|Year : 2015 | Volume
| Issue : 1 | Page : 128
Preventive role of endothelin antagonist on kidney ischemia: Reperfusion injury in male and female rats
Nazgol Esmalian Afyouni1, Hanieh Halili1, Fatemeh Moslemi1, Mehdi Nematbakhsh2, Ardeshir Talebi3, Soheila Shirdavani1, Maryam Maleki1
1 Water and Electrolytes Research Center, Isfahan University of Medical Sciences, Isfahan, Iran
2 Water and Electrolytes Research Center; Department of Physiology, Faculty of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran
3 Department of Clinical Pathology, Faculty of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran
|Date of Submission||14-Jul-2015|
|Date of Acceptance||22-Sep-2015|
|Date of Web Publication||23-Dec-2015|
Department of Physiology, Water and Electrolytes Research Center, Isfahan University of Medical Sciences, Isfahan
Source of Support: None, Conflict of Interest: None
Background: Renal ischemia/reperfusion injury (RIRI) is the most common cause of acute kidney injury. We tested the protective role of endothelin-1 receptor blocker; bosentan (BOS) in animal model of RIRI in two different genders.
Methods: Male and female Wistar rats were assigned as sham operated (sham), control group (ischemia), and case group (ischemia + BOS) treated with BOS (50 mg/kg) 2 h before bilateral kidney ischemia induced by clamping renal vessels for 45 min followed by 24 h of renal reperfusion.
Results: The RIRI significantly increased the serum levels of blood urea nitrogen and creatinine in both genders (P < 0.05). These values were significantly decreased by BOS in both genders. In male rats, the serum levels of malondialdehyde in the ischemia + BOS group were decreased significantly when compared with ischemia group (P < 0.05).
Conclusions: BOS can be used in both genders to attenuate kidney ischemia injury possibly due to its effect in the renal vascular system.
Keywords: Bosentan, endothelin, gender, ischemia-reperfusion, kidney
|How to cite this article:|
Afyouni NE, Halili H, Moslemi F, Nematbakhsh M, Talebi A, Shirdavani S, Maleki M. Preventive role of endothelin antagonist on kidney ischemia: Reperfusion injury in male and female rats. Int J Prev Med 2015;6:128
|How to cite this URL:|
Afyouni NE, Halili H, Moslemi F, Nematbakhsh M, Talebi A, Shirdavani S, Maleki M. Preventive role of endothelin antagonist on kidney ischemia: Reperfusion injury in male and female rats. Int J Prev Med [serial online] 2015 [cited 2020 Jul 12];6:128. Available from: http://www.ijpvmjournal.net/text.asp?2015/6/1/128/172549
| Introduction|| |
Acute renal failure (ARF) or acute kidney injury includes a wide range of disturbance in kidney function.  Renal ischemia/reperfusion injury (RIRI) which occurs mainly in different clinical circumstances such as renal transplantation  is responsible for high morbidity and mortality rate. , Reactive oxygen species in RIRI increase renal vascular resistance and reduce renal blood flow.  Endothelin-1 (ET-1) as one of the most known potent vasoconstrictors  is up-regulated during renal ischemia,  so it increases renal vascular resistance and causes reduction in renal blood flow after reperfusion. ET-1 seems to exacerbate RIRI, because administration of bosentan (BOS) - a dual ET-1 receptor antagonist  demonstrated protective effects on experimental RIRI.  ET-1 and its receptors are effected by sex hormones. Therefore, ET-1 is probably a key mediator in the maintenance of gender-mediated differences after RIRI.  In this short study, we tested the role of BOS RIRI in both male and female rats.
| Methods|| |
Thirty-six age-matched male and female Wistar anesthetized rats were used. Groups 1 (male, n = 6) and 2 (female, n = 6) as sham-operated were subjected to surgery without ischemia. The groups 3 (male, n = 6) and 4 (female, n = 6) received saline as vehicle 2 h before induction of ischemia. Groups 5 (male, n = 6) and 6 (female, n = 6) were treated similar to groups 3 and 4 except they received BOS (50 mg/kg) instead of vehicle. Bilateral kidney ischemia was induced by clamping renal vessels for 45 min. Then, the clamps were removed to induce renal reperfusion. Twenty-four hours postreperfusion, the animal was anesthetized again, and blood samples were obtained via heart puncture. After sacrificing the animals, the kidneys were removed and weighted. Right kidneys were homogenized and centrifuged to obtain supernatant for the measurement.
The serum levels of creatinine (Cr) and blood urea nitrogen (BUN) were measured by quantitative diagnostic kits (Pars Azmoon, Tehran, Iran). Serum and tissue levels of nitrite were measured by an assay kit (Promega Corporation, USA) that involves the Griess reaction. Malondialdehyde (MDA) levels in serum and tissue were also measured manually.
The data were expressed as mean ± standard error of mean The groups were compared in terms of the serum levels of BUN, Cr, nitrite, and MDA; and tissue nitrite and MDA levels and kidney weight (KW) using the one-way analysis of variance followed by the least significant difference test.
| Results|| |
Effects of bosentan on serum levels of blood urea nitrogen and creatinine
RIRI significantly increased the serum levels of BUN and Cr in both genders when compared with the sham operated groups. These observations were significantly decreased by BOS in both genders [Figure 1].
|Figure 1: Serum levels of blood urea nitrogen and creatinine, and kidney weight per 100 g body weight in three experimental groups of sham, ischemia and ischemia treated with bosentan in male and female rats. Star (*) and (#) indicate significant differences (P < 0.05) from sham or ischemia group, respectively. N = 6 in each group|
Click here to view
Effects of bosentan on kidney and body weight
KW was significantly increased by the RIRI, and it was significantly decreased by BOS in female rats [Figure 1]. Body weight change indicated no significant difference between the males and females groups [Table 1].
|Table 1: SN (ìmole/l) and SMDA (ìmole/l), KN (μmole/g tissue) and KMDA(nanomole/g tissue), and ΔW (g) in three experimental groups of sham, ischemia and ischemia treated with BOS in male and female rats |
Click here to view
Effects of bosentan on serum and kidney levels of nitrite and malondialdehyde
There were significant differences in serum and kidney levels of MDA in male but not those of female. Neither male nor female groups demonstrated no significant changes in serum and kidney levels of nitrite [Table 1].
| Discussion|| |
RIRI is the first most common cause of inpatient's ARF. , Over the past years, studies have identified a variety of methods to treat renal RIRI. , In the present study, we attempted to investigate the gender-related effect of BOS as ET-1 blocker on renal RIRI. Our results showed that renal ischemia-reperfusion induced renal failure that characterized by increasing BUN and Cr as well as KW in both genders. These observations were in agreement with others. ,,, There are some reports suggesting that ET-1 is involved in the development of postischemic ARF in clinical transplantation. ,
Recent studies suggest the importance of enhanced renal production of ET-1 in the pathogenesis of ischemic ARF. For example, plasma ET-1 levels are elevated in ARF, and renal ischemia increases renal ET-1 content , and ET receptor affinity. , Infusion of ET-1 decreases glomerular filtration rate, renal blood flow, sodium excretion and increases filtration fraction and renal vascular resistant while co-injection of VML 588 (ET-A antagonist) will reduce all sequences except glomerular filtration rate.  Herrero et al. showed that ET-1 contributes to experimental renal cold ischemia-reperfusion injury, and BOS can attenuate this injury.  The present study showed that pretreatment with BOS as a nonselective dual ETA/ETB receptor antagonist could attenuate post-ischemic renal injury in both genders. Moreover, several investigators have noted that exogenous monoclonal or polyclonal antibody to ET, ,,, ETA receptor antagonists, ,,,, or ETA/ETB dual receptor antagonists ,, ameliorated declines in glomerular filtration rate and tubular damage in ischemia-reperfusion injury. BOS was shown to have a beneficial effect on experimental ischemia/reperfusion injury in the spinal cord, , testis,  heart  and kidney. It was showed that BOS treated rats had higher renal blood flow, Cr clearance, glomerular filtration rate and lower plasma Cr after RIRI.  Also, administration of an ET receptor antagonist 24 h after the ischemic damage was highly effective in reversing ARF. , Despite sexual dimorphism in the majority of physiological and pathophysiological conditions, the majority of experiments on ischemia-induced ARF have been conducted in male animals only. The course of post-ischemic renal failure has not been systematically compared between males and females. Therefore in this study to gain further insights into the role of gender in ischemic renal damage, we compared this process in male and female rats.  There is identified that female mice were more resistant to renal insulin receptor (IR) injury compared with male mice.  Furthermore, the prevalence of ARF is noticeably greater in males than females. ,,, Williams et al. measured the serum level of BUN and Cr levels for 0, 0.5, 1, 2, 4, 6, 9, and 24 h and 1 week post-RIRI, and reported that the earliest renal injury started at the 4 th h following ischemia and peaked at the 1 st day.  Commonly used definitions of ARF include an increase in serum Cr, a reduction in the calculated Cr clearance of 50%, or a decrease in renal function that results in the need for dialysis. ,, However, serum Cr production changes significantly according to age, sex, muscle mass and dietary intake.  The possible effect of sex on serum Cr level might be one of the reasons that we did not infer any significant difference in Cr level among different genders. By contrast, Müller et al. suggested that gender has a major impact on ischemia-induced renal damage and sex hormones play a crucial role in this difference.  Sex hormones have been reported to have an important role in I/R-induced inflammatory processes in the kidneys.  Previous studies have demonstrated that testosterone has an important role in increasing the susceptibility to ischemic renal injury. , By contrast, other experimental results suggest that estrogen has a protective effect in ischemic renal injury in female  via suppression of ET-1 production and activation of the phopsphatidylinositol-3 kinase/protein kinase B signaling pathway. , Our result showed no difference in nitrite level between the male and female groups, and kidney MDA level reduced only in male, but other study showed that kidney MDA level decreased in male and female ischemic animals,  and IR decreased serum and kidney level of nitrite. 
| Conclusions|| |
We conclude that BOS can be used in both genders to attenuate injury induced in kidney ischemia possibly due to its effect in the renal vascular system.
| Acknowledgements|| |
This research was supported by Isfahan University of Medical Sciences.
| References|| |
Kellum JA, Unruh ML, Murugan R. Acute kidney injury. BMJ Clin Evid 2011;2011. pii: 2001.
Abernethy VE, Lieberthal W. Acute renal failure in the critically ill patient. Crit Care Clin 2002;18:203-22, v.
Xing L, Cui R, Peng L, Ma J, Chen X, Xie RJ, et al.
Mesenchymal stem cells, not conditioned medium, contribute to kidney repair after ischemia-reperfusion injury. Stem Cell Res Ther 2014;5:101.
Wang P, Isaak CK, Siow YL, O K. Downregulation of cystathionine ß-synthase and cystathionine?-lyase expression stimulates inflammation in kidney ischemia-reperfusion injury. Physiol Rep 2014;2. pii: E12251.
Cristol JP, Thiemermann C, Mitchell JA, Walder C, Vane JR. Support of renal blood flow after ischaemic-reperfusion injury by endogenous formation of nitric oxide and of cyclo-oxygenase vasodilator metabolites. Br J Pharmacol 1993;109:188-94.
Yanagisawa M, Kurihara H, Kimura S, Tomobe Y, Kobayashi M, Mitsui Y, et al.
A novel potent vasoconstrictor peptide produced by vascular endothelial cells. Nature 1988;332:411-5.
Wilhelm SM, Simonson MS, Robinson AV, Stowe NT, Schulak JA. Endothelin up-regulation and localization following renal ischemia and reperfusion. Kidney Int 1999;55:1011-8.
Clozel M, Breu V, Gray GA, Kalina B, Löffler BM, Burri K, et al.
Pharmacological characterization of bosentan, a new potent orally active nonpeptide endothelin receptor antagonist. J Pharmacol Exp Ther 1994;270:228-35.
Jerkic M, Miloradovic Z, Jovovic D, Mihailovic-Stanojevic N, Elena JV, Nastic-Miric D, et al.
Relative roles of endothelin-1 and angiotensin II in experimental post-ischaemic acute renal failure. Nephrol Dial Transplant 2004;19:83-94.
Müller V, Losonczy G, Heemann U, Vannay A, Fekete A, Reusz G, et al.
Sexual dimorphism in renal ischemia-reperfusion injury in rats: Possible role of endothelin. Kidney Int 2002;62:1364-71.
Ferenbach DA, Kluth DC, Hughes J. Hemeoxygenase-1 and renal ischaemia-reperfusion injury. Nephron Exp Nephrol 2010;115:e33-7.
Lee HT, Emala CW. Protective effects of renal ischemic preconditioning and adenosine pretreatment: Role of A (1) and A (3) receptors. Am J Physiol Renal Physiol 2000;278:F380-7.
Hu L, Yang C, Zhao T, Xu M, Tang Q, Yang B, et al.
Erythropoietin ameliorates renal ischemia and reperfusion injury via inhibiting tubulointerstitial inflammation. J Surg Res 2012;176:260-6.
Henry SD, Guarrera JV. Protective effects of hypothermic ex vivo
perfusion on ischemia/reperfusion injury and transplant outcomes. Transplant Rev (Orlando) 2012;26:163-75.
Azarkish F, Nematbakhsh M, Fazilati M, Talebi A, Pilehvarian AA, Pezeshki Z, et al.
N-acetylcysteine prevents kidney and lung disturbances in renal ischemia/reperfusion injury in rat. Int J Prev Med 2013;4:1139-46.
Moeini M, Nematbakhsh M, Fazilati M, Talebi A, Pilehvarian AA, Azarkish F, et al
. Protective role of recombinant human erythropoietin in kidney and lung injury following renal bilateral ischemia-reperfusion in rat model. Int J Prev Med 2013;4:648.
Malek M, Nematbakhsh M. The preventive effects of diminazene aceturate in renal ischemia/reperfusion injury in male and female rats. Adv Prev Med 2014;2014:740647.
Iran-Nejad A, Nematbakhsh M, Eshraghi-Jazi F, Talebi A. Preventive role of estradiol on kidney injury induced by renal ischemia-reperfusion in male and female rats. Int J Prev Med 2015;6:22.
Yamada K, Gunji Y, Hishikawa E, Kashiwabara H, Sakamoto K, Arita S, et al
. Possible involvement of endothelin in posttransplant acute tubular necrosis. I: Studies in renal transplant patients. Transplantation 1994;57:1137-9.
Tomita K, Ujiie K, Nakanishi T, Tomura S, Matsuda O, Ando K, et al.
Plasma endothelin levels in patients with acute renal failure. N Engl J Med 1989;321:1127.
Mino N, Kobayashi M, Nakajima A, Amano H, Shimamoto K, Ishikawa K, et al.
Protective effect of a selective endothelin receptor antagonist, BQ-123, in ischemic acute renal failure in rats. Eur J Pharmacol 1992;221:77-83.
Shibouta Y, Suzuki N, Shino A, Matsumoto H, Terashita Z, Kondo K, et al.
Pathophysiological role of endothelin in acute renal failure. Life Sci 1990;46:1611-8.
Ruschitzka F, Shaw S, Gygi D, Noll G, Barton M, Lüscher TF. Endothelial dysfunction in acute renal failure: Role of circulating and tissue endothelin-1. J Am Soc Nephrol 1999;10:953-62.
Nambi P, Pullen M, Jugus M, Gellai M. Rat kidney endothelin receptors in ischemia-induced acute renal failure. J Pharmacol Exp Ther 1993;264:345-8.
Vuurmans JL, Boer P, Koomans HA. Effects of endothelin-1 and endothelin-1-receptor blockade on renal function in humans. Nephrol Dial Transplant 2004;19:2742-6.
Herrero I, Torras J, Riera M, Condom E, Coll O, Cruzado JM, et al.
Prevention of cold ischaemia-reperfusion injury by an endothelin receptor antagonist in experimental renal transplantation. Nephrol Dial Transplant 1999;14:872-80.
Kon V, Yoshioka T, Fogo A, Ichikawa I. Glomerular actions of endothelin in vivo
. J Clin Invest 1989;83:1762-7.
López-Farré A, Gómez-Garre D, Bernabeu F, López-Novoa JM. A role for endothelin in the maintenance of post-ischaemic renal failure in the rat. J Physiol 1991;444:513-22.
Espinosa G, López Farré A, Cernadas MR, Manzarbeitia F, Tan D, Digiuni E, et al.
Role of endothelin in the pathophysiology of renal ischemia-reperfusion in normal rabbits. Kidney Int 1996;50:776-82.
Büyükgebiz O, Aktan AO, Haklar G, Yalçin AS, Yegen C, Yalin R, et al.
BQ-123, a specific endothelin (ETA) receptor antagonist, prevents ischemia-reperfusion injury in kidney transplantation. Transpl Int 1996;9:201-7.
Birck R, Knoll T, Braun C, Kirchengast M, Münter K, van der Woude FJ, et al.
Improvement of postischemic acute renal failure with the novel orally active endothelin-A receptor antagonist LU 135252 in the rat. J Cardiovasc Pharmacol 1998;32:80-6.
Chan L, Chittinandana A, Shapiro JI, Shanley PF, Schrier RW. Effect of an endothelin-receptor antagonist on ischemic acute renal failure. Am J Physiol 1994;266:F135-8.
Brooks DP, dePalma PD, Gellai M, Nambi P, Ohlstein EH, Elliott JD, et al.
Nonpeptide endothelin receptor antagonists. III. Effect of SB 209670 and BQ123 on acute renal failure in anesthetized dogs. J Pharmacol Exp Ther 1994;271:769-75.
Gellai M, Jugus M, Fletcher T, Nambi P, Ohlstein EH, Elliott JD, et al
. Nonpeptide endothelin receptor antagonists. V: Prevention and reversal of acute renal failure in the rat by SB 209670. J Pharmacol Exp Ther 1995;275:200-6.
Kusumoto K, Kubo K, Kandori H, Kitayoshi T, Sato S, Wakimasu M, et al.
Effects of a new endothelin antagonist, TAK-044, on post-ischemic acute renal failure in rats. Life Sci 1994;55:301-10.
Gong S, Peng L, Yan B, Dong Q, Seng Z, Wang W, et al.
Bosentan reduces neuronal apoptosis following spinal cord ischemic reperfusion injury. Spinal Cord 2014;52:181-5.
Gong S, Seng Z, Wang W, Lv J, Dong Q, Yan B, et al.
Bosentan protects the spinal cord from ischemia reperfusion injury in rats through vascular endothelial growth factor receptors. Spinal Cord 2015;53:19-23.
Turkili B, Kurcer Z, Dengiz GO, Kandemir NO, Mungan G, Ozacmak VH, et al.
Role of angiotensin and endothelin in testicular ischemia reperfusion injury. Int J Urol 2012;19:257-63.
Gupta SK, Saxena A, Singh U, Arya DS. Bosentan, the mixed ETA-ETB endothelin receptor antagonist, attenuated oxidative stress after experimental myocardial ischemia and reperfusion. Mol Cell Biochem 2005;275:67-74.
Gellai M, Jugus M, Fletcher T, DeWolf R, Nambi P. Reversal of postischemic acute renal failure with a selective endothelinA receptor antagonist in the rat. J Clin Invest 1994;93:900-6.
Kang KP, Lee JE, Lee AS, Jung YJ, Kim D, Lee S, et al.
Effect of gender differences on the regulation of renal ischemia-reperfusion-induced inflammation in mice. Mol Med Rep 2014;9:2061-8.
Paganini EP, Halstenberg WK, Goormastic M. Risk modeling in acute renal failure requiring dialysis: The introduction of a new model. Clin Nephrol 1996;46:206-11.
Chertow GM, Lazarus JM, Paganini EP, Allgren RL, Lafayette RA, Sayegh MH. Predictors of mortality and the provision of dialysis in patients with acute tubular necrosis. The Auriculin Anaritide Acute Renal Failure Study Group. J Am Soc Nephrol 1998;9:692-8.
Mehta RL, Pascual MT, Gruta CG, Zhuang S, Chertow GM. Refining predictive models in critically ill patients with acute renal failure. J Am Soc Nephrol 2002;13:1350-7.
Soljancic A, Ruiz AL, Chandrashekar K, Maranon R, Liu R, Reckelhoff JF, et al.
Protective role of testosterone in ischemia-reperfusion-induced acute kidney injury. Am J Physiol Regul Integr Comp Physiol 2013;304:R951-8.
Williams P, Lopez H, Britt D, Chan C, Ezrin A, Hottendorf R. Characterization of renal ischemia-reperfusion injury in rats. J Pharmacol Toxicol Methods 1997;37:1-7.
Moore RD, Smith CR, Lipsky JJ, Mellits ED, Lietman PS. Risk factors for nephrotoxicity in patients treated with aminoglycosides. Ann Intern Med 1984;100:352-7.
Zanardo G, Michielon P, Paccagnella A, Rosi P, Caló M, Salandin V, et al.
Acute renal failure in the patient undergoing cardiac operation. Prevalence, mortality rate, and main risk factors. J Thorac Cardiovasc Surg 1994;107:1489-95.
Solomon R, Werner C, Mann D, D'Elia J, Silva P. Effects of saline, mannitol, and furosemide to prevent acute decreases in renal function induced by radiocontrast agents. N Engl J Med 1994;331:1416-20.
Shemesh O, Golbetz H, Kriss JP, Myers BD. Limitations of creatinine as a filtration marker in glomerulopathic patients. Kidney Int 1985;28:830-8.
Kher A, Meldrum KK, Wang M, Tsai BM, Pitcher JM, Meldrum DR. Cellular and molecular mechanisms of sex differences in renal ischemia-reperfusion injury. Cardiovasc Res 2005;67:594-603.
Park KM, Kim JI, Ahn Y, Bonventre AJ, Bonventre JV. Testosterone is responsible for enhanced susceptibility of males to ischemic renal injury. J Biol Chem 2004;279:52282-92.
Kim J, Kil IS, Seok YM, Yang ES, Kim DK, Lim DG, et al.
Orchiectomy attenuates post-ischemic oxidative stress and ischemia/reperfusion injury in mice. A role for manganese superoxide dismutase. J Biol Chem 2006;281:20349-56.
Takaoka M, Yuba M, Fujii T, Ohkita M, Matsumura Y. Oestrogen protects against ischaemic acute renal failure in rats by suppressing renal endothelin-1 overproduction. Clin Sci (Lond) 2002;103 Suppl 48:434S-7S.
Satake A, Takaoka M, Nishikawa M, Yuba M, Shibata Y, Okumura K, et al.
Protective effect of 17beta-estradiol on ischemic acute renal failure through the PI3K/Akt/eNOS pathway. Kidney Int 2008;73:308-17.
|This article has been cited by|
||Endothelin-1 in hypertensive patients with ischemic heart disease
| ||Carlo Moroni,Stefano Tolone,Francesco Bondanini,Orazio Schillaci,Cesare Affricano,Rosario Cassone,Achille Gaspardone,Carlo Gaudio |
| ||Internal and Emergency Medicine. 2019; |
|[Pubmed] | [DOI]|
||Involvement of Endothelin 1 in Remote Preconditioning-Induced Cardioprotection through connexin 43 and Akt/GSK-3ß Signaling Pathway
| ||Min Zhang,Wei Wei Gu,Xing yu Hong |
| ||Scientific Reports. 2018; 8(1) |
|[Pubmed] | [DOI]|
||Gender Differences in the Acute Kidney Injury to Chronic Kidney Disease Transition
| ||Ixchel Lima-Posada,Cinthya Portas-Cortés,Rosalba Pérez-Villalva,Francesco Fontana,Roxana Rodríguez-Romo,Rodrigo Prieto,Andrea Sánchez-Navarro,Guadalupe L. Rodríguez-González,Gerardo Gamba,Elena Zambrano,Norma A. Bobadilla |
| ||Scientific Reports. 2017; 7(1) |
|[Pubmed] | [DOI]|