柴胡皂苷-d对体外培养的大鼠肾小球系膜细胞增殖和细胞外基质分泌的影响

目的探讨柴胡皂苷-d（saikosaponin—d，SSd）对脂多糖（1ipopolysaccharides，LPS）作用后肾小球系膜细胞（mesangialcell，MC）增殖及细胞外基质（extracellular matrix，ECM）的异常增生的影响，为肾小球硬化的防治提供实验依据。方法体外培养、鉴定大鼠肾小球MC。利用MTT、LDH释放实验、流式细胞技术观察SSd对LPS刺激后MC增殖的作用，ELISA法检测细胞外基质Ⅳ型胶原（type1Vcollagen，ColⅣ）、纤维连接蛋白（fibronectin，FN）及转化生长因子（transforminggrowthfactor-β1，TGF-β1）。细胞免疫化学法检测细胞素依赖性蛋白激酶4（cyclin—dependentkinase4，CDK4）、c—Jun、c—Fos。结果SSd对MC增殖有抑制作用，可使Go／G1期细胞增多，并诱导细胞凋亡；SSd可抑制MC分泌ColⅣ、FN、TGF-β1，并抑制了CDK4、c-Jun、C—Fos的表达。结论SSd对肾小球硬化（glomerulosclerosis，GS）的抑制作用是通过抑制CDK4、c—Jun、c-Fos的表达实现的。

Effect of Saikosaponin- d on Rat's Glomerular Mesangial Cell Proliferation and Extracellular Matrix Hyperplasia in vitro

OBJECTIVE: To investigate the effects of saikosaponin-d (SSd) on glomerular mesangial cells (MCs) proliferation and hyperplastic extracellular matrix (ECM) induced by lipopolysaccharide (LPS) to provide experimental proof for its use in prevention and treatment of glomerulosclerosis. METHODS: Rat's MCs were cultivated and identified. The cultured MCs were stimulated by LPS and incubated with different concentrations of SSd. Cell proliferation was determined by MTT assay, LDH assay and flow cytometry, respectively. Type IV collagen (Col IV), fibronectin (FN) and transforming growth factor beta1 (TGF-beta1) in the conditioned medium were measured by ELISA. The expressions of cyclin-dependent kinase 4 (CDK4), c-Jun and c-Fos were detected by immunohistochemistry. RESULTS: After treated by SSd, MC proliferation was inhibited, cells in G0/G1 phase increased, and apoptosis induced. Moreover, secretion of Col IV, FN and TGF-beta1 and the expressions of CDK4, c-Jun and c-Fos in MC were inhibited. CONCLUSION: The inhibitory action of SSd on glomerulosclerosis was realized through inhibiting the expressions of CDK4, c-J un and c-Fos.