雷帕霉素对大鼠缺血再灌注肾Fas，bcl-2蛋白表达的影响

Effect of RPM on bcl-2 and Fas protein expression in renal ischemia-reperfusion injury of rats

目的:探讨雷帕霉素(Rapamycin,RPM)对肾缺血再灌注不同时间Fas,bcl-2蛋白表达的影响。方法:Wistar大鼠54只。随机分为3组:假手术组(切除右肾,分离左肾动脉,不阻断血流,缺血前灌胃给等量生理盐水)、手术组(切除右肾,分离左肾动脉,阻断血流,缺血前灌胃给等量生理盐水)、药物组(切除右肾,分离左肾动脉,阻断血流,缺血前灌胃给RPM 4mg/(kg·d)×3d,术日术前2h灌胃,术后给药至各个观察日)。采用HE染色分析肾组织病理损伤程度,免疫组化SABC法检测bcl-2.Fas蛋白表达的变化。结果:Fas蛋白在假手术组呈阴性表达,手术组再灌注0h有少量表达,再灌注24、48及72h呈阳性表达,以48h时达高峰(P<0.05),主要表达在皮质肾小管;药物组肾组织Fas蛋白水平显著低于手术组(P<0.05)。bcl-2蛋白在假手术组呈弱阳性表达,手术组再灌注0h有少量表达,再灌注24、48及72h呈阳性表达,48h达高峰(P<0.05);皮质肾小管表达为主;药物组肾组织bcl-2蛋白水平显著高于手术组(P<0.05)。HE染色可见手术组肾小管上皮细胞有不同程度的片状坏死灶.肾小管中可见大量死亡脱落细胞残留物质;药物组肾组织损伤明显减轻。结论:RPM可能通过下调Fas,上调bcl-2蛋白表达水平,减轻肾缺血再灌注损伤。

Objective：To investigate the effect of RPM on bcl-2 and Fas protein expressions at different ischemia-reperfusion points. Methods：Acute ischemic renal injury model was established （right kidney ectomized,45 min of left renal ischemia and reperfusion for 0h,24h,48h and 72h）. Fifty four normal male Wistar rats were randomly divided into 3 groups：sham-operated group （sham, without renal ischemia）,ischemia-reperfusion group （IR, without RPM ）, RPM and ischemia-reperfusion group （RPM 4mg/（kg·d）·3d ,reperfusion at 2h before operation and each observation day）. Pathomorphological changes of renal ischemiareperfusion injury were observed by HE stain and SABC immunohistochemical methods were used to detect the changes of expressions of bcl-2 and Fas. Results ：bcl-2 and Fas concentration were significantly higher in IR group than those in sham group（P〈0.05）. As compared with ischemia-reperfusion group（P〈0.05）,bcl-2 expression increased and Fas expression decreased significantly in RPM ＋ IR group. In IR group local necrosis of renal tubule epithelium cells was found by HE stain and large quantity of apoptototis residues were observed, while similar changes were scarcely visible in RPM ＋ IR group. Conelusion：RPM has significant effect on the up-regulation of bcl-2 expression and down-regulation of Fas expression in renal ischemia-reperfusion injury to achieve the purpose of alleviating the injury.