海风藤对人肾小球系膜细胞TGF-β/Smad信号通路影响

Effect of Kadsura Pepper Stem on TGF-β/Smad Pathway of Human Glomerular Mesangial Cells

目的:探讨海风藤对血管紧张素Ⅱ诱导的人肾小球系膜细胞TGF-β/smad传导通路的影响。方法:培养人肾小球系膜细胞,分为正常组,模型组,海风藤高剂量(10mg/mL)、中剂量(1mg/mL)、低剂量(0.1mg/mL)组。经预实验筛选结果,采用10^-8 mol/L血管紧张素Ⅱ诱导模型组及海风藤各组人肾小球系膜细胞增殖,采用MTT法检测正常组、模型组及海风藤药液培养的经血管紧张素Ⅱ诱导的人肾小球系膜细胞于24、48、72h等时间点的细胞活性。收集培养48h时间点各组细胞,采用westem blot法及Real-time-PCR法分别检测smad2、3蛋白表达及TGF-β、smad7 mRNA的表达。结果:模型组与正常组比较,培养24、48、72h内,系膜细胞显著增殖;海风藤各组能够抑制其增殖;于48h时间点收集的系膜细胞中smad2、smad3含量及TGF-β mRNA的表达显著上升(P<0.05),smad7 mRNA的表达显著降低(P<0.05);海风藤高、中、低各组与模型组比较,均能逆转上述变化,但量-效依赖趋势并不显著。结论:血管紧张素Ⅱ能够有效刺激肾小球系膜细胞增殖,海风藤可能通过多靶点抑制TGF-β/Smad信号通路的传导,从而抑制延缓肾小球硬化。

Objective:To explore effect of kadsura pepper stem on human glomerular mesangial cells induced by AngⅡ on TGF-β/Smad pathway. Methods:Cultivate human glomerular mesangial cells,divided into normal group,model group,kadsura pepper stem high dose(10 mg/mL),middle dose(1 mg/mL),low dose group(0.1 mg/mL). For the preliminary screening experiment results,using 10-8 mol/L angiotensinⅡ induced model group and kadsura pepper stem each human glomerular mesangial cells proliferation,using MTT method to detect the normal group,model group,and the kadsura pepper stem induced by angiotensin Ⅱ cane liquid culture of human glomerular mesangial cells in 24 h,48 h,72 h time points such as cell activity. Collected training 48 h point groups of cells,using Western blot method and Real-time-PCR method respectively detected Smad2,Smad3 protein expression and TGF-β and Smad7 mRNA expression. Results:Compared with normal group,model group in training within 24 h,48 h,72 h mesangial cells of value-added activities,kadsura pepper stem groups could inhibit the proliferation,48 h time cultured mesangial cells Smad2 and Smad3 content,TGF-β mRNA expression significantly increased(P<0.05),The expression of Smad7 mRNA significantly reduced(P<0.05);Compared with model group,kadsura pepper stem high,medium and low dose groups could reverse the above changes,and the inhibitory effect of(promote)is a certain amount of time dependencies,but,there was no significant dose-response trend. Conclusion:AngⅡ can effectively stimulate the glomerular mesangial cell proliferation,kadsura pepper stem may through the multiple targets for inhibition of TGF-β/Smad pathway conduction,thus inhibiting delay glomerular sclerosis.