蟾蜍灵对大鼠肾小球系膜细胞增殖和细胞外基质分泌的影响

Effect of Bufalin on rat's glomerular mesangial cell proliferation and extracellular matrix hyperplasia

目的:探讨蟾蜍灵(bufalin)对脂多糖(LPS)诱导的大鼠肾小球系膜细胞(GMC)增殖及细胞外基质(ECM)分泌的影响.方法:体外培养大鼠肾小球系膜细胞,分为对照组、LPS刺激组(LPS5mg/L)和蟾蜍灵干预组(LPS5mg/L和蟾蜍灵1×10-8mol/L),应用台盼蓝拒染法检测蟾蜍灵对GMC的细胞毒性作用,通过MTT、流式细胞技术检测GMC增殖变化,RT-PCR检测转化生长因子-β1(TGF-β1)的mRNA的表达,ELISA法检测GMC培养上清中纤维连接蛋白(FN)及TGF-β1的表达.结果:MTT结果显示蟾蜍灵浓度在1×10-8mol/L及以上时呈剂量依赖性抑制GMC增殖(P<0.01).细胞周期分析显示蟾蜍灵组S期细胞比例较脂多糖组降低(P<0.01).蟾蜍灵干预组与脂多糖组相比TGF-β1的mRNA相对表达量减少(P<0.05),FN,TGF-β1蛋白分泌量减少(P<0.01).结论:蟾蜍灵对LPS作用后肾小球系膜细胞增殖及细胞外基质分泌具有抑制作用.

AIM：To investigate the effect of Bufalin on rat＇s glomerular mesangial cells（GMC）proliferation and hyperplastic extracellular matrix（EMC）induced by lipopolysaccharide（LPS）.METHODS：Rat＇s GMC lines were cultivated.GMCs in vitro were divided into 3 groups：control group,LPS group（LPS 5 mg/L）and Bufalin group（LPS 5 mg/L and Bufalin 1×10-8mol/L）.The proliferation of GMC was assessed by MTT assay and flow cytometry.mRNA of transforming growth factor β1（TGF-β1）was measured by reverse transcriptase polymerase chain reaction（RT-PCR）and the expression of Fibronectin（FN）and TGF-β1 in the conditioned medium was measured by enzyme linked immunosorbent assay（ELISA）.RESULTS：The GMC proliferation was induced in LPS group,but it was inhibited in Bufalin group in a dose-dependent manner.The percentage of cells in S phase in Bufalin group was significantly lower than that in LPS group（P〈0.01）.The expression of TGF-β1 mRNA was significantly inhibited in Bufalin group（P〈0.05）.The expressions of FN and TGF-β1 protein in Bufalin group were significantly less than those in LPS group（P〈0.01）.CONCLUSION：Bufalin inhibits the GMC proliferation and LPA-induced hyperplastic ECM components.