing fish oil with alternative fatty acid sources and their effect on the immune system. However, little is known about the effect of these oils on immune parameters such as the fish interferon system. In this study we evaluate the effect of two vegetable oils (linseed and soybean) on gilthead sea bream Mx expression and other innate immune parameters. Experimental diets were formulated where fish oil was totally replaced by vegetable oils or for a mixture of them (50% linseed and 50% soybean). Another diet prepared with pure fish oil was used as a control. Two experiments were carried out in order to evaluate growth, feed utilization, serum alternative complement pathway activity, serum lysozyme and phagocytic activity of head kidney leucocytes as well as Mx expression in the liver. In the first experiment fish were fed with experimental diets for 6 months and then, growth and feed utilization as well as immune parameters were analyzed. In the second experiment, fish from the previous feeding trial were injected with either a sub-lethal dose of Photobacterium damselae subsp. piscicida (94/99) or a synthetic dsRNA (Poly I:C) in order to stimulate an Mx response. The results show that total substitution of fish oil by vegetable oils decreased the growth of gilthead sea bream juveniles. Furthermore, both phagocytic activity and serum alternative complement pathway activity were significantly reduced by the inclusion of either vegetable oil individually in the sea bream diets, but the diet with mixed vegetable oils had no significant effect. There was no effect on serum lysozyme levels but the basal constitutive levels of Mx transcript expression in the liver were elevated in the fish fed the vegetable oil diets. The time-course of the Mx response to injection of Poly I:C was shorter in the fish fed the fish oil diet and the fish fed the diet based on a mixture of both vegetable oils showed a faster Mx response to bacterial injection. Following stimulation with Poly I:C or PDP the fish fed the vegetable oil based diets still maintained higher basal levels of hepatic Mx expression than the fish fed the fish oil diet which returned to undetectable levels.
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Affiliation(s)
- D Montero
- Grupo de Investigación en Acuicultura, ICCM-IUSA, Telde, Las Palmas, Canary Islands, Spain.
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Jeong SC, Aikata H, Katamura Y, Azakami T, Kawaoka T, Saneto H, Uka K, Mori N, Takaki S, Kodama H, Waki K, Imamura M, Shirakawa H, Kawakami Y, Takahashi S, Chayama K. Effects of a 24-week course of interferon-α therapy after curative treatment of hepatitis C virus-associated hepatocellular carcinoma. World J Gastroenterol 2007; 13:5343-50. [PMID: 17879404 PMCID: PMC4171324 DOI: 10.3748/wjg.v13.i40.5343] [Citation(s) in RCA: 37] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 02/06/2023] Open
Abstract
AIM: To assess whether a 24-wk course of interferon (IFN) could prevent hepatocellular carcinoma (HCC) recurrence and worsening of liver function in patients with hepatitis C virus (HCV)-infected patients after receiving curative treatment for primary HCC.
METHODS: Outcomes in 42 patients with HCV infection treated with IFN-α, after curative treatment for primary HCC (IFN group), were compared with 42 matched curatively treated historical controls not given IFN (non-IFN group).
RESULTS: Although the rate of initial recurrence did not differ significantly between IFN group and non-IFN group (0%, 44%, 61%, and 67% vs 4.8%, 53%, 81%, and 87% at 1, 3, 5, and 7 years, P = 0.153, respectively), IFN group showed a lower rate than the non-IFN group for second recurrence (0%, 10.4%, 28%, and 35% vs 0%, 30%, 59%, and 66% at 1, 3, 5 and 7 years, P = 0.022, respectively). Among the IFN group, patients with sustained virologic response (SVR) were less likely to have a second HCC recurrence than IFN patients without an SVR, or non-IFN patients. Multivariate analysis identified the lack of SVR as the only independent risk factor for a second recurrence, while SVR and Child-Pugh class A independently favored overall survival.
CONCLUSION: Most intrahepatic recurrences of HCV-related HCC occurred during persistent viral infection. Eradication of HCV is essential for the prevention of HCC recurrence and improvement of survival.
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Affiliation(s)
- Soo Cheol Jeong
- Department of Medicine and Molecular Science, Graduate School of Biomedical Sciences, Hiroshima University, 1-2-3, Kasumi, Minami-ku, Hiroshima 734-8551, Japan
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Cheng Z, Sheldrick EL, Marshall E, Wathes DC, Abayasekara DRE, Flint APF. Control of cyclic AMP concentration in bovine endometrial stromal cells by arachidonic acid. Reproduction 2007; 133:1017-26. [PMID: 17616730 DOI: 10.1530/rep-06-0220] [Citation(s) in RCA: 9] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/08/2022]
Abstract
Second messenger signalling through cyclic AMP (cAMP) plays an important role in the response of the endometrium to prostaglandin (PG) E(2) during early pregnancy. Arachidonic acid, which is a by-product of the luteolytic cascade in ruminants, is a potential paracrine signal from the epithelium to the stroma. We investigated the effects of arachidonic acid on the response of the stroma to PGE(2). cAMP was measured in bovine endometrial stromal cells treated with agents known to activate or inhibit adenylyl cyclase, protein kinase C (PKC) or phosphodiesterase (PDE). PGE(2) increased the intracellular cAMP concentration within 10 min, and this effect was attenuated by arachidonic acid and the PKC activator, 4beta-phorbol myristate acetate (PMA). The inhibitory effect of arachidonic acid on PGE(2)-induced cAMP accumulation was prevented by the PKC inhibitor, RO318425, and was absent in cells in which PKC had been downregulated by exposure to PMA for 24 h. The effect of arachidonic acid was also prevented by the PDE inhibitor, 3-isobutyl-1-methylxanthine. Arachidonic acid was shown by immunoblotting to prevent induction of cyclooxygenase-2 by PGE(2), forskolin or dibutyryl cAMP. The results indicate that arachidonic acid activates PDE through a mechanism involving PKC, counteracting a rise in intracellular cAMP in response to PGE(2). The data suggest that arachidonic acid antagonizes PGE(2) signalling through cAMP in the bovine endometrium, possibly acting to ensure a rapid return to oestrus in the case of failure of the maternal recognition of pregnancy.
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Affiliation(s)
- Z Cheng
- School of Biosciences, University of Nottingham, Sutton Bonington Campus, Loughborough, UK
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Jung CR, Choi S, Im DS. The NS5A protein of hepatitis C virus represses gene expression of hRPB10alpha, a common subunit of host RNA polymerases, through interferon regulatory factor-1 binding site. Virus Res 2007; 129:155-65. [PMID: 17714821 DOI: 10.1016/j.virusres.2007.07.005] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/21/2007] [Revised: 07/09/2007] [Accepted: 07/12/2007] [Indexed: 11/22/2022]
Abstract
The nonstructural (NS) 5A protein of hepatitis C virus (HCV) plays important roles in both viral RNA replication and modulation of the physiology of the host cell. Here we report that NS5A repressed gene expression of hRPB10alpha, a common subunit of host RNA polymerases (Pol), in hepatoma cell lines and Huh-7 cells harboring HCV replicon. Analysis of the hRPB10alpha promoter region revealed that interferon regulatory factor-1 binding element (IRF-E) was essential for its transcription. The IRF-E was responsible for the NS5A-mediated repression of the hRPB10alpha transcription and its induction by IRF-1 that is known to be induced by interferon-alpha. Electrophoretic mobility shift assay showed that IRF-1 bound to the IRF-E and the binding reduced when NS5A was expressed. NS5A appeared to negatively regulate IRF-1 expression, which might be partly responsible for the decrease of hRPB10alpha expression. NS5A expression moderately decreased promoter-independent Pol activity in vitro. Transcription of adenoviral genes that are dependent on Pol II or III and propagation of adenoviral genome were impaired in HeLa cells with stable NS5A expression. The results suggest that NS5A may partly modulate host cell transcription by the down-regulation of hRPB10alpha.
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Affiliation(s)
- Cho-Rok Jung
- Gene Therapy Research Unit, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Yusong, Daejeon 305-806, Republic of Korea
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Bahrami H, Daryani NE, Haghpanah B, Moayyeri A, Moghadam KF, Mirmomen S, Kamangar F. Effects of indomethacin on viral replication markers in asymptomatic carriers of hepatitis B: a randomized, placebo-controlled trial. Am J Gastroenterol 2005; 100:856-61. [PMID: 15784032 DOI: 10.1111/j.1572-0241.2005.41144.x] [Citation(s) in RCA: 12] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 12/11/2022]
Abstract
OBJECTIVES Previous studies have suggested some benefits of nonsteroidal antiinflammatory drugs (NSAIDs) use in patients with chronic viral hepatitis. We evaluated potential effects of indomethacin in asymptomatic carriers of hepatitis B surface antigen (HBsAg). DESIGN Randomized, placebo-controlled, double-masked clinical trial. METHODS One hundred and twelve patients who were confirmed to be HBsAg carriers for at least 6 months and had normal liver function tests, normal abdominal sonography, and no sign of cirrhosis were randomly assigned into two groups. One group (56 participants, mean age (+/-SD) 31.7 (+/-9.6) yr, 29 male, mean serum alanine aminotransferase (ALT) (+/-SD) 24.9 (+/-9.2)) received indomethacin capsules (25 mg) three times daily and the other group (56 participants, mean age (+/-SD) 33.8 (+/-10.2) yr, 33 male, mean serum ALT (+/-SD) 24.5 (+/-8.7)) took placebo capsules with identical package and appearance. All participants were under treatment for 6 months and were followed 3 months thereafter. Statistical analyses were performed both by intention-to-treat and on-treatment methods. RESULTS Nine participants in the indomethacin group (16%) and 8 in the placebo group (14%) did not complete the trial. HBsAg seroconversion did not differ by treatment group (2 subjects in each group became seronegative). Hepatitis B virus DNA (HBV-DNA) became negative in sera of 7 participants in the indomethacin group but only in 1 in the placebo group (intention-to-treat p= 0.06; on-treatment p= 0.03). Seroconversion of HBeAg to anti-HBe occurred only in 5 participants in the indomethacin group (intention-to-treat p= 0.06; on-treatment p= 0.03). Adverse events included one case of hepatotoxicity and two cases of gastritis in the indomethacin group and one suspected gastritis in the placebo group. CONCLUSIONS We suggest use of indomethacin only in the subgroup of asymptomatic HBsAg carriers who have detectable HBV-DNA or HBeAg in their sera.
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Affiliation(s)
- Hossein Bahrami
- Department of Epidemiology, The Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland 21205, USA
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Andreone P, Gramenzi A, Loggi E, Favarelli L, Cursaro C, Margotti M, Biselli M, Lorenzini S, Bernardi M. In vitro effect of indomethacin and interferon-α on Th1 and Th2 cytokine synthesis in patients with chronic hepatitis C. Cytokine 2004; 26:95-101. [PMID: 15135802 DOI: 10.1016/j.cyto.2003.08.014] [Citation(s) in RCA: 9] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/10/2003] [Revised: 08/16/2003] [Accepted: 08/25/2003] [Indexed: 11/29/2022]
Abstract
Current evidences suggest that non-steroidal anti-inflammatory drugs could enhance the antiviral activity of interferon-alpha in chronic HCV infection. In this study, we investigated the effect of indomethacin, a non-steroidal anti-inflammatory drug, and interferon-alpha on cytokine production by peripheral blood mononuclear cells from 12 untreated patients with chronic hepatitis C. We evaluated the effect of incubation with indomethacin, interferon-alpha or both on synthesis of Th1- (interleukin-2, interferon-gamma) and Th2-associated cytokines (interleukin-4, interleukin-10), and of the antiviral protein 2',5'-oligoadenylate synthetase. Interferon-alpha induced a significant increase in production of interleukin-2. Smaller increases were also seen in the presence of indomethacin, while incubation with both indomethacin and interferon-alpha leads to a synergistic effect. Incubation with indomethacin decreased both interleukin-4 and interleukin-10, whereas interferon-alpha increased these cytokines. The addition of indomethacin to interferon-alpha significantly reversed this interferon-induced increase. Finally, both indomethacin and the association interferon-alpha plus indomethacin determined a significant increase in 2',5'-oligoadenylate synthetase production compared to both baseline and interferon-alpha alone. In conclusion, indomethacin was able to enhance the antiviral activity of interferon-alpha and to modulate the interferon-induced Th1 and Th2 cytokine response by increasing the Th1-response, fundamental for sustained clearance of HCV, and by decreasing the Th-2 type response, associated with HCV persistence.
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Affiliation(s)
- Pietro Andreone
- Semeiotica Medica, Dipartimento di Medicina Interna, Cardioangiologia ed Epatologia, Università di Bologna, Via Massarenti, 9, 40138 Bologna, Italy.
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Yao JK, Sistilli CG, van Kammen DP. Membrane polyunsaturated fatty acids and CSF cytokines in patients with schizophrenia. Prostaglandins Leukot Essent Fatty Acids 2003; 69:429-36. [PMID: 14623497 DOI: 10.1016/j.plefa.2003.08.015] [Citation(s) in RCA: 22] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 11/30/2022]
Abstract
Findings to date provide evidence that altered membrane structure and function are present in patients with either first-episode or chronic schizophrenia, suggesting defects in phospholipid metabolism and cell signaling in schizophrenia. The purpose of this investigation is to test whether decreased membrane polyunsaturated fatty acids (PUFAs) were associated with an increased secretion of proinflammatory cytokines. Thus, we measured interleukin 6 (IL-6) and interleukin 10 (IL-10) in cerebrospinal fluid (CSF) of patients with chronic schizophrenia as well as PUFAs of red blood cell (RBC) membranes from the same individuals. A significant and inverse correlation was found between CSF IL-6 (not IL-10) and RBC membrane PUFAs levels in both haloperidol-treated and medication-free patients with schizophrenia. Specifically, such an association was found in the n-6 (18:2, 20:4, and 22:4) and, to a lesser extent, the n-3 fatty acids. Taken together, the present findings suggest that decreased membrane PUFAs may be related to an immune disturbance in schizophrenia, possibly resulting from an increased phospholipase A2 activity mediated through the proinflammatory cytokines.
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Affiliation(s)
- J K Yao
- VA Pittsburgh Healthcare System and Western Psychiatric Institute and Clinic, University of Pittsburgh Medical Center, 7180 Highland Drive, Building 13, Pittsburgh, PA 15206, USA.
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Andreone P, Gramenzi A, Cursaro C, Biselli M, Lorenzini S, Loggi E, Felline F, Fiorino S, Di Giammarino L, Porzio F, Galli S, Bernardi M. Interferon-alpha combined with ketoprofen as treatment of naïve patients with chronic hepatitis C: a randomized controlled trial. J Viral Hepat 2003; 10:306-9. [PMID: 12823598 DOI: 10.1046/j.1365-2893.2003.00449.x] [Citation(s) in RCA: 13] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 12/09/2022]
Abstract
In this randomized controlled study, we evaluated the efficacy and safety of interferon-alpha combined with ketoprofen to that of interferon-alpha alone in naïve patients with chronic hepatitis C. Forty patients were randomized to receive Interferon-alpha2a (3 million units three times a week) and ketoprofen (150 mg twice a day) and 40 to receive only interferon-alpha2a at the same dose. Patients were treated for 6 months and followed up for 6 months. Response was defined by undetectable HCV-RNA in serum at the end-of-treatment and after 6 months from the completion of therapy (long term response). At the end of treatment the response was similar in the two group. However, combination treatment showed significantly higher efficacy than monotherapy in achieving long term response (10%vs 32.5%; P = 0.014). Overall adverse events were similar in the two groups. 'Flu-like syndrome was significantly less common in the ketoprofen plus interferon group which experienced a significantly higher incidence of epigastric pain'. Our results indicate that the combination of ketoprofen plus interferon is significantly more effective than interferon alone in the treatment of naïve patients with chronic hepatitis C and is well tolerated. However this combined treatment appears to be less effective than the association of pegylated IFN and ribavirin which represent the current standard treatment. Thus, the role of ketoprofen in the treatment of chronic hepatitis C needs to be further evaluated against such a treatment.
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Affiliation(s)
- P Andreone
- Semeiotica Medica, Dipartimento di Medicina Interna, Cardioangiologia ed Epatologia, Università di Bologna, Italy.
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16
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Tang S, Bhatia B, Maldonado CJ, Yang P, Newman RA, Liu J, Chandra D, Traag J, Klein RD, Fischer SM, Chopra D, Shen J, Zhau HE, Chung LWK, Tang DG. Evidence that arachidonate 15-lipoxygenase 2 is a negative cell cycle regulator in normal prostate epithelial cells. J Biol Chem 2002; 277:16189-201. [PMID: 11839751 DOI: 10.1074/jbc.m111936200] [Citation(s) in RCA: 92] [Impact Index Per Article: 4.0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/06/2022] Open
Abstract
15-Lipoxygenase 2 (15-LOX2) is a recently cloned human lipoxygenase that shows tissue-restricted expression in prostate, lung, skin, and cornea. The protein level and enzymatic activity of 15-LOX2 have been shown to be down-regulated in prostate cancers compared with normal and benign prostate tissues. The biological function of 15-LOX2 and the role of loss of 15-LOX2 expression in prostate tumorigenesis, however, remain unknown. We report the cloning and functional characterization of 15-LOX2 and its three splice variants (termed 15-LOX2sv-a, 15-LOX2sv-b, and 15-LOX2sv-c) from primary prostate epithelial cells. Western blotting with multiple primary prostate cell strains and prostate cancer cell lines reveals that the expression of 15-LOX2 is lost in all prostate cancer cell lines, accompanied by decreased enzymatic activity revealed by liquid chromatography/tandem mass spectrometry analyses. Further experiments show that the loss of 15-LOX2 expression results from transcriptional repression caused by mechanism(s) other than promoter hypermethylation or histone deacetylation. Subsequent functional studies indicate the following: 1) the 15-LOX2 product, 15(S)-hydroxyeicosatetraenoic acid, inhibits prostate cancer cell cycle progression; 2) 15-LOX2 expression in primary prostate epithelial cells is inversely correlated with cell cycle; and 3) restoration of 15-LOX2 expression in prostate cancer cells partially inhibits cell cycle progression. Taken together, these results suggest that 15-LOX2 could be a suppressor of prostate cancer development, which functions by restricting cell cycle progression.
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Affiliation(s)
- Shaohua Tang
- Department of Carcinogenesis, the University of Texas MD Anderson Cancer Center, Science Park Research Division, Smithville, Texas 78957, USA
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17
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Abstract
It is well established that fatty acid metabolites of cyclooxygenase, lipoxygenase (LOX), and cytochrome P450 are implicated in essential aspects of cellular signaling including the induction of programmed cell death. Here we review the roles of enzymatic and non-enzymatic products of polyunsaturated fatty acids in controlling cell growth and apoptosis. Also, the spontaneous oxidation of polyunsaturated fatty acids yields reactive aldehydes and other products of lipid peroxidation that are potentially toxic to cells and may also signal apoptosis. Significant conflicting data in terms of the role of LOX enzymes are highlighted, prompting a re-evaluation of the relationship between LOX and prostate cancer cell survival. We include new data showing that LNCaP, PC3, and Du145 cells express much lower levels of 5-LOX mRNA and protein compared with normal prostate epithelial cells (NHP2) and primary prostate carcinoma cells (TP1). Although the 5-LOX activating protein inhibitor MK886 killed these cells, another 5-LOX inhibitor AA861 hardly showed any effect. These observations suggest that 5-LOX is unlikely to be a prostate cancer cell survival factor, implying that the mechanisms by which LOX inhibitors induce apoptosis are more complex than expected. This review also suggests several mechanisms involving peroxisome proliferator activated receptor activation, BCL proteins, thiol regulation, and mitochondrial and kinase signaling by which cell death may be produced in response to changes in non-esterified and non-protein bound fatty acid levels. Overall, this review provides a context within which the effects of fatty acids and fatty acid oxidation products on signal transduction pathways, particularly those involved in apoptosis, can be considered in terms of their overall importance relative to the much better studied protein or peptide signaling factors.
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Affiliation(s)
- Dean G Tang
- Division of Pharmacology and Toxicology, College of Pharmacy, The University of Texas, Austin 78712, USA
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18
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Gutman H, Schachter J, Stopel E, Gutman R, Lahav J. Impaired platelet aggregation in melanoma patients treated with interferon-alpha-2b adjuvant therapy. Cancer 2002; 94:780-5. [PMID: 11857313 DOI: 10.1002/cncr.10261] [Citation(s) in RCA: 15] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/09/2022]
Abstract
BACKGROUND High-dose interferon (INF)- alpha-2b is the only Food and Drug Administration-approved adjuvant treatment for patients with melanoma who are at high risk of recurrence. Although circumstantial evidence points to a potentially harmful effect of INF-alpha-2b on platelet function, to the authors' knowledge this has never been studied in humans. METHODS The study group was comprised of patients who had undergone surgery for melanoma and were free of disease but at a high risk of recurrence. All patients were candidates for adjuvant INF treatment (high-dose) and were undergoing routine evaluation to which platelet aggregation was added. Aggregation was triggered in standard fashion with adenosine diphosphate, epinephrine, collagen, thrombin, arachidonic acid, and ristocetin. Blood samples were drawn immediately before treatment, during the intravenous loading phase, during the subcutaneous maintenance phase, and 3-6 weeks after cessation of treatment. Patients receiving low-dose, long-standing INF-alpha-2b treatment also were tested. All results at each phase were compared with those of normal controls. RESULTS In those patients receiving high-dose INF-alpha-2b, ristocetin-induced aggregation did not appear to be affected. However, the response to > or = 1 of the other agonists was impaired in 5 of 6 samples during loading, 14 of 15 samples during the maintenance phase, and 8 of 13 samples after treatment, compared with only 1 of 8 samples before treatment (P = 0.025, P = 0.002, and P = 0.067, respectively). During treatment with low-dose INF, platelet function was affected to a lesser extent. CONCLUSIONS INF treatment in melanoma patients appears to be associated with severe impairment of platelet aggregation, which appears to be dose-dependent and cumulative-dose-dependent. This is not detectable by the standard coagulation profile. This effect has significant implications in the event of accidental injury or elective surgery. The antiaggregation activity may be the mechanism by which INF delays, reduces, or prevents the formation of melanoma metastases.
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Affiliation(s)
- Haim Gutman
- Department of Surgery B, Rabin Medical Center, Beilinson Campus, Petah Tiqva, and Sackler School of Medicine, Tel Aviv University, Tel Aviv, Israel.
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19
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Affiliation(s)
- Maria Teresa Rizzo
- Signal Transduction Laboratory, Methodist Research Institute, Indianapolis, Indiana 46202, USA.
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20
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Doualla-Bell F, Koromilas AE. Induction of PG G/H synthase-2 in bovine myometrial cells by interferon-tau requires the activation of the p38 MAPK pathway. Endocrinology 2001; 142:5107-15. [PMID: 11713203 DOI: 10.1210/endo.142.12.8524] [Citation(s) in RCA: 11] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 11/19/2022]
Abstract
PGs are regulators of a plethora of uterine functions during reproductive processes, including uterine contractility. In bovine uterus, the rate-limiting step in PG synthesis is catalyzed by the PG endoperoxide G/H synthase (PGHS) enzymes. It has previously been established that PGHS-2 isoform expression is affected by the ruminant-specific interferon (IFN)-tau in bovine endometrial cells. Here, we show that PGHS-2 mRNA and protein levels are induced by IFN-tau in primary cell cultures from bovine myometrium. Treatment with recombinant bovine IFN-tau induces the activation of the JAK-STAT and p38 MAPK pathways in bovine myometrial cells. Inhibition of the p38 pathway by the specific inhibitor SB203580 strongly decreases PGHS-2 mRNA and protein expression without affecting the phosphorylation and DNA-binding of transcription factors STAT-1 and STAT-2. The p38 pathway regulates PGHS-2 expression at the posttranscriptional level, because the presence of SB203580 results in the destabilization of IFN-tau-induced PGHS-2 mRNA. Taken together, these data demonstrate the ability of IFN-tau to induce the activation of the JAK-STAT pathway in a manner similar to other types of IFN (i.e. alpha, beta, and gamma) and to regulate PGHS-2 mRNA stability through the activation of the p38 pathway. These findings provide new insights into the physiological function of IFN-tau, in regard to regulation of specific genes associated with myometrial contractility.
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Affiliation(s)
- F Doualla-Bell
- Perinatal Research and Developmental Pharmacology Unit, Lady Davis Institute for Medical Research, Sir Mortimer B. Davis Jewish General Hospital, Montréal, Québec H3T 1E2, Canada.
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21
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Lechner J, Pfaller W. Interferon alpha2b increases paracellular permeability of renal proximal tubular LLC-PK1 cells via a mitogen activated protein kinase signaling pathway. Ren Fail 2001; 23:573-88. [PMID: 11499571 DOI: 10.1081/jdi-100104739] [Citation(s) in RCA: 12] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/03/2022] Open
Abstract
The therapeutic administration of Interferon alpha2b (IFNalpha) is often accompanied by impaired renal function, i.e. reduced glomerular filtration rate and sometimes a so-called "capillary leak syndrome". To clarify the mechanism behind the renal dysfunction, confluent monolayers of LLC-PK1 cells were used as a model system to analyze the effects of IFNalpha on renal tubular epithelium. Examination of epithelial barrier function via measurement of transepithelial resistance (TER) revealed a dose dependent increase in paracellular permeability by IFNalpha treatment. The effect was reversible upon removal of IFNalpha at doses up to 5 x 10(3) U/mL. Apical or basolateral application of IFNalpha yielded the same decrease in TER. Tyrphostin A25, an inhibitor of phosphotyrosine kinases, ameliorated the IFNalpha induced decrease of TER. In order to unravel intracellular signal transduction pathways that may mediate IFNalpha induced changes of epithelial barrier function, we inhibited IFNalpha signaling through a mitogen activated protein kinase pathway by the Mek1 inhibitor PD98059. The inhibitor could be shown to prevent IFNalpha induced decrease of transepithelial resistance. Inhibitors of the p38 mitogen activated protein kinase pathway did not affect IFNalpha mediated changes of epithelial barrier function, indicating a highly specific role for the Mek/Erk pathway. Activation of mitogen activated protein kinase pathways by epidermal growth factor or anisomycin could not, per se, imitate the effect of IFNalpha on the paracellular permeability of LLC-PK1 monolayers. These findings provide evidence that IFNalpha can affect barrier function in renal epithelial cells via activation of the Mek/Erk pathway.
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Affiliation(s)
- J Lechner
- Institute of Physiology, University of Innsbruck, Austria
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22
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Abstract
Dendritic cells are important antigen-presenting cells of the immune system that induce and modulate immune responses. They interact with T and B lymphocytes as well as with natural killer cells to promote activation and differentiation of these cells. Dendritic cells generated in vitro from monocytes by use of the cytokines GM-CSF and IL-4 are increasingly used clinically to enhance antitumor immunity in cancer patients. However, recent studies revealed that the functional repertoire of monocyte-derived dendritic cells may be incomplete. Important functions of monocyte-derived dendritic cells such as migration or the ability to induce natural killer cell activation or type 2 T helper cell differentiation appear to be impaired. We propose that all these deficiencies relate to a single biochemical deficiency of monocyte-derived dendritic cells. IL-4, which is used to generate monocyte-derived dendritic cells, suppresses phospholipase A2, the enzyme that liberates arachidonic acid from membrane phospholipids and contributes to the synthesis of platelet-activating factor. Monocyte-derived dendritic cells must therefore fail to generate platelet-activating factor as well as arachidonic acid derivatives such as prostaglandins, leukotrienes, and lipoxins, collectively referred to as eicosanoids. Since eicosanoids and platelet-activating factor are known to play an important role in processes such as leukocyte migration, natural killer cell activation, and type 2 T helper cell differentiation, the deficiency in eicosanoid and platelet-activating factor biosynthesis may be responsible for the observed handicaps of monocyte-derived dendritic cells.
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Affiliation(s)
- M Thurnher
- Department of Urology, University of Innsbruck, A-6020 Innsbruck, Austria.
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23
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Affiliation(s)
- Martin Thurnher
- Department of Urology University of Innsbruck A-6020 Innsbruck Austria
| | | | - Reinhold Ramoner
- Department of Urology University of Innsbruck A-6020 Innsbruck Austria
| | - Georg Bartsch
- Department of Urology University of Innsbruck A-6020 Innsbruck Austria
| | - Lorenz Höoltl
- Department of Urology University of Innsbruck A-6020 Innsbruck Austria
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24
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Goodbourn S, Didcock L, Randall RE. Interferons: cell signalling, immune modulation, antiviral response and virus countermeasures. J Gen Virol 2000; 81:2341-2364. [PMID: 10993923 DOI: 10.1099/0022-1317-81-10-2341] [Citation(s) in RCA: 723] [Impact Index Per Article: 28.9] [Reference Citation Analysis] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/15/2022] Open
Affiliation(s)
- S Goodbourn
- Department of Biochemistry and Immunology, St George's Hospital Medical School, University of London, London SW17 0RE, UK1
| | - L Didcock
- Biomolecular Sciences Building, North Haugh, University of St Andrews, Fife KY16 9TS, UK2
| | - R E Randall
- Biomolecular Sciences Building, North Haugh, University of St Andrews, Fife KY16 9TS, UK2
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25
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Nie D, Tang K, Szekeres K, Trikha M, Honn KV. The role of eicosanoids in tumor growth and metastasis. ERNST SCHERING RESEARCH FOUNDATION WORKSHOP 2000:201-17. [PMID: 10943334 DOI: 10.1007/978-3-662-04047-8_10] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 02/17/2023]
Affiliation(s)
- D Nie
- Department of Radiation Oncology, Wayne State University, Detroit, MI 48202, USA
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26
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Andreone P, Gramenzi A, Cursaro C, Bernardi M. Inhibition of the cyclooxygenase/lipoxygenase pathways to improve interferon alpha efficacy in chronic hepatitis C: don't lose the tract! Dig Liver Dis 2000; 32:542-3. [PMID: 11057930 DOI: 10.1016/s1590-8658(00)80012-2] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.0] [Reference Citation Analysis] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 12/11/2022]
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27
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Nie D, Tang K, Szekeres K, Li L, Honn KV. Eicosanoid regulation of angiogenesis in human prostate carcinoma and its therapeutic implications. Ann N Y Acad Sci 2000; 905:165-76. [PMID: 10818452 DOI: 10.1111/j.1749-6632.2000.tb06548.x] [Citation(s) in RCA: 20] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/26/2022]
Abstract
Cancer of the prostate is the most commonly diagnosed cancer in America. There are several lines of evidence implicating the involvement of arachidonate 12-lipoxygenase, an enzyme metabolizing arachidonic acid to form 12(S)-hydroxyeicosatetraenoic acid (HETE), in prostate cancer progression. First, as prostate cancer reaches a more advanced stage, the level of 12-lipoxygenase expression is increased. Second, overexpression of 12-lipoxygenase in human prostate cancer cells stimulates angiogenesis and tumor growth. Third, an inhibitor of 12-lipoxygenase has been found effective against metastatic prostate tumor growth, and the inhibition of 12-lipoxygenase is related with the reduction of tumor angiogenesis. Collectively, these studies suggest that 12-lipoxygenase regulates tumor angiogenesis in prostate cancer and that inhibition of 12-lipoxygenase is a novel therapeutic approach for the treatment of prostate cancers.
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Affiliation(s)
- D Nie
- Department of Radiation Oncology, Wayne State University, Detroit, Michigan 48202, USA
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28
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Muñoz AE, Levi D, Podestá A, Gorín JM, González J, Bartellini MA, Munne MS, Cabanne A, Flichman D, Terg R. Interferon-alpha 2b combined with daily ketoprofen administration improves virological response in chronic hepatitis C: a prospective and randomised trial. Gut 2000; 46:427-31. [PMID: 10673309 PMCID: PMC1727838 DOI: 10.1136/gut.46.3.427] [Citation(s) in RCA: 17] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 01/22/2023]
Abstract
BACKGROUND Less than 15% of patients with chronic hepatitis C show a sustained virological response to interferon treatment. AIM To evaluate the efficacy and safety of different doses of ketoprofen combined with interferon-alpha 2b in the treatment of chronic hepatitis C. PATIENTS/METHODS Seventy compensated patients with chronic hepatitis C received interferon-alpha 2b 3 million units three times a week for six months. They were randomly assigned to: group 1 (n = 23), interferon-alpha 2b alone; group 2 (n = 23), interferon-alpha 2b plus 200 mg ketoprofen three times a week; group 3 (n = 24), interferon-alpha 2b plus 200 mg ketoprofen twice a day. Complete and sustained responses were defined as normal serum alanine aminotransferase levels and negative serum hepatitis C virus RNA at six and 12 months respectively. RESULTS Complete and sustained responses were similar in groups 1 and 2: 10% v 5% and 5% v 0% respectively. In group 3, complete response was 29% (p = 0.13 v group 1 and p = 0.04 v group 2) and sustained response was 26% (p = 0.07 v group 1 and p = 0.01 v group 2). Overall, adverse events were similar in the three groups. However, 'flu-like syndrome was less common in group 2 (30%) and group 3 (37%) than in group 1 (77%) (p = 0.01). CONCLUSIONS Twice daily ketoprofen administration combined with interferon-alpha 2b produced an increase in complete and sustained responses. Although the combination of interferon-alpha 2b with ketoprofen was well tolerated and decreased the incidence of 'flu-like syndrome, it is advisable to monitor possible non-steroid anti-inflammatory drug hepatotoxicity.
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Affiliation(s)
- A E Muñoz
- Unidad de Hepatología, Hospital Dr Bonorino Udaondo, School of Medicine, University of Salvador, Buenos Aires, Argentina
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29
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Bastie A, Castera L, Roudot-Thoraval F, Dhumeaux D, Pawlotsky JM. Inhibition of the cyclooxygenase/lipoxygenase pathways to improve interferon alfa efficacy in chronic hepatitis C: a wrong track. Hepatology 2000; 31:547-8. [PMID: 10691381 DOI: 10.1002/hep.510310248] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 12/07/2022]
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30
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Romano-Fontes LG, Curi R, Peres CM, Nishiyama-Naruke A, Brunaldi K, Abdulkader F, Procopio J. Fatty acid transport across lipid bilayer planar membranes. Lipids 2000; 35:31-4. [PMID: 10695921 DOI: 10.1007/s11745-000-0491-4] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/29/2022]
Abstract
The transport of palmitic acid (PA) across planar lipid bilayer membranes was measured using a high specific activity [14C]palmitate as tracer for PA. An all-glass trans chamber was employed in order to minimize adsorbance of PA onto the surface. Electrically neutral (diphytanoyl phosphatidylcholine) and charged (Azolectin) planar bilayers were maintained at open electric circuit. We found a permeability to PA of (8.8 +/- 1.9) x 10(-6) cm s(-1) (n = 15) in neutral and of (10.3 +/- 2.2) x 10(-6) cm s(-1) (n = 5) in charged bilayers. These values fall within the order of magnitude of those calculated from desorption constants of PA in different vesicular systems. Differences between data obtained from planar and vesicular systems are discussed in terms of the role of solvent, radius of curvature, and pH changes.
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Affiliation(s)
- L G Romano-Fontes
- Departamento de Fisiologia e Biofísica, Instituto de Ciencias Biomédicas, Universidade de São Paulo, Brazil
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31
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Abayasekara DR, Wathes DC. Effects of altering dietary fatty acid composition on prostaglandin synthesis and fertility. Prostaglandins Leukot Essent Fatty Acids 1999; 61:275-87. [PMID: 10670689 DOI: 10.1054/plef.1999.0101] [Citation(s) in RCA: 151] [Impact Index Per Article: 5.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 02/02/2023]
Abstract
Several studies over the past 20 years have demonstrated that subjects on diets composed of substances with high levels of n-3 polyunsaturated fatty acids (PUFAs) (e.g. fish) have a decreased incidence of heart disease. On this basis, a recent report from the Department of Health has advised UK consumers to decrease the proportion of saturated as opposed to unsaturated fats in their diet and to increase the ratio of n-3 to n-6 PUFAs. This could be achieved by altering the amounts of these constituents in milk and meat. n-3 Fatty acids can most easily be added to animal feed as either fish oil or linseed oil and can be increased in the blood and milk of ruminants following protection to avoid hydrogenation in the rumen. In western countries the ratio of consumption of n-6 to n-3 PUFAs is greater than 10 and current evidence tends to suggest that a ratio nearer 5 would be more desirable and compatible with cardiovascular well being. As fertility in the UK dairy herd is already poor, it is important to establish whether alterations in dietary n-3 and n-6 PUFAs affects herd fertility before widespread changes in animal diets are recommended. Therefore, this review considers the role played by PUFAs and eicosanoids in fertility, with particular reference to the implications for farm livestock production. The evidence reviewed shows that alteration of the concentration and ratio of n-6 and n-3 PUFAs in feeds can influence prostaglandin synthesis/metabolism in a number of mammalian systems. The changed patterns of prostaglandin synthesis can as a consequence, affect the diverse functions (e.g. hormone secretion) that are normally mediated via prostaglandins. Similarly, changes in prostaglandin synthesis effected through manipulation of PUFAs has a major bearing on fertility (as PGs affect many reproductive parameters, e.g. ovulation). Several studies in cattle and other mammals, show that feeding or infusing different types of fat with varying PUFA content to females can alter: the number and size of ovarian follicles, the ovulation rate, progesterone production by the corpus luteum, the timing of luteolysis and gestational length. In the male most recent work has focussed on sperm production and experiments in fowl have demonstrated clear effects of dietary PUFAs on both the sperm membrane phospholipid composition and on fertilizing ability.
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Affiliation(s)
- D R Abayasekara
- Reproduction and Development Group, Department of Veterinary Basic Sciences, Royal Veterinary College, London, UK.
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32
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Ruthig DJ, Meckling-Gill KA. Both (n-3) and (n-6) fatty acids stimulate wound healing in the rat intestinal epithelial cell line, IEC-6. J Nutr 1999; 129:1791-8. [PMID: 10498749 DOI: 10.1093/jn/129.10.1791] [Citation(s) in RCA: 84] [Impact Index Per Article: 3.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/11/2022] Open
Abstract
The control of proliferation and epithelial restitution are processes that are poorly understood. The effects of (n-3), (n-6) and trans fatty acids on proliferation of subconfluent IEC-6 cultures and restitution of wounded IEC-6 monolayers were investigated. Incorporation of supplemented fatty acids into cellular phospholipid was also assessed. Sulforhodamine B protein dye binding assay was utilized to assess the proliferative effects of fatty acids on growth of IEC-6 cultures. Incorporation of supplemental fatty acids into cellular phospholipid was examined by thin-layer chromatography combined with gas chromatography. The modulation of epithelial restitution was examined by razor blade wounding confluent IEC-6 monolayers grown in media supplemented with various fatty acids. Inhibition of eicosanoid synthesis by indomethacin during the wounding assay was also assessed. Both (n-3) and (n-6) fatty acids significantly inhibited growth of this intestinal epithelial cell model at concentrations above 125 micromol/L. The trans fatty acid, linoelaidate 18:2(n-6)trans, inhibited growth of IEC-6 cells at concentrations above 250 micromol/L. Another trans fatty acid, elaidate 18:1(n-9)trans, was well-tolerated at concentrations as high as 500 micromol/L. Eicosapentanoic 20:5(n-3), linoleic 18:2(n-6), alpha-linolenic 18:3(n-3), gamma-linolenic 18:3(n-6) and arachidonic 20:4(n-6) acids all significantly enhanced cellular migration in the IEC-6 model of wound healing. Eicosapentanoate, linoleate, alpha-linolenate, gamma-linolenate and arachidonate are all capable of improving reconstitution of epithelial integrity following mucosal injury. Inhibition of eicosanoid synthesis reduced the enhancement of restitution by n-6 fatty acids back to control levels.
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MESH Headings
- Animals
- Cell Line
- Cell Membrane/drug effects
- Cell Membrane/metabolism
- Cells, Cultured
- Chromatography, Gas
- Chromatography, Liquid
- Fatty Acids, Omega-3/metabolism
- Fatty Acids, Omega-3/pharmacology
- Fatty Acids, Omega-3/therapeutic use
- Fatty Acids, Omega-6
- Fatty Acids, Unsaturated/metabolism
- Fatty Acids, Unsaturated/pharmacology
- Fatty Acids, Unsaturated/therapeutic use
- Intestinal Mucosa/drug effects
- Intestinal Mucosa/metabolism
- Phospholipids/metabolism
- Rats
- Wound Healing/drug effects
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Affiliation(s)
- D J Ruthig
- Department of Human Biology and Nutritional Sciences, University of Guelph, Guelph, Ontario, Canada N1G 2W1
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33
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Fabris P, Tositti G, Negro F, Marranconi F, Infantolino D, Rassu M, De Lalla F. Interferon alfa-2b alone or in combination with ketoprofen as treatment for interferon-naive chronic hepatitis C patients. Aliment Pharmacol Ther 1999; 13:1329-34. [PMID: 10540048 DOI: 10.1046/j.1365-2036.1999.00613.x] [Citation(s) in RCA: 10] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 12/08/2022]
Abstract
BACKGROUND Non-steroidal anti-inflammatory drugs may amplify the anti-viral effect of alpha-interferon in vitro but in vivo data are still controversial. AIM : To test the hypothesis that ketoprofen may increase the rate of response to alpha-interferon of chronic hepatitis C patients. METHODS Fifty patients with chronic hepatitis C who had never received alpha-interferon were randomly assigned to receive 3-8 MU of alpha2b-interferon, three times weekly for 6 months, alone or in association with ketoprofen at a dose of 200 mg/day five times weekly. The virological response to treatment (undetectable HCV RNA in serum) was evaluated after 3 months and at the end of treatment, and 6 and 12 months after therapy withdrawal. RESULTS One patient under combination therapy stopped the ketoprofen for persisting epigastric pain. Complete response under treatment was observed in 15 out of 24 (62.5%) patients receiving alpha2b-interferon alone and in 14 out of 26 (53.8%) patients under combination therapy (P=N.S.). One year after the end of treatment, a sustained response was seen in 4 out of 24 (16.2%) patients treated with alpha2b-interferon and in 5 out of 26 (19.2%) patients having received the combination (P=N.S.). CONCLUSION Administration of ketoprofen does not increase either the primary or the sustained response to alpha2b-interferon therapy of interferon-naive chronic hepatitis C patients.
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Affiliation(s)
- P Fabris
- Department of Infectious Diseases, S. Bortolo Hospital, Vicenza, Italy.
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Giambartolomei S, Artini M, Almerighi C, Moavero SM, Levrero M, Balsano C. Nonsteroidal anti-inflammatory drug metabolism potentiates interferon alfa signaling by increasing STAT1 phosphorylation. Hepatology 1999; 30:510-6. [PMID: 10421661 DOI: 10.1002/hep.510300224] [Citation(s) in RCA: 32] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 12/07/2022]
Abstract
A sustained response to standard interferon therapy for chronic hepatitis C has been demonstrated in no more than 25% of patients. To improve interferon alfa (IFN-alpha) antiviral effect, a number of combination therapies with IFNs plus other drugs have been proposed for both relapser and nonresponder hepatitis C virus (HCV)-infected patients. Although the causes of IFN resistance in subsets of HCV-infected patients are unknown, both viral and host factors have been involved, including defects in IFN signal transduction and IFN-alpha/beta receptor down-regulation. Here, we report that nonsteroidal anti-inflammatory drugs (NSAIDs), which have been proposed for IFN-alpha combination therapy in nonresponders, potentiate IFN-alpha signaling. We found that, in the hepatoma cell lines, CCL13/Chang and HepG2, indomethacin, a selective cyclo-oxygenase 1 and 2 (COX-1 and COX-2) inhibitor, increases IFN-alpha stimulation of interferon-stimulated response element (ISRE)-dependent transcription in a dose-dependent manner. Interestingly, maximal potentiation was observed with suboptimal IFN-alpha concentrations. Indomethacin exerts its effects by synergizing with IFN-alpha in inducing STAT1 activation by phosphorylation, without affecting concurrent Jak1 phosphorylation. Our data indicate that blockade of arachidonic acid (AA) metabolism by indomethacin activates a signaling pathway that converges on STAT1 activation to potentiate IFN-alpha-dependent gene activation.
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Affiliation(s)
- S Giambartolomei
- Department of Internal Medicine, University of L'Aquila, L'Aquila, Italy
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Lechner J, Krall M, Netzer A, Radmayr C, Ryan MP, Pfaller W. Effects of interferon alpha-2b on barrier function and junctional complexes of renal proximal tubular LLC-PK1 cells. Kidney Int 1999; 55:2178-91. [PMID: 10354267 DOI: 10.1046/j.1523-1755.1999.00487.x] [Citation(s) in RCA: 25] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/13/2022]
Abstract
BACKGROUND Interferon alpha-2b (IFNalpha) treatment of diseases can be accompanied by impaired renal function and capillary leak syndrome. To explore potential mechanisms of IFNalpha-induced renal dysfunction, an in vitro cell culture model system was established to investigate the effects of IFNalpha on barrier function and junctional complexes. METHODS LLC-PK1 cells were cultured on microporous membranes. Transepithelial resistance (TER) was measured, and the dose- and time-dependent effects of IFNalpha were assessed. The expression patterns of junctional proteins were examined by Western blot analysis and by confocal immunofluorescence microscopy. RESULTS IFNalpha produced a dose- and time-dependent decrease in TER. The effect was reversible on removal of IFNalpha at doses up to 5 x 103 U/ml. Tyrphostin, an inhibitor of phosphotyrosine kinases, ameliorated the IFNalpha-induced decrease in TER. Increased expression of occludin and E-cadherin was detected by Western blot analysis after IFNalpha treatment. Immunofluorescence confocal microscopy revealed a broader staining of occludin and E-cadherin following IFNalpha treatment, with prominent staining at the basal cell pole in addition to localization at the junctional region. A marked increase in phosphotyrosine staining along the apico-lateral cell border was detected after IFNalpha treatment. CONCLUSIONS These findings provide evidence that IFNalpha can directly affect barrier function in renal epithelial cells. The mechanisms involve enhanced tyrosine phosphorylation and overexpression and possibly displacement or missorting of the junctional proteins occludin and E-cadherin.
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Affiliation(s)
- J Lechner
- Institute of Physiology, University of Innsburck, Austria
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Okanoue T, Itoh Y, Minami M, Sakamoto S, Yasui K, Sakamoto M, Nishioji K, Murakami Y, Kashima K. Interferon therapy lowers the rate of progression to hepatocellular carcinoma in chronic hepatitis C but not significantly in an advanced stage: a retrospective study in 1148 patients. Viral Hepatitis Therapy Study Group. J Hepatol 1999; 30:653-9. [PMID: 10207807 DOI: 10.1016/s0168-8278(99)80196-2] [Citation(s) in RCA: 173] [Impact Index Per Article: 6.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 12/29/2022]
Abstract
BACKGROUND/AIM Hepatocellular carcinoma frequently develops during the advanced stages of chronic hepatitis C. We examined whether interferon prevents the development of hepatocellular carcinoma in chronic hepatitis C patients. METHODS Japanese patients with chronic hepatitis C (n = 1.148; 117 with portal fibrous expansion (F1), 636 with bridging fibrosis (F2), 355 with bridging fibrosis and architectural distortion (F3)) and 40 cirrhotic (F4) patients were treated with interferon. These patients were followed from 1 to 7 years after interferon therapy. Blood tests and image analysis were serially performed to assess response to interferon and to detect hepatocellular carcinoma. Fifty-five cirrhotic type C patients (control F4) not receiving interferon were enrolled in this study. RESULTS Sustained (SR: 27.5%) and transient (TR: 23.0%) responders totaled 50.5%, while 49.5% did not respond to interferon. SR showed an improvement in disease stage reflected by increased platelet counts. Fifty-two patients (9 F2, 36 F3, and 7 F4) developed hepatocellular carcinoma in the follow-up period; 3 SR, 8 TR, and 41 non-responders (NR). The cumulative incidence of hepatocellular carcinoma in F2 was significantly lower (p = 0.019) in SR compared with NR, but not in SR in F3 and F4 patients. However, the cumulative incidence of hepatocellular carcinoma was significantly decreased in all SR (p = 0.0001) and TR (p = 0.0397) compared with all NR. CONCLUSION These results indicate that interferon therapy in chronic hepatitis C patients lowered the rate of progression of hepatocellular carcinoma in sensitive cases but not in patients in an advanced stage.
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Affiliation(s)
- T Okanoue
- Third Department of Internal Medicine, Kyoto Prefectural University of Medicine, Japan.
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Brandt R, Ebert AD. Growth inhibitors for mammary epithelial cells. PROGRESS IN MOLECULAR AND SUBCELLULAR BIOLOGY 1999; 20:197-248. [PMID: 9928532 DOI: 10.1007/978-3-642-72149-6_10] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 01/10/2023]
Affiliation(s)
- R Brandt
- Novartis Pharma Inc., Basel, Switzerland
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38
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Affiliation(s)
- D W Leaman
- Gemini Technologies Inc., Cleveland, OH 44106, USA
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39
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Zarski JP, Maynard-Muet M, Chousterman S, Baud M, Barnoud R, Abergel A, Bacq Y, Combis JM, Causse X, Tran A, Oberti F, Minello A, Bresson-Hadni S, Bailly F, Raabe JJ, Leroy V, Hamici L, Hicham T, Girardin MF. Tenoxicam, a non-steroid anti-inflammatory drug, is unable to increase the response rate in patients with chronic hepatitis C treated by alpha interferon. Hepatology 1998; 27:862-7. [PMID: 9500719 DOI: 10.1002/hep.510270332] [Citation(s) in RCA: 26] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 01/09/2023]
Abstract
The purpose of this study is to compare a combination of interferon (IFN)-alpha2a (Roferon) + Tenoxicam with IFN-alpha2a alone in the treatment of chronic hepatitis C. This prospective, randomized double-blind study included 149 patients, all of whom were diagnosed with active chronic hepatitis C but non-cirrhotic (ALT > or = 1.5 upper limit of normal, anti-hepatitis C virus (HCV) positive by enzyme-linked immunosorbant assay2 and RIBA3). The patients were randomized in two groups, as follows: G1 (n = 76): IFNalpha2a 3 million units times per week during 6 months + placebo; and G2 (n = 73): IFNalpha2a 3 million units three times per week + Tenoxicam (20 mg/day) during 6 months. Alanine aminotransferase (ALT) and HCV RNA were determined before and at months 6 and 12 of treatment. 2'5' oligoadenylate synthetase activity (2'5' AS) was dosed in mononuclear cells before and at 3-month treatment intervals in 28 patients. Liver biopsy was performed before and 6 months after the end of therapy. Parameters were similar before therapy for both groups. Biochemical and virological responses were similar for both groups at month 6 (49.3% vs. 42.9% and 43.3% vs. 38.3%, respectively) and month 12 (28.3% vs. 23.8% and 17.2% vs. 17.5%, respectively). HCV RNA level significantly decreased in both groups at month 6, with no difference whatever the therapy; however, the HCV RNA level returned to initial values at month 12 and was the only significant prognostic factor of a sustained response. No peak of 2'5' AS activity was observed during treatment in patients with dual therapy. A histological improvement was also noted in both groups without difference, regardless of therapy. The percentage of adverse events was identical for both groups. Paracetamol intake, assessed in 80 patients, was 49.1 g per 6 months in the G1 group and 22.5 g per 6 months in the G2 group (not significant). In conclusion, the non-steroid anti-inflammatory drug, Tenoxicam, does not increase IFNalpha efficacy in the treatment of chronic hepatitis C. This combination is well tolerated and partially lowers Paracetamol intake, but not preexisting alpha-IFN adverse events.
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Affiliation(s)
- J P Zarski
- Department of Gastroenterology and Hepatology Grenoble, France
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Tang DG, Porter AT, Honn KV. Critical role of arachidonate lipoxygenases in regulating apoptosis. ADVANCES IN EXPERIMENTAL MEDICINE AND BIOLOGY 1997; 407:405-11. [PMID: 9321984 DOI: 10.1007/978-1-4899-1813-0_61] [Citation(s) in RCA: 6] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 02/05/2023]
Affiliation(s)
- D G Tang
- Department of Radiation Oncology, Wayne State University, Detroit, Michigan 48202, USA
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Wolf FI, Di Francesco A, Covacci V, Cittadini A. Regulation of magnesium efflux from rat spleen lymphocytes. Arch Biochem Biophys 1997; 344:397-403. [PMID: 9264554 DOI: 10.1006/abbi.1997.0199] [Citation(s) in RCA: 25] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/05/2023]
Abstract
Rat spleen lymphocytes (RSL) incubated at 37 degrees C in Mg-free medium (O-trans conditions) exibited Mg2+ efflux with apparent velocity of 0.2 nmol/mg protein/min. After 30 min, this process accounted for the mobilization of about 15% of cell total Mg2+. Half of the Mg2+ efflux depended on extracellular Na+ and was stimulated by cAMP. IFN-alpha significantly enhanced Mg2+ efflux under O-trans conditions as well as in the presence of physiological extracellular Mg2+. Pretreatment of RSL with indomethacin completely abolished IFN-alpha-induced Mg2+ efflux, suggesting a crucial role for cyclooxygenase-dependent arachidonate metabolism. On the other hand, pretreatment of RSL with the PKA inhibitor (Rp)8-Br-cAMPS prevented IFN-alpha stimulation of Mg2+ efflux, indicating the involvement of cAMP. Consistently, both IFN-alpha and exogenous PGE1 increased cAMP from 50 to 125 pmol/mg protein. Altogether these results show that IFN-alpha stimulates Mg2+ efflux by activating arachidonate metabolism and synthesis of prostaglandins. By influencing adenylcyclase activity, PGEs can eventually promote cAMP-dependent Mg2+ efflux, possibly through the activity of a Na-Mg antiport. In RSL, therefore, magnesium movements can be under the control of IFN-alpha and, perhaps, of other cytokines, suggesting the involvement of Mg2+ in cell response to receptor-mediated stimuli.
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Affiliation(s)
- F I Wolf
- Institute of General Pathology and Giovanni XXIII Cancer Research Centre, Università Cattolica del Sacro Cuore, Rome, Italy
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Herrmann JL, Menter DG, Beham A, von Eschenbach A, McDonnell TJ. Regulation of lipid signaling pathways for cell survival and apoptosis by bcl-2 in prostate carcinoma cells. Exp Cell Res 1997; 234:442-51. [PMID: 9260915 DOI: 10.1006/excr.1997.3653] [Citation(s) in RCA: 33] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/05/2023]
Abstract
Compelling evidence indicates that activation of the JNK/SAPK signaling pathway is obligatory for apoptosis induction by multiple cell stresses that activate the sphingomyelin cycle. Moreover, ectopic expression of bcl-2 can impair apoptosis signaling by most of the cell stresses that activate the ceramide/JNK pathway. Here we show that enforced expression of bcl-2 protects prostate carcinoma cells against the induction of apoptosis by exogenous C2-ceramide. Moreover, enforced bcl-2 expression blocked the capacity of C2-ceramide to activate JNK1, indicating bcl-2 functions at the level of JNK1 or upstream of JNK1 in the ceramide/JNK pathway. The contribution of bcl2 to the regulation of the arachidonate pathway for prostate carcinoma cell survival was also investigated using highly selective inhibitors of arachidonate metabolism. Our results indicate bcl-2 can protect cells against diminished availability of arachidonic acid, 12-HETE, and 15-HETE. Finally, arachidonic acid substantially suppresses the induction of apoptosis by C2-ceramide, providing evidence for the opposing influences of these lipid signaling pathways in the mediation of prostate carcinoma cell survival. These results provide evidence for opposing influences of the ceramide and arachidonate signaling pathways in the mediation of cell death and cell survival, respectively, in prostate carcinoma cells and suggest a dual role for bcl-2 in this context.
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Affiliation(s)
- J L Herrmann
- Department of Molecular Pathology, The University of Texas, M. D. Anderson Cancer Center, Houston 77030, USA
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Massey JB, Bick DH, Pownall HJ. Spontaneous transfer of monoacyl amphiphiles between lipid and protein surfaces. Biophys J 1997; 72:1732-43. [PMID: 9083677 PMCID: PMC1184367 DOI: 10.1016/s0006-3495(97)78819-2] [Citation(s) in RCA: 68] [Impact Index Per Article: 2.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/04/2023] Open
Abstract
The kinetics of transfer of natural and fluorescent nonesterified fatty acids (NEFA) and lysolecithins (lysoPC) from phospholipid and protein surfaces were measured. The kinetics of transfer of 12-(1-pyrenyl)dodecanoic acid, from liquid crystalline and gel phase single unilamellar phospholipid vesicles, very low, low, and high density lipoproteins, human serum albumin, and rat liver fatty acid-binding protein, were first-order and characterized by similar rate constants. The halftimes (t1/2) of NEFA transfer from lipids and proteins were dependent on the acyl chain structure according to log t1/2 = -0.62n + 0.59m + 12.0, where n and m, respectively, are the numbers of carbon atoms and double bonds. The structure of the donor surface had a measurable but smaller effect on transfer rates. The kinetics of NEFA and lysoPC transfer are slow relative to the lipolytic processes that liberate them. Therefore, one would predict a transient accumulation of NEFA and lysoPC during lipolysis and an attendant modulation of many metabolic processes within living cells and within the plasma compartment of blood. These data will be useful in the refinement of current models of membrane and lipoprotein function and in the selection of fluorescent NEFA analogs for studying transport in living cells.
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Affiliation(s)
- J B Massey
- Department of Medicine, Baylor College of Medicine, Houston, Texas, USA
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Arachidonic Acid Mediates Interferon-γ–Induced Sphingomyelin Hydrolysis and Monocytic Marker Expression in HL-60 Cell Line. Blood 1997. [DOI: 10.1182/blood.v89.1.81] [Citation(s) in RCA: 32] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/22/2023] Open
Abstract
AbstractThe biochemical signaling mechanisms involved in transducing the effects of interferon-γ (IFN-γ) on human leukemia-derived HL-60 cell differentiation are not completely understood. Recent studies established the existence of a sphyngomyelin (SM) cycle that operates in response to the action of IFN-γ on HL-60 cells, but the mechanisms by which IFN-γ induces the SM hydrolysis remain unexplored. In this study, biochemical events mediating IFN-γ effects on SM turnover and their specificity and role in HL-60 differentiation were investigated. The activation of the SM cycle by IFN-γ occurred rapidly, with a decrease of approximately 20% in the SM level observed after 60 minutes with a concomitant increase in ceramide level. Treatment of HL-60 cells with IFN-γ did not influence the 1,2-diacylglycerol concentration, intracellular Ca2+ concentration, or phospholipase D activity. IFN-γ stimulated a rapid release of arachidonic acid (AA) from HL-60 cells; the effect was abolished by the pretreatment of cells with pertussis toxin, suggesting a role for a pertussis-toxin–sensitive G protein in IFN-γ–mediated activation of phospholipase A2 (PLA2 ). At 4 to 120 hours after the stimulation of the cells with IFN-γ, a significant increase in the particulate and soluble PLA2 activity was observed, corresponding to an increase in the level of immunoreactive cPLA2 in both cytosol and membrane fractions. The treatment of cells with tyrosine kinase inhibitor herbimycin A completely abolished the effect of IFN-γ on PLA2 activity in membrane and cytosolic fractions, but had no effect on IFN-γ–mediated early AA release suggesting dual mechanism of PLA2 activation. Melittin, potent activator of PLA2 , and AA mimicked the effect of IFN-γ on SM hydrolysis. Pretreatment of HL-60 cells with the PLA2 inhibitor, bromophenacyl bromide (BPB), or pertussis toxin abolished the effect of IFN-γ on SM hydrolysis; exogenous addition of AA overcame the effects of BPB and pertussis toxin. Long-term exposure (5 days) of HL-60 cells to IFN-γ caused an increase in nitroblue tetrazolium (NBT)-reducing and nonspecific esterase (NSE) activity and induced expression of FcγRI (CD64) without significant effects on cell number, adherence, or fagocytic activity. The treatment of cells with AA or melittin induced NBT, NSE, and CD64 expression to the level similar to that observed with IFN-γ, and no further increase was observed with the combination of IFN-γ and AA or IFN-γ and melittin. Treatment of HL-60 cells with indomethacin, an inhibitor of cyclo-oxygenase, and nordihydroguaiaretic acid (NDGA), an inhibitor of lipoxygenase, had no effects on IFN-γ–mediated induction of CD64 expression. These studies indicate a key role for the phospholipase A2/AA pathway, as an early biochemical signal elicited by the occupation of IFN-γ–receptor, in mediating IFN-γ induction of the SM cycle and phenotypic changes associated with differentiation of HL-60 along monocytic lineage.
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Arachidonic Acid Mediates Interferon-γ–Induced Sphingomyelin Hydrolysis and Monocytic Marker Expression in HL-60 Cell Line. Blood 1997. [DOI: 10.1182/blood.v89.1.81.81_81_91] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/20/2022] Open
Abstract
The biochemical signaling mechanisms involved in transducing the effects of interferon-γ (IFN-γ) on human leukemia-derived HL-60 cell differentiation are not completely understood. Recent studies established the existence of a sphyngomyelin (SM) cycle that operates in response to the action of IFN-γ on HL-60 cells, but the mechanisms by which IFN-γ induces the SM hydrolysis remain unexplored. In this study, biochemical events mediating IFN-γ effects on SM turnover and their specificity and role in HL-60 differentiation were investigated. The activation of the SM cycle by IFN-γ occurred rapidly, with a decrease of approximately 20% in the SM level observed after 60 minutes with a concomitant increase in ceramide level. Treatment of HL-60 cells with IFN-γ did not influence the 1,2-diacylglycerol concentration, intracellular Ca2+ concentration, or phospholipase D activity. IFN-γ stimulated a rapid release of arachidonic acid (AA) from HL-60 cells; the effect was abolished by the pretreatment of cells with pertussis toxin, suggesting a role for a pertussis-toxin–sensitive G protein in IFN-γ–mediated activation of phospholipase A2 (PLA2 ). At 4 to 120 hours after the stimulation of the cells with IFN-γ, a significant increase in the particulate and soluble PLA2 activity was observed, corresponding to an increase in the level of immunoreactive cPLA2 in both cytosol and membrane fractions. The treatment of cells with tyrosine kinase inhibitor herbimycin A completely abolished the effect of IFN-γ on PLA2 activity in membrane and cytosolic fractions, but had no effect on IFN-γ–mediated early AA release suggesting dual mechanism of PLA2 activation. Melittin, potent activator of PLA2 , and AA mimicked the effect of IFN-γ on SM hydrolysis. Pretreatment of HL-60 cells with the PLA2 inhibitor, bromophenacyl bromide (BPB), or pertussis toxin abolished the effect of IFN-γ on SM hydrolysis; exogenous addition of AA overcame the effects of BPB and pertussis toxin. Long-term exposure (5 days) of HL-60 cells to IFN-γ caused an increase in nitroblue tetrazolium (NBT)-reducing and nonspecific esterase (NSE) activity and induced expression of FcγRI (CD64) without significant effects on cell number, adherence, or fagocytic activity. The treatment of cells with AA or melittin induced NBT, NSE, and CD64 expression to the level similar to that observed with IFN-γ, and no further increase was observed with the combination of IFN-γ and AA or IFN-γ and melittin. Treatment of HL-60 cells with indomethacin, an inhibitor of cyclo-oxygenase, and nordihydroguaiaretic acid (NDGA), an inhibitor of lipoxygenase, had no effects on IFN-γ–mediated induction of CD64 expression. These studies indicate a key role for the phospholipase A2/AA pathway, as an early biochemical signal elicited by the occupation of IFN-γ–receptor, in mediating IFN-γ induction of the SM cycle and phenotypic changes associated with differentiation of HL-60 along monocytic lineage.
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Scarpa S, Giuffrida A, Palumbo C, Vasaturo F, Signorelli P, Forni G, Modesti M, Ferrantini M, Belardelli F, Musiani P, Modesti A. Extracellular matrix remodelling in a murine mammary adenocarcinoma transfected with the interferon-alpha 1 gene. J Pathol 1997; 181:116-23. [PMID: 9072013 DOI: 10.1002/(sici)1096-9896(199701)181:1<116::aid-path116>3.0.co;2-x] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/04/2023]
Abstract
The rejection of interferon alpha 1 gene-transfected mammary adenocarcinoma cells (TSA-IFN alpha) injected into syngeneic BALB/c mice was accompanied by an unusual stromal reaction and marked CD8-positive T-lymphocyte involvement. To investigate the biological background of this reaction, the possibility was evaluated that an interaction between TSA-IFN alpha and stromal cells might remodel the extracellular matrix (EM). When fibroblasts were co-cultured with TSA-IFN alpha or treated with exogenous IFN alpha, there was no change in their replication rate or collagen synthesis. By contrast, their fibronectin (FN) production and release were increased, resulting in enhanced fibroblast chemotaxis. These findings were mirrored by increased FN staining in the peritumoural and tumoural areas in vivo. IFN alpha thus determines increased FN production and hence massive local recruitment and activation of fibroblasts, with a modification of the EM. The several activities of IFN alpha should thus be considered prior to its employment in clinical trials.
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Affiliation(s)
- S Scarpa
- Department of Experimental Medicine and Pathology, Universitá La Sapienza, Roma, Italy
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Venuta A, Spanò C, Laudizi L, Bettelli F, Beverelli A, Turchetto E. Essential fatty acids: the effects of dietary supplementation among children with recurrent respiratory infections. J Int Med Res 1996; 24:325-30. [PMID: 8854285 DOI: 10.1177/030006059602400402] [Citation(s) in RCA: 13] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/02/2023] Open
Abstract
The impact of dietary supplementation with essential fatty acids (EFA) on recurrent respiratory infections (RRI) in children was evaluated by means of a randomized cross-over double-blind study. Linoleic acid (596 mg/day) and alpha-linolenic acid [855 mg/day] as a commercial preparation or placebo (olive oil) were administered for two consecutive winter seasons (November to February, T0-T120) to 20 children affected by RRI, aged between 36 and 49 months. Plasma levels of n-3 and n-6 metabolites increased after the administration of EFA. The number of infective episodes, days' fever and days' absence from school were reduced significantly during the observation period (extended from T120 to T180) in children receiving EFA supplementation. Our results suggest that n-3 and n-6 polyunsaturated fatty acids may play a favourable role in the defence mechanism of these subjects.
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Affiliation(s)
- A Venuta
- Department of Gynaecology, Obstetrics and Paediatrics, University of Modena, Italy
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Abstract
Mixed cryoglobulinemia is a systemic vasculitis with clinical manifestations ranging from the characteristic benign-appearing syndrome of palpable purpura, arthrologies, and fatigue to severe vasculitis involving vital organs. A strong association of the disease with hepatitis C virus infection and the demonstration of the specific concentration of the virus in the cryoglobulins have implicated hepatitis C virus in the etiopathogenesis of the disease. The increase in illicit intravenous drug use in the past 30 years seems to have raised the occurrence in the United States of this once uncommon disease and changed the demographics: there seem to be more male intravenous drug users in their forties with the disease than women without risk factors for hepatitis C virus infection in their fifties and sixties. Pathogenesis, therapy, and the hypothesis on the etiologic role of hepatitis C virus are reviewed, and the implications of recent studies and new concepts for treatment of this often benign-appearing but deceptive and potentially life-threatening disease are discussed.
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Affiliation(s)
- V Agnello
- Lahey Hitchcock Medical Center, Burlington, Massachusetts, USA
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Toniato E, Flati V, Cifone MG, Del Grosso E, Roncaioli P, Cilenti L, Tessitore A, Lista F, Frati L, Gulino A, Martinotti S. Involvement of an arachidonic-acid-dependent pathway in the interferon-beta-mediated expression of C202 gene in Ehrlich-ascites-tumor cells. EUROPEAN JOURNAL OF BIOCHEMISTRY 1996; 235:91-6. [PMID: 8631372 DOI: 10.1111/j.1432-1033.1996.00091.x] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 02/01/2023]
Abstract
We have investigated the signal transduction mechanism of the expression of the C202 gene mediated by interferon beta (IFN-beta) in the murine Ehrlich's ascites tumor cell line. We have shown that treatment of cells with IFN-beta transiently enhances within minutes the release of free arachidonic acid through membrane phospholipase activity. Furthermore, prior treatment with either p-bromophenacyl bromide, an antagonist of both cytosolic and secretory phospholipase A2, or neomycin, which blocks phospholipase C activity, significantly decreased the activation of the murine IFN-beta-inducible gene, C202. Moreover, an increase of the expression of the C202 gene was observed after blocking of both the cyclooxygenase and lipoxygenase pathways. This suggests that further metabolism of arachidonic acid to epoxides via epoxygenase-catalysed pathways may be a mechanism by which second messengers for IFN-beta-mediated effects on C202 gene expression are generated. Taken together, these results indicate that lipids as second messengers may be important mediators in the IFN-beta-based activation of C202 gene expression.
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Affiliation(s)
- E Toniato
- Department of Experimental Medicine, University of L'Aquila, Italy
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Kalvakolanu DV, Borden EC. An overview of the interferon system: signal transduction and mechanisms of action. Cancer Invest 1996; 14:25-53. [PMID: 8597888 DOI: 10.3109/07357909609018435] [Citation(s) in RCA: 108] [Impact Index Per Article: 3.7] [Reference Citation Analysis] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/31/2023]
Affiliation(s)
- D V Kalvakolanu
- Department of Microbology, University of Maryland School of Medicine, Baltimore, Maryland, USA
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