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Treatment With Edaravone Improves the Survival Rate in Renal Warm Ischemia-Reperfusion Injury Using Rat Model
Treatment With Edaravone Improves the Survival Rate in Renal Warm Ischemia-Reperfusion Injury Using Rat Model M. Matsuyama, T. Hayama, K. Funao, K. Tsuchida, Y. Takemoto, K. Sugimura, Y. Kawahito, H. Sano, T. Nakatani, and R. Yoshimura ABSTRACT Renal ischemia-reperfusion (I/R) injury during renal transplantation is a significant cause of renal dysfunction. The pathological role of free radicals in this process is a major concern. We investigated the effect of a free radical scavenger, edaravone (MCI-186), in renal I/R injury. Male Lewis rats (270 to 320 g) were used for the model. The right kidney was harvested and left renal artery and vein were clamped as laparotomy. The kidney was reperfused after 90 minutes of ischemia. Edaravone (10 mg/kg) was delivered intravenously before ischemia and after reperfusion to prevent the neutrophil activation. In the nontreatment I/R group, no rat survived beyond 4 days. However, in the edaravone I/R treatment group, one among five rats survived more than 7 days. These results suggested that treatment with edaravone ameliorated renal I/R injury, and that the agent has the potential to ameliorate preservation injury in renal transplantation.
R
ENAL TRANSPLANTATION is an acceptable therapeutic approach for patients with end-stage renal disease. Renal ischemia-reperfusion (I/R) injury is a main cause of delayed graft function after kidney transplantation. Recent studies of I/R injury have focused on the function of neutrophils, inflammatory cytokines, tissue factor, intercellular adhesion molecule-1, free oxygen radicals, vascular plugging, edema, and other complications.1 MCI-186 (edaravone) is a synthetic radical scavenger that has been used for protection against I/R injury in patients with cerebral infarction. Some papers have demonstrated that edaravone may ameliorate organ I/R injury in the brain, heart, lung, liver, intestine, and kidney. However, no report has demonstrated the treatment with edaravone could improve the survival rate after renal warm I/R injury. In this study, we examined whether treatment with edaravone improved the survival rate in a renal warm I/R injury using a rat model.
MATERIALS AND METHODS Male Lewis rats (270 to 320 g) were used for the model. The abdomen was dissected under anesthesia with ether. The right kidney was harvested and then the left renal artery and vein were clamped with a hemostasis clip for 90 minutes. The clip was subsequently removed to permit reperfusion. Edaravone (10 mg/kg) was delivered intravenously before ischemia and after reperfusion to prevent activation of neutrophils into the inferior vena cava. The abdomen was closed during I/R. We studied two groups: group A, nontreatment
I/R group (n ⫽ 5), versus group B, treatment with edaravone I/R group (n ⫽ 5).
RESULTS
Survival rates were examined after ischemic times of 90 minutes (n ⫽ 5). In the nontreatment I/R group (group A), 5/5 rats survived beyond 1 day; 3/5 rats, 2 days; 2/5 rats, 3 days; and 0/5 rats survived 4 days. However, after treatment with edaravone I/R group (group B), 5/5 rats survived 1 day; 5/5, 2 days; 3/5, 3 days; and 1/5, 4 days and to 7 days (Table 1). In all experimental groups, neither intestinal congestion nor declamping shock was observed. DISCUSSION
In renal transplantation, the problem is the onset of acute tubular necrosis when the transplantation requires a long From the Department of Urology (M.M., T.H., K.F., K.T., Y.T., K.S., T.N., R.Y.), Osaka City University Graduate School of Medicine, Osaka, Japan; Inflammation and Immunology (Y.K.), Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto, Japan; and Division of Rheumatology and Clinical Immunology (H.S.), Department of Internal Medicine, Hyogo College of Medicine, Nishinomiya, Japan. Address reprint requests to Masahide Matsuyama, MD, PhD, Department of Urology, Osaka City University Graduate School of Medicine, 1-4-3 Asahimachi, Abeno-ku, Osaka 545-8585, Japan. E-mail: [email protected]
© 2006 by Elsevier Inc. All rights reserved. 360 Park Avenue South, New York, NY 10010-1710
0041-1345/06/$–see front matter doi:10.1016/j.transproceed.2006.06.077
Transplantation Proceedings, 38, 2199 –2200 (2006)
2199
2200
MATSUYAMA, HAYAMA, FUNAO ET AL Table 1. Effect of Edaravone on Survival Days Over a 7-Day Period Group
Day 0
Day 1
Day 2
Day 3
Day 4
Day 5
Day 6
Day 7
Nontreatment I/R group (n ⫽ 5) Treatment with edaravone I/R group (n ⫽ 5)
5 5
5 5
3 5
2 3
0 1
0 1
0 1
0 1
Note: The survival days over a 7-day period after the ischemic time of 90 minutes. Survival days over a 5-day period were examined after the ishemic time 90 minutes. None (0%) of the five rats in the nontreatment I/R group (group A) survived 4 days. However, 1 (20%) of five rats in the treatment with edaravone I/R group (group B) survived 7 days.
ischemic interval as a consequence of using a cardiac arrest donor’s kidney. It has been reported that the slighter the I/R injury, the milder and shorter the ATN, the better the renal function, and higher postoperative survival rate.2 The priority in renal transplantation is to prevent I/R injury and reduce ATN. Edaravone is a free radical scavenger, which is an antioxidant that has been shown to reduce brain edema in cerebral ischemia through inhibition of the lipoxygenase (LOX) pathway of arachidonic acid. LOX occurs in various isoforms (5-, 8-, 12-, and 15-LOX), and the 5-LOX enzyme synthesizes the leukotrienes (LTs) from the arachidonic acid. 5-LOX induced by cytokines produces various kinds of LTs. The LTA4 is poor in biologic activity, but the LTB4 promotes neutrophilic extravascular migration and venular constriction. The LTC4, D4, and E4 cause smooth muscle contraction, augmented blood vessel permeability, increased production and accentuation of this action. Thromboxane A2, and platelet activating factor. In such activities, we think the LTs serve the purpose of biophylaxis, although their overreaction can raise organ injury to the extreme. It is surmised that 5-LOX is involved in I/R injury.3 Patel et al reported peroral administration of zileuton (3 mg/kg), a 5-LOX inhibitor, reduced renal function and expression of ICAM-1 and associated polymorphonuclear leukocytes at 24 hours after reperfusion in mice at 30 minutes of warm I/R injury.4 We also reported that they augmented immunohistochemical staining in the renal I/R injury model verifying strong expression of 5-LOX several hours after I/R injury.5 Tahara et al reported this in a 72-hour dog renal cold-I/R injury model, edaravone (3 mg/kg) improve a survival.6 Doi et al reported that administration of edara-
vone significantly attenuated renal function and pathological findings by immunohistochemistry and Western analysis in a 45-minute rat renal warm-I/R injury model. Furthermore, edaravone reduce a in vitro oxidative stress using cultured human renal tubular cell.7 These reports support the role of edaravone to ameliorate renal I/R injury through reducing ATN. In conclusion, edaravone may be a potent renal protective therapeutic agent against renal I/R injury. REFERENCES 1. Matsuyama M, Yoshimura R, Akioka K, et al: Tissue factor antisense oligonucleotides prevent renal ischemia-reperfusion injury. Transplantation 76:786, 2003 2. Ojo AO, Wolfe RA, Held PJ, et al: Delayed graft function: risk factors and implications for renal alograft survival. Transplantation 63:968, 1997 3. Matsuyama M, Yoshimura R: Prospects of antisense oligodeoxynucleotides to alleviate renal ischaemia-reperfusion injury. Expert Opin Biol Ther 4:1931, 2004 4. Patel NS, Cuzzocrea S, Chatterjee PK, et al: Reduction of renal ischemia-reperfusion injury in 5-lipoxygenase knockout mice and by the 5-lipoxygenase inhibitor zileuton. Mol Pharmacol 66: 220, 2004 5. Matsuyama M, Nakatani T, Hase T, et al: The expression of cyclooxygenases and lipoxygenases in renal ischemia-reperfusion injury. Transplant Proc 36:1939, 2004 6. Tahara M, Nakayama M, Jin MB, et al: A radical scavenger, edaravone, protects canine kidneys from ischemia-reperfusion injury after 72 hours of cold preservation and autotransplantation. Transplantation 80:213, 2005 7. Doi K, Suzuki Y, Nakao A, et al: Radical scavenger edaravone developed for clinical use ameliorates ischemia/reperfusion injury in rat kidney. Kidney Int 65:1714, 2004
R
ENAL TRANSPLANTATION is an acceptable therapeutic approach for patients with end-stage renal disease. Renal ischemia-reperfusion (I/R) injury is a main cause of delayed graft function after kidney transplantation. Recent studies of I/R injury have focused on the function of neutrophils, inflammatory cytokines, tissue factor, intercellular adhesion molecule-1, free oxygen radicals, vascular plugging, edema, and other complications.1 MCI-186 (edaravone) is a synthetic radical scavenger that has been used for protection against I/R injury in patients with cerebral infarction. Some papers have demonstrated that edaravone may ameliorate organ I/R injury in the brain, heart, lung, liver, intestine, and kidney. However, no report has demonstrated the treatment with edaravone could improve the survival rate after renal warm I/R injury. In this study, we examined whether treatment with edaravone improved the survival rate in a renal warm I/R injury using a rat model.
MATERIALS AND METHODS Male Lewis rats (270 to 320 g) were used for the model. The abdomen was dissected under anesthesia with ether. The right kidney was harvested and then the left renal artery and vein were clamped with a hemostasis clip for 90 minutes. The clip was subsequently removed to permit reperfusion. Edaravone (10 mg/kg) was delivered intravenously before ischemia and after reperfusion to prevent activation of neutrophils into the inferior vena cava. The abdomen was closed during I/R. We studied two groups: group A, nontreatment
I/R group (n ⫽ 5), versus group B, treatment with edaravone I/R group (n ⫽ 5).
RESULTS
Survival rates were examined after ischemic times of 90 minutes (n ⫽ 5). In the nontreatment I/R group (group A), 5/5 rats survived beyond 1 day; 3/5 rats, 2 days; 2/5 rats, 3 days; and 0/5 rats survived 4 days. However, after treatment with edaravone I/R group (group B), 5/5 rats survived 1 day; 5/5, 2 days; 3/5, 3 days; and 1/5, 4 days and to 7 days (Table 1). In all experimental groups, neither intestinal congestion nor declamping shock was observed. DISCUSSION
In renal transplantation, the problem is the onset of acute tubular necrosis when the transplantation requires a long From the Department of Urology (M.M., T.H., K.F., K.T., Y.T., K.S., T.N., R.Y.), Osaka City University Graduate School of Medicine, Osaka, Japan; Inflammation and Immunology (Y.K.), Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto, Japan; and Division of Rheumatology and Clinical Immunology (H.S.), Department of Internal Medicine, Hyogo College of Medicine, Nishinomiya, Japan. Address reprint requests to Masahide Matsuyama, MD, PhD, Department of Urology, Osaka City University Graduate School of Medicine, 1-4-3 Asahimachi, Abeno-ku, Osaka 545-8585, Japan. E-mail: [email protected]
© 2006 by Elsevier Inc. All rights reserved. 360 Park Avenue South, New York, NY 10010-1710
0041-1345/06/$–see front matter doi:10.1016/j.transproceed.2006.06.077
Transplantation Proceedings, 38, 2199 –2200 (2006)
2199
2200
MATSUYAMA, HAYAMA, FUNAO ET AL Table 1. Effect of Edaravone on Survival Days Over a 7-Day Period Group
Day 0
Day 1
Day 2
Day 3
Day 4
Day 5
Day 6
Day 7
Nontreatment I/R group (n ⫽ 5) Treatment with edaravone I/R group (n ⫽ 5)
5 5
5 5
3 5
2 3
0 1
0 1
0 1
0 1
Note: The survival days over a 7-day period after the ischemic time of 90 minutes. Survival days over a 5-day period were examined after the ishemic time 90 minutes. None (0%) of the five rats in the nontreatment I/R group (group A) survived 4 days. However, 1 (20%) of five rats in the treatment with edaravone I/R group (group B) survived 7 days.
ischemic interval as a consequence of using a cardiac arrest donor’s kidney. It has been reported that the slighter the I/R injury, the milder and shorter the ATN, the better the renal function, and higher postoperative survival rate.2 The priority in renal transplantation is to prevent I/R injury and reduce ATN. Edaravone is a free radical scavenger, which is an antioxidant that has been shown to reduce brain edema in cerebral ischemia through inhibition of the lipoxygenase (LOX) pathway of arachidonic acid. LOX occurs in various isoforms (5-, 8-, 12-, and 15-LOX), and the 5-LOX enzyme synthesizes the leukotrienes (LTs) from the arachidonic acid. 5-LOX induced by cytokines produces various kinds of LTs. The LTA4 is poor in biologic activity, but the LTB4 promotes neutrophilic extravascular migration and venular constriction. The LTC4, D4, and E4 cause smooth muscle contraction, augmented blood vessel permeability, increased production and accentuation of this action. Thromboxane A2, and platelet activating factor. In such activities, we think the LTs serve the purpose of biophylaxis, although their overreaction can raise organ injury to the extreme. It is surmised that 5-LOX is involved in I/R injury.3 Patel et al reported peroral administration of zileuton (3 mg/kg), a 5-LOX inhibitor, reduced renal function and expression of ICAM-1 and associated polymorphonuclear leukocytes at 24 hours after reperfusion in mice at 30 minutes of warm I/R injury.4 We also reported that they augmented immunohistochemical staining in the renal I/R injury model verifying strong expression of 5-LOX several hours after I/R injury.5 Tahara et al reported this in a 72-hour dog renal cold-I/R injury model, edaravone (3 mg/kg) improve a survival.6 Doi et al reported that administration of edara-
vone significantly attenuated renal function and pathological findings by immunohistochemistry and Western analysis in a 45-minute rat renal warm-I/R injury model. Furthermore, edaravone reduce a in vitro oxidative stress using cultured human renal tubular cell.7 These reports support the role of edaravone to ameliorate renal I/R injury through reducing ATN. In conclusion, edaravone may be a potent renal protective therapeutic agent against renal I/R injury. REFERENCES 1. Matsuyama M, Yoshimura R, Akioka K, et al: Tissue factor antisense oligonucleotides prevent renal ischemia-reperfusion injury. Transplantation 76:786, 2003 2. Ojo AO, Wolfe RA, Held PJ, et al: Delayed graft function: risk factors and implications for renal alograft survival. Transplantation 63:968, 1997 3. Matsuyama M, Yoshimura R: Prospects of antisense oligodeoxynucleotides to alleviate renal ischaemia-reperfusion injury. Expert Opin Biol Ther 4:1931, 2004 4. Patel NS, Cuzzocrea S, Chatterjee PK, et al: Reduction of renal ischemia-reperfusion injury in 5-lipoxygenase knockout mice and by the 5-lipoxygenase inhibitor zileuton. Mol Pharmacol 66: 220, 2004 5. Matsuyama M, Nakatani T, Hase T, et al: The expression of cyclooxygenases and lipoxygenases in renal ischemia-reperfusion injury. Transplant Proc 36:1939, 2004 6. Tahara M, Nakayama M, Jin MB, et al: A radical scavenger, edaravone, protects canine kidneys from ischemia-reperfusion injury after 72 hours of cold preservation and autotransplantation. Transplantation 80:213, 2005 7. Doi K, Suzuki Y, Nakao A, et al: Radical scavenger edaravone developed for clinical use ameliorates ischemia/reperfusion injury in rat kidney. Kidney Int 65:1714, 2004