Objective: To observe the effect of traditional Chinese medicine for intervention of phlegm and blood stasis in regulating TGF-β1/Smad3 signaling and relieving nephropathy in diabetic rats.
Methods: SD rats were divided into blank group (NC), diabetic model group (MC group), intervention of phlegm and blood stasis (RPDBS) group, phlegm-removing (RP) group and blood-removing (DBS) group. Diabetic models were established in all the rats except for those in the blank group. After 4 weeks of feeding, the rats in RPDBS group, RP group and DBS group were given corresponding drug intervention for 8 weeks. HE staining was used to observe the changes in renal histopathology. Western blotting and real-time fluorescence quantitative PCR were used to detect the expression levels of transforming growth factor-β1 (TGF-β1) and Smad3.
Results: The structure and arrangement of the glomeruli and renal tubules improved significantly in the treatment groups in comparison with those in the MC group. The expression levels of TGF-β1, Smad3 and p-Smad3 were significantly downregulated at both the protein and mRNA levels in the treatment groups (P < 0.05), and the down-regulation was more obvious in RPDBS group than in RP group and DBS group (P < 0.05).
Conclusions: Intervention of phlegm and blood stasis may inhibit the activation of TGF-β1/Smad3 signaling pathway and delay diabetic nephropathy and fibrosis to protect the renal function in diabetic rats.
目的: 观察痰瘀同治法通过调节TGF-β1/Smad3信号通路对糖尿病大鼠肾脏病变的影响。
方法: 选取SD大鼠随机分为:空白组(NC)、模型组(MC)、痰瘀同治组(RPDBS)、化痰组(RP)、化瘀组(DBS), 予以单次注射链脲佐菌素溶液制备糖尿病模型, 喂养4周后, RPDBS组、RP组和DBS组分别予以相应的药物干预8周, 麻醉状态下取材。采用HE染色法观察肾脏组织病理学变化; Western blot法和实时荧光定量PCR法检测肾脏组织转化生长因子-β1(TGF-β1)及其信号蛋白Smad3蛋白和基因表达水平。
结果: 与MC组比较, 光镜下各治疗组肾小球和肾小管结构和排列改善明显; 各治疗组TGF-β1、Smad3和p-Smad3蛋白和基因表达水平均明显下调(P < 0.05);其中RPDBS组TGF-β1、Smad3和p-Smad3蛋白和基因表达水平较RP组和DBS组下调更明显(P < 0.05)。
结论: 痰瘀同治可能通过抑制TGF-β1/Smad3信号通路激活, 延缓糖尿病肾病纤维化, 发挥肾脏保护作用。
Keywords: TGF-β1/Smad3 signal pathway; diabetic nephropathy; intervention of phlegm and blood stasis.