肾平颗粒对1-磷酸鞘氨醇刺激大鼠肾小球系膜细胞增生的调节作用研究

Regulatory Role of the Shenping Particles on Sphingosine Phosphate-1 Stimulation of Rat Mesangial Cell Proliferation

目的:观察肾平颗粒对1-磷酸鞘氨醇(S1P)刺激大鼠肾小球系膜细胞(RBMC)增生的调节作用。方法:S1P刺激RGMC后,用MTT法观察RGMC增生情况的变化,用Real-Time PCR法观察结缔组织生长因子(CTGF)mRNA水平的变化,用Western blot法观察内皮分化基因受体3/5(endothelial differentiation gene3/5,EDG3、EDG5)、CTGF蛋白水平的变化;制备EDG3、EDG5质粒。EDG3、EDG5质粒同时转染RGMC后,观察肾平颗粒对RGMC增生及EDG3、EDG5、CTGF表达的影响。结果:(1)肾平颗粒能抑制S1P刺激引起的RGMC增生。转染EDG3和EDG5后,抑制作用明显减弱。(2)肾平颗粒能抑制S1P刺激引起的RGMC表面EDG3和EDG5蛋白水平的过表达,转染EDG3和EDG5后,肾平颗粒的抑制作用明显减弱。(3)肾平颗粒能抑制S1P刺激引起的RGMC内S1P通路下游CTGF蛋白的过表达,转染EDG3和EDG5后,肾平颗粒对CTGF蛋白和mRNA的抑制作用明显减弱。结论:肾平颗粒能通过抑制S1P刺激引起的RGMC表面的EDG3、EDG5过度表达,来实现抑制RGMC增生及S1P通路下游CTGF表达的作用。

Objective：Understanding of the regulatory role of Shenping particles on Sphingosine Phosphate - 1 （S1 P） stim- ulation of rat mesangial cells （RBMC） proliferation. Methods： After S1P stimulation, RGMCs were treated with or without Shenp- ingkeli. We observed RGMCs proliferation by MTI＇, connective tissue growth factor （CTGF） mRNA level by real time PCR and Endo- thelial Differentiation Gene 3/5 （ EDG3, EDG5） and CTGF protein level by western blot. Then after EDG3/EDG5 were simultaneous- ly overexpressed ~n RGMCs, RGMCs were treated with Shenpingkeli, we examined the RGMCs proliferation, the expression of EDG3, EDG5 and CTGF. Results： （1）Shenping particles can inhibit RGMC proliferation caused by S1P stimulation. The inhibitive effect was deceased after EDG3 and EDG5 transfection. （2） Shenping particles can inhibit the overexpression of RGMC surface marker EDG3and EDG5 at the protein level caused by S1P stimulation. Similarly, the inhibitive effect was significantly reduced after EDG3 and EDG5 transfection. （3）Shenping particles can inhibit the overexpression of S1P downstream effector CTGF in RGMC at the protein and mRNA level. The inhibitive effect was deceased after EDG3 and EDG5 transfection. Conclusion： Shenping particles inhibit the overexpression of RGMC surface marker EDG3, EDG5 caused by S1P. Hence, Shenping particles can inhibit RGMC proliferation and the expression of S1P downstream effector CTGF.