|
1
|
Fang W, Wang G, Tang L, Su H, Chen H, Liao W, Xu J. Hydrogen gas inhalation protects against cutaneous ischaemia/reperfusion injury in a mouse model of pressure ulcer. J Cell Mol Med 2018; 22:4243-4252. [PMID: 29921037 PMCID: PMC6111801 DOI: 10.1111/jcmm.13704] [Citation(s) in RCA: 26] [Impact Index Per Article: 3.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/19/2018] [Accepted: 04/25/2018] [Indexed: 12/13/2022] Open
Abstract
Pressure ulcer formation depends on various factors among which repetitive ischaemia/reperfusion(I/R) injury plays a vital role. Molecular hydrogen (H2) was reported to have protective effects on I/R injuries of various internal organs. In this study, we investigated the effects of H2 inhalation on pressure ulcer and the underlying mechanisms. H2 inhalation significantly reduced wound area, 8‐oxo‐dG level (oxidative DNA damage) and cell apoptosis rates in skin lesions. H2 remarkably decreased ROS accumulation and enhanced antioxidant enzymes activities by up‐regulating expression of Nrf2 and its downstream components in wound tissue and/or H2O2‐treated endothelia. Meanwhile, H2 inhibited the overexpression of MCP‐1, E‐selectin, P‐selectin and ICAM‐1 in oxidant‐induced endothelia and reduced inflammatory cells infiltration and proinflammatory cytokines (TNF‐α, IL‐1, IL‐6 and IL‐8) production in the wound. Furthermore, H2 promoted the expression of pro‐healing factors (IL‐22, TGF‐β, VEGF and IGF1) and inhibited the production of MMP9 in wound tissue in parallel with acceleration of cutaneous collagen synthesis. Taken together, these data indicated that H2 inhalation suppressed the formation of pressure ulcer in a mouse model. Molecular hydrogen has potentials as a novel and alternative therapy for severe pressure ulcer. The therapeutic effects of molecular hydrogen might be related to its antioxidant, anti‐inflammatory, pro‐healing actions.
Collapse
Affiliation(s)
- Wei Fang
- Department of Dermatology, Huashan Hospital, Fudan University, Shanghai, China.,The Shanghai Institute of Dermatology, Shanghai, China.,Department of Dermatology and Venereology, Changzheng Hospital, Shanghai Key Laboratory of Molecular Medical Mycology, Shanghai, China
| | - Guizhen Wang
- Emergency room, Shanghai Tenth People's Hospital of Tongji University, Shanghai, China
| | - Luyan Tang
- Department of Dermatology, Huashan Hospital, Fudan University, Shanghai, China.,The Shanghai Institute of Dermatology, Shanghai, China
| | - Huilin Su
- Department of Dermatology, Huashan Hospital, Fudan University, Shanghai, China
| | - Huyan Chen
- Department of Dermatology, Huashan Hospital, Fudan University, Shanghai, China
| | - Wanqing Liao
- Department of Dermatology and Venereology, Changzheng Hospital, Shanghai Key Laboratory of Molecular Medical Mycology, Shanghai, China
| | - Jinhua Xu
- Department of Dermatology, Huashan Hospital, Fudan University, Shanghai, China.,The Shanghai Institute of Dermatology, Shanghai, China
| |
Collapse
|
|
2
|
Abstract
Hepatic ischemia/reperfusion (I/R) injury is a major complication of liver surgery, including liver resection, liver transplantation, and trauma surgery. Much has been learned about the inflammatory injury response induced by I/R, including the cascade of proinflammatory mediators and recruitment of activated leukocytes. In this review, we discuss the complex network of events that culminate in liver injury after I/R, including cellular, protein, and molecular mechanisms. In addition, we address the known endogenous regulatory mediators that function to maintain homeostasis and resolve injury. Finally, we cover more recent insights into how the liver repairs and regenerates after I/R injury, a setting in which physical mass remains unchanged, but functional liver mass is greatly reduced. In this regard, we focus on recent work highlighting a novel role of CXC chemokines as important regulators of hepatocyte proliferation and liver regeneration after I/R injury.
Collapse
Affiliation(s)
- Takanori Konishi
- Department of Surgery, College of Medicine, University of Cincinnati, Cincinnati, OH, USA
| | - Alex B. Lentsch
- Department of Surgery, College of Medicine, University of Cincinnati, Cincinnati, OH, USA
| |
Collapse
|
|
3
|
van Golen RF, Stevens KM, Colarusso P, Jaeschke H, Heger M. Platelet aggregation but not activation and degranulation during the acute post-ischemic reperfusion phase in livers with no underlying disease. J Clin Transl Res 2015; 1:107-115. [PMID: 26925465 DOI: 10.18053/jctres.201502.001] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Key Words] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/10/2023] Open
Abstract
BACKGROUND Platelets and P-selectin (CD62P) play an unequivocal role in the pathology of hepatic ischemia/reperfusion (I/R) injury. Inhibition or knock-out of P-selectin or immunodepletion of platelets results in amelioration of post-ischemic inflammation, reduced hepatocellular damage, and improved survival. However, P-selectin expression on platelets and endothelial cells, which concurs with platelet activation, has never been clearly demonstrated in I/R-subjected livers. AIMS To determine whether platelets become activated and degranulate in the acute phase of liver I/R and whether the platelets interact with neutrophils. METHODS Hepatic I/R was induced in male C57BL/6J mice (N = 12) using 37.5-min ischemia time. Platelets, endothelial cells, and neutrophils were fluorescently labeled by systemic administration of non-blocking antibodies. Cell kinetics were monitored by intravital spinning disk confocal microscopy during 90 min of reperfusion. Image analysis and quantification was performed with dedicated software. RESULTS Platelets adhered to sinusoids more extensively in post-ischemic livers compared to livers not subjected to I/R and formed aggregates, which occurred directly after ischemia. Platelets and endothelial cells did not express P-selectin in post-ischemic livers. There was no interaction between platelets and neutrophils. CONCLUSIONS Platelets aggregate but do not become activated and do not degranulate in post-ischemic livers. There is no platelet-neutrophil interplay during the early reperfusion phase in a moderate model of hepatic I/R injury. The mechanisms underlying the biological effects of platelets and P-selectin in this setting warrant further investigation. RELEVANCE FOR PATIENTS I/R in surgical liver patients may compromise outcome due to post-ischemic oxidative stress and sterile inflammation. Both processes are mediated in part by platelets. Understanding platelet function during I/R is key to developing effective interventions for I/R injury and improving clinical outcomes.
Collapse
Affiliation(s)
- Rowan F van Golen
- Department of Experimental Surgery, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands
| | - Katarzyna M Stevens
- Live Cell Imaging Facility, Snyder Institute for Chronic Diseases, University of Calgary, Calgary, Alberta, Canada
| | - Pina Colarusso
- Live Cell Imaging Facility, Snyder Institute for Chronic Diseases, University of Calgary, Calgary, Alberta, Canada
| | - Hartmut Jaeschke
- Department of Pharmacology, Toxicology and Therapeutics, University of Kansas Medical Center, Kansas City, USA
| | - Michal Heger
- Department of Experimental Surgery, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands
| |
Collapse
|
|
4
|
Marques PE, Oliveira AG, Chang L, Paula-Neto HA, Menezes GB. Understanding liver immunology using intravital microscopy. J Hepatol 2015; 63:733-42. [PMID: 26055800 DOI: 10.1016/j.jhep.2015.05.027] [Citation(s) in RCA: 38] [Impact Index Per Article: 3.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 04/01/2015] [Revised: 05/21/2015] [Accepted: 05/21/2015] [Indexed: 12/15/2022]
Abstract
The liver has come a long way since it was considered only a metabolic organ attached to the gastrointestinal tract. The simultaneous ascension of immunology and intravital microscopy evidenced the liver as a central axis in the immune system, controlling immune responses to local and systemic agents as well as disease tolerance. The multiple hepatic cell populations are organized in a vascular environment that promotes intimate cellular interactions, including initiation of innate and adaptive immune responses, rapid leukocyte recruitment, pathogen clearance and production of a variety of immune mediators. In this review, we focus on the advances in liver immunology supported by intravital microscopy in diseases such as isquemia/reperfusion, acute liver injury and infections.
Collapse
Affiliation(s)
- Pedro Elias Marques
- Laboratório de Imunobiofotônica, Departamento de Morfologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Brazil
| | - André Gustavo Oliveira
- Laboratório de Imunobiofotônica, Departamento de Morfologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Brazil
| | | | - Heitor Affonso Paula-Neto
- Departamento de Biotecnologia Farmacêutica, Faculdade de Farmácia, Universidade Federal do Rio de Janeiro, Brazil
| | - Gustavo Batista Menezes
- Laboratório de Imunobiofotônica, Departamento de Morfologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Brazil.
| |
Collapse
|
|
5
|
Tian XX, Wang BL, Cao YZ, Zhong YX, Tu YY, Xiao JB, He QF, Zhai LN. Comparison of protective effects of safflor injection and extract of Ginkgo biloba on lung ischemia/reperfusion injury in rabbits. Chin J Integr Med 2015; 21:229-233. [PMID: 24142260 DOI: 10.1007/s11655-013-1513-8] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/28/2011] [Indexed: 02/05/2023]
Abstract
OBJECTIVE To observe the protective effects of safflor Injection (SI) and extract of Ginkgo biloba (EGB) on lung ischemia-reperfusion injury (LIRI) and investigate its mechanism. METHODS In vivo rabbit model of LIRI was reconstructed. Forty rabbits were randomly and equally divided into four groups: sham-operation group (sham group), ischemia-reperfusion group (model group), ischemia-reperfusion plus SI group (safflor group) and ischemia-reperfusion plus EGB injection group (EGB group). Malondialdehyde (MDA) content, superoxide dismutase (SOD) and xanthine oxidase (XO) activity in serum were measured. The wet/dry weight ratio (W/D) of the lung tissue and activity of myeloperoxidase (MPO) were also tested. Ultrastructure change of the lung tissue was observed by the electron microscope. The expression of intercellular adhesion molecule-1 (ICAM-1) was measured by immunohistochemistry (IHC). RESULTS In the model group, MDA and XO increased and SOD decreased in serum compared with the sham group (P<0.01). The values of W/D, MPO and ICAM-1 of the model group were higher than those of the sham group (P<0.01), but those of the safflor group and EGB group were significantly lower than those of the model group (P<0.01). The IHC demonstrated that ICAM-1 expression in lung tissue of the model group was significantly higher than those of the safflor group (P<0.01). Compared with safflor group, in the EGB group MDA, XO, MPO decreased, SOD and ICAM-1 expression increased (P<0.05), but the change of W/D was not statistically significant (P>0.05). CONCLUSIONS SI and EGB may attenuate LIRI through antioxidation, inhibition of neutrophil aggregation and down-regulation of ICAM-1 expression. But EGB had more effect on the antioxidation, while SI did better on regulating ICAM-1 expression.
Collapse
Affiliation(s)
- Xiao-xi Tian
- Emergency Department, Tangdu Hospital, Fourth Military Medical University, Xi'an, 710038, China
| | | | | | | | | | | | | | | |
Collapse
|
|
6
|
Datta G, Fuller BJ, Davidson BR. Molecular mechanisms of liver ischemia reperfusion injury: Insights from transgenic knockout models. World J Gastroenterol 2013; 19:1683-98. [PMID: 23555157 PMCID: PMC3607745 DOI: 10.3748/wjg.v19.i11.1683] [Citation(s) in RCA: 152] [Impact Index Per Article: 12.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 02/13/2012] [Revised: 06/29/2012] [Accepted: 07/09/2012] [Indexed: 02/06/2023] Open
Abstract
Ischemia reperfusion injury is a major obstacle in liver resection and liver transplantation surgery. Understanding the mechanisms of liver ischemia reperfusion injury (IRI) and developing strategies to counteract this injury will therefore reduce acute complications in hepatic resection and transplantation, as well as expanding the potential pool of usable donor grafts. The initial liver injury is initiated by reactive oxygen species which cause direct cellular injury and also activate a cascade of molecular mediators leading to microvascular changes, increased apoptosis and acute inflammatory changes with increased hepatocyte necrosis. Some adaptive pathways are activated during reperfusion that reduce the reperfusion injury. IRI involves a complex interplay between neutrophils, natural killer T-cells cells, CD4+ T cell subtypes, cytokines, nitric oxide synthases, haem oxygenase-1, survival kinases such as the signal transducer and activator of transcription, Phosphatidylinositol 3-kinases/Akt and nuclear factor κβ pathways. Transgenic animals, particularly genetic knockout models, have become a powerful tool at elucidating mechanisms of liver ischaemia reperfusion injury and are complementary to pharmacological studies. Targeted disruption of the protein at the genetic level is more specific and maintained than pharmacological inhibitors or stimulants of the same protein. This article reviews the evidence from knockout models of liver IRI about the cellular and molecular mechanisms underlying liver IRI.
Collapse
|
|
7
|
Lentsch AB. Regulatory mechanisms of injury and repair after hepatic ischemia/reperfusion. SCIENTIFICA 2012; 2012:513192. [PMID: 24278708 PMCID: PMC3820555 DOI: 10.6064/2012/513192] [Citation(s) in RCA: 12] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Abstract] [Track Full Text] [Subscribe] [Scholar Register] [Received: 08/01/2012] [Accepted: 09/12/2012] [Indexed: 06/02/2023]
Abstract
Hepatic ischemia/reperfusion injury is an important complication of liver surgery and transplantation. The mechanisms of this injury as well as the subsequent reparative and regenerative processes have been the subject of thorough study. In this paper, we discuss the complex and coordinated responses leading to parenchymal damage after liver ischemia/reperfusion as well as the manner in which the liver clears damaged cells and regenerates functional mass.
Collapse
Affiliation(s)
- Alex B. Lentsch
- Department of Surgery, College of Medicine, University of Cincinnati, 231 Albert Sabin Way, ML 0558, Cincinnati, OH 45267-0558, USA
| |
Collapse
|
|
8
|
Klune JR, Tsung A. Molecular biology of liver ischemia/reperfusion injury: established mechanisms and recent advancements. Surg Clin North Am 2010; 90:665-77. [PMID: 20637940 DOI: 10.1016/j.suc.2010.04.003] [Citation(s) in RCA: 147] [Impact Index Per Article: 9.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/31/2022]
Abstract
Hepatic ischemia/reperfusion (I/R) injury occurs in a variety of clinical contexts, including transplantation, liver resection surgery, trauma, and hypovolemic shock. The mechanism of organ damage after I/R has been studied extensively and consists of complex interactions of multiple inflammatory pathways. The major contributors to I/R injury include production of reactive oxygen species, release of proinflammatory cytokines and chemokines, and activation of immune cells to promote inflammation and tissue damage. Recent research has focused on the mechanisms by which these immune responses are initially activated through signaling molecules and their cellular receptors. Thorough understanding of the pathophysiology of liver I/R may yield novel therapeutic strategies to reduce I/R injury and lead to improved clinical outcomes.
Collapse
Affiliation(s)
- John R Klune
- Department of Surgery, F675 UPMC Presbyterian Hospital, University of Pittsburgh Medical Center, 200 Lothrop Street, Pittsburgh, PA 15213, USA
| | | |
Collapse
|
|
9
|
Clarke CN, Kuboki S, Tevar A, Lentsch AB, Edwards M. CXC chemokines play a critical role in liver injury, recovery, and regeneration. Am J Surg 2009; 198:415-9. [PMID: 19716886 DOI: 10.1016/j.amjsurg.2009.01.025] [Citation(s) in RCA: 42] [Impact Index Per Article: 2.6] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/22/2008] [Revised: 01/27/2009] [Accepted: 01/27/2009] [Indexed: 12/31/2022]
Abstract
BACKGROUND Hepatic ischemia/reperfusion (I/R) injury is a principal consideration of trauma, resectional liver surgery, and transplantation. Despite improvements in supportive care, hepatic I/R injury continues to negatively impact patient outcomes because of significant tissue damage and organ dysfunction. CXC chemokines have been implicated as key mediators in the deleterious inflammatory cascade after hepatic I/R and also as important, beneficial regulators of liver recovery and regeneration. As such, their potential to mediate both beneficial and detrimental effects on hepatocytes makes them a key target for therapy. Herein, we provide a review of the inflammatory mechanisms of hepatic I/R injury, with a focus on the divergent functions of CXC chemokines in this response compared with other liver insults, and offer an explanation of this apparent paradox. DATA SOURCES MEDLINE and PubMed. CONCLUSIONS CXC chemokines are key mediators of both the inflammatory response to hepatic I/R as well as the recovery from this injury. Their contrasting functions in the regeneration of liver mass after an ischemic insult indicates that therapeutic manipulation of these mediator pathways should differ depending on the surgical milieu.
Collapse
Affiliation(s)
- Callisia N Clarke
- Department of Surgery, University of Cincinnati, Cincinnati, OH 45267-0558, USA
| | | | | | | | | |
Collapse
|
|
10
|
Yao XM, Chen H, Li Y. Protective effect of bicyclol on liver injury induced by hepatic warm ischemia/reperfusion in rats. Hepatol Res 2009; 39:833-42. [PMID: 19473433 DOI: 10.1111/j.1872-034x.2009.00504.x] [Citation(s) in RCA: 26] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 12/21/2022]
Abstract
AIM Bicyclol is a synthetic anti-hepatitis drug with anti-oxidative property. The purpose of this study was to investigate the effect of bicyclol on hepatic ischemia/reperfusion (I/R) injury and related mechanisms. METHODS Rats were subjected to 90 min of hepatic ischemia followed by reperfusion for 1, 3, 6 and 24 h. Three doses of bicyclol were orally administered before ischemia. Liver injury was evaluated by biochemical and histopathological examinations. Liver malondialdehyde (MDA), superoxide dismutase (SOD), nitric oxide (NO), inducible nitric oxide synthase (iNOS) and myeloperoxidase (MPO) contents, and plasma endotoxin levels were spectrophotometrically measured. The expressions of inflammatory and anti-inflammatory cytokines were detected by reverse transcription polymerase chain reaction and enzyme-linked immunosorbent assay. Liver intercellular adhesion molecule-1 (ICAM-1) was examined by immunohistochemistry, and the expressions of nuclear factor (NF)-kappaB, inhibitor of NF-kappaB (IkappaB) and toll-like receptor 4 (TLR4) were determined by western blot. RESULTS Bicyclol significantly inhibited the elevations of serum alanine aminotransferase, total bilirubin and plasma endotoxin levels, alleviated the formation of liver MDA and nitrite/nitrate, restored impaired antioxidant SOD, attenuated hepatic necrosis and neutrophil infiltration, and also improved the 7-day survival in I/R rats. Additionally, bicyclol markedly downregulated the overexpression of ICAM-1, modulated the expression of inflammatory/anti-inflammatory cytokines and inhibited the expression of NF-kappaB and TLR4 in I/R rats. CONCLUSION Bicyclol had a remarkable protective effect on hepatic I/R injury, which was partially due to inhibiting the expression of TLR4 and NF-kappaB via its ability to attenuate oxidative stress and endotoxin.
Collapse
Affiliation(s)
- Xiao-Min Yao
- Department of New Drug Development, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China
| | | | | |
Collapse
|
|
11
|
Miura S, Kubes P, Granger DN. Gastrointestinal and Liver Microcirculations: Roles in Inflammation and Immunity. Compr Physiol 2008. [DOI: 10.1002/cphy.cp020414] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/10/2022]
|
|
12
|
Dold S, Laschke MW, Lavasani S, Menger MD, Thorlacius H. Cholestatic liver damage is mediated by lymphocyte function antigen-1-dependent recruitment of leukocytes. Surgery 2008; 144:385-93. [PMID: 18707037 DOI: 10.1016/j.surg.2008.05.010] [Citation(s) in RCA: 11] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/09/2007] [Accepted: 05/14/2008] [Indexed: 12/27/2022]
Abstract
BACKGROUND The role of specific adhesion molecules in cholestasis-induced leukocyte recruitment in the liver is not known. Therefore, the aim of our experimental study was to evaluate the role of lymphocyte function antigen-1 (LFA-1) in cholestatic liver injury. METHODS C57BL/6 mice underwent bile duct ligation for 12 hours. Mice were pretreated with an anti-LFA-1 antibody or control antibody. Subsequently, hepatic accumulation of leukocytes and sinusoidal perfusion were determined by means of intravital fluorescence microscopy. Hepatocellular damage was monitored by measuring serum levels of alanine aminotransferase and aspartate aminotransferase. CXC chemokines in the liver were determined by enzyme-linked immunosorbent assay. RESULTS Bile duct ligation provoked clear-cut recruitment of leukocytes and liver damage, as indicated by increased serum activities of liver enzymes and sinusoidal perfusion failure. Neutrophils expressed greater levels of LFA-1 and inhibition of LFA-1 significantly decreased serum activity of alanine aminotransferase and aspartate aminotransferase levels in cholestatic mice. Immunoneutralization of LFA-1 reduced leukocyte adhesion in postsinusoidal venules that had been induced by bile duct ligation, whereas leukocyte rolling and sinusoidal accumulation were not changed. Moreover, blocking LFA-1 function restored sinusoidal perfusion in cholestatic animals. CONCLUSION These findings demonstrate an important role of LFA-1 in supporting cholestasis-induced leukocyte recruitment in the liver. Thus, targeting LFA-1 may help to protect against pathologic inflammation and liver damage in cholestatic liver diseases.
Collapse
Affiliation(s)
- Stefan Dold
- Department of Surgery, Malmö University Hospital, Lund University, Malmö, Sweden
| | | | | | | | | |
Collapse
|
|
13
|
Miura S, Kubes P, Granger DN. Gastrointestinal and Liver Microcirculations. Microcirculation 2008. [DOI: 10.1016/b978-0-12-374530-9.00016-4] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/27/2022]
|
|
14
|
Platelet-dependent accumulation of leukocytes in sinusoids mediates hepatocellular damage in bile duct ligation-induced cholestasis. Br J Pharmacol 2007; 153:148-56. [PMID: 18026126 DOI: 10.1038/sj.bjp.0707578] [Citation(s) in RCA: 78] [Impact Index Per Article: 4.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/30/2023] Open
Abstract
BACKGROUND Although it is well known that extrahepatic cholestasis induces liver damage, the mechanisms are still not completely understood. The aim of the present study was to evaluate the role of platelets and P-selectin in cholestasis-induced liver injury. EXPERIMENTAL APPROACH C57BL/6 mice underwent bile duct ligation (BDL) and pretreatment with an anti-GP1balpha antibody, which depletes platelets, an anti-P-selectin antibody or a control antibody. Hepatic platelet and leukocyte recruitment as well as microvascular perfusion were determined by intravital fluorescence microscopy. KEY RESULTS BDL caused significant liver damage and sinusoidal perfusion failure. BDL further induced hepatic platelet accumulation with widespread intravascular platelet aggregates, increased platelet adhesion in postsinusoidal venules and massive platelet accumulation in liver sinusoids. Administration of the anti-GP1balpha antibody reduced systemic platelet count by 90%. Depletion of platelets in BDL mice not only abolished accumulation and adhesion of platelets in sinusoids and venules but also restored sinusoidal perfusion and reduced liver enzymes by more than 83%. Platelet depletion further reduced BDL-associated sinusoidal leukocyte accumulation by 48% although leukocyte-endothelium interactions in venules were not affected. Immunoneutralization of P-selectin also inhibited hepatic microvascular accumulation of platelets and leukocytes, and protected against cholestasis-provoked hepatocellular damage. CONCLUSIONS AND IMPLICATIONS Platelets play an important role in BDL-induced liver injury by promoting leukocyte recruitment and deteriorating microvascular perfusion. Moreover, our findings demonstrate that cholestasis-induced accumulation of platelets is mediated by P-selectin. Thus, targeting platelet accumulation may be a useful strategy against liver damage associated with obstructive jaundice.
Collapse
|
|
15
|
Abstract
Reperfusion injury occurring in the transplanted liver is a complex lesion and has been the focus of considerable research over the past decade. This section will review recent major developments in understanding the mechanisms involved and their application to clinical transplantation.
Collapse
Affiliation(s)
- K Tanigawa
- Department of Emergency and Critical Care Medicine, Fukuoka University, Fukuoka, Japan.
| |
Collapse
|
|
16
|
Wang YI, Li G, Zhang Y, Gao W, Yao Z. The Expression of von Willebrand Factor, Soluble Thrombomodulin, and Soluble P-Selectin During Orthotopic Liver Transplantation. Transplant Proc 2007; 39:172-5. [PMID: 17275499 DOI: 10.1016/j.transproceed.2006.10.027] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/16/2006] [Indexed: 11/17/2022]
Abstract
BACKGROUND We evaluated von Willebrand factor (vWF), soluble thrombomodulin (sTM), and soluble P-selectin (sP-selectin) levels in ischemia/reperfusion injury during orthotopic liver transplantation (OLT). METHODS The vWF, sTM, and sP-selectin were analyzed in 20 patients who underwent liver transplantation. Blood samples were drawn from the radial artery at serial times during surgery. Plasma levels of sTM and sP-selectin were detected by enzyme-linked immunosorbent assay (ELISA). The wWF activity was measured using the immuno-turbidimetric method. Plasma aspartate transaminase (AST) and alanine transaminase (ALT) were assayed by routine clinical chemistry testing. RESULTS Marked elevation levels of plasma AST and ALT were released during the 15 minutes after reperfusion phase, with a peak on the first postoperative day (P < .01). The sTM level remained unchanged from preoperative to anhepatic phase (P > .05). In contrast, a mean 2.5-fold increase of sTM was observed during the 15-minute reperfusion stage compared with the preoperative value (P < .01). The vWF activity only showed significant increase during the 60-minute reperfusion stage compared with the preoperative value (P < .05). No significant increase occurred in sP-seletin levels during the reperfusion phase. Platelet count showed significant decrease during the entire observation period compared with the preoperative value (P < .01). CONCLUSION The endothelial reperfusion injury after OLT is characterized by increased vWF and sTM but not by sP-selectin in peripheral blood.
Collapse
Affiliation(s)
- Y-I Wang
- Department of Immunology, Tianjin Medical University, and Tianjin Institute of Thrombosis and Hemostasis, Organ Transplantation Center of Tianjin First Central Hospital, Tianjin, China
| | | | | | | | | |
Collapse
|
|
17
|
Tsuchihashi S, Kaldas F, Chida N, Sudo Y, Tamura K, Zhai Y, Qiao B, Busuttil RW, Kupiec-Weglinski JW. FK330, a novel inducible nitric oxide synthase inhibitor, prevents ischemia and reperfusion injury in rat liver transplantation. Am J Transplant 2006; 6:2013-22. [PMID: 16796718 DOI: 10.1111/j.1600-6143.2006.01435.x] [Citation(s) in RCA: 37] [Impact Index Per Article: 1.9] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/06/2023]
Abstract
Nitric oxide (NO), produced via inducible NO synthase (iNOS), is implicated in the pathophysiology of liver ischemia/reperfusion injury (IRI). We examined the effects of a novel iNOS inhibitor, FK330 (FR260330), in well-defined rat liver IRI models. In a model of liver cold ischemia followed by ex vivo reperfusion, treatment with FK330 improved portal venous flow, increased bile production and decreased hepatocellular damage. FK330 prevented IRI in rat model of 40-h cold ischemia followed by syngeneic orthotopic liver transplantation (OLT), as evidenced by: (1) increased OLT survival (from 20% to 80%); (2) decreased hepatocellular damage (serum glutamic oxaloacetic transaminase/glutamic pyruvic transaminase levels); (3) improved histological features of IRI; (4) reduced intrahepatic leukocyte infiltration, as evidenced by decreased expression of P-selectin/intracellular adhesion molecule 1, ED-1/CD3 cells and neutrophils; (5) depressed lymphocyte activation, as evidenced by expression of pro-inflammatory cytokine (TNF-alpha, IL-1beta, IL-6) and chemokine (IP-10, MCP-1, MIP-2) programs; (6) prevented hepatic apoptosis and down-regulated Bax/Bcl-2 ratio. Thus, by modulating leukocyte trafficking and cell activation patterns, treatment of rats with FK330, a specific iNOS inhibitor, prevented liver IRI. These results provide the rationale for novel therapeutic approaches to maximize organ donor pool through the safer use of liver grafts despite prolonged periods of cold ischemia.
Collapse
Affiliation(s)
- S Tsuchihashi
- The Dumont-UCLA Transplant Center, Division of Liver and Pancreas Transplantation, Department of Surgery, David Geffen School of Medicine at UCLA, Los Angeles, California, USA
| | | | | | | | | | | | | | | | | |
Collapse
|
|
18
|
Luh SP, Yang PC. Organ preconditioning: the past, current status, and related lung studies. J Zhejiang Univ Sci B 2006; 7:331-41. [PMID: 16615162 PMCID: PMC1462933 DOI: 10.1631/jzus.2006.b0331] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/08/2006] [Accepted: 03/23/2006] [Indexed: 12/13/2022]
Abstract
Preconditioning (PC) has emerged as a powerful method for experimentally and clinically attenuating various types of organ injuries. In this paper related clinical and basic research issues on organ preconditioning issues were systemically reviewed. Since lung injuries, including ischemia-reperfusion and others, play important roles in many clinical results, including thromboembolism, trauma, thermal injury, hypovolemic and endotoxin shock, reimplantation response after organ transplantation, and many respiratory diseases in critical care. It is of interest to uncover methods, including the PCs, to protect the lung from the above injuries. However, related studies on pulmonary PC are relatively rare and still being developed, so we will review previous literature on experimental and clinical studies on pulmonary PC in the following paragraphs.
Collapse
Affiliation(s)
- Shi-ping Luh
- Department of Cardiothoracic Surgery, Taipei Tzu-Chi Medical University Hospital, Taiwan 231, China.
| | | |
Collapse
|
|
19
|
Tsuchihashi SI, Fondevila C, Shaw GD, Lorenz M, Marquette K, Benard S, Shen XD, Ke B, Busuttil RW, Kupiec-Weglinski JW. Molecular characterization of rat leukocyte P-selectin glycoprotein ligand-1 and effect of its blockade: protection from ischemia-reperfusion injury in liver transplantation. THE JOURNAL OF IMMUNOLOGY 2006; 176:616-24. [PMID: 16365457 DOI: 10.4049/jimmunol.176.1.616] [Citation(s) in RCA: 47] [Impact Index Per Article: 2.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 12/26/2022]
Abstract
P-selectin glycoprotein ligand-1 (PSGL-1) mediates the initial tethering of leukocytes to activated platelets and endothelium. We report molecular cloning and characterization of the rat PSGL-1 gene. A neutralizing Ab was generated, and its binding epitope was mapped to the N-terminal binding region of rat PSGL-1. We examined the effects of early PSGL-1 blockade in rat liver models of cold ischemia, followed by ex vivo reperfusion or transplantation (orthotopic liver transplantation (OLT)) using an anti-PSGL-1 Ab with diminished Fc-mediated effector function. In the ex vivo hepatic cold ischemia and reperfusion model, pretreatment with anti-PSGL-1 Ab improved portal venous flow, increased bile production, and decreased hepatocellular damage. Rat pretreatment with anti-PSGL-1 Ab prevented hepatic insult in a model of cold ischemia, followed by OLT, as assessed by 1) decreased hepatocellular damage (serum glutamic oxaloacetic transaminase/glutamic-pyruvic transaminase levels), and ameliorated histological features of ischemia/reperfusion injury, consistent with extended OLT survival; 2) reduced intrahepatic leukocyte infiltration, as evidenced by decreased expression of P-selectin, ED-1, CD3, and OX-62 cells; 3) inhibited expression of proinflammatory cytokine genes (TNF-alpha, IL-1beta, IL-6, IFN-gamma, and IL-2); and 4) prevented hepatic apoptosis accompanied by up-regulation of antiapoptotic Bcl-2/Bcl-xL protective genes. Thus, targeting PSGL-1 with a blocking Ab that has diminished Fc-mediated effector function is a simple and effective strategy that provides the rationale for novel therapeutic approaches to maximize the organ donor pool through the safer use of liver transplants despite prolonged periods of cold ischemia.
Collapse
Affiliation(s)
- Sei-ichiro Tsuchihashi
- Dumont-University of California Transplant Center, Division of Liver and Pancreas Transplantation, Department of Surgery, David Geffen School of Medicine, University of California, Los Angeles, CA 90095, USA
| | | | | | | | | | | | | | | | | | | |
Collapse
|
|
20
|
Abstract
Microvascular dysfunction mediates many of the local and systemic consequences of ischemic-reperfusion (I/R) injury, with a spectrum of changes specific to arterioles, capillaries, and venules. This review discusses the specific changes in the endothelium during I/R injury; describes the differential responses of the various levels of the vasculature including arterioles, capillaries, and venules; and explores mechanisms for remote organ injury. Vascular dysfunction is largely a consequence of changes in the endothelial cells themselves, affecting the integrity of barrier function, cytokine and adhesion molecule expression, and vascular tone. The bioavailability of nitric oxide, an important mediator of vasodilation, is profoundly decreased during the reperfusion period, resulting in impaired vasodilation of arterioles. Release of inflammatory mediators and increased expression of adhesion molecules initiate inflammatory and coagulation cascades that culminate in the occlusion of capillaries, known as the "no-reflow''" phenomenon. In postcapillary venules, the recruitment and transmigration of leukocytes further compromise the integrity of the endothelial barrier and increase the oxidative burden, resulting in leakage and tissue edema. I/R injury can have significant and untoward consequences beyond the affected tissue, with such conditions as systemic inflammatory response syndrome. This review highlights recent progress in understanding of the varied phenomena of vascular dysfunction in I/R injury and some promising advances in the understanding and application of ischemic preconditioning and other potential therapies.
Collapse
Affiliation(s)
- John B Seal
- Pritzker School of Medicine, University of Chicago, Chicago, IL 60637, USA
| | | |
Collapse
|
|
21
|
Thorlacius H, Klintman D. Response to the letter by Dr. Jaeschke. J Leukoc Biol 2004. [DOI: 10.1189/jlb.0704397] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/24/2022] Open
|
|
22
|
Kruskal JB, Thomas P, Kane RA, Goldberg SN. Hepatic perfusion changes in mice livers with developing colorectal cancer metastases. Radiology 2004; 231:482-90. [PMID: 15128993 DOI: 10.1148/radiol.2312030160] [Citation(s) in RCA: 39] [Impact Index Per Article: 1.9] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/23/2022]
Abstract
PURPOSE To evaluate whether intrahepatic flow alterations occur during formation of hepatic colorectal cancer metastases and to identify possible causes of these alterations. MATERIALS AND METHODS Intravital imaging of exteriorized livers was performed in 72 live mice. Three groups of mice were studied: a sham-operated control group (n = 24), a group with nonmetastasizing subcutaneous gliomas (n = 24), and a group with developing hepatic CX-1 colon cancer metastases (n = 24). Microvascular flow parameters, leukocyte-endothelial interactions, and wall shear stress were directly measured in hepatic sinusoids and postsinusoidal venules at 2-day intervals prior to and during the development of metastases. The Kruskal-Wallis test was used initially to test for overall equality of medians in each data group. Single posttest comparisons of independent samples were performed with the Mann-Whitney test, with an overall statistical significance of .05. RESULTS Prior to the development of visible colorectal cancer metastases, significant (P <.05) reductions occurred in sinusoidal and postsinusoidal flow and wall shear rates, coupled with increased leukocyte rolling and adherence. With tumor growth, flow was further compromised in 92% of tumors larger than 0.5 mm in diameter by extrinsic compression of sinusoids and portal venules and narrowing caused by adherent leukocytes. CONCLUSION Significant intrahepatic flow alterations occur in mouse livers prior to growth of visible metastases and provide a rational explanation for elevation in the Doppler perfusion index that occurs prior to tumor formation.
Collapse
Affiliation(s)
- Jonathan B Kruskal
- Department of Radiology, Beth Israel Deaconess Medical Center and Harvard Medical School, 1 Deaconess Rd, West Campus 302B, Boston, MA 02215, USA.
| | | | | | | |
Collapse
|
|
23
|
Loi P, Paulart F, Pajak B, Nagy N, Salmon I, Moser M, Goldman M, Flamand V. The fate of dendritic cells in a mouse model of liver ischemia/reperfusion injury. Transplant Proc 2004; 36:1275-9. [PMID: 15251311 DOI: 10.1016/j.transproceed.2004.05.052] [Citation(s) in RCA: 24] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/31/2022]
Abstract
Ischemia/reperfusion during liver transplantation triggers a complex cascade of inflammatory events that may lead to organ dysfunction. Herein, we investigated the consequences of hepatic ischemia/reperfusion on liver dendritic cells. Liver damage was documented by increased levels of serum alanine aminotransferase and by histopathology showing large areas of hepatocyte cytolysis. MHC class II+ CD45-B220 F4/80 dendritic cells were detected in necrotic areas 20 hours after reperfusion. Dendritic cells freshly isolated from reperfused livers displayed a mature phenotype characterized by upregulated expression of B7 costimulatory molecules; MHC-class II, and CD1d molecules. As shown by real-time PCR, IL-10, and TGF-beta mRNA accumulated in liver dendritic cells isolated after reperfusion, whereas IL-12p40 mRNA levels were decreased and IFN-gamma mRNA levels were unchanged. These results suggest that hepatic ischemia/reperfusion results in maturation of dendritic cells, which preferentially produce inhibitory cytokines.
Collapse
Affiliation(s)
- P Loi
- Institute for Medical Immunology-Laboratory of Experimental Immunology, Universite Libre de Bruxelles, Brussels
| | | | | | | | | | | | | | | |
Collapse
|
|
24
|
Teoh NC, Farrell GC. Hepatic ischemia reperfusion injury: pathogenic mechanisms and basis for hepatoprotection. J Gastroenterol Hepatol 2003; 18:891-902. [PMID: 12859717 DOI: 10.1046/j.1440-1746.2003.03056.x] [Citation(s) in RCA: 299] [Impact Index Per Article: 13.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 12/13/2022]
Abstract
This review highlights recent advances in our understanding of mechanisms underlying reperfusion injury to the liver after warm hepatic ischemia. Sinusoidal endothelial cells and hepatocytes are targets of injury in the early 'cytotoxic' phase, although participation of apoptosis in the cell-death process remains contentious. Kupffer cells may play an important role as the initial cytotoxic cell type and are likely a source of reactive oxygen species and proinflammatory mediators, particularly tumor necrosis factor (TNF)-alpha. The latter are involved with subsequent neutrophil activation and recruitment. Microcirculatory disruption results from an imbalance between the actions of vasoconstrictors and vasodilators, such as nitric oxide, and also has a major impact on reperfusion injury. There is growing evidence that a brief prior ischemia-reperfusion period, termed 'ischemic preconditioning', is hepatoprotective. This can be mimicked by drugs that produce oxidative stress, and by interleukin-6 and TNF-alpha; both these cytokines are involved with priming hepatocytes to enter the cell cycle. Several mechanisms have been implicated including mobilization of adenosine and activation of adenosine type 2 receptors, nitric oxide, abrogation of TNF synthesis, preservation of energy metabolism, protection of the microcirculation and accelerated cell-cycle entry. A better understanding of preconditioning mechanisms will lead to novel approaches to improve outcomes of liver surgery.
Collapse
Affiliation(s)
- Narci C Teoh
- Storr Liver Unit, Westmead Millennium Institute, University of Sydney at Westmead Hospital, Westmead, New South Wales, Australia
| | | |
Collapse
|
|
25
|
Nedrebø T, Reed RK, Berg A. Effect of alpha-trinositol on interstitial fluid pressure, edema generation, and albumin extravasation after ischemia-reperfusion injury in rat hind limb. Shock 2003; 20:149-53. [PMID: 12865659 DOI: 10.1097/01.shk.0000072128.33223.15] [Citation(s) in RCA: 17] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/25/2023]
Abstract
Reperfusion of ischemic tissue often leads to an acute inflammatory response, which acts directly to aggravate the injury in the reperfused zone, characterized by adhesion and subsequent infiltration of inflammatory cells that injure the tissue through the generation of oxygen-derived free radicals and release of various inflammatory mediators. The rapid edema formation associated with reperfusion injury is induced by increased microvascular permeability to plasma proteins and/or increased net filtration pressure across the microvascular wall, and the latter is at least in part induced by lowering of the interstitial fluid pressure (P(if)). We investigated the anti-inflammatory effect of alpha-trinositol (D-myo-inositol-1,2,6-trisphosphate) on edema formation, microvascular protein leakage, and P(if) in rat hind limb after ischemia-reperfusion (I/R) injury. There was significant increase of both albumin extravasation from 0.02 +/- 0.02 to 0.41 +/- 0.21 mL g dry weight-1 (P < 0.05) and total tissue water from 1.08 +/- 0.07 to 1.65 +/- 0.55 mL g dry weight(-1) (P < 0.05) in the skin of paws undergoing I/R injury. P(if) was significantly lowered from -0.51 +/- 0.34 to -5.00 +/- 1.53 mmHg (P < 0.05) concomitant with substantial edema formation. The edema formation, and lowering of P(if) during I/R injury was significantly lowered and nearly totally abolished in the animals treated with alpha-trinositol 30 min before reperfusion. We conclude that alpha-trinositol limits the increased vascular permeability and edema formation by preventing the decrease in P(if) as well as acting protective on the microvascular wall.
Collapse
|
|
26
|
Abstract
The shortage of organs has led centers to expand their criteria for the acceptance of marginal donors. The combination of multiple marginal factors seems to be additive on graft injury. In this review, the utility of various marginal donors in patients requiring liver transplantation will be described, including older donors, steatotic livers, non-heart-beating donors, donors with viral hepatitis, and donors with malignancies. The pathophysiology of the marginal donor will be discussed, along with strategies for minimizing the ischemia reperfusion injury experienced by these organs. Finally, new strategies for improving the function of the marginal/expanded donor liver will be reviewed.
Collapse
Affiliation(s)
- Ronald W Busuttil
- Department of Surgery, Division of Liver and Pancreas Transplantation, Dumont-UCLA Transplant Center, Los Angeles, CA 90095, USA.
| | | |
Collapse
|
|
27
|
Maier M, Ströbele H, Voges J, Bauer C, Marzi I. Attenuation of leukocyte adhesion by recombinant TNF-binding protein after hemorrhagic shock in the rat. Shock 2003; 19:457-61. [PMID: 12744490 DOI: 10.1097/01.shk.0000056961.48384.f2] [Citation(s) in RCA: 19] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/26/2022]
Abstract
Ischemia/reperfusion injury involves a large number of humoral and cellular mediators that activate leukocytes that subsequently migrate to local tissues. Tumor necrosis factor (TNF)-alpha may be one of the most important mediators of this post-shock inflammatory response. In this study, we investigated the influence of a recombinant Type I (55 kDa) TNF-binding protein (TNF-BP) on leukocyte-endothelial interactions in the liver after hemorrhagic shock. Hemorrhagic shock was induced in female Sprague-Dawley rats (40 mmHg for 90 min) and a standardized resuscitation regimen was applied. At the time of resuscitation, animals were treated intravenously with either TNF-BP 4 mg/kg or placebo. The liver microcirculation was investigated using intravital fluorescence microscopy and immunohistochemistry at 5 h and 48 h after reperfusion. At 5 h, treatment with TNF-BP significantly reduced temporary leukocyte adhesion in the liver sinusoids as well as mean adhesion time of leukocytes in the hepatic central vein. In contrast, after 48 h, permanent leukocyte adhesion in the central hepatic vein was significantly reduced in the group receiving TNF-BP, whereas temporary leukocyte adhesion and mean adhesion time did not differ between the two groups. Both types of leukocyte adhesion, rolling adhesion after 5 h and firm adhesion after 48 h, were reduced in the group treated with TNF-BP, thereby suggesting a long-lasting anti-inflammatory effect.
Collapse
MESH Headings
- Animals
- Carrier Proteins/therapeutic use
- Cell Adhesion/drug effects
- Chemotaxis, Leukocyte/drug effects
- Chemotaxis, Leukocyte/physiology
- Disease Models, Animal
- Female
- Hematocrit
- Intercellular Adhesion Molecule-1/metabolism
- Leukocyte Count
- Leukocytes/drug effects
- Leukocytes/physiology
- Liver/drug effects
- Liver/pathology
- Rats
- Rats, Sprague-Dawley
- Receptors, Tumor Necrosis Factor
- Receptors, Tumor Necrosis Factor, Type I
- Recombinant Proteins/therapeutic use
- Resuscitation
- Shock, Hemorrhagic/blood
- Shock, Hemorrhagic/immunology
- Shock, Hemorrhagic/pathology
- Tumor Necrosis Factor Decoy Receptors
- Tumor Necrosis Factor-alpha
Collapse
Affiliation(s)
- Marcus Maier
- Department of Trauma Surgery, Johann Wolfgang Goethe-University, 60590 Frankfurt, Germany
| | | | | | | | | |
Collapse
|
|
28
|
Wan MX, Wang Y, Liu Q, Schramm R, Thorlacius H. CC chemokines induce P-selectin-dependent neutrophil rolling and recruitment in vivo: intermediary role of mast cells. Br J Pharmacol 2003; 138:698-706. [PMID: 12598424 PMCID: PMC1573702 DOI: 10.1038/sj.bjp.0705094] [Citation(s) in RCA: 36] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/09/2022] Open
Abstract
1. Based on in vitro chemotaxis experiments, it is widely held that CC chemokines, such as macrophage inflammatory protein-1alpha (MIP-1alpha) and macrophage chemotactic protein-1 (MCP-1) mainly support lymphocyte trafficking. 2. The objective of the present study was to examine the role of MIP-1alpha and MCP-1 in neutrophil recruitment in vivo by use of intravital microscopy of the mouse cremaster microcirculation. 3. MIP-1alpha and MCP-1 caused a dose-dependent increase in leukocyte rolling, adhesion and recruitment. Indeed, neutrophils comprised more than 85% of the leukocyte response to MIP-1alpha and MCP-1. An anti-P-selectin antibody reduced MIP-1alpha and MCP-1-provoked leukocyte rolling by more than 94%. Concomitantly, firm adhesion and extravasation of neutrophils in response to MIP-1alpha and MCP-1 challenge were significantly decreased by more than 78 and 84%, respectively. In contrast, an anti-E-selectin antibody had no influence on CC chemokine-induced neutrophil recruitment. 4. Flow cytometric analysis revealed that MIP-1alpha and MCP-1 had no effect on P-selectin expression on endothelial cells, suggesting that neutrophil recruitment elicited by CC chemokines in vivo is not mediated via a direct effect on the endothelium but rather via an indirect effect involving activation of an intermediary tissue cell. Indeed, it was found that MIP-1alpha-induced neutrophil accumulation was significantly decreased by 58% in mast cell-deficient mice. 5. These findings demonstrate that CC chemokines trigger P-selectin-dependent rolling and tissue recruitment of neutrophils via tissue mast cells in vivo and suggest that CC chemokines may also be important targets in neutrophil-mediated tissue damage in multicellular organs.
Collapse
Affiliation(s)
- Ming Xiu Wan
- Department of Surgery, Malmö University Hospital, Lund University, 20502 Malmö, Sweden
| | - Yusheng Wang
- Department of Surgery, Malmö University Hospital, Lund University, 20502 Malmö, Sweden
| | - Qing Liu
- Department of Surgery, Malmö University Hospital, Lund University, 20502 Malmö, Sweden
| | - Rene Schramm
- Department of Surgery, Malmö University Hospital, Lund University, 20502 Malmö, Sweden
| | - Henrik Thorlacius
- Department of Surgery, Malmö University Hospital, Lund University, 20502 Malmö, Sweden
- Author for correspondence:
| |
Collapse
|
|
29
|
Jaeschke H. Molecular mechanisms of hepatic ischemia-reperfusion injury and preconditioning. Am J Physiol Gastrointest Liver Physiol 2003; 284:G15-26. [PMID: 12488232 DOI: 10.1152/ajpgi.00342.2002] [Citation(s) in RCA: 611] [Impact Index Per Article: 27.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 01/31/2023]
Abstract
Ischemia-reperfusion injury is, at least in part, responsible for the morbidity associated with liver surgery under total vascular exclusion or after liver transplantation. The pathophysiology of hepatic ischemia-reperfusion includes a number of mechanisms that contribute to various degrees in the overall injury. Some of the topics discussed in this review include cellular mechanisms of injury, formation of pro- and anti-inflammatory mediators, expression of adhesion molecules, and the role of oxidant stress during the inflammatory response. Furthermore, the roles of nitric oxide in preventing microcirculatory disturbances and as a substrate for peroxynitrite formation are reviewed. In addition, emerging mechanisms of protection by ischemic preconditioning are discussed. On the basis of current knowledge, preconditioning or pharmacological interventions that mimic these effects have the greatest potential to improve clinical outcome in liver surgery involving ischemic stress and reperfusion.
Collapse
Affiliation(s)
- Hartmut Jaeschke
- Department of Pharmacology and Toxicology, University of Arkansas for Medical Sciences, Little Rock 72205, USA
| |
Collapse
|
|
30
|
Moore TM, Shirah WB, Khimenko PL, Paisley P, Lausch RN, Taylor AE. Involvement of CD40-CD40L signaling in postischemic lung injury. Am J Physiol Lung Cell Mol Physiol 2002; 283:L1255-62. [PMID: 12388354 DOI: 10.1152/ajplung.00016.2002] [Citation(s) in RCA: 16] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/22/2022] Open
Abstract
Our studies show that ischemia-reperfusion (I/R) in the isolated rat lung causes retention of lymphocytes, which is associated with increased microvascular permeability, as determined by quantitative measurement of the microvascular filtration coefficient (K(f,c)). Immunoneutralization of either CD40 or CD40L, cell surface proteins important in lymphocyte-endothelial cell proinflammatory events, results in significantly lower postischemic K(f,c) values. Antagonism of CD40-CD40L signaling also results in attenuation of I/R-elicited macrophage inflammatory protein-2 production. Rat lymphocytes activated ex vivo with phorbol 12-myristate, 13-acetate increased K(f,c) in isolated lungs independently of I/R, and this increase was prevented by pretreating lungs with anti-CD40. In addition to lymphocyte involvement via CD40-CD40L interactions, our studies also show that I/R injury is potentiated by antagonism of IL-10 produced locally within the postischemic lung, whereas exogenous, rat recombinant IL-10 provided protection against I/R-induced microvascular damage. Thus acute lymphocyte involvement in lung I/R injury involves CD40-CD40L signaling mechanisms, and these events may be influenced by local IL-10 generation.
Collapse
Affiliation(s)
- Timothy M Moore
- Department of Physiology, University of Alabama College of Medicine, Mobile, Alabama 36688-0002, USA
| | | | | | | | | | | |
Collapse
|
|
31
|
Khandoga A, Biberthaler P, Enders G, Teupser D, Axmann S, Luchting B, Hutter J, Messmer K, Krombach F. P-selectin mediates platelet-endothelial cell interactions and reperfusion injury in the mouse liver in vivo. Shock 2002; 18:529-35. [PMID: 12462561 DOI: 10.1097/00024382-200212000-00008] [Citation(s) in RCA: 50] [Impact Index Per Article: 2.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/13/2023]
Abstract
Platelets are suggested to participate in the pathogenesis of hepatic ischemia-reperfusion (I/R) injury. This study was designed to analyze platelet-endothelial cell interactions in the postischemic mouse liver in vivo and to define the role of endothelial versus platelet P-selectin for these interactions. Platelet-endothelial cell interactions were quantitatively analyzed using intravital fluorescence microscopy after lobar hepatic I/R in C57BL/6 wild-type and P-selectin-deficient mice after infusion of ex vivo rhodamine-6G-labeled wild-type and P-selectin-deficient platelets. Reperfusion injury and apoptosis were assessed by established methods. In wild-type animals, hepatic I/R caused significantly enhanced platelet-endothelial cell interactions in terminal arterioles and postsinusoidal venules as well as platelet stagnation in sinusoids. Concomitantly, transaminase and caspase-3 activities were elevated and sinusoidal perfusion was impaired. In contrast, platelet-endothelial cell interactions were nearly absent in arterioles and venules of mice lacking endothelial P-selectin, irrespective of the presence of P-selectin on infused platelets, but still significantly elevated in sinusoids. Simultaneously, sinusoidal perfusion failure was ameliorated, and transaminase- and caspase-3 activities were significantly reduced in P-selectin-deficient mice as compared with wild-type animals. The present intravital microscopic study provides, for the first time, quantitative analyses of platelet-endothelial cell interactions in the postischemic hepatic microcirculation. Our in vivo data show that endothelial P-selectin is critical for postischemic platelet-endothelial cell interactions within hepatic presinusoidal arterioles and postsinusoidal venules. P-selectin deficiency prevents microvascular injury and apoptosis after warm hepatic I/R.
Collapse
Affiliation(s)
- Andrej Khandoga
- Institute for Surgical Research, University of Munich, Germany
| | | | | | | | | | | | | | | | | |
Collapse
|
|
32
|
Riaz AA, Wan MX, Schäfer T, Dawson P, Menger MD, Jeppsson B, Thorlacius H. Allopurinol and superoxide dismutase protect against leucocyte-endothelium interactions in a novel model of colonic ischaemia-reperfusion. Br J Surg 2002; 89:1572-80. [PMID: 12445069 DOI: 10.1046/j.1365-2168.2002.02279.x] [Citation(s) in RCA: 39] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/14/2022]
Abstract
BACKGROUND Leucocyte recruitment is a key feature in ischaemia-reperfusion (I/R)-triggered tissue injury. However, the mechanisms underlying leucocyte-endothelium interactions in the large bowel remain elusive because of a previous lack of models to examine the colonic microcirculation. The aim of this study was to develop and validate a novel method for studying reperfusion-induced leucocyte-endothelium interactions in the colon. METHODS The superior mesenteric artery was occluded for 30 min in male C57/Bl6 mice and leucocyte responses were analysed in colonic venules after 30-240 min of reperfusion. Analysis of leucocyte rolling and adhesion in colonic venules was made possible by an inverted approach using intravital fluorescence microscopy. RESULTS Thirty minutes of ischaemia and 120 min of reperfusion induced the strongest and most reproducible increase in leucocyte rolling and adhesion. This was associated with a significant increase in colonic levels of malondialdehyde (MDA). Administration of allopurinol and superoxide dismutase reduced I/R-induced leucocyte responses in a dose-dependent manner. Pretreatment with allopurinol attenuated the tissue content MDA in the colon by more than 60 per cent. CONCLUSION A new method for examining I/R-induced leucocyte responses in the colonic microcirculation is described. Oxygen free radicals play an important role in triggering leucocyte rolling and adhesion in colonic venules.
Collapse
Affiliation(s)
- A A Riaz
- Department of Surgery, Malmö University Hospital, Lund University, Malmö, Sweden, Imperial College School of Medicine, Hammersmith Hospital, London, UK
| | | | | | | | | | | | | |
Collapse
|
|
33
|
Riaz AA, Wan MX, Schaefer T, Schramm R, Ekberg H, Menger MD, Jeppsson B, Thorlacius H. Fundamental and distinct roles of P-selectin and LFA-1 in ischemia/reperfusion-induced leukocyte-endothelium interactions in the mouse colon. Ann Surg 2002; 236:777-84; discussion 784. [PMID: 12454516 PMCID: PMC1422644 DOI: 10.1097/00000658-200212000-00010] [Citation(s) in RCA: 34] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/26/2023]
Abstract
OBJECTIVE To study the adhesive mechanisms underlying ischemia/reperfusion (I/R)-induced leukocyte-endothelium interactions in the colon. SUMMARY BACKGROUND DATA Leukocyte recruitment is a key feature in I/R-induced tissue injury, but the mechanisms regulating leukocyte rolling and adhesion in the colon are not known. The authors recently developed a new model to study the molecular mechanisms of I/R-provoked leukocyte-endothelium interactions in the colon microcirculation using inverted intravital fluorescence microscopy. METHODS The superior mesenteric artery was occluded for 30 minutes and leukocyte responses were analyzed after 120 minutes of reperfusion in colonic venules in mice. The adhesive mechanisms underlying I/R-induced leukocyte rolling and adhesion were investigated using monoclonal antibodies against L-, E- and P-selectin, and CD11a gene-targeted mice were used to examine the role of lymphocyte function antigen-1 (LFA-1, CD11a/CD18). RESULTS Reperfusion provoked a clear-cut increase in leukocyte rolling and adhesion in colonic venules compared to negative controls. Both P- and E-selectin mRNA were expressed in the colon after this I/R insult. Pretreatment with an anti-P-selectin antibody reduced leukocyte rolling and adhesion by 88% and 85%, respectively, whereas antibodies against L- and E-selectin had no effect. Moreover, I/R-induced leukocyte adhesion in LFA-1-deficient mice was reduced by more than 95%. CONCLUSIONS This study provides evidence that leukocyte rolling is exclusively and nonredundantly mediated by P-selectin and that firm adhesion is supported by LFA-1 in I/R-induced leukocyte recruitment in the colon. Taken together, both P-selectin and LFA-1 may be important targets to control pathologic inflammation in I/R-induced tissue injury in the colon.
Collapse
Affiliation(s)
- Amjid Ali Riaz
- Department of Surgery, Malmö University Hospital, Lund University, Malmö, Sweden
| | | | | | | | | | | | | | | |
Collapse
|
|
34
|
Zhou T, Chen JL, Song W, Wang F, Zhang MJ, Ni PH, Geng JG. Effect of N-desulfated heparin on hepatic/renal ischemia reperfusion injury in rats. World J Gastroenterol 2002; 8:897-900. [PMID: 12378638 PMCID: PMC4656583 DOI: 10.3748/wjg.v8.i5.897] [Citation(s) in RCA: 12] [Impact Index Per Article: 0.5] [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 investigate the effect of N-desulfated heparin on hepatic/renal ischemia and reperfusion injury in rats.
METHODS: Using rat models of 60 min hepatic or renal ischemia followed by 1 h, 3 h, 6 h and 24 h reperfusion, animals were randomly divided into following groups, the sham operated controls, ischemic group receiving only normal saline, and treated group receiving N-desulfated heparin at a dose of 12 mg/kg at 5 min before reperfusion. P-selectin expression was detected in hepatic/renal tissues with immunohistochemistry method.
RESULTS: P-selectin expression, serum ALT, AST, BUN and Cr levels were significantly increased during 60 minute ischemia and 1 h, 3 h, 6 h and 24 h reperfusion, while the increment was significantly inhibited, and hepatic/renal pathology observed by light microscopy was remarkably improved by treatment with the N-desulfated heparin. Furthermore, the heparin was found no effects on PT and KPTT.
CONCLUSION: P-selectin might mediate neutrophil infiltration and contribute to hepatic/renal ischemia and reperfusion. The N-desulfated heparin might prevent hepatic/renal damage induced by ischemia and reperfusion injury without significant anticoagulant activity.
Collapse
Affiliation(s)
- Tong Zhou
- Department of Nephrology, Ruijin Hospital, Shanghai Second Medical University, Shanghai 200025,China.
| | | | | | | | | | | | | |
Collapse
|
|
35
|
Tawadrous MN, Zimmermann A, Zhang XY, Wheatley AM. Persistence of impaired hepatic microcirculation after nonarterialized liver transplantation in the rat. Microcirculation 2002; 9:363-75. [PMID: 12375174 DOI: 10.1038/sj.mn.7800153] [Citation(s) in RCA: 13] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/20/2001] [Accepted: 04/08/2002] [Indexed: 11/09/2022]
Abstract
OBJECTIVES The nonarterialized orthotopic rat liver transplant (NOLT) is a frequently used model in transplantation research that was recently used to investigate microcirculatory alterations during acute rejection, which occurs within 7 days. The present study sought to establish whether NOLT represents a reasonable model for the study of the hepatic microcirculation beyond the immediate reperfusion phase. METHODS Three groups of animals were studied: sham-operated control (n = 8), NOLT (n = 7) and arterialized orthotopic liver transplant (AOLT; n = 8). The hepatic microcirculation was investigated by intravital fluorescence microscopy and laser Doppler flowmetry (LDF) on day 7 postoperatively. Liver histology and plasma levels of liver enzymes were also assessed. RESULTS Plasma levels of aspartate aminotransferase, alanine aminotransferase, and bilirubin were significantly higher in NOLT than in AOLT and control animals. The low LDF signal recorded from the surface of the NOLT liver (92 +/- 25 vs. 210 +/- 25 and 172 +/- 14 PU in AOLT and control liver, respectively; p < 0.05) was associated with heterogeneous perfusion at both the lobular and sinusoidal levels (density of perfused sinusoids (n/40,000 microm(2)): 5.8 +/- 0.8, 8.1 +/- 0.3, 8.0 +/- 0.3, respectively; p < 0.05). The percentage of hyperfluorescent Ho342-stained hepatocytes (apoptotic) ranged between 2 and 5% and was not significantly different between groups. The lack of post-transplant arterial supply was associated with an increased hepatic cord width-to-sinusoid diameter ratio (3.77 +/- 0.3, 2.02 +/- 0.04, and 1.72 +/- 0.06 in NOLT, AOLT, and control animals, respectively; p < 0.001 vs. control and AOLT) and increased temporary leukocyte adherence to the walls of the terminal hepatic venules. Intense vitamin A autofluorescence around shunt- and large-diameter, slow-velocity vessels was occasionally encountered in the NOLT liver, which coincided with mild fibrosis and bile ductular proliferation. In the well-perfused areas, both AOLT and NOLT were associated with a significant rise in sinusoidal RBC(vel), which was more marked in the NOLT group. CONCLUSIONS Our data indicate that NOLT represents an inappropriate model for the long-term study of the hepatic microcirculation. Lack of a post-transplant arterial supply may lead to persistent microvascular perfusion failure, hepatocellular/endothelial cell swelling, and microvascular anomalies related to bile duct injury. Recovery from microcirculatory alterations induced by cold preservation/reperfusion injury appears to depend on an intact hepatic arterial blood supply.
Collapse
Affiliation(s)
- Michael N Tawadrous
- Microcirculation Research Laboratory, Department of Physiology, University of Otago, Dunedin, New Zealand
| | | | | | | |
Collapse
|
|
36
|
Scott JR, Fox-Robichaud AE. Hepatic leukocyte recruitment in a model of acute colitis. Am J Physiol Gastrointest Liver Physiol 2002; 283:G561-6. [PMID: 12181168 DOI: 10.1152/ajpgi.00462.2001] [Citation(s) in RCA: 8] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 01/31/2023]
Abstract
There is a close relationship between inflammatory bowel disease (IBD) and various hepatobiliary disorders. The objective of this study was to determine whether hepatic leukocyte recruitment occurs in experimental colitis. We used the murine model of colitis induced by 2,4-dinitrobenezenesulfonic acid (DNBS). Male C57Bl/6 mice received an intrarectal injection of 4 mg DNBS in 100 microl 50% ethanol. Controls received 100 microl 50% ethanol. The hepatic microcirculation was examined at 3 and 14 days post-DNBS by intravital video microscopy. Three days post-DNBS, when mice had developed acute colitis, there was associated hepatic leukocyte recruitment. Within the postsinusoidal venules there was a fourfold increase in the flux of rolling leukocytes that was P-selectin dependent but not alpha(4)-integrin dependent. There was also an increase in stationary leukocytes within the sinusoids, although this was not associated with an increase in serum alanine transaminase. By 14 days post-DNBS when macroscopic evidence of colonic inflammation was resolved, rolling within the postsinusoidal venules had returned to control levels. In this murine model of colitis, we describe a link between acute colonic inflammation and remote hepatic leukocyte recruitment that is P-selectin dependent. Active IBD may lead to remote hepatic inflammation.
Collapse
Affiliation(s)
- Jeffrey R Scott
- Intestinal Disease Research Programme, Department of Medicine, McMaster University, Hamilton, Ontario, Canada L8N 3Z5
| | | |
Collapse
|
|
37
|
Kubes P, Payne D, Woodman RC. Molecular mechanisms of leukocyte recruitment in postischemic liver microcirculation. Am J Physiol Gastrointest Liver Physiol 2002; 283:G139-47. [PMID: 12065301 DOI: 10.1152/ajpgi.00058.2002] [Citation(s) in RCA: 29] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 01/31/2023]
Abstract
Evidence shows that leukocyte recruitment into inflamed liver sinusoids does not require selectins, with one notable exception: ischemia-reperfusion (I/R). We used intravital microscopy to directly visualize the liver microcirculation during I/R and localized endotoxemia (liver superfused with lipopolysaccharide). General anti-selectin therapy (fucoidan) or anti-adhesion therapy with an antithrombin inhibitor (hirudin) was also used. Many neutrophils rolled and adhered in postsinusoidal vessels and sequestered in the sinusoids during I/R and local endotoxin superfusion. Although fucoidan blocked rolling in both forms of inflammation, leukocyte recruitment into sinusoids was only blocked in I/R. Adhesion was also inhibited in postischemic sinusoids with a second anti-adhesive agent (hirudin). Because liver I/R inevitably induces ischemia upstream in the intestine, anti-selectin therapy may prevent intestinal injury, which could prevent downstream liver inflammation. To test this hypothesis, we completely removed the intestine and rerouted blood flow from the superior mesenteric artery to the superior mesenteric vein. I/R was induced in the liver microcirculation, and many leukocytes rolled and adhered in postsinusoidal venules and adhered in sinusoids. Although fucoidan significantly reduced the rolling in postsinusoidal vessels, adhesion persisted in the sinusoids. Our data suggest that anti-adhesion therapy is effective in liver I/R in the sinusoids and postsinusoidal venules, perhaps in part due to its beneficial effect on the intestine.
Collapse
Affiliation(s)
- Paul Kubes
- Immunology Research Group, Department of Physiology and Biophysics and Department of Medicine, University of Calgary Health Sciences Center, Calgary, Alberta, Canada T2N 4N1.
| | | | | |
Collapse
|
|
38
|
Peralta C, Perales JC, Bartrons R, Mitchell C, Gilgenkrantz H, Xaus C, Prats N, Fernández L, Gelpí E, Panés J, Roselló-Catafau J. The combination of ischemic preconditioning and liver Bcl-2 overexpression is a suitable strategy to prevent liver and lung damage after hepatic ischemia-reperfusion. THE AMERICAN JOURNAL OF PATHOLOGY 2002; 160:2111-22. [PMID: 12057915 PMCID: PMC1850813 DOI: 10.1016/s0002-9440(10)61160-1] [Citation(s) in RCA: 38] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 12/21/2022]
Abstract
The present study evaluates the effectiveness of ischemic preconditioning and Bcl-2 overexpression against the liver and lung damage that follow hepatic ischemia-reperfusion and investigates the underlying protective mechanisms. Preconditioning and Bcl-2, respectively, reduced the increased tumor necrosis factor (TNF) and macrophage inflammatory protein-2 (MIP)-2 levels observed after hepatic reperfusion. Bcl-2 overexpression or anti-MIP-2 pretreatment seems to be more effective than preconditioning or anti-TNF pretreatment against inflammatory response, microcirculatory disorders, and subsequent hepatic ischemia-reperfusion injury. Furthermore, each one of these strategies individually was unable to completely inhibit hepatic injury. The combination of preconditioning and Bcl-2 overexpression as well as the combined anti-TNF and anti-MIP-2 pretreatment totally prevented hepatic injury, whereas the benefits of preconditioning and Bcl-2 were abolished by TNF and MIP-2. In contrast to preconditioning, Bcl-2 did not modify lung damage induced by hepatic reperfusion. This could be explained by the differential effect of both treatments on TNF release. Anti-TNF therapy or preconditioning, by reducing TNF release, reduced pulmonary inflammatory response, whereas the benefits of preconditioning on lung damage were abolished by TNF. Thus, the induction of both Bcl-2 overexpression in liver and preconditioning, as well as pharmacological strategies that simulated their benefits, such as anti-TNF and anti-MIP-2 therapies, could be new strategies aimed to reduce lung damage and inhibit the hepatic injury associated with hepatic ischemia-reperfusion.
Collapse
Affiliation(s)
- Carmen Peralta
- Department of Medical Bioanalysis, Instituto de Investigaciones Biomédicas de Barcelona, Consejo Superior de Investigaciones Científicas, Institut d'Investigacions Biomèdiques August Pi i Sunyer, CSIC-IDIBAPS, Barcelona, Spain
| | | | | | | | | | | | | | | | | | | | | |
Collapse
|
|
39
|
Klintman D, Schramm R, Menger MD, Thorlacius H. Leukocyte recruitment in hepatic injury: selectin-mediated leukocyte rolling is a prerequisite for CD18-dependent firm adhesion. J Hepatol 2002; 36:53-9. [PMID: 11804664 DOI: 10.1016/s0168-8278(01)00226-4] [Citation(s) in RCA: 40] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 12/04/2022]
Abstract
BACKGROUND/AIMS This study was designed to examine the role of selectins and CD18 in leukocyte recruitment in hepatic injury induced by tumor necrosis factor-alpha (TNF-alpha) and galactosamine (Gal) in vivo. METHODS Intravital fluorescence microscopy of the hepatic microcirculation was used to quantify leukocyte-endothelium interactions provoked by 24 h of systemic TNF-alpha/Gal challenge in rats. Hepatic injury was evaluated with liver enzymes. RESULTS When administered after 24 h of TNF-alpha/Gal challenge, fucoidan, a selectin-function inhibitor, reduced leukocyte rolling by 69%, whereas firm adhesion was unaltered. In contrast, passive immunization against CD18 decreased leukocyte adhesion by 60%, whereas rolling remained unchanged. Notably, when administered prior to TNF-alpha/Gal, fucoidan attenuated both leukocyte rolling and adhesion, by 57 and 69%, respectively. Pretreatment with an anti-CD18 antibody decreased TNF-alpha/Gal-induced rolling and firm adhesion by 25 and 90%, respectively. Moreover, pretreatment with fucoidan and the anti-CD18 antibody both protected against TNF-alpha/Gal-induced increases in liver enzymes. For example, the pretreatments reduced alanine aminotransferase by 59 and 87%, respectively. CONCLUSIONS Our data suggest that TNF-alpha/Gal-induced leukocyte rolling is selectin-mediated and a precondition for CD18-dependent firm adhesion in hepatic venules. Thus, reducing leukocyte recruitment by inhibition of selectins or CD18 may be useful to control TNF-alpha-induced liver injury.
Collapse
Affiliation(s)
- Daniel Klintman
- Department of Surgery, Malmö University Hospital, Lund University, S-205 02 Malmo, Sweden
| | | | | | | |
Collapse
|
|
40
|
Singbartl K, Forlow SB, Ley K. Platelet, but not endothelial, P-selectin is critical for neutrophil-mediated acute postischemic renal failure. FASEB J 2001; 15:2337-44. [PMID: 11689459 DOI: 10.1096/fj.01-0199com] [Citation(s) in RCA: 134] [Impact Index Per Article: 5.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/11/2022]
Abstract
In a neutrophil-dependent model of acute postischemic renal failure (APRF), eliminating or blocking P-selectin reduces postischemic neutrophil infiltration and preserves kidney function. This study was designed to identify the role of platelet vs. endothelial P-selectin in APRF. Using wild-type (wt) and P-selectin-deficient (P-/-) mice, we generated chimeric mice by bone marrow transplantation. Chimeric mice exclusively expressed either platelet (Plt-P) or endothelial P-selectin (EC-P). APRF was induced by bilateral renal ischemia in situ (32 min), followed by reperfusion; 48 h after reperfusion, EC-P had significantly lower creatinine concentrations (twofold over sham) than Plt-P (eightfold over sham). Compared with wt, protection from renal failure in EC-P was similar to that observed in P-/-. Plt-P and EC-P demonstrated similar overall postischemic neutrophil infiltration as measured by renal myeloperoxidase activity. However, Plt-P showed massive neutrophil infiltration into outer and inner medulla, similar to that in wt. EC-P had only patchy, more diffuse neutrophil influx. Our study identifies platelet P-selectin as crucial for postischemic neutrophil recruitment into outer and inner medulla, which is detrimental to the development of APRF. This suggests that novel therapeutic strategies for postischemic organ failure could be aimed at neutrophil-platelet interactions.
Collapse
Affiliation(s)
- K Singbartl
- Department of Biomedical Engineering, University of Virginia, Charlottesville, Virginia, USA
| | | | | |
Collapse
|
|
41
|
Bajt ML, Farhood A, Jaeschke H. Effects of CXC chemokines on neutrophil activation and sequestration in hepatic vasculature. Am J Physiol Gastrointest Liver Physiol 2001; 281:G1188-95. [PMID: 11668027 DOI: 10.1152/ajpgi.2001.281.5.g1188] [Citation(s) in RCA: 99] [Impact Index Per Article: 4.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 01/31/2023]
Abstract
The initiating step of neutrophil-induced cytotoxicity in the liver is the recruitment of these phagocytes into sinusoids. The aim of our study was to compare the efficacy of systemic exposure with individual inflammatory mediators on neutrophil activation and sequestration in the hepatic vasculature of C3Heb/FeJ mice as assessed by flow cytometry and histochemistry, respectively. The CXC chemokine macrophage inflammatory protein-2 (MIP-2; 20 microg/kg) induced a time-dependent upregulation of Mac-1 (318% at 4 h) and shedding of L-selectin (41% at 4 h). MIP-2 treatment caused a temporary increase of sinusoidal neutrophil accumulation at 0.5 h [97 +/- 6 polymorphonuclear leukocytes (PMN)/50 high-power fields (HPF)], which declined to baseline (8 +/- 2) at 4 h. The CXC chemokine KC was largely ineffective in activating neutrophils or recruiting them into the liver. Cytokines (tumor necrosis factor-alpha and interleukin-1alpha) and cobra venom factor substantially increased Mac-1 expression and L-selectin shedding on neutrophils and caused stable sinusoidal neutrophil accumulation (170-220 PMN/50 HPF). Only cytokines induced venular neutrophil margination. Thus CXC chemokines in circulation are less effective than cytokines or complement in activation of neutrophils and their recruitment into the hepatic vasculature in vivo.
Collapse
Affiliation(s)
- M L Bajt
- Department of Pharmacology and Toxicology, University of Arkansas for Medical Sciences, Little Rock, Arkansas 72205, USA
| | | | | |
Collapse
|
|
42
|
Granger DN, Stokes KY, Shigematsu T, Cerwinka WH, Tailor A, Krieglstein CF. Splanchnic ischaemia-reperfusion injury: mechanistic insights provided by mutant mice. ACTA PHYSIOLOGICA SCANDINAVICA 2001; 173:83-91. [PMID: 11678730 DOI: 10.1046/j.1365-201x.2001.00888.x] [Citation(s) in RCA: 28] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 01/07/2023]
Abstract
Reperfusion of ischaemic tissues often leads to microvascular dysfunction that is manifested as impaired endothelium-dependent dilation of arterioles, enhanced fluid filtration and leucocyte plugging in capillaries, and the trafficking of leucocytes and plasma protein extravasation in postcapillary venules. Efforts to define the mechanisms that underlie these microvascular responses to ischaemia and reperfusion have largely relied on pharmacological agents and monoclonal antibodies. Gene-targeting technology has been applied to the production of transgenic and knockout mice that are rapidly gaining acceptance as tools for mechanistic studies of ischaemia-reperfusion (I/R) injury that obviate some of the concerns (e.g. specificity) raised about previously employed experimental strategies. This review summarizes some of our efforts to apply gene-targeted mice to the study of I/R injury in the splanchnic vascular bed. A role for endothelial cell adhesion molecules (CAMs) and reactive oxygen metabolites is supported by results from mutant mice. Low density lipoprotein receptor mice also reveal that the microvascular and inflammatory responses to I/R are greatly exaggerated during chronic hypercholesterolaemia. The wide variety of mutant mice that have been produced for inflammation-related research makes this experimental strategy particularly promising for mechanistic investigations of the tissue responses to I/R.
Collapse
Affiliation(s)
- D N Granger
- Department of Molecular and Cellular Physiology, Louisiana State University Health Sciences Center, Shreveport, LA 71130-3932, USA
| | | | | | | | | | | |
Collapse
|
|
43
|
Dammers R, Wehrens XH, oude Egbrink MG, Slaaf DW, Kurvers HA, Ramsay G. Microcirculatory effects of experimental acute limb ischaemia-reperfusion. Br J Surg 2001; 88:816-24. [PMID: 11412251 DOI: 10.1046/j.0007-1323.2001.01794.x] [Citation(s) in RCA: 11] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/10/2023]
Abstract
BACKGROUND The object of this study was to develop an animal model in which changes in microvascular haemodynamics and leucocyte-vessel wall interactions due to acute limb ischaemia-reperfusion (I/R) can be measured in the skin. Furthermore, it was investigated whether these changes are related to local muscle injury. METHODS Male Lewis rats were subjected to unilateral limb ischaemia for 1 h (n = 8) or 2 h (n = 8) by cuff inflation, or to a sham protocol (n = 6). Intravital video microscopic measurements of leucocyte-vessel wall interactions, venular diameter, red blood cell velocity and reduced velocity (which is proportional to wall shear rate) were performed in skin venules before ischaemia and at 0.5, 1, 2, 3 and 4 h after the start of reperfusion. Oedema and leucocyte infiltration of ischaemic/reperfused skeletal muscle were quantified histologically. RESULTS In skin venules, both 1 and 2 h of ischaemia induced a significant increase in leucocyte rolling (six and five times baseline, respectively; P < 0.05) and adherence during reperfusion (eight and four times baseline; P < 0.05). No significant increase in muscular leucocyte infiltration was detected. After an initial hyperaemic response of 180 per cent of baseline values (P < 0.05), blood flow decreased to about 60 per cent after 4 h of reperfusion in skin venules of both experimental groups. I/R induced tibial muscle oedema, the severity of which depended on the ischaemic interval (wet to dry ratio: control, 4.0; 1 h, 4.5 (P not significant); 2 h, 5.8 (P < 0.05)). CONCLUSION A non-invasive animal model was developed that enables investigation of the consequences of acute limb I/R.
Collapse
Affiliation(s)
- R Dammers
- Department of General Surgery, University Hospital Maastricht, Maastricht, The Netherlands
| | | | | | | | | | | |
Collapse
|
|
44
|
Sindram D, Porte RJ, Hoffman MR, Bentley RC, Clavien PA. Synergism between platelets and leukocytes in inducing endothelial cell apoptosis in the cold ischemic rat liver: a Kupffer cell-mediated injury. FASEB J 2001; 15:1230-2. [PMID: 11344097 DOI: 10.1096/fj.00-0554fje] [Citation(s) in RCA: 79] [Impact Index Per Article: 3.3] [Reference Citation Analysis] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/13/2022]
Affiliation(s)
- D Sindram
- Hepatobiliary and Liver Transplant Laboratory, Department of Surgery, Duke University Medical Center, Durham North Carolina, USA
| | | | | | | | | |
Collapse
|
|
45
|
Taut FJ, Schmidt H, Zapletal CM, Thies JC, Grube C, Motsch J, Klar E, Martin E. N-acetylcysteine induces shedding of selectins from liver and intestine during orthotopic liver transplantation. Clin Exp Immunol 2001; 124:337-41. [PMID: 11422213 PMCID: PMC1906050 DOI: 10.1046/j.1365-2249.2001.01531.x] [Citation(s) in RCA: 10] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/28/2023] Open
Abstract
In orthotopic liver transplantation (OLT), N-acetylcysteine (NAC) reduces ischaemia/reperfusion (I/R) injury, improves liver synthesis function and prevents primary nonfunction of the graft. To further elucidate the mechanisms of these beneficial effects of NAC, we investigated influence of high-dose NAC therapy on the pattern of adhesion molecule release from liver and intestine during OLT. Nine patients receiving allograft OLT were treated with 150 mg NAC/kg during the first hour after reperfusion; 10 patients received the carrier only. One hour after reperfusion, samples of arterial, portal venous and hepatic venous plasma were taken and blood flow in the hepatic artery and the portal vein was measured. Absolute concentrations of sICAM-1, sVCAM-1, sP-selectin and sE-selectin were not markedly different. However, balance calculations showed release of selectins from NAC-treated livers as opposed to net uptake in controls (P < or = 0.02 for sP-selectin). This shedding of selectins might be a contributing factor to the decrease in leucocyte adherence and improved haemodynamics found experimentally with NAC-treatment.
Collapse
Affiliation(s)
- F J Taut
- Department of Anaesthesiology, University of Heidelberg, Heidelberg, Germany.
| | | | | | | | | | | | | | | |
Collapse
|
|
46
|
Olinde JG, Zibari GB, Brown MF, Howell JG, Akgur FM, Granger DN, McDonald JC. Persantine Attenuates Hemorrhagic Shock-Induced P-Selectin Expression. Am Surg 2000. [DOI: 10.1177/000313480006601202] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/03/2022]
Abstract
Ischemia/reperfusion (I/R), a phenomenon that is associated with conditions such as organ transplantation, trauma, vascular disease, and stroke, involves the recruitment of activated and adherent leukocytes that subsequently mediate tissue injury. Endothelial cell adhesion molecules such as P-selectin mediate I/R-induced leukocyte recruitment and allow the adherent leukocytes to damage the vascular wall and parenchymal cells. This study examines the influence of dypiridamole (persantine) on hemorrhagic shock (H/S)-induced P-selectin expression. H/S was induced in C57BL/6 mice by withdrawing blood to drop the mean arterial blood pressure to 30 to 35 mm Hg for 45 minutes. The mice were resuscitated by infusing the shed blood and Ringer's lactate (50% shed blood volume). In vivo P-selectin expression was determined using a dual monoclonal antibody technique in the heart, lung, liver, kidneys, stomach, small bowel, and colon of a control group, a hemorrhagic shock group, and a hemorrhagic shock group that was pretreated with Persantine (Boehringer, Ingelheim, Ingelheim, Germany). H/S significantly ( P < 0.01) increased P-selectin expression in all regional vascular beds of untreated mice. Persantine treatment largely prevented the H/S-induced P-selectin expression in the same vascular beds. Persantine significantly attenuates the upregulation of P-selectin in the hemorrhagic shock model.
Collapse
Affiliation(s)
- John G. Olinde
- Department of Surgery, Louisiana State University Medical Center-Shreveport, Shreveport, Louisiana
| | - Gazi B. Zibari
- Department of Surgery, Louisiana State University Medical Center-Shreveport, Shreveport, Louisiana
| | - Mark F. Brown
- Department of Surgery, Louisiana State University Medical Center-Shreveport, Shreveport, Louisiana
| | - James G. Howell
- Department of Surgery, Louisiana State University Medical Center-Shreveport, Shreveport, Louisiana
| | - Feza M. Akgur
- Department of Surgery, Louisiana State University Medical Center-Shreveport, Shreveport, Louisiana
| | - D. Neil Granger
- Department of Physiology, Louisiana State University Medical Center-Shreveport, Shreveport, Louisiana
| | - John C. McDonald
- Department of Surgery, Louisiana State University Medical Center-Shreveport, Shreveport, Louisiana
| |
Collapse
|
|
47
|
Martinez-Mier G, Toledo-Pereyra LH, Ward PA. Adhesion molecules in liver ischemia and reperfusion. J Surg Res 2000; 94:185-94. [PMID: 11104660 DOI: 10.1006/jsre.2000.6006] [Citation(s) in RCA: 53] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/22/2022]
Affiliation(s)
- G Martinez-Mier
- Surgery Research Sciences and Molecular Biology, Borgess Research Institute, Kalamazoo, Michigan 49001, USA
| | | | | |
Collapse
|
|
48
|
Mori N, Horie Y, Nimura Y, Wolf R, Granger DN. Hepatic microvascular responses to ischemia-reperfusion in low-density lipoprotein receptor knockout mice. Am J Physiol Gastrointest Liver Physiol 2000; 279:G1257-64. [PMID: 11093949 DOI: 10.1152/ajpgi.2000.279.6.g1257] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 01/31/2023]
Abstract
The overall objective of this study was to determine whether genetically induced hypercholesterolemia alters the inflammatory and microvascular responses of mouse liver to ischemia-reperfusion (I/R). The accumulation of rhodamine 6G-labeled leukocytes and the number of nonperfused sinusoids (NPS) were monitored (by intravital microscopy) in the liver of wild-type (WT) and low-density lipoprotein receptor-deficient (LDLr(-/-)) mice for 1 h after a 30-min period of normothermic ischemia. Plasma alanine transaminase (ALT) levels were used to monitor hepatocellular injury. Microvascular leukostasis as well as increases in NPS and plasma ALT were observed at 60 min after hepatic I/R in both WT and in LDLr(-/-) mice; however, these responses were greatly exaggerated in LDLr(-/-) mice. Pretreatment of LDLr(-/-) mice with gadolinium chloride, which reduces Kupffer cell function, attenuated the hepatic leukostasis, NPS, and hepatocellular injury elicited by I/R. Similar protection against I/R was observed in LDLr(-/-) mice pretreated with antibodies directed against tumor necrosis factor-alpha, intercellular adhesion molecule-1 (ICAM-1), or P-selectin. These findings indicate that chronic hypercholesterolemia predisposes the hepatic microvasculature to the deleterious effects of I/R. Kupffer cell activation and the leukocyte adhesion receptors ICAM-1 and P-selectin appear to contribute to the exaggerated inflammatory responses observed in the postischemic liver of LDLr(-/-) mice.
Collapse
Affiliation(s)
- N Mori
- Departments of Molecular and Cellular Physiology and Medicine, Center of Excellence in Arthritis and Rheumatology, Lousiana State University Health Sciences Center, Shreveport, Louisiana 71130, USA
| | | | | | | | | |
Collapse
|
|
49
|
|
|
50
|
Lentsch AB, Kato A, Yoshidome H, McMasters KM, Edwards MJ. Inflammatory mechanisms and therapeutic strategies for warm hepatic ischemia/reperfusion injury. Hepatology 2000; 32:169-73. [PMID: 10915720 DOI: 10.1053/jhep.2000.9323] [Citation(s) in RCA: 342] [Impact Index Per Article: 13.7] [Reference Citation Analysis] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 12/21/2022]
Affiliation(s)
- A B Lentsch
- Department of Surgery, University of Louisville School of Medicine, Louisville, KY, USA.
| | | | | | | | | |
Collapse
|