雷公藤内酯醇对足细胞EMT相关蛋白mRNA表达的影响

Effect of triptolide on expression of EMT-related protein mRNA in podocytes

目的通过观察雷公藤内酯醇(TP)对高糖刺激下ILK、MMP9、NEPH1蛋白表达的影响,探讨TP抑制足细胞上皮-间充质转分化(EMT)治疗糖尿病肾病的可能机制。方法将培养分化成熟的足细胞分为对照组(D-葡萄糖5 mmol/L)、高糖组(D-葡萄糖25 mmol/L)、低TP组(25 mmol/L D-葡萄糖+8 ng/m L TP)、中TP组(25 mmol/L D-葡萄糖+16 ng/m L TP)以及高TP组(25 mmol/L D-葡萄糖+32 ng/m L TP)。体外培养48 h后,倒置显微镜观察细胞形态,并采用PCR半定量分析技术检测ILK、MMP9、NEPH1的表达。结果高糖刺激下足细胞NEPH1的表达较对照组显著减少(P<0.01),而ILK、MMP9的m RNA表达较对照组显著升高(P<0.01)。各剂量TP组足细胞NEPH1 m RNA的表达均较高糖组明显上调(P<0.01),ILK、MMP9 m RNA的表达较高糖组明显下降(P<0.01),其中以中TP组的干预作用最显著(P<0.01)。结论 TP可能通过抑制ILK信号通路来抑制足细胞EMT。

Objective To observe the effect of triptolide（TP） on the expression of ILK, MMP9 and NEPH1 protein in- duced by high glucose and to explore the possible mechanism of TP inhibiting podocyte-mesenchymal transdifferentia- tion（EMT） in the treatment of diabetic nephropathy. Methods The podocytes which were cultured and maturely differ- entiated were assigned to the control group（D-glucose 5 mmol/L）, high glucose group（D-glucose 25 mmol/L）, low TP group （25 mmol/L D-glucose＋8 ng/mL TP）, medium glucose group （25 mmol/L D-glucose＋16 ng/mL TP） and high TP group （25 mmol/L D-glucose＋32 ng/mL TP）. The cells were cultured in vitro for 48 hours, and the forms of cells were ob- served by inverted microscope. The expression of ILK, MMP9 and NEPH1 was detected by PCR semi-quantitative analysis. Results The expression of NEPH1 in podocytes stimulated by high glucose was significantly lower than that in the control group（P〈0.01）, while the mRNA expression of ILK, MMP9 was significantly higher than that in the control group（P〈0.01）. The mRNA expression of NEPH1 in each dosage TP group was significantly higher than that in the high glucose group（P〈0.01）. The mRNA expression of ILK and MMP9 was significantly lower than that in the high glucose group, and the intervention effect in TP group was the most significant（P〈0.01）. Conclusion TP may inhibit EMT of podocytes by inhibiting ILK signaling pathway.