摘要
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(LPS 5 mg/L)和蟾蜍灵干预组(LPS 5 mg/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MTY、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.
Abstract
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 × 10-8 mol/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.
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维普
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