Hydrogen sulfide treatment protects against renal ischemia-reperfusion injury via induction of heat shock proteins in rats

Document Type : Original Article


1 Department of Urology, Renmin Hospital of Wuhan University, Hubei, China

2 Physician, Department of Urology, Renmin Hospital of Wuhan University, Hubei, China


Objective(s): Hydrogen sulfide (H2S) attenuates ischemia-reperfusion injury (IRI) in different organs. However, its mechanism of action in renal IRI remains unclear. The present study investigated the hypothesis that H2S attenuates renal IRI via the induction of heat shock proteins (HSPs).
Materials and Methods: Adult Wistar rats were subjected to unilateral renal ischemia for 45 min followed by reperfusion for 6 hr. One group of rats underwent I/R without treatment, one group was administered 150 μmol/l sodium hydrosulfide (NaHS) prior to I/R, one group  was injected with 100 mg/kg quercetin (an HSP inhibitor) intraperitoneally prior to I/R, and another group received quercetin prior to I/R and treatment with NaHS following I/R. Two other groups underwent a sham operation and one of them received 150 μmol/l NaHS following the sham operation whereas the other received no treatment. Renal function and histological changes were compared and relevant indices of oxidative stress, apoptosis, and inflammation were examined.
Results: IRI increased serum creatinine and blood urea nitrogen concentrations, promoted lipid peroxidation by elevating malondialdehyde levels, suppressed superoxide dismutase activity, stimulated inflammation by inducing NF-kB, IL-2, and TLR-4 expression, and increased renal apoptosis. Levels of HSP70, heme-oxygenase-1 (HO-1) and HSP 27 were increased following IRI and reversed following H2S treatment. H2S attenuated changes observed in pathology, lipid peroxidation, inflammation, and apoptosis following IRI. The administration of quercetin reversed all protective effects of H2S.
Conclusion: The present study indicated that H2S protected renal tissue against IRI induced lipid peroxidation, inflammation, and apoptosis, which may be attributed to the upregulation of HSP 70, HO-1, and HSP 27.


Main Subjects

1. Kosieradzki M, Rowinski W. Ischemia/reperfusion injury in kidney transplantation: mechanisms and prevention. Transplant Proc 2008;40:3279-3288.
2. Pruchnicki MC, Dasta JF. Acute renal failure in hospitalized patients: part I. Ann Pharmacother 2002;36:1261-1267.
3. Kunduzova OR BPPN. Regulation of JNK/ERK activation, cell apoptosis, and tissue regeneration by monoamine oxidases after renal ischemia‑reperfusion. FASEB J 2002:16: 1129‑1131.
4. Akoh JA. Transplant nephrectomy. World J Transplantation 2011:1: 4‑12.
5. Kimura H. The physiological role of hydrogen sulfide and beyond. Nitric Oxide 2014;41:4-10.
6. Purandhar K, Jena PK, Prajapati B, Rajput P, Seshadri S. Understanding the role of heat shock protein isoforms in male fertility, aging and apoptosis. World J Mens Health 2014;32:123-132.
7. Zhang PL, Lun M, Schworer CM, Blasick TM, Masker KK, Jones JB, et al.. Heat shock protein expression is highly sensitive to ischemia-reperfusion injury  in rat kidneys. Ann Clin Lab Sci 2008;38:57-64.
8. Guo Q, Du X, Zhao Y, Zhang D, Yue L, Wang Z. Ischemic postconditioning prevents renal ischemia reperfusion injury through the  induction of heat shock proteins in rats. Mol Med Rep 2014;10:2875-2881.
9. Stricher F, Macri C, Ruff M, Muller S. HSPA8/HSC70 chaperone protein: structure, function, and chemical targeting. Autophagy 2013;9:1937-1954.
10.    Wang CF, Wang ZY, Li JY. Dual protective role of HO-1 in transplanted liver grafts: a review of experimental and clinical studies. World J Gastroenterol 2011;17:3101-3108.
11.    Ziemann E, Zembron-Lacny A, Kasperska A, Antosiewicz J, Grzywacz T, Garsztka T, et al.. Exercise training-induced changes in inflammatory mediators and heat shock proteins in young tennis players. J Sports Sci Med 2013;12:282-289.
12.    Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 2001;25:402-408.
13.    Arya R, Mallik M, Lakhotia SC. Heat shock genes - integrating cell survival and death. J Biosci 2007;32:595-610.
14.    Seemampillai B, Germack R, Felkin LE, McCormack A, Rose ML. Heat shock protein-27 delays acute rejection after cardiac transplantation: an experimental model. Transplantation 2014;98:29-38.
15.    Riehle KJ, Hoagland V, Benz W, Campbell JS, Liggitt DH, Langdale LA. Hepatocellular heme oxygenase-1: a potential mechanism of erythropoietin-mediated protection after liver ischemia-reperfusion injury. Shock 2014;42:424-431.
16.    Zhang L, Gan W, An G. Influence of Tanshinone IIa on heat shock protein 70, Bcl-2 and Bax expression in rats with spinal ischemia/reperfusion injury. Neural Regen Res 2012;7:2882-2888.
17.    Li C, Hu M, Wang Y, Lu H, Deng J, Yan X. Hydrogen sulfide preconditioning protects against myocardial ischemia/reperfusion injury in rats through inhibition of endo/sarcoplasmic reticulum stress. Int J Clin Exp Pathol 2015;8:7740-7751.
18.    Zhang Q, Fu H, Zhang H, Xu F, Zou Z, Liu M, et al.. Hydrogen sulfide preconditioning protects rat liver against ischemia/reperfusion  injury by activating Akt-GSK-3beta signaling and inhibiting mitochondrial permeability transition. PLoS One 2013;8:e74422.
19.    Gurbuxani S, Bruey JM, Fromentin A, Larmonier N, Parcellier A, Jaattela M, et al.. Selective depletion of inducible HSP 70 enhances immunogenicity of rat colon cancer cells. Oncogene 2001;20:7478-7485.
20.    Liao W, Li X, Mancini M, Chan L. Proteasome inhibition induces differential heat shock protein response but not unfolded protein response in HepG2 cells. J Cell Biochem 2006;99:1085-1095.
21.    Lepore DA, Knight KR, Anderson RL, Morrison WA. Role of priming stresses and HSP 70 in protection from ischemia-reperfusion injury in cardiac and skeletal muscle. Cell Stress Chaperones 2001;6:93-96.
22.    Cao YA, Kusy S, Luong R, Wong RJ, Stevenson DK, Contag CH. Heme oxygenase-1 deletion affects stress erythropoiesis. PLoS One 2011;6:e20634.
23.    Yang Y, Song HL, Zhang W, Wu BJ, Fu NN, Dong C, et al.. Heme oxygenase-1-transduced bone marrow mesenchymal stem cells in reducing acute  rejection and improving small bowel transplantation outcomes in rats. Stem Cell Res Ther 2016;7:164-172.
24.    Xiong J, Wang K, Yuan C, Xing R, Ni J, Hu G, et al.. Luteolin protects mice from severe acute pancreatitis by exerting HO-1-mediated anti-inflammatory and antioxidant effects. Int J Mol Med 2017;39:113-125.
25.    Jia XM, Zhou ZX, Huang JJ, Chu W, Guan QH. [Protective effects of the induction of heme oxygenase-1 on ischemia reperfusion  lung injury: in vivo experiment with rats]. Zhonghua Yi Xue Za Zhi 2007;87:1211-1213.
26.    Katavetin P, Inagi R, Miyata T, Shao J, Sassa R, Adler S, et al.. Erythropoietin induces heme oxygenase-1 expression and attenuates oxidative stress. Biochem Biophys Res Commun 2007;359:928-934.
27.    Steel R, Doherty JP, Buzzard K, Clemons N, Hawkins CJ, Anderson RL. Hsp72 inhibits apoptosis upstream of the mitochondria and not through interactions with Apaf-1. J Biol Chem 2004;279:51490-51499.
28.    Braunersreuther V, Jaquet V. Reactive oxygen species in myocardial reperfusion injury: from physiopathology to therapeutic approaches. Curr Pharm Biotechnol 2012;13:97-114.
29.    Wang Y, Zhang ZZ, Wu Y, Zhan J, He XH, Wang YL. Honokiol protects rat hearts against myocardial ischemia reperfusion injury by reducing oxidative stress and inflammation. Exp Ther Med 2013;5:315-319.
30.    Ma JQ, Liu CM, Qin ZH, Jiang JH, Sun YZ. Ganoderma applanatum terpenes protect mouse liver against benzo(alpha)pyren-induced oxidative stress and inflammation. Environ Toxicol Pharmacol 2011;31:460-468.
31.    Wong ET, Tergaonkar V. Roles of NF-kappaB in health and disease: mechanisms and therapeutic potential. Clin Sci (Lond) 2009;116:451-465.