活血解毒中药下调8-OhdG表达抑制梗阻性肾病大鼠细胞凋亡

摘要

Aim To observe the effect of eplerenone(EPL) and Chinese decoction on cell apoptosis in obstructive nephropathy rats.Methods Sixty male Wistar rats were randomly divided into sham group, UUO group, EPL group and ZY group(n=15).Except sham group, the rats in the other groups were ligated with unilateral ureteral obstruction(UUO) for renal interstitial fibrosis model.The rats were treated with eplerenone at 100 mg·kg-1·d-1 added to diet in EPL group, and orally 13.7 g·kg-1·d-1 decoction of Chinese medicine in ZY group.The kidneys were harvested on 14th day, the number of renal cell apoptosis were detected by TUNEL, and serum aldosterone and 8-OhdG were detected with radioimmunoassay and ELISA.Caspase-12, caspase-9, Bax and Bcl-2 were examined by immunohistochemistry and Western blot.Results The levels of serum aldosterone, serum and urine 8-OhdG and the number of positive apoptotic cells increased significantly in UUO rats compared with Sham group.The overexpression of caspase-9, caspase-12 and Bax and down-regulated Bcl-2 were obvious in UUO group(P<0.01).The level of 8-OhdG, expression of caspase-9, caspase-12 and Bax were down-regulated, and Bcl-2 expression was up-regulated in eplerenone and Chinese decoction treated rats(P<0.01).Conclusion Eplerenone and Chinese decoction could inhibit cell apoptosis induced by oxidative damage after UUO via caspases and(or) Bax pathway.%目的 观察依普利酮及活血解毒中药对梗阻性肾病大鼠肾小管上皮细胞凋亡的影响及作用机制.方法 将60只清洁级♂ Wistar大鼠随机分为假手术组、模型组、依普利酮组、中药组,每组15只.除假手术组外,其余大鼠结扎单侧输尿管复制肾间质纤维化动物模型.依普利酮组给予依普利酮100 mg·kg-1·d-1加入饲料喂养,中药组给予活血解毒中药方煎剂13.7 g·kg-1·d-1灌胃.14 d后摘取梗阻侧肾脏,TUNEL法检测肾脏细胞凋亡,放射免疫分析法检测血清醛固酮含量,ELISA检测血清及尿液8-OhdG含量,免疫组化、Western blot检测caspase-12、caspase-9、Bax以及Bcl-2蛋白表达.结果 UUO大鼠血清醛固酮、血清及尿液8-OhdG含量及肾脏阳性凋亡细胞明显升高,caspase-9、caspase-12、Bax蛋白表达明显增强,Bcl-2表达下降(P<0.01);依普利酮及中药可降低血尿8-OhdG水平(P<0.01),下调caspase-9、caspase-12、Bax蛋白水平的表达(P<0.01).结论 依普利酮及活血解毒中药可以通过抑制氧化损伤,阻断caspase和(或)Bax信号通路,减少输尿管梗阻后肾小管上皮细胞凋亡.