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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。
OBJECTIVE: To observe the effect of different concentrations of bufalin on the expression of FN and CTGF induced by high glucose in rat mesangial cells (MC), and to explore its possible role in prevention and treatment of diabetic nephropathy. Methods: MCs were cultured in vitro and divided into 5 groups: normal control group, high glucose group, high concentration of glucose and low concentration of bufalin (5 × 10-9 mol / L) 10-8 mol / L) and high concentration of high concentration of bufalin (5 × 10-7 mol / L). Forty-eight hours later, the expression of FN, CTGF mRNA and CTGF protein and the changes of CTGF protein in MC were detected by RT-PCR and Western-blot respectively. Results: Bufalin at a concentration of 5 × 10-9 ~ 5 × 10-7 mol / L had no significant cytotoxic effect on MC. After 48 h, compared with the control group, the expression of CTGF mRNA and protein and FN mRNA in high glucose group were significantly increased (P <0.05); high, medium and low concentrations of bufalin could significantly reduce the high glucose-induced expression of the above indicators, And the concentration-dependent effect (P <0.05). CONCLUSION: Toad deprivation partially inhibits the high glucose-induced MC overexpression FN in a concentration-dependent manner, which may be partially mediated by the reduction of CTGF synthesis, which suggests that toad is further studied in the field of DN fibrosis prevention value.