排毒保肾丸对肾纤维化大鼠肾组织TGF-β1/Smad信号通路的影响

Effect of Paidu Baoshen Wan( 排毒保肾丸) on TGF-β1 / Smad Signal Pathway of Nephridial Tissue in Renal Fibrosis Rats

目的观察排毒保肾丸对5/6肾切除大鼠肾纤维化的影响并探讨其可能作用机制。方法将50只SD大鼠随机分为正常组、假手术组、模型组、排毒保肾丸组、尿毒清颗粒组,每组10只。除正常组、假手术组外,其余各组大鼠均采用5/6肾切除方法建立肾纤维化模型。排毒保肾丸组和尿毒清颗粒组分别予排毒保肾丸1.0 g/(kg·d)和尿毒清颗粒3.33 g/(kg·d)灌胃,正常组、假手术组、模型组予2 ml生理盐水灌胃,每日1次。各组均灌胃4周后处死大鼠,取残肾组织行HE、PAS、PASM、Masson染色病理观察,检测肾组织转化生长因子β1(TGF-β1)、Smad2、Smad7蛋白及其mRNA的表达水平。结果病理结果显示,模型组大鼠局灶节段性肾小球缺血性皱缩,较多炎性细胞浸润伴轻至中度纤维化;与模型组比较,排毒保肾丸组残肾组织病变程度减轻。与正常组、假手术组比较,模型组大鼠TGF-β1蛋白及其mRNA、Smad2蛋白及其mRNA表达水平均显著升高,Smad7蛋白及其mRNA表达水平均显著下降(P<0.01);与模型组比较,排毒保肾丸组和尿毒清颗粒组TGF-β1蛋白及其mRNA、Smad2蛋白及其mRNA表达水平均显著降低,而Smad7蛋白及其mRNA表达水平均显著升高(P<0.01)。结论排毒保肾丸有抗肾纤维化的作用,下调肾组织TGF-β1、Smad2的表达和上调Smad7的表达可能是其作用机制之一。

Objective To observe effect of Paidu Baoshen Wan （PDBSW） on renal fibrosis of rats with 5/6 nephrectomy and to discuss the underlying mechanism. Methods Fifty SD rats were randomized into normal group, sham-operation group, model group, PDBSW group and Niaoduqing Granules （尿素清颗粒, NDQG） group, with 10 rats in each group. Excluding normal group and sham-operation group, the others received 5/6 nephrectomy to make renal fibrosis model. The PDBSW group and the NDQG group were respectively given PDBSW 1.0 g/ （ kg·d） and NDQG 3.33 g/ （kg ·d） by garage, while the normal group, the sham-operation group and the model group were given normal saline 2 ml/d by gavage. Four weeks later, killed rats and collected remnant nephridial tissue to observe pathology stained by HE, PAS, PASM, Masson, and then to test expression level of protein and mRNA of transforming growth factor/31 （TGF-β1） , Smad2 and Smad7. Results In the model group, topo-segmental glomerulus ischemic atrophy occurred, with much inflammatory cell infiltration and mild to moderate fibrosis. Compared with the model group, the degree of remnant nephridial lesion decreased in PDBSW group. Compared with the normal group and the sham-operation group, expression levels of protein and mRNA of TGF-β1, Smad2 increased markedly in the model group, while those of Smad7 decreased markedly （P 〈 0. 01 ）. Compared with the model group, expression levels of protein and mRNA of TGF-β1, Smad2 in the PDBSW group and the NDQG group decreased markedly, while those of Smad7 increased markedly （P 〈 0. 01 ）. Conclusion PDBSW may help resist renal fibrosis. These effects may be due to reducing expression of TGF-β1 and Smad2, meanwhile increasing expression of Smad7 in nephridial tissue.