益气活血散结法对糖尿病肾病大鼠肾组织中MCP-1、TGF-β的影响

摘要

目的:通过动物实验,基于免疫炎性反应因子MCP-1、TGF-β,探索益气活血散结法改善肾小球硬化的分子机制.方法:24只GK大鼠,随机分为模型组、假手术组、益气活血散结组.通过高脂高能量喂养加单肾切除术复制糖尿病肾病模型.检测各组大鼠的24h尿蛋白,观察大鼠在干预后肾脏组织的病理改变,同时采用Western blotting以及免疫组化法检测肾脏组织的MCP-1和TGF-β的表达情况,采用RT-PCR法检测MCP-1mRNA以及TGF-βmRNA的表达水平.结果:PASM染色和Masson染色显示:益气活血散结组阳性率明显低于模型组(P＜0.01).益气活血散结组中24 h尿蛋白在药物干预后明显减少(P＜0.01).Western blotting、免疫组化结果均显示:益气活血散结组MCP-1 (P ＜0.05)、TGF-β(P＜0.01)的表达程度明显低于模型组.同样,RT-PCR结果也显示:MCP-1mRNA(P＜0.01)、TGF-βmRNA(P＜0.05)水平在益气活血散结组明显低于模型组.结论:益气活血散结法可以有效地减少24 h尿蛋白定量、延缓肾小球纤维化,其机制可能与抑制肾脏组织中MCP-1和TGF-β的表达有关.%Objective:To study the mechanism of Qi-tonifying blood-activating and mass-dissipating method in treating diabetic nephropathy based on MCP-1 and TGF-β through animal experiment.Methods:A total of 24 GK rats were randomly divided into model group,sham-operated group and Qi-tonifying blood-activating and mass-dissipating group.Each group had 8 rats.The DN model was established by feeding with high-fat diet and unilateral nephrectomy.24-hour urine protein was detected and the renal pathological changes were observed.Expressions of MCP-1 and TGF-β in the renal tissue were detected by Western blotting and immunohistochemistry,while RT-PCR was used to detect the expression of MCP-1mRNA and TGF-β mRNA.Results:Results of PASM staining and Masson staining show that the positive rate of Qi-tonifying blood-activating and mass-dissipating group was significantly lower than that of model group(P ＜ 0.01).The 24-hour urinary protein in Qi-tonifying blood-activating and mass-dissipating group decreased significantly after drug intervention(P ＜ 0.01).Results of Western blotting and immunohistochemistry showed that expression levels of MCP-1 (P ＜ 0.05) and TGF-β (P ＜ 0.01) were lower in Qi-tonifying blood-activating and mass-dissipating group than those in model group.Results of RT-qPCR showed that expression levels of MCP-1 mRNA(P ＜ 0.01) and TGF-β mRNA(P ＜0.05) in Qi-tonifying blood-activating and mass-dissipating group were significantly lower than those in model group.Conclusion:The treatment of diabetic nephropathy with Qi-tonifying blood-activating and mass-dissipating method can decrease the proteinuria and delay the glomerular fibrosis.The molecular mechanism may be related to the inhibition of MCP-1 and TGF-β.