益气通阳方对单侧输尿管梗阻大鼠肾间质纤维化的影响

Effects of Yiqi Tongyang Decoction on Renal Interstitial Fibrosis in Unilateral Ureteral Obstruction Rats

目的观察益气通阳方对单侧输尿管梗阻(UUO)大鼠肾间质纤维化及TGF-β1/Smad7信号通路的影响,探讨其可能的作用机制。方法将40只SD大鼠随机分为假手术组、模型组、中药组和西药组各10只。除假手术组外,其余各组建立UUO模型。西药组予依那普利溶液0.18mg/mL灌胃,中药组予益气通阳方浓缩液1.188g/mL灌胃,假手术组和模型组予等量生理盐水灌胃,每日1次。14d后心脏取血检测大鼠血清肌酐(SCr)和尿素氮(BUN)水平,处死大鼠取其肾组织行HE和Masson染色,免疫组化检测肾组织转化生长因子-β1(TGF-β1)和纤维连接蛋白(FN)的表达,Western blot和RT-PCR分别检测Smad7蛋白和m RNA的表达。结果与假手术组比较,模型组大鼠血清SCr、BUN明显升高(P<0.01),病理改变明显,TGF-β1、FN蛋白表达明显升高(P<0.01),Smad7蛋白和m RNA表达明显降低(P<0.01);与模型组比较,中药组和西药组大鼠血清SCr、BUN明显降低(P<0.05),病理改变减轻,TGF-β1、FN蛋白表达明显降低(P<0.05),Smad7蛋白和m RNA表达明显升高(P<0.05)。结论益气通阳方可能通过上调Smad7表达,抑制UUO大鼠肾组织TGF-β1的高表达,进而延缓肾间质纤维化进展。

Objective To investigate the effects of Yiqi Tongyang Decoction on renal interstitial fibrosis in unilateral ureteral obstruction (UUO) rats and the role of TGF-β 1/Smad7 signaling pathway;To explore possible the mechanism of action. Methods Totally 40 SD rats were randomly divided into sham-operation group, model group, TCM group and Western medicine group, with 10 rats in each group. Excluding the sham-operation group, UUO models were established in the others. Then the Western medicine group was given enalapril liquid 0.18 mg/mL for gavage;the TCM group was given Yiqi Tongyang Decoction concentrated liquid 1.188 g/mL for gavage;the other groups were given the same amount of normal saline for gavage once a day. After 14 days, serum was collected for detecting the levels of SCr and BUN. Then the rats were killed for taking the renal tissue and conducting HE and Masson staining. The expressions of TGF-β 1 and FN protein were detected by using immunohistochemistry method. Western blot and RT-PCR were used to detect the expressions of Smad7 protein and mRNA. Results Compared with the sham-operation group, the levels of SCr and BUN in the model group significantly increased ( P <0.01), and the pathological changes were obvious, the expression of TGF-β 1 and FN were significantly increased ( P <0.01), and the expression of Smad7 and its mRNA decreased ( P <0.01). Compared with the model group, the levels of SCr and BUN in the TCM group and Western medicine group significantly decreased ( P <0.05), the degree of tubulointerstitial injury decreased, also the expressions of TGF-β 1 and FN protein significantly decreased ( P <0.05), while Smad7 and its mRNA expression significantly increased (P<0.05). Conclusion Yiqi Tongyang Decoction can reduce renal interstitial fibrosis possibly by inhibiting the expression of TGF-β1 and up-regulating the expression of Smad7 in UUO rats.