益气活血解毒中药下调SGK-1表达抑制梗阻性肾病炎性损伤的研究

Study on Yiqi Huayu Jiedu Recipe ameliorate the inflammatory injury of obstructive nephropathy by down-regulating the expressions of SGK-1

目的:探讨益气活血解毒中药下调血清糖皮质激素诱导蛋白激酶1(SGK-1)表达抑制梗阻性肾病炎性损伤的作用机制。方法:将48只清洁级雌性Wistar大鼠随机分成假手术组、单侧输尿管结扎组(UUO组)、依普利酮组和益气活血解毒中药组(中药组),每组12只。采用单侧输尿管结扎方式制备梗阻性肾病实验动物模型,两给药组分别予依普利酮(100mg·kg^-1·d^-1)和益气活血解毒中药(13.7g·kg^-1·d^-1)治疗10d。采用免疫组化及Western Blot方法检测肾组织SGK-1、单核细胞趋化蛋白-1(MCP-1)、转化生长因子-β1(TGF-β1)以及小鼠含生长因子样模体黏液样激素样受体(EMR1,又称F4/80)的表达情况。结果:免疫组化结果显示,与假手术组比较,UUO组大鼠肾脏SGK-1、MCP-1和TGF-β1表达明显增强,F4/80阳性细胞明显增多;与UUO组比较,依普利酮组及中药组SGK-1、MCP-1、TGF-β1表达明显减弱,F4/80阳性细胞数目明显减少。Western Blot检测结果显示UUO组大鼠肾脏的SGK-1、MCP-1、TGF-β1蛋白表达较假手术组显著增强(P<0.01),依普利酮组及中药组可显著下调其表达(P<0.01,P<0.05)。结论:益气活血解毒中药可通过下调SGK-1的表达,减少炎性介质MCP-1的分泌,进而下调致纤维化因子TGF-β1的表达,抑制细胞外基质沉积,从而减缓肾间质纤维化的进程。

Objective:To study the effect of Yiqi Huayu Jiedu Recipe ameliorate the inflammatory injury of obstructive nephropathy by down-regulating the expression of serumand glucocorticoid-inducible kinase 1(SGK-1).Methods:Forty-eight female Wistar rats were randomly divided into sham group,unilateral ureter obstruction(UUO)group,eplerenon(EPL)group,and Yiqi Huayu Jiedu Recipe(ZY)group,12 rats in each group.The rat models of obstructive nephropathy were established by unilateral ureter obstruction and rats from EPL and ZY groups were then treated with Eplerenone(100 mg·kg^-1·d^-1)or Chinese materia medica(13.7 g·kg^-1·d^-1)for 10 days.The expression of SGK-1,monocyte chemotactic protein 1(MCP-1),transforming growth factor-β1(TGF-β1)and mouse EGF-like module-containing mucin-like hormone receptor-like 1(F4/80)in kidney were detected by the immunohistochemistry and Western Blot.Results:The immunohistochemistry results showed that compared with the sham group,the expression of SGK-1,MCP-1,TGF-β1 enhanced markedly and F4/80 increased markedly in UUO group;compared with the UUO group,the expression of SGK-1,MCP-1,TGF-β1 reduced markedly and F4/80 decreased markedly in the EPL and ZY group respectively.Western Blot results showed that compared with the sham group,protein expression of SGK-1,MCP-1 and TGF-β1 increased significantly in the UUO group(P<0.01),while EPL and ZY group could down-regulating the expression(P<0.05,P<0.01).Conclusion:Yiqi Huayu Jiedu Recipe could ameliorate renal interstitial fibrosis by reducing the expression of SGK-1 and MCP-1,which could further result in down-regulating expression of TGF-β1,thus decreasing the deposition of extracellular matrix.