内皮素受体拮抗剂CPU0213 拮抗的对NADPH 氧化酶和PKCe介导的应激性肾损害

摘要/Abstract

摘要： 目的：应激所致的早期肾功能不全, 可能与内皮素-1(ET-1) -蛋白激酶C (PKCε) 通路的异常有关。探讨内皮素受体拮抗剂CPU0213 抑制ET系统和PKCε, 从而改善应激所致的肾功能损伤。方法：SD 大鼠, (240 ±20) g, 随机分成3 组:正常对照组、异丙肾上腺素(ISO 1 mg/kg, s.c., ×10 d) 组、ISO +CPU0213(20 mg/kg, s.c., d 6 -10)治疗。检测血清肌酐、24 h 尿白蛋白、肾皮质GSH-Px 、SOD 和MDA, 及ETA R、PKCε、NAPDH 氧化酶-p67^phox 的mRNA 和蛋白表达。结果：与正常组相比, 血清肌酐和24 h 尿白蛋白浓度明显增高(P <0.01), 肾皮质GSH-Px 、SOD 活力降低, MDA增加, ET_AR 、PKCε、NAPDH 氧化酶-phox67 的mRNA和蛋白表达均上调(P <0.05) 。经CPU0213干预后, 上述指标均趋于正常。结论：应激所致早期肾脏损伤中, NADPH 氧化酶表达增加和PKCε过磷酸化, 由ET_AR 所介导。内皮素受体拮抗剂CPU0213 抑制氧化应激, 改善应激所致的早期肾损伤。

关键词: 内皮素受体拮抗剂剂, PKCε, NAPDH 氧化酶-p67^phox, 应激, 肾损害

Abstract: AIM: Stress likely contributes to oxidative stress initiating early renal insufficiency, that may relate to abnormal endothelin-1 (ET-1) -PKC(protein kinase C) εpathway.It is aimed to test if the endothelin receptor antagonist CPU0213 could reverse stress induced renal early lesion by suppressing abnormal ET and pPKCε. METHODS: Sprague-Dawley rats were subdivided into 3 groups:(1) normal, (2) isoproterenol (ISO) 1 mg/kg, s.c.for 10 days and (3) the ISO injected rats were intervened with CPU0213 20mg/kg, s.c., at day 6 -10.Biochemical measurements and expressions of ETAR (endothelin receptor A), NADPH oxidase p67^phox and PKCεwere conducted. RESULTS: Serum creatinine and 24 h urinary albumin were increased significantly (P < 0.01), compared with normal.The activities of GSHPx and SOD were decreased and the content of MDA was increased in the renal cortex associating with upregulated mRNA and protein of ETAR, PKC (, and NADPH oxidase p67^phox in ISO treated rats.CPU0213 was effective to reverse the renal function by suppressing the abnormalities. CONCLUSION: Activated ETAR, NADPH oxidase and pPKCεare assocaited with renal dysfunction in response to ISO.Endothelin receptor antagonist of CPU0213 is effective in reversing these changes.

Key words: endothelin receptor antagonist, NADPH oxidase-p67^phox, pPKCε, stress, renal injury

中图分类号:

R965.2

徐瑾, 戴德哉, 彭洪军, 戴茵. 内皮素受体拮抗剂CPU0213 拮抗的对NADPH 氧化酶和PKCe介导的应激性肾损害[J]. 中国临床药理学与治疗学, 2009, 14(11): 1247-1252.

XU Jin, DAI De-zai, PENG Hong-jun, DAI Yin. Stress induced early renal injury by upregulating NADPH oxidase, hyperphosphorylation of PKCε is mitigated by endothelin receptor antagonist CPU0213[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2009, 14(11): 1247-1252.

参考文献

[1] Zhang GX, Kimura S, Nishiyama A.Cardiac oxidative stress in acute and chronic Isoproterenol-infused rats[J]. Cardiovascr Res, 2005, 65(1):230-238.
[2] Sorokin A, Kohan DE.Physiology and pathology of endothelin- 1 in renal mesangium[J].Am J Physiol, 2003, 285(4):F579-589.
[3] Anand R, Harry D, Holt S, et al.Endothelin is an important determinant of renal function in a rat model of acute liver and renal failure[J].Gut, 2002, 50(1):111-117.
[4] NittiM, Pronzato MA, Marinari UM, et al.PKC signaling in oxidative hepatic damage[J].Mol AspMed, 2008, 29(1 2):36-42.
[5] Rathore R, Zheng YM, Li XQ, et al.Mitochondrial ROSPKCεsignaling axis is uniquely involved in hypoxic increase in[Ca2+] i in pulmonary artery smooth muscle cells[J].Biochem Biophys Res Commun, 2006, 351(3): 784-790.
[6] Yan F, Li W, Jono H, et al.Reactive oxygen species regulate seudomonas aeruginosa lipopolysaccharide-induced MUC5AC mucin expression via PKC-NADPH oxidase- ROS-TGF-a signaling pathways in human airway epithelial cells[J].Biochem Biophys Res Commun, 2008, 366(2):513-519.
[7] Li N, Jia N, Dai DZ, et al.Endothelin receptor antagonist CPU0213 and vitamin E reverse down regulation of FKBP12.6 and SERCA2a:A role of hyperphosphorylation of PKCepsilon[J].Eur J Pharmacol, 2008, 591(1/2/3): 211-218.
[8] Neuhofer W, Pittrow D.Role of endothelin and endothelin receptor antagonists in renal disease[J].Eur J Clin In-vest, 2006, 36 Suppl 3:78-88.
[9] Qi MY, Xia HJ, Dai DZ, et al.A novel endothelin receptor antagonist CPU0213 improves diabetic cardiac insufficiency attributed to up-regulation of the expression of FKBP12.6, SERCA2a, and PLB in rats[J].J Cardiovasc Pharmacol, 2006, 47(6):729-735.
[10] Xu J, Li N, Dai DZ, et al.The endothelin receptor antagonist CPU0213 is more effective than aminoguanidine to attenuate isoproterenol-induced vascular abnormality by suppressing overexpression of NADPH oxidase, ETA, ETB, and MMP9 in the vasculature[J].J Cardiovasc Pharmacol, 2008, 52(1):42-48.
[11] Xu M, Dai DZ, Dai Y.Normalizing NADPH oxidase contributes to attenuating diabetic nephropathy by a dual endothelin receptor antagonist CPU0213 in rats[J].Am J Nephrol, 2008, 29(3):252-256.
[12] Zhang GX, Ohmori K, Nagai Y, et al.Role of AT1 receptor in isoproterenol-induced cardiac hypertrophy and oxidative stress in mice[J].J Mol Cell Cardiol, 2007, 42(4):804-811.
[13] Xu M, Ji H, Dai DZ, et al.Protective effect of the endothelin antagonist CPU0213 against isoprenaline-induced heart failure by suppressing abnormal expression of leptin, calcineurin and SERCA2a in rats[J].J Pharm Pharmacol, 2008, 60(6):739-745.
[14] Duda M, KoniorA, Klemenska E, et al.Preconditioning protects endothelium by preventing ET-1-induced activation of NADPH oxidase and xanthine oxidase in post-ischemic heart[J].J Mol Cell Cardiol, 2007, 42(2):4000-4041.
[15] Feng Y, Tang XY, Dai DZ, et al.Reversal of isoproterenol- induced downregulation of phospholamban and FKBP12.6 by CPU0213-mediated antagonism of endothelin receptors[J].Acta Pharmacol Sin, 2007, 28(11):1746-1754.
[16] Barton M, Yanagisawa M.Endothelin:20 years from discovery to therapy[J].Can J Physiol Pharmacol, 2008, 86(8):485-498.
[17] Barton M, Medscape.Reversal of proteinuric renal disease and the emerging role of endothelin[J].Nat Clin Pract Nephrol, 2008, 4(9):490-501.
[18] Ortmann J, Amann K, Brandes RP, et al.Role of podocytes for reversal of glomerulosclerosis and proteinuria in the aging kidney after endothelin inhibition[J].Hypertension, 2004, 44(6):974-981.
[19] Imai E, Horio M, Yamagata K, et al.A longitudinal 10- year follow-up study slower decline of glomerular filtration rate in the Japanese general population:a longitudinal 10- year follow-up study[J].Hypertens Res, 2008, 31(3): 433-441.
[20] Zanatta CM, Canani LH, Silveiro SP, et al.Endothelin system function in diabetic nephropathy[J].Arq Bras EndocrinolMetabol, 2008, 52(4):581-588.
[21] Das Evcimen N, King GL.The role of protein kinase C activation and the vascular complications of diabetes[J]. Pharmacol Res, 2007, 55(6):498-510.