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FR167653 ameliorates ischemia-reperfusion injury of the rat liver through P38 mitogen-activated protein kinase pathway
FR167653 Ameliorates Ischemia-Reperfusion Injury of the Rat Liver Through P38 Mitogen-Activated Protein Kinase Pathway M. Kobayashi, I. Takeyoshi, D. Yoshinari, Y. Koibuchi, T. Koyama, Y. Kawashima, S. Ohwada, K. Matsumoto, and Y. Morishita
W
E PREVIOUSLY reported that FR167653(FR), a potent suppressant of interleukin-1 (IL-1) and tumor necrosis factor-␣ (TNF-␣), ameliorated ischemiareperfusion injury of the canine lung and liver.1,2 Several studies have implicated the mitogen-activated protein kinases (MAPKs) signal pathway in ischemia-reperfusion of organs. However, the role of p38 MAPK in hepatic ischemia-reperfusion injury remains unclear. This study aimed to investigate the role of p38 MAPK on hepatic ischemiareperfusion injury.
MATERIALS AND METHODS Male Sprague-Dawley rats were used. Liver warm ischemia was induced for 30 minutes using the Pringle’s maneuver. Animals were divided into two groups; the control group and the FR-treated group, which received FR (0.1 mg/kg per hour) from 30 minutes prior to ischemia to 2 hours after reperfusion. Serum levels of asparate transaminase (AST), alanine transaminase (ALT), lactate dehydrogenase (LDH), TNF-␣, and IL-1 were measured 2 hours after reperfusion. The liver tissue blood flow (LTBF) and histologic findings were investigated. The expression of total p38 MAPK and phosphorylated p38 MAPK in the liver tissue were measured using Western blot analysis.
RESULTS
Serum levels of AST, ALT, LDH, TNF-␣, and IL-1 were significantly lower in the FR-treated group than in the control group. LTBF was significantly higher in the FRtreated group than in the control group. Histopathologically, tissue damage was mild in the FR-treated group. Western blot analysis revealed that the expression of total p38 MAPK did not differ between the control and the FR-treated group. Meanwhile, the expression of phosphorylated p38 MAPK after 30 minutes of reperfusion was dramatically suppressed in the FR-treated group compared to the control group.
© 2001 by Elsevier Science Inc. 655 Avenue of the Americas, New York, NY 10010
DISCUSSION
IL-1 and TNF-␣ derived from activated Kuppfer cells play a pivotal role in the pathogenesis of hepatic ischemiareperfusion injury.3 FR is a potent suppressant of IL-1 and TNF-␣ in a model of disseminated intravascular coagulation.4 In the MAPKs, p38 MAPK and c-Jun N-terminal kinase (JNK) are reported to be activated by a variety of cellular stresses, such as inflammatory cytokines, lipopolysaccharides, heat shock, and ischemia-reperfusion, and also reported to mediate apoptosis and production of inflammatory cytokines.5 However, the role of p38 MAPK in hepatic ischemia-reperfusion injury remains unclear. In the present study, FR administration ameliorated ischemia-reperfusion injury of the rat liver with suppressing IL-1, TNF-␣, and inhibited phosphorylation of p38 MAPK. These results suggest that p38 MAPK inhibition attenuates ischemiareperfusion injury of the rat liver. REFERENCES 1. Kamoshita N, Takeyoshi I, Ohwada S, et al: J Heart Lung Transplant 16:1062, 1997 2. Kobayashi J, Takeyoshi I, Ohwada S, et al: Hepatology 28:459, 1998 3. Andus T, Bauer J, Gerock W: Hepatology 13:364, 1991 4. Yamamoto N, Sakai F, Yamazaki H, et al: Eur J Pharmacol 314:137, 1996 5. Clerk A, Fuller SJ, Michael A, et al: J Biol Chem 273:7228, 1998 From the Second Department of Surgery, Gunma University School of Medicine (M.K., I.T., D.Y., Y.K., T.K., Y.K., S.O., Y.M.), Maebashi, Japan; and Department of Pathology, Nippon Medical School (K.M.), Kawasaki, Japan. Address reprint requests to Dr I. Takeyoshi, Gunma University School of Medicine, 2nd Department of Surgery, 3-39-15 Showa-machi, Maebashi, Gunma, 371-8511 Japan.
0041-1345/01/$–see front matter PII S0041-1345(00)02353-8 865
Transplantation Proceedings, 33, 865 (2001)
W
E PREVIOUSLY reported that FR167653(FR), a potent suppressant of interleukin-1 (IL-1) and tumor necrosis factor-␣ (TNF-␣), ameliorated ischemiareperfusion injury of the canine lung and liver.1,2 Several studies have implicated the mitogen-activated protein kinases (MAPKs) signal pathway in ischemia-reperfusion of organs. However, the role of p38 MAPK in hepatic ischemia-reperfusion injury remains unclear. This study aimed to investigate the role of p38 MAPK on hepatic ischemiareperfusion injury.
MATERIALS AND METHODS Male Sprague-Dawley rats were used. Liver warm ischemia was induced for 30 minutes using the Pringle’s maneuver. Animals were divided into two groups; the control group and the FR-treated group, which received FR (0.1 mg/kg per hour) from 30 minutes prior to ischemia to 2 hours after reperfusion. Serum levels of asparate transaminase (AST), alanine transaminase (ALT), lactate dehydrogenase (LDH), TNF-␣, and IL-1 were measured 2 hours after reperfusion. The liver tissue blood flow (LTBF) and histologic findings were investigated. The expression of total p38 MAPK and phosphorylated p38 MAPK in the liver tissue were measured using Western blot analysis.
RESULTS
Serum levels of AST, ALT, LDH, TNF-␣, and IL-1 were significantly lower in the FR-treated group than in the control group. LTBF was significantly higher in the FRtreated group than in the control group. Histopathologically, tissue damage was mild in the FR-treated group. Western blot analysis revealed that the expression of total p38 MAPK did not differ between the control and the FR-treated group. Meanwhile, the expression of phosphorylated p38 MAPK after 30 minutes of reperfusion was dramatically suppressed in the FR-treated group compared to the control group.
© 2001 by Elsevier Science Inc. 655 Avenue of the Americas, New York, NY 10010
DISCUSSION
IL-1 and TNF-␣ derived from activated Kuppfer cells play a pivotal role in the pathogenesis of hepatic ischemiareperfusion injury.3 FR is a potent suppressant of IL-1 and TNF-␣ in a model of disseminated intravascular coagulation.4 In the MAPKs, p38 MAPK and c-Jun N-terminal kinase (JNK) are reported to be activated by a variety of cellular stresses, such as inflammatory cytokines, lipopolysaccharides, heat shock, and ischemia-reperfusion, and also reported to mediate apoptosis and production of inflammatory cytokines.5 However, the role of p38 MAPK in hepatic ischemia-reperfusion injury remains unclear. In the present study, FR administration ameliorated ischemia-reperfusion injury of the rat liver with suppressing IL-1, TNF-␣, and inhibited phosphorylation of p38 MAPK. These results suggest that p38 MAPK inhibition attenuates ischemiareperfusion injury of the rat liver. REFERENCES 1. Kamoshita N, Takeyoshi I, Ohwada S, et al: J Heart Lung Transplant 16:1062, 1997 2. Kobayashi J, Takeyoshi I, Ohwada S, et al: Hepatology 28:459, 1998 3. Andus T, Bauer J, Gerock W: Hepatology 13:364, 1991 4. Yamamoto N, Sakai F, Yamazaki H, et al: Eur J Pharmacol 314:137, 1996 5. Clerk A, Fuller SJ, Michael A, et al: J Biol Chem 273:7228, 1998 From the Second Department of Surgery, Gunma University School of Medicine (M.K., I.T., D.Y., Y.K., T.K., Y.K., S.O., Y.M.), Maebashi, Japan; and Department of Pathology, Nippon Medical School (K.M.), Kawasaki, Japan. Address reprint requests to Dr I. Takeyoshi, Gunma University School of Medicine, 2nd Department of Surgery, 3-39-15 Showa-machi, Maebashi, Gunma, 371-8511 Japan.
0041-1345/01/$–see front matter PII S0041-1345(00)02353-8 865
Transplantation Proceedings, 33, 865 (2001)