蟾蜍灵对高糖诱导大鼠系膜细胞过表达纤维连接蛋白和结缔组织生长因子的影响

目的:观察不同浓度蟾蜍灵对高糖诱导的大鼠系膜细胞(MC)过表达FN及CTGF的影响,拟探讨其防治糖尿病肾病的可能作用?方法:体外培养MC,将细胞分为5组:正常对照组?高糖组?高糖 + 低浓度蟾蜍灵(5 × 10-9 mol/L)组?高糖 + 中浓度蟾蜍灵(5 × 10-8 mol/L)?高糖+高浓度蟾蜍灵组(5 × 10-7 mol/L)?以台盘兰拒染法检测蟾蜍灵对MC的毒性作用,48 h后,采用RT-PCR法和Western-blot法检测各组MC中FN?CTGF mRNA和CTGF蛋白的表达及其变化?结果:蟾蜍灵在5 × 10-9 ~5 × 10-7 mol/L浓度间对MC无明显细胞毒作用?48 h后,与对照组相比,高糖组CTGF mRNA和蛋白以及FN mRNA表达均显著增加(P < 0.05);高?中?低浓度蟾蜍灵干预能显著降低高糖诱导的上述指标表达,且呈浓度依赖效应(P < 0.05)?结论:蟾蜍灵能部分抑制高糖诱导的MC过表达FN,呈浓度依赖效应,其作用可能部分是通过降低促纤维化因子CTGF合成而实现,提示蟾蜍灵在DN纤维化防治领域具有进一步研究的价值?

Effect of bufalin on overexprssing FN and CTGF in glomerular mesangial cell of rat induced by high glucose

Objective: To observe the effects of bufalin on overexpression of FN and CTGF in glomerular mesangial cells (MCs) of rat induced by high glucose,in order to explore the potential protective role of bufalin against diabetic nephropathy (DN). Methods: Cultured rat MCs were divided into 5 groups:control, high glucose(30 mmol/L),high glucose plus 5 × 10-9 mol/L bufalin,high glucose plus 5 × 10-8 mol/L bufalin,high glucose plus 5 × 10-7 mol/L bufalin. The cell toxicity of bufalin on MC were analyzed by Trypan blue method,expression of FN and CTGF was measured by RT-PCR and Western blot,respectively. Results: No cell toxicity was observed when bufalin was applied at the concentrations of 5 × 10-9 -5 × 10-7 mol/L. The expression of CTGF mRNA(P < 0.05) and protein(P < 0.05),FN mRNA level (P < 0.05) was significantly increased in the high glucose group compared with that of the control group. While the above-mentioned over-expression value induced by high glucose can be down-regulated by high,median,and low concentration bufalin in a concentration-dependent manner. Conclusion: Bufalin can inhibit the over-expression of FN caused by high glucose in MCs in a concentration-dependent maner,and the mechanism may be related to downregulating synthesis of CTGF,which indicates that bufalin deserves more investigation against DN.