益肾通络方对肾纤维化模型大鼠肾组织转化生长因子-β1、Smad7表达的影响

Effects of Yishen Tongluo Fang( 益肾通络方) on Expression of Transforming Growth Factor-β1 and Smad7 in Renal Tissue of Renal Fibrosis Model Rats

目的探讨益肾通络方治疗肾纤维化的可能作用机制。方法采用阳离子牛血清白蛋白皮下和尾静脉注射建立肾纤维化大鼠模型,将造模成功的50只大鼠随机分为模型组、对照组和益肾通络方低、中、高剂量组,每组各10只,并设10只同批正常大鼠为正常组。益肾通络方低、中、高剂量组分别予益肾通络方生药4.58、9.16、18.32 g/(kg·d)灌胃,对照组以肾炎四味片1.58 g/(kg·d)灌胃,模型组和正常组以等容量(1 ml/100 g)的蒸馏水灌胃,每天灌胃1次,连续4周。采用免疫组织化学方法结合图像分析系统测定大鼠肾组织转化生长因子-β1(TGF-β1)、Smad7的平均灰度值、平均光密度值、阳性细胞数、阳性率,并进行相关性分析。结果光镜下模型组肾组织TGF-β1表达明显增多,Smad7表达不明显。与模型组比较,益肾通络方各剂量组和对照组肾组织TGF-β1平均灰度值、阳性率、阳性细胞数明显减少,Smad7平均灰度值、阳性率、阳性细胞数明显提高,并且以益肾通络方中、高剂量作用更明显(P<0.05或P<0.01)。相关分析显示,TGF-β1与Smad7呈线性负相关(P<0.01)。结论益肾通络方治疗肾纤维化可能机制是调节TGF-β1与Smad7的相互作用关系,促进Smad7表达,抑制TGF-β1表达。

Objective To study the possible mechanism of Yishen Tongluo Fang（ 益肾通络方） in treating renal fibrosis. Methods Built rat models with renal fibrosis by subcutaneous and caudal vein injecting cationic bovine serum albumin. Fifty successful models were randomly divided into model group,control group and the low,medium and high dose groups of Yishen Tongluo Fang group,10 rats in each group,and the same batch of 10 normal rats were picked out into normal group. The low,medium and high dose groups of Yishen Tongluo Fang group were respectively given Yishen Tongluo Fang 4. 58,9. 16 and 18. 32 g /（ kg·d） to gavage. The control group was given Shenyan Siwei Pian（ 肾炎四味片） 1. 58 g /（ kg·d） to gavage and the model group and the normal group were given distilled water（ 1 ml/100 g） with the same capacity to gavage,once a day for four weeks. Measure the transforming growth factor-β1（ TGF-β1）,average grey value of Smad7,average optical density value,number of positive cells and positive rate in the renal tissue of rats using immunohistochemical method combining with system of image analysis and made correlation analysis. Results The expression of TGF-β1 of the renal tissue in the model group under optical microscope was obviously increased,but Smad7 was unobviously expressed. Compared with the model group,TGF-β 1 average grey value,positive rate and the number of positive cells of renal tissue decreased significantly in each dose group of Yishen Tongluo Fang and the control group. The average grey value of Smad7,positive rate and the number of positive cells obviously increased and effects were more remarkable in medium and high dose of Yishen Tongluo Fang group（ P〈0. 05 or P〈0. 01）. The correlation analysis showed that the correlation between TGF-β 1 and Smad7 was linear negative（ P〈0. 01）. Conclusion The mechanisms of Yishen Tongluo Fang treating renal fibrosis may adjust the interaction relations between TGF-β 1 and Smad7,promote the expression of Smad7 and inhibit the expression of TGF-β1.