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Muthuraman A, Ramesh M, Mustaffa F, Nadeem A, Nishat S, Paramakrishnan N, Lim KG. In Silico and In Vitro Methods in the Characterization of Beta-Carotene as Pharmaceutical Material via Acetylcholine Esterase Inhibitory Actions. Molecules 2023; 28:molecules28114358. [PMID: 37298835 DOI: 10.3390/molecules28114358] [Citation(s) in RCA: 5] [Impact Index Per Article: 2.5] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/03/2023] [Revised: 05/17/2023] [Accepted: 05/24/2023] [Indexed: 06/12/2023] Open
Abstract
Molecular docking is widely used in the assessment of the therapeutic potential of pharmaceutical agents. The binding properties of beta-carotene (BC) to acetylcholine esterase (AChE) proteins were characterized using the molecular docking method. The mechanism of AChE inhibition was assessed by an experimental in vitro kinetic study. In addition, the role of BC action was tested by the zebrafish embryo toxicity test (ZFET). The results of the docking ability of BC to AChE showed significant ligand binding mode. The kinetic parameter, i.e., the low AICc value shown as the compound was the competitive type of inhibition of AChE. Further, BC also showed mild toxicity at a higher dose (2200 mg/L) in ZFET assessment with changes in biomarkers. The LC50 value of BC is 1811.94 mg/L. Acetylcholine esterase (AChE) plays a pivotal role in the hydrolysis of acetylcholine, which leads to the development of cognitive dysfunction. BC possesses the regulation of acetylcholine esterase (AChE) and acid phosphatase (AP) activity to prevent neurovascular dysfunction. Therefore, the characterization of BC could be used as a pharmaceutical agent for the treatment of cholinergic neurotoxicity-associated neurovascular disorders such as developmental toxicity, vascular dementia, and Alzheimer's disease due to its AChE and AP inhibitory actions.
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Affiliation(s)
| | - Muthusamy Ramesh
- Department of Pharmaceutical Analysis, Omega College of Pharmacy, Hyderabad 501301, India
| | - Fazlina Mustaffa
- Pharmacology Unit, Faculty of Pharmacy, AIMST University, Bedong 08100, Malaysia
| | - Ahmed Nadeem
- Department of Pharmacology and Toxicology, College of Pharmacy, King Saud University, Riyadh 11451, Saudi Arabia
| | - Shamama Nishat
- Comprehensive Cancer Center, Wexner Medical Centre, Ohio State University, Columbus, OH 43210, USA
| | - Nallupillai Paramakrishnan
- Department of Pharmacognosy, JSS College of Pharmacy, Mysore, JSS Academy of Higher Education and Research, Mysore 570015, India
| | - Khian Giap Lim
- Pharmacology Unit, Faculty of Pharmacy, AIMST University, Bedong 08100, Malaysia
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Liu Y, Myojin T, Li K, Kurita A, Seto M, Motoyama A, Liu X, Satoh A, Munemasa S, Murata Y, Nakamura T, Nakamura Y. A Major Intestinal Catabolite of Quercetin Glycosides, 3-Hydroxyphenylacetic Acid, Protects the Hepatocytes from the Acetaldehyde-Induced Cytotoxicity through the Enhancement of the Total Aldehyde Dehydrogenase Activity. Int J Mol Sci 2022; 23:ijms23031762. [PMID: 35163684 PMCID: PMC8836260 DOI: 10.3390/ijms23031762] [Citation(s) in RCA: 8] [Impact Index Per Article: 2.7] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/24/2022] [Revised: 01/31/2022] [Accepted: 02/01/2022] [Indexed: 12/20/2022] Open
Abstract
Aldehyde dehydrogenases (ALDHs) are the major enzyme superfamily for the aldehyde metabolism. Since the ALDH polymorphism leads to the accumulation of acetaldehyde, we considered that the enhancement of the liver ALDH activity by certain food ingredients could help prevent alcohol-induced chronic diseases. Here, we evaluated the modulating effects of 3-hydroxyphenylacetic acid (OPAC), the major metabolite of quercetin glycosides, on the ALDH activity and acetaldehyde-induced cytotoxicity in the cultured cell models. OPAC significantly enhanced the total ALDH activity not only in mouse hepatoma Hepa1c1c7 cells, but also in human hepatoma HepG2 cells. OPAC significantly increased not only the nuclear level of aryl hydrocarbon receptor (AhR), but also the AhR-dependent reporter gene expression, though not the nuclear factor erythroid-2-related factor 2 (Nrf2)-dependent one. The pretreatment of OPAC at the concentration required for the ALDH upregulation completely inhibited the acetaldehyde-induced cytotoxicity. Silencing AhR impaired the resistant effect of OPAC against acetaldehyde. These results strongly suggested that OPAC protects the cells from the acetaldehyde-induced cytotoxicity, mainly through the AhR-dependent and Nrf2-independent enhancement of the total ALDH activity. Our findings suggest that OPAC has a protective potential in hepatocyte models and could offer a new preventive possibility of quercetin glycosides for targeting alcohol-induced chronic diseases.
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Affiliation(s)
- Yujia Liu
- School of Biological Engineering, Dalian Polytechnic University, Dalian 116034, China;
- School of Food Science and Technology, Dalian Polytechnic University, Dalian 116034, China; (K.L.); (X.L.)
- Graduate School of Environmental and Life Science, Okayama University, Okayama 700-8530, Japan; (T.M.); (A.K.); (M.S.); (A.M.); (S.M.); (Y.M.); (T.N.)
| | - Takumi Myojin
- Graduate School of Environmental and Life Science, Okayama University, Okayama 700-8530, Japan; (T.M.); (A.K.); (M.S.); (A.M.); (S.M.); (Y.M.); (T.N.)
| | - Kexin Li
- School of Food Science and Technology, Dalian Polytechnic University, Dalian 116034, China; (K.L.); (X.L.)
- Graduate School of Environmental and Life Science, Okayama University, Okayama 700-8530, Japan; (T.M.); (A.K.); (M.S.); (A.M.); (S.M.); (Y.M.); (T.N.)
| | - Ayuki Kurita
- Graduate School of Environmental and Life Science, Okayama University, Okayama 700-8530, Japan; (T.M.); (A.K.); (M.S.); (A.M.); (S.M.); (Y.M.); (T.N.)
| | - Masayuki Seto
- Graduate School of Environmental and Life Science, Okayama University, Okayama 700-8530, Japan; (T.M.); (A.K.); (M.S.); (A.M.); (S.M.); (Y.M.); (T.N.)
| | - Ayano Motoyama
- Graduate School of Environmental and Life Science, Okayama University, Okayama 700-8530, Japan; (T.M.); (A.K.); (M.S.); (A.M.); (S.M.); (Y.M.); (T.N.)
| | - Xiaoyang Liu
- School of Food Science and Technology, Dalian Polytechnic University, Dalian 116034, China; (K.L.); (X.L.)
- Graduate School of Environmental and Life Science, Okayama University, Okayama 700-8530, Japan; (T.M.); (A.K.); (M.S.); (A.M.); (S.M.); (Y.M.); (T.N.)
| | - Ayano Satoh
- Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University, Okayama 700-8530, Japan;
| | - Shintaro Munemasa
- Graduate School of Environmental and Life Science, Okayama University, Okayama 700-8530, Japan; (T.M.); (A.K.); (M.S.); (A.M.); (S.M.); (Y.M.); (T.N.)
| | - Yoshiyuki Murata
- Graduate School of Environmental and Life Science, Okayama University, Okayama 700-8530, Japan; (T.M.); (A.K.); (M.S.); (A.M.); (S.M.); (Y.M.); (T.N.)
| | - Toshiyuki Nakamura
- Graduate School of Environmental and Life Science, Okayama University, Okayama 700-8530, Japan; (T.M.); (A.K.); (M.S.); (A.M.); (S.M.); (Y.M.); (T.N.)
| | - Yoshimasa Nakamura
- Graduate School of Environmental and Life Science, Okayama University, Okayama 700-8530, Japan; (T.M.); (A.K.); (M.S.); (A.M.); (S.M.); (Y.M.); (T.N.)
- Correspondence: ; Tel.: +81-86-251-8300; Fax: +81-86-251-8388
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Mozaheb N, Arefian E, Aliyan A, Amoozegar MA. Induction of the antioxidant defense system using long-chain carotenoids extracted from extreme halophilic archaeon, Halovenus aranensis. Int Microbiol 2021; 25:165-175. [PMID: 34487298 DOI: 10.1007/s10123-021-00198-6] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/01/2021] [Revised: 07/05/2021] [Accepted: 07/28/2021] [Indexed: 11/28/2022]
Abstract
The field of microbial pigments is an emerging area in natural products science. Carotenoids form a major class of such pigments and are found to be diversely synthesized by microorganisms that reside in hypersaline ecosystems to provide resistance against oxidative stress. Human cells can benefit from compounds such as carotenoids as antioxidant agents through either their capability to quench free radicals or their effect on promoting the antioxidant defense pathway. In this study, the antioxidant effectiveness of carotenoid extract from an extremely halophilic archaeon Halovenus aranensis strain EB27T has been evaluated using different approaches. Finally, the ability of the extracted pigment to induce the antioxidant defense pathway of human primary skin fibroblast cells was studied. Hvn. aranensis carotenoid extract exhibited strong effectiveness such that at 2 µg/ml, the carotenoid extract fully neutralized the oxidative stress of hydrogen peroxide at its EC50 based on MTT assay. Results from real-time PCR of relevant genes, luciferase bioreporter of oxidative stress, and the western blot analysis further confirmed the antioxidant capability of the carotenoids. It was also shown the carotenoid extract had more antioxidant activity compared to β-carotene the same concentration. Results suggest the carotenoid extract from this archaeon to have high potential for clinical and industrial applications.
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Affiliation(s)
- Negar Mozaheb
- Cellular & Molecular Pharmacology Unit (FACM), Université Catholique de Louvain (UCL), Louvain Drug Research Institute (LDRI), 1200, Brussels, Belgium.,Department of Microbiology, School of Biology and Center of Excellence in Phylogeny of Living Organisms, College of Science, University of Tehran, 1417466191, Tehran, Iran
| | - Ehsan Arefian
- Department of Microbiology, School of Biology and Center of Excellence in Phylogeny of Living Organisms, College of Science, University of Tehran, 1417466191, Tehran, Iran.
| | - Amir Aliyan
- Pasargad Institute for Advanced Innovative Solutions (PIAIS), Tehran, 1991633361, Iran.,Khatam University, Tehran, 1991633356, Iran
| | - Mohammad Ali Amoozegar
- Department of Microbiology, School of Biology and Center of Excellence in Phylogeny of Living Organisms, College of Science, University of Tehran, 1417466191, Tehran, Iran.
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Application of extracted β-glucan from oat for β-carotene encapsulation. Journal of Food Science and Technology 2020; 58:2641-2650. [PMID: 34194099 DOI: 10.1007/s13197-020-04770-2] [Citation(s) in RCA: 8] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Subscribe] [Scholar Register] [Revised: 08/26/2020] [Accepted: 08/28/2020] [Indexed: 10/23/2022]
Abstract
Abstract The cell walls of cereals are rich sources of polysaccharide β-glucan. In this study, the β-glucan was extracted from oat bran using the hot-water extraction method and dried in a pure powder form. The concentration of the β-glucan in the extract was determined using the l-cysteine sulfuric acid method. The results showed that the yield of β-glucan using the hot-water extraction method is the highest compared to its yield achieved by enzymatic, acid, and alkaline methods. In this paper, the usage of the β-glucan as a coating material for a water-insoluble carotenoid is considered. This study demonstrates for the first time the encapsulation of β-carotene with modified octanoic acid β-glucan. It implements to obtain a stable encapsulated polysaccharide-carotenoid system, which has been studied by a set of physicochemical methods and a cytotoxic analysis was performed on the HCT-116 cell line. The SEM image of the resulting encapsulated system is perfectly correlated with the DLS data, which has determined the size of MG capsules at 200 nm. The cytotoxic analysis demonstrates that the cell viability was more than 70%, which indicates its potential using in the food industry. Graphic abstract
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Shi J, Wang X, Lyu L, Jiang H, Zhu HJ. Comparison of protein expression between human livers and the hepatic cell lines HepG2, Hep3B, and Huh7 using SWATH and MRM-HR proteomics: Focusing on drug-metabolizing enzymes. Drug Metab Pharmacokinet 2018; 33:133-140. [PMID: 29610054 DOI: 10.1016/j.dmpk.2018.03.003] [Citation(s) in RCA: 43] [Impact Index Per Article: 6.1] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/01/2018] [Revised: 02/07/2018] [Accepted: 02/13/2018] [Indexed: 12/25/2022]
Abstract
Human hepatic cell lines are widely used as an in vitro model for the study of drug metabolism and liver toxicity. However, the validity of this model is still a subject of debate because the expressions of various proteins in the cell lines, including drug-metabolizing enzymes (DMEs), can differ significantly from those in human livers. In the present study, we first conducted an untargeted proteomics analysis of the microsomes of the cell lines HepG2, Hep3B, and Huh7, and compared them to human livers using a sequential window acquisition of all theoretical mass spectra (SWATH) method. Furthermore, high-resolution multiple reaction monitoring (MRM-HR), a targeted proteomic approach, was utilized to compare the expressions of pre-selected DMEs between human livers and the cell lines. In general, the SWATH quantifications were in good agreement with the MRM-HR analysis. Over 3000 protein groups were quantified in the cells and human livers, and the proteome profiles of human livers significantly differed from the cell lines. Among the 101 DMEs quantified with MRM-HR, most were expressed at substantially lower levels in the cell lines. Thus, appropriate caution must be exercised when using these cell lines for the study of hepatic drug metabolism and toxicity.
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Affiliation(s)
- Jian Shi
- Department of Clinical Pharmacy, University of Michigan, Ann Arbor, MI 48109, United States
| | - Xinwen Wang
- Department of Clinical Pharmacy, University of Michigan, Ann Arbor, MI 48109, United States
| | - Lingyun Lyu
- Department of Biostatistics, University of Pittsburgh, Pittsburgh, PA 15213, United States
| | - Hui Jiang
- Department of Biostatistics, University of Michigan, Ann Arbor, MI 48109, United States
| | - Hao-Jie Zhu
- Department of Clinical Pharmacy, University of Michigan, Ann Arbor, MI 48109, United States.
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Želježić D, Mladinić M, Žunec S, Lucić Vrdoljak A, Kašuba V, Tariba B, Živković T, Marjanović AM, Pavičić I, Milić M, Rozgaj R, Kopjar N. Cytotoxic, genotoxic and biochemical markers of insecticide toxicity evaluated in human peripheral blood lymphocytes and an HepG2 cell line. Food Chem Toxicol 2016; 96:90-106. [DOI: 10.1016/j.fct.2016.07.036] [Citation(s) in RCA: 29] [Impact Index Per Article: 3.2] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/23/2015] [Revised: 07/27/2016] [Accepted: 07/29/2016] [Indexed: 10/21/2022]
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Kanyong P, Hughes G, Pemberton RM, Jackson SK, Hart JP. Amperometric Screen-Printed Galactose Biosensor for Cell Toxicity Applications. ANAL LETT 2015. [DOI: 10.1080/00032719.2015.1070166] [Citation(s) in RCA: 9] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/16/2022]
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8
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Sansone RA, Sansone LA. Carrot man: a case of excessive beta-carotene ingestion. Int J Eat Disord 2012; 45:816-8. [PMID: 22431270 DOI: 10.1002/eat.22015] [Citation(s) in RCA: 8] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Accepted: 02/11/2012] [Indexed: 11/07/2022]
Abstract
In this case report, the authors describe a 48-year-old male who complained to his primary care physician of abdominal discomfort and yellow/orange skin discoloration. Physical examination was normal except for some mild mid-abdominal discomfort (no observed skin color changes). An abdominal CT scan indicated a colon that was full of stool. Laboratory studies indicated elevated liver enzymes. Upon further questioning, the patient reported ingesting 6-7 pounds of carrots per week to facilitate his dieting effort. The patient was diagnosed with constipation, hypercarotinemia, and possible vitamin A toxicity. Following the cessation of excessive carrot ingestion, his liver enzymes normalized within 1 month.
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Affiliation(s)
- Randy A Sansone
- Department of Psychiatry, Wright State University School of Medicine, Dayton, OH, USA.
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9
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Scientific Opinion on the re‐evaluation of mixed carotenes (E 160a (i)) and beta‐carotene (E 160a (ii)) as a food additive. EFSA J 2012. [DOI: 10.2903/j.efsa.2012.2593] [Citation(s) in RCA: 27] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/11/2022] Open
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10
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Assessment of tryptophol genotoxicity in four cell lines in vitro: a pilot study with alkaline comet assay. Arh Hig Rada Toksikol 2011; 62:41-9. [PMID: 21421532 DOI: 10.2478/10004-1254-62-2011-2090] [Citation(s) in RCA: 11] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/20/2022] Open
Abstract
Tryptophol is an aromatic alcohol and secondary metabolite of the opportunistic fungus Candida albicans. Although its toxicity profile at cell level has been poorly investigated, recent data point to cytotoxic, cytostatic, and genotoxic effects in lymphocytes and the induction of apoptosis in leukaemic blood monocytes. In this pilot study we evaluated the genotoxicity of tryptophol in vitro on four permanent cell lines of animal and human origin: ovary cells, alveolar epithelium, liver cells, and blood monocytes using the alkaline comet assay. We selected cells that might be principal targets of tryptophol and other low-molecular geno(toxins) secreted by Candida albicans during host invasion. Our results suggest that tryptophol applied in vitro at 2 mmol L(-1) for 24 h damages DNA in HepG2, A549 and THP-1 cells, obviously due to bioactivation and/or decomposition of the parent compound, which results in the formation of more genotoxic compound(s) and production of reactive species that additionally damage DNA. On the other hand, notably lower levels of primary DNA damage were recorded in CHO cells, which lack metabolic activity. Future studies with tryptophol should look further into mechanisms involved in its toxic action and should focus on other cell types prone to infection with Candida spp. such as vaginal epithelial cells or keratinocytes of human origin.
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Hong J, Kandasamy K, Marimuthu M, Choi CS, Kim S. Electrical cell-substrate impedance sensing as a non-invasive tool for cancer cell study. Analyst 2010; 136:237-45. [PMID: 20963234 DOI: 10.1039/c0an00560f] [Citation(s) in RCA: 110] [Impact Index Per Article: 7.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/21/2022]
Abstract
Cell-substrate interactions are investigated in a number of studies for drug targets including angiogenesis, arteriosclerosis, chronic inflammatory diseases and carcinogenesis. One characteristic of malignant cancerous cells is their ability to invade tissue. Cell adhesion and cytoskeletal activity have served as valuable indicators for understanding the cancer cell behaviours, such as proliferation, migration and invasion. This review focuses on bio-impedance based measurement for monitoring the behaviours in real time and without using labels. Electric cell-substrate impedance sensing (ECIS) provides rich information about cell-substrate interactions, cell-cell communication and cell adhesion. High sensitivity of the ECIS method allows for observing events down to single-cell level and achieving nanoscale resolution of cell-substrate distances. Recently, its miniaturization and integration with fluorescent detection techniques have been highlighted as a new tool to deliver a high-content platform for anticancer drug development.
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Affiliation(s)
- Jongin Hong
- Department of Chemistry and Institute of Biomedical Engineering, Imperial College London, South Kensington Campus, London, SW7 2AZ United Kingdom.
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Wright M, Albanes D. Potential Adverse Effects of beta-Carotene Supplementation in Cigarette Smokers and Heavier Drinkers. ACTA ACUST UNITED AC 2010. [DOI: 10.1201/9780203026649.ch25] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 03/27/2023]
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13
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Acetaldehyde-induced mitochondrial dysfunction sensitizes hepatocytes to oxidative damage. Cell Biol Toxicol 2009; 25:599-609. [PMID: 19137438 DOI: 10.1007/s10565-008-9115-5] [Citation(s) in RCA: 63] [Impact Index Per Article: 3.9] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/12/2008] [Accepted: 12/15/2008] [Indexed: 12/28/2022]
Abstract
Acetaldehyde (Ac), the main metabolite of ethanol oxidation, is a very reactive compound involved in alcohol-induced liver damage. In the present work, we studied the effect of Ac in mitochondria functionality. Mitochondria from Wistar rats were isolated and treated with Ac. Ac decreased respiratory control by 50% which was associated with a decrease in adenosine triphosphate content (28.5%). These results suggested that Ac could be inducing changes in cell redox status. We determined protein oxidation, superoxide dismutase (SOD) activity, and glutathione ratio, indicating that Ac induced an enhanced oxidation of proteins and a decrease in SOD activity (90%) and glutathione/oxidized GSH ratio (36%). The data suggested that Ac-induced oxidative stress mediated by mitochondria dysfunction can lead to cell sensitization and to a second oxidative challenge. We pretreated hepatocytes with Ac followed by treatment with antimycin A, and this experiment revealed a noticeable decrease in cell viability, determined by neutral red assay, in comparison with cells treated with Ac alone. Our data demonstrate that Ac impairs mitochondria functionality generating oxidative stress that sensitizes cells to a second damaging signal contributing to the development of alcoholic liver disease.
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Kosalec I, Šafranić A, Pepeljnjak S, Bačun-Družina V, Ramić S, Kopjar N. Genotoxicity of Tryptophol in a Battery of Short-Term Assays on Human White Blood Cells in vitro. Basic Clin Pharmacol Toxicol 2008; 102:443-52. [DOI: 10.1111/j.1742-7843.2007.00204.x] [Citation(s) in RCA: 8] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/23/2023]
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Kun Y, Ssonko Lule U, Xiao-Lin D. Lycopene: Its Properties and Relationship to Human Health. FOOD REVIEWS INTERNATIONAL 2006. [DOI: 10.1080/87559120600864753] [Citation(s) in RCA: 33] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 10/24/2022]
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Yeon JH, Park JK. Cytotoxicity test based on electrochemical impedance measurement of HepG2 cultured in microfabricated cell chip. Anal Biochem 2005; 341:308-15. [PMID: 15907877 DOI: 10.1016/j.ab.2005.03.047] [Citation(s) in RCA: 66] [Impact Index Per Article: 3.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/07/2005] [Indexed: 10/25/2022]
Abstract
This paper presents the use of electrochemical impedance measurement on a cell chip to monitor cell growth as a consequence of treatment with potentially cytotoxic agents. The cell chip consists of an eight-well cell culture chamber incorporated with a three-electrode system on each well. The gold electrode for impedance measurements is fabricated by sputtering on polycarbonate film. Human hepatocellular carcinoma cell (HepG2) is adapted to cytotoxicity test using the cell chip. Although the relatively small quantity of cells on the electrode has been measured indirectly, the cell chip can monitor toxic effects on the HepG2 cells cultured in the cell chip continuously and detect cellular behavior without multiple reagents. The cells in the stationary phase after plating are used for the cytotoxicity experiment and the impedance is decreased after treatments with several toxicants, such as tamoxifen and menadione, indicating the detachment of dead cells. These results reveal that the microfabricated cell chip system provides an easy and real-time monitoring method for cytotoxicity test.
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Affiliation(s)
- Ju Hun Yeon
- Department of BioSystems, Korea Advanced Institute of Science and Technology (KAIST), 373-1 Guseong-dong, Yuseong-gu, Daejeon 305-701, Korea
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Moreb JS, Gabr A, Vartikar GR, Gowda S, Zucali JR, Mohuczy D. Retinoic acid down-regulates aldehyde dehydrogenase and increases cytotoxicity of 4-hydroperoxycyclophosphamide and acetaldehyde. J Pharmacol Exp Ther 2005; 312:339-45. [PMID: 15470086 DOI: 10.1124/jpet.104.072496] [Citation(s) in RCA: 66] [Impact Index Per Article: 3.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/22/2022] Open
Abstract
Multiple prior studies have identified aldehyde dehydrogenases (ALDH) that are capable of oxidizing retinal to retinoic acid. In this study, we test the hypothesis that the accumulation of intracellular retinoic acid may lead to the suppression of ALDH expression and thus increase cytotoxicity to 4-hydroperoxycyclophosphamide (4-HC) in vitro. Mainly A549, but also other lung cancer cell lines, were used in our experiments, with the former having high levels of two ALDH isozymes expressed. Dose-response and time-course experiments were performed by incubating the cells with all-trans retinoic acid (ATRA) as well as other commercially available retinoids. The results show that incubation of A549 cells with any of the retinoids at pharmacologic doses for > or =48 h results in a significant decrease in ALDH-1A1 and ALDH-3A1 enzyme activity and protein levels but not the corresponding mRNAs. Such a decrease in ALDH activity was seen in all cell lines tested and results in a significant increase in toxicity of 4-HC and acetaldehyde, both of which are substrates for the enzymes. Prior incubation with ATRA also results in increased cytotoxicity, although to a lesser degree, of phenylketophosphamide and melphalan, neither of which is a substrate for ALDHs. These results suggest a post-translational mechanism through which retinoids decrease both ALDH expression, which results in increased cytotoxicity of 4-HC and acetaldehyde, although other previously described effects of these retinoids may contribute to the slight increase in cytotoxicity seen with other chemotherapy agents. These results may have clinical implications in regard to the use of retinoids in lung cancer prevention and treatment.
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Affiliation(s)
- Jan S Moreb
- University of Florida, College of Medicine, Department of Medicine, Division of Hematology/Oncology, P.O. Box 100277, 1600 SW Archer Road, Room R4-220, Gainesville, FL 32610-0277, USA.
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Gumpricht E, Dahl R, Devereaux MW, Sokol RJ. Beta-carotene prevents bile acid-induced cytotoxicity in the rat hepatocyte: Evidence for an antioxidant and anti-apoptotic role of beta-carotene in vitro. Pediatr Res 2004; 55:814-21. [PMID: 14764912 DOI: 10.1203/01.pdr.0000117845.23762.6b] [Citation(s) in RCA: 24] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 12/24/2022]
Abstract
Hydrophobic bile acids are implicated in the pathogenesis of cholestatic liver disorders through mechanisms involving oxidative stress and mitochondrial dysfunction. Antioxidants ameliorate bile acid-induced cytotoxicity in rat hepatocyte suspensions. The purpose of the current study was to evaluate the potential protective role of beta-carotene (betaC), a putative fat-soluble antioxidant that is reduced in patients with cholestasis, against bile acid-induced hepatotoxicity. In freshly isolated rat hepatocyte suspensions that were exposed to the toxic hydrophobic bile acid glycochenodeoxycholic acid (100 or 500 microM), betaC (100 microM) decreased generation of reactive oxygen species by >50%, similar to the inhibition afforded by alpha-tocopherol. Commensurate with this antioxidant effect, 100 microM betaC also protected hepatocytes against both glycochenodeoxycholic acid-induced cellular necrosis and apoptosis, which was associated with reduction in caspase 3 activation, inhibition of mitochondrial cytochrome c release in rat hepatocytes, and prevention of the mitochondrial permeability transition in both liver mitochondria and rat hepatocytes. A lower concentration of betaC (50 microM) produced similar antioxidant and anti-apoptotic protection but with less inhibition against cell necrosis, suggesting that the higher concentration of betaC may have conferred additional cytoprotection not directly related to its antioxidant function. These results demonstrate that the antioxidant effects of betaC may provide hepatoprotection against cholestatic liver injury by preventing bile acid-induced oxidative stress and mitochondrial perturbations.
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Affiliation(s)
- Eric Gumpricht
- Section of Pediatric Gastroenterology, Hepatology and Nutrition, Department of Pediatrics, University of Colorado School of Medicine and The Children's Hospital, Denver, Colorado 80262, USA
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Wu J, Cheng ML, Zhang GH, Zhai RW, Huang NH, Li CX, Luo TY, Lu S, Yu ZQ, Yao YM, Zhang YY, Ren LZ, Ye L, Li L, Zhang HN. Epidemiological and histopathological study of relevance of Guizhou Maotai liquor and liver diseases. World J Gastroenterol 2002; 8:571-4. [PMID: 12046095 PMCID: PMC4656446 DOI: 10.3748/wjg.v8.i3.571] [Citation(s) in RCA: 13] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 02/06/2023] Open
Abstract
AIM: To explore the relevance of Maotai liquor and liver diseases.
METHODS: Epidemiological study was conducted on groups of subjects, each consisting of 3 subjects from the Maotai liquor group consisting of 99 individuals and one from the non-alcoholic control group consisting of 33 individuals. Liver biopsy was performed on 23 volunteers from Guizhou Maotai Distillery who had a constant and long history of drinking Maotai liquor. Experimental histopathological study was conducted as follows: sixty male Wistar rats were divided into 3 groups randomly and fed with Maotai liquor, ordinary white wine, and physiological saline respectively for a period of 8 and 12 weeks. The rats were sacrificed in batches, then serum ALT, AST, TBil, and AKP were measured. Rat livers were harvested to measure the liver indexes, GSH, and MDA. Histopathological examinations were also performed. Another eighty mice were randomly divided into 4 groups and fed with Maotai (at different dosages of 10 mL·kg-1 and 20 mL·kg-1), ethanol, and physiological saline. The animals were sacrificed after 4 weeks and serum ALT was determined. Then the livers were harvested and liver indexes and MDA were measured.
RESULTS: The incidence rate of hepatic symptoms, splenomegaly, liver function impairment, reversal of Albumin/Globulin and increased diameter of portal veins in the Maotai liquor group were 1.0% (1/99), 1.0% (1/99), 1.0% (1/99), 1.0% (1/99), 0 (0/99) and 0 (0/99), 0 (0/99), 0 (0/99), 0 (0/99), 0 (0/99), respectively. There was no significant difference between the Maotai group and the non-alcoholic control group (P > 0.05). Various degree of fatty infiltration of hepatocytes was found in the 23 volunteers receiving liver biopsy, but there was no obvious hepatic fibrosis or cirrhosis. A comparison was made between the Maotai liquor group and the ordinary white wine group. It was found that hepatic MDA in rats and mice were 0.33 ± 0.10 and 0.49 ± 0.23 respectively in Maotai group and 0.61 ± 0.22 and 0.66 ± 0.32 in the ordinary white wine group; MDA had an obvious decrease in the Maotai liquor group (P < 0.05); hepatic GSH were 0.12 mg·g-1± 0.06 mg·g-1 in rats of the Maotai liquor group and (0.08 ± 0.02) mg·g-1 in white wine group, it was obviously increased in the Maotai liquor group (P < 0.05). After the 20 rats had been fed with ordinary white wine for 8 weeks consecutively, disarranged hepatocyte cords, fatty infiltration of hepatocytes, and fibrous septa of varying widths due to hepatic connective tissues proliferation were observed; after 12 weeks, the fibrous tissue proliferation continued and early cirrhosis appeared. Compared with the ordinary white wine group, fatty infiltration was observed in the 8-week and 12-week groups, but no necrosis or fibrosis or cirrhosis was found in the Maotai liquor group (P < 0.05).
CONCLUSION: Maotai liquor may cause fatty liver but not hepatic fibrosis or cirrhosis, and it can strengthen lipid peroxidation in the liver.
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MESH Headings
- Adult
- Alcoholic Beverages/adverse effects
- Animals
- China/epidemiology
- Fatty Liver, Alcoholic/epidemiology
- Fatty Liver, Alcoholic/etiology
- Fatty Liver, Alcoholic/pathology
- Female
- Humans
- Liver Cirrhosis, Alcoholic/epidemiology
- Liver Cirrhosis, Alcoholic/etiology
- Liver Cirrhosis, Alcoholic/pathology
- Liver Diseases, Alcoholic/epidemiology
- Liver Diseases, Alcoholic/etiology
- Liver Diseases, Alcoholic/pathology
- Male
- Mice
- Middle Aged
- Rats
- Rats, Wistar
- Wine/adverse effects
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Affiliation(s)
- Jun Wu
- Department of Infectious Diseases,Affiliated Hospital, Guiyang Medical College, Guizyang 550004, Guizhou Province, China.
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Cheng ML, Wu J, Wang HQ, Xue LM, Tan YZ, Ping L, Li CX, Huang NH, Yao YM, Ren LZ, Ye L, Li L, Jia ML. Effect of Maotai liquor in inducing metallothioneins and on hepatic stellate cells. World J Gastroenterol 2002; 8:520-3. [PMID: 12046083 PMCID: PMC4656434 DOI: 10.3748/wjg.v8.i3.520] [Citation(s) in RCA: 13] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 05/02/2002] [Revised: 05/13/2002] [Accepted: 05/25/2002] [Indexed: 02/06/2023] Open
Abstract
AIM To explore the possible mechanism why drinking Maotai liquor dose not cause hepatic fibrosis. METHODS After being fed with Maotai for 56 days consecutively, the male SD rats were decollated for detecting the biological indexes, and the livers were harvested to examine the liver indexes and the level of hepatic metallothioneins (MT). Hepatic stellate cells (HSC) proliferation and collagen generation were also observed. RESULTS Hepatic MT contents were 216.0 ng.g(-1)+/-10.8 ng.g(-1) in the rats of Maotai group and 10.0 ng.g(-1)+/-2.8 ng.g(-1) in the normal control group, which was increased obviously in Maotain group (P<0.05). In the rats with grade CCL(2) poisoning induced by Maotai, hepatic MT content was 304.8 ng.g(-1)+/-12.1 ng.g(-1) whereas in the controls with grade CCL(4) poisoning, it was 126.4 ng.g(-1)+/-4.8 ng.g(-1) (P<0.05). MDA was 102.0 nmol.g(-1)+/-3.4 nmol.g(-1) in Maotai group and 150.8 nmol.g(-1)+/-6.7 nmol.g(-1) in the control group (P<0.05). When both of the groups were suffering from grade CCL(4) poisoning, hepatic MT contents was negatively correlated with MDA (r=-0.8023, n=20, P<0.01). The 570 nmA values of each tube with HSC regeneration at concentrations of 0, 10, 50, 100, and 200 g.L(-1) of Maotai were 0.818, 0.742, 0.736, 0.72, 0.682, and 0.604, respectively. From the concentration of 10 g.L(-1), Maotai began to show obvious inhibitory effects against HSC, and the inhibition was concentration-dependent (P<0.05, P<0.01). Type I collagen contents in HSC were 61.4, 59.9, 50.1, 49.2, 48.7, 34.4 microg.g(-1) at concentrations of 0, 10, 50, 100, and 200 g.L(-1) of Maotai. At the concentration of 100-200 g.L(-1), Maotai had obvious inhibitory effect against the secretion of type I collagen (P<0.05). Gene expression analysis was conducted on cells with Maotai concentrations of 0, 50, 100g.L(-1) respectively and the ash values of beta-actin gene expression were 0.88, 0.74, and 0.59, respectively,suggesting that at the concentration of 100g.L(-1), Maotai could obviously inhibit gene expression of type I procollagen (P<0.05), but the effect was not obvious at the concentration of 50 g.L(-1) (P>0.05). At the concentration of 10 g.L(-1), HSC growth in vitro inhibition rates were 16.4+/-2.3 in Maotai group and -8.4+/-2.3 in the control group (P<0.05). CONCLUSION Maotai liquor can increase metallothioneins in the liver and inhibit the activation of HSC and the synthesis of collagen in many aspects, which might be the mechanism that Maotai liquor interferes in the hepatic fibrosis.
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Affiliation(s)
- Ming-Liang Cheng
- Department of Infectious Diseases, Affiliated Hospital, Guiyang Medical College, Guizyang 550004, Guizhou Province, China.
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