芪归益肾方延缓单侧输尿管梗阻小鼠肾脏纤维化进展机制的探讨

Anti-Fibrosis Effect of Qigui Yishen Formula in Kidney of Mice with Unilateral Ureteral Obstruction

目的:利用单侧输尿管梗阻(unilateral ureteral obstruction,UUO)肾病模型研究芪归益肾方通过调控细胞信号通路TGF-β/Smad/ILK的表达水平影响肾脏纤维化进展的程度。方法:昆明种小鼠32只,随机分为正常组、模型组、贝那普利组和芪归益肾方组,每组8只,除正常组外,其余各组制备UUO肾病模型后分别进行药物干预,第10天处死后取肾组织,分别采用苏木素-伊红(HE)染色观察小鼠肾组织的病理变化,免疫组化观察小鼠肾组织转化生长因子-β1(TGF-β1),细胞信号转导分子2(Smad2),Ⅰ型胶原(Col-Ⅰ)和整合素连接激酶(ILK)蛋白的表达,逆转录-聚合酶链式反应(RT-PCR)和蛋白质免疫印迹(Western blot)检测小鼠肾组织TGF-β1,Col-Ⅰ和ILK mRNA及蛋白的表达情况。结果:与正常组比较,免疫组化结果显示模型组小鼠肾组织TGF-β1,Smad2,Col-Ⅰ,ILK蛋白表达量明显升高,RT-PCR和Western blot检测结果显示小鼠肾组织TGF-β1,Col-Ⅰ和ILK mRNA及蛋白表达明显升高(P<0.05),模型组小鼠肾脏纤维化程度较为明显;与模型组比较,贝那普利组和芪归益肾方组治疗后肾脏纤维化程度均有减轻,明显降低小鼠肾组织TGF-β1,Smad2,Col-Ⅰ,ILK蛋白表达,明显降低小鼠肾组织TGF-β1,Col-Ⅰ和ILK mRNA及蛋白的表达(P<0.05),其中芪归益肾方治疗组比较贝那普利组效果更为明显(P<0.05)。结论:芪归益肾方对于UUO肾病模型的纤维化病变程度具有抑制作用,其作用主要与调控细胞信号通路TGF-β/Smad/ILK表达相关。

Objective： To study the effect of Qigui Yishen formula（ QGYS） on delaying the renal fibrosis by regulating the expression of cell signaling pathway TGF-β/Smad/ILK in unilateral ureteral obstruction（ UUO） nephropathy models. Method： The 32 Kunming mice were randomly divided into normal group,model group,benazepril group,and QGYS group,n = 8 in each group. All the mice except those in normal group were used to prepare UUO nephropathy models. Corresponding drugs were used in various groups then. Renal tissues were taken after sacrificing on day 10 and were used for HE staining to observe the pathological changes of renal tissues in mice; immunohistochemistry was used to observe transforming growth factor-1（ TGF-β1） of renal tissues,cell signal transduction molecules-2（ Smad2）,type I collagen（ Col-I） and integrin linked kinase（ ILK） protein expressions; RT-PCR and Western blot were used to observe the mRNA and protein expression levels of TGF-β1,Col-I and ILK. Result： As compared with the normal group,immunohistochemistry results showed that the protein expression levels of TGF-β1,Smad2,Col-I and ILK were significantly increased in renal tissues of model group mice; RT-PCR and Western blot results showed that the mRNA and protein expression levels of TGF-β1,Col-I and ILK were significantly increased in renal tissues of model group mice（ P〈0. 05）,with more obvious renal fibrosis.As compared with the model group,renal fibrosis was relieved in benazepril group,and QGYS group; protein expression levels of TGF-β1,Smad2,Col-I and ILK were significantly reduced（ P〈0. 05）,and the effect in QGYS group was more obvious in that of benazepril group（ P〈0. 05）. Conclusion： QGYS could decrease the level of renal fibrosis in UUO nephropathy models,and the action mechanism may be associated with regulating the expression of the cell signaling pathway of TGF-β/Smad/ILK.