也页目的：探讨糖肾平对高糖环境下脂多糖( lipopolysaccharide,LPS)刺激足细胞上皮间质转分化的影响,并探讨其作用机制。方法：以体外培养大鼠肾小球足细胞为研究对象,以高糖(25 mmol/L)、LPS(1μg/mL)刺激足细胞建立模型,分为正常组、高糖组、高糖+LPS组、厄贝沙坦组、抑制剂组、糖肾平小、中、大剂量组。采用Western blotting及RT-PCR方法检测足细胞中转化生长因子-β1( transforming growth factor-β1,TGF-β1)、Smad2/3、整合素连接激酶( integrin-linked kinase, ILK)、CD2相关蛋白(CD2AP)、α-平滑肌肌动蛋白(α-Smooth muscle actin,α-SMA)的表达水平。结果：与正常组比较,高糖组和高糖+LPS组足细胞TGF-β1、ILK、α-SMA蛋白及其mRNA表达明显增加(P<0.01),P-Smad2/3蛋白及Smad2/3 mRNA表达明显增加(P<0.01),CD2AP蛋白及其mRNA表达明显减少(P<0.01)；与高糖+LPS组比较,厄贝沙坦组足细胞P-Smad2/3、α-SMA蛋白表达明显减少(P<0.01),TGF-β1蛋白表达减少(P<0.05),TGF-β1,Smad2/3,α-SMAmRNA表达明显减少(P<0.01),ILK mRNA表达减少(P<0.05),CD2AP蛋白及其mRNA表达明显增加(P<0.01)；糖肾平大、中、小各剂量组足细胞TGF-β1蛋白表达明显减少(P<0.01),糖肾平大剂量组足细胞TGF-β1 mRNA表达减少(P<0.05),小、中剂量组表达明显减少(P<0.01)；糖肾平小、中、大各剂量组足细胞P-Smad2/3蛋白及Smad2/3mRNA表达均明显减少(P<0.01)；糖肾平小、大剂量组足细胞ILK mRNA表达减少(P<0.05),中剂量组表达明显减少(P<0.01)；糖肾平大剂量组足细胞α-SMA蛋白及其mRNA表达减少(P<0.05)；糖肾平小、中、大各剂量组足细胞CD2AP蛋白及其mRNA表达均明显增加(P<0.01)。结论：糖肾平能够降低足细胞TGF-β1、ILK蛋白及mRNA表达,降低P-Smad2/3蛋白及Smad2/3 mRNA表达,升高足细胞标志物CD2AP蛋白及mRNA表达,降低间充质细胞标志物α-SMA蛋白及mRNA表达,通过抑制TGF-β1-Smad2/3-ILK信号通路的激活减少足细胞转分化,保护足细胞,可能是其防治糖尿病肾病的作用机制之一。%Objective:To observe the influence of the capsule of Tangshenping on LPS-stimulated podocytes epithelial-mesenchymal transition ( EMT) under high glucose conditions,and explore its possible mechanism. Methods:Using cultured rat glo-merular podocytes as the object of study, making model with high glucose(25 mmol/L)、LPS(1 μg/ml) stimulating podocytes, and divide them into 8 groups:control group, high glucose group, high glucose plus LPS group, irbesartan group, low-dose group of Tan-gshenping capsule, moderate dose group of Tangshenping capsule, high-dose group of Tangshenping, inhibitor group. Detect podo-cyte transforming growth factor-β1(TGF-β1), Smad2/3, integrin-linked kinase(ILK), CD2AP and α-smooth muscle actin (α-SMA) mRNA expression of each group by Western blotting and RT-PCR. Results:Compared with the control group, the ex-pression of podocyte TGF-β1、ILK、α-SMA protein and mRNA increased definitely(P<0. 01), P-Smad2/3 protein and Smad2/3 mRNA expression increased definitely (P<0. 01), CD2AP protein and mRNA decreased definitely(P<0. 01) in the high glucose group and high glucose plus LPS group. Compared with the high glucose plus LPS group, the expression of podocyte P-Smad2/3、α-SMA protein decreased definitely(P<0. 01), TGF-β1 protein decreased (P<0. 05), TGF-β1,Smad2/3,α-SMAmRNA de-creased definitely(P<0. 01), ILKmRNA decreased(P<0. 05), CD2AP protein and mRNA increased definitely(P<0. 01) in the irbesartan group;The expression of podocyte TGF-β1 protein decreased definitely(P<0. 01) in the low, moderate and high dose group of Tangshenping, TGF-β1 mRNA decreased in the high dose group of Tangshenping(P<0. 05)and decreased definitely in the moderate,low dose group of Tangshenping (P<0. 01); In the low, moderate and high dose group of Tangshenping, podocyte P-Smad2/3 protein and Smad2/3mRNA expression decreased definitely(P<0. 01);The expression of podocyte ILKmRNA decreased in the low and high dose group of Tangshenping(P<0. 05), decreased definitely in the moderate dose group(P<0. 01); In the high dose group of Tangshenping, podocyte α-SMA protein and mRNA expression decreased (P<0. 05);In the low,moderate and high dose group of Tangshenping, podocyte CD2AP protein and mRNA expression increased definitely(P<0. 01). Conclusion:Tangshenp-ing can reduce podocyte TGF-β1, ILK protein and mRNA expression, reduce P-Smad2/3 protein and Smad2/3mRNA expression, increase podocytes marker CD2AP protein and mRNA expression and reduce mesenchymal cell marker α-SMA protein and mRNA expression, Tangshenping can relieve podocytes epithelial-mesenchymal transition by inhibiting TGF-β1 -Smad2/3 -ILK signal transduction pathway, this mechanism may be considered as a means of prevention and treatment for diabetic nephropathy.
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