祛风通络方对UUO大鼠肾间质纤维化相关蛋白α-SMA及vimentin 表达的影响

Effects of Qufeng Tongluo Prescription on Renal Interstitial Fibrosis-associated Protein α-SMA and Vimentin Expression of UUO Rats

目的:观察祛风通络方对单侧输尿管结扎术(unilateral urethral occlusion,UUO)大鼠肾间质纤维化相关蛋白α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)、波形蛋白(vimentin)的影响。方法:50只雄性成年SD大鼠随机分为假手术组10只和UUO组40只,其中UUO组根据干预方法不同随机平均分为模型组、阳性对照组、祛风通络方高剂量组、祛风通络方低剂量组。术后第1天开始灌胃相应药物,术后第21天将大鼠全部处死,取术侧肾脏组织,Masson染色观察肾间质纤维化程度,RT-PCR和Western blotting检测肾脏组织α-SMA、vimentin mRNA水平及蛋白表达。结果:①UUO后21 d,模型组及治疗各组大鼠梗阻侧肾体积较对侧增大,剖面肾实质不同程度萎缩,肾盂扩张呈囊状,模型组最为明显。②模型组大鼠近端肾小管上皮细胞萎缩,集合管和远曲小管管腔明显扩张,周围大量炎性细胞浸润伴间质胶原成分显著增加、纤维组织增生,肾小球有硬化或数目减少,各干预组上述病理情况均不同程度改善,其中祛风通络方高剂量组改善最为明显。③与假手术组比较,各组大鼠肾组织α-SMA、vimentin mRNA及蛋白表达均明显升高(P<0.05),以模型组升高最为显著(P<0.01);与模型组比较,各治疗组α-SMA、vimentin mRNA及蛋白表达均不同程度下降(P<0.05),以祛风通络方高剂量组下降最为显著(P<0.01),且祛风通络方高剂量组与祛风通络方低剂量组比较,差异亦有统计学意义(P<0.05)。结论:大剂量祛风通络方可显著改善肾纤维化病理、下调肾纤维化相关蛋白α-SMA、vimentin的表达。

Objective:To observe the the effect of Qufeng Tongluo Prescription on unilateral ureteral ligationa-smooth muscle actin(α-SMA),a protein of renal interstitial fibrosis associated with unilateral urethral occlusion(UUO)in rats with unilateral urethral occlusion(UUO)and vimentin.Methods:Fifty male adult SD rats were randomly divided into 10 sham-operated groups and 40 UUO groups.The UUO group was randomly divided into model group,positive control group and high-dose group of Qufeng Tongluo Prescription,and low-dose group of Qufeng Tongluo Prescription according to different intervention methods.On the first day after operation,the corresponding drugs were administered.The rats were sacrificed on the 21 st day after operation.The kidney tissues were taken from the operation side.The degree of renal interstitial fibrosis was observed by Masson staining.Theα-SMA,vimentin mRNA levels and protein expression of renal tissues was detected with RT-PCR and Western blotting.Results:(1)On the21 st day after UUO,the renal volume of the obstructed side of the model group and the treated group increased compared with the contralateral side.The renal parenchyma of the section was atrophied to varying degrees,and the renal pelvis was cystic,and the model group was the most obvious.(2)In the model group,the proximal renal tubular epithelial cells atrophied,the collecting duct and the distal convoluted tubules were obviously dilated,and a large number of inflammatory cell infiltration accompanied by interstitial collagen components increased significantly,fibrous tissue hyperplasia,glomerular sclerosis or decreased number The above pathological conditions of each intervention group were improved to varying degrees,and the high dose group of Qufeng Tongluo was the most obvious.(3)Compared with the sham operation group,the expressions ofα-SMA,vimentin mRNA and protein in the kidney of each group were significantly increased(P<0.05),and the increase was the most significant in the model group(P<0.01).Compared with the model group,The expressions ofα-SMA,vimentin mRNA and protein in different treatment groups were decreased in different degrees(P<0.05),and the highest in the high dose group of Qufeng Tongluo Recipe(P<0.01),and the high dose group of Qufeng Tongluo Recipe The difference was also statistically significant in the low-dose group of Qufeng Tongluo Recipe(P<0.05).Conclusion:High-dose Qifeng Tongluo Prescription can significantly improve the pathology of renal fibrosis and down-regulate the expression ofα-SMA and vimentin related to renal fibrosis.