百令胶囊联合还原型谷胱甘肽保护单侧输尿管梗阻大鼠肾纤维化的机制探究

Mechanisms of Bailing Capsule Combined with Reduced Glutathione in Protecting against Renal Fibrosis in Rats with Unilateral Ureteral Obstruction

目的探究百令胶囊联合还原型谷胱甘肽保护单侧输尿管梗阻(UUO)大鼠肾纤维化的机制。方法40只SD大鼠随机分为假手术组、UUO组、还原型谷胱甘肽组及百令胶囊联合还原型谷胱甘肽组(联合组),每组10只,采用HE和Masson染色观察大鼠肾组织病理变化,测定血肌酐(Scr)和尿素(BUN),制备肾皮质匀浆液测定活性氧(ROS)、超氧化物歧化酶(SOD)和丙二醛(MDA)含量,蛋白免疫印迹法检测肾组织中Nrf2和Keap1蛋白表达。结果与假手术组比较,UUO组血清Scr、BUN和梗阻侧肾脏指数明显升高,肾组织匀浆液中ROS和MDA含量明显升高、SOD含量明显降低,肾组织中Nrf2蛋白表达明显降低、Keap1蛋白表达明显升高(P<0.05);与UUO组比较,还原型谷胱甘肽组和联合组Scr、BUN和梗阻侧肾脏指数明显降低,ROS和MDA含量明显降低、SOD含量明显升高,Nrf2蛋白表达明显升高、Keap1蛋白表达明显降低,且联合组改善效果优于还原型谷胱甘肽组(P<0.05)。结论百令胶囊联合还原型谷胱甘肽可明显减轻UUO大鼠肾纤维化,可能与通过调节Nrf2/Keap1信号通路减轻氧化应激有关。

Objective To explore the mechanism of Bailing Capsule combined with reduced glutathione in protecting against renal fibrosis in rats with unilateral ureteral obstruction(UUO).Methods Forty SD rats were randomly divided into sham operation group,UUO group,reduced glutathione group and Bailing Capsule combined with reduced glutathione group(combination group,n=10 in each group).HE and Masson staining were used to observe pathological changes of renal tissues,and serum levels of creatinine(Scr)and urea(BUN)were measured.Renal cortex homogenate was prepared to detect contents of reactive oxygen species(ROS),superoxide dismutase(SOD)and malondialdehyde(MDA).Protein expressions of Nrf2 and Keap1 in kidney tissues were detected by Western immunoblotting method.Results Compared with those in sham operation group,serum levels of Scr and BUN and renal index of the obstructive side were significantly increased,and ROS and MDA levels in renal tissue homogenate were significantly increased,but SOD content and expression of Nrf2 protein in renal tissue were significantly decreased,while expression of Keap1 protein was significantly increased in UUO group(P<0.05).Compared with those in UUO group,levels of Scr,BUN and renal index of the obstructive side as well as contents of ROS and MDA were significantly decreased,while SOD content and Nrf2 protein expression were significantly increased,while Keap1 protein expression was significantly decreased in reduced glutathione and combination groups;the improvement effects in combination group were better than those in reduced glutathione group(P<0.05).Conclusion Bailing Capsule combined with reduced glutathione may significantly ameliorate renal fibrosis in UUO rats,which may be related to reducing oxidative stress by regulating Nrf2/Keap1 signaling pathway.