中文摘要：

目的观察新型免疫抑制剂FTY720对体外培养的正常大鼠肾小球系膜细胞（GMC）的促凋亡作用及其对细胞周期调节蛋白基因表达谱的影响，以探讨其作用机制。方法体外培养大鼠GMC，加入20μmol／LFTY720，分别作用6h、12h、24h、48h后，以MTT法检测GMC增殖情况；流式细胞术检测GMC凋亡；Hoechst33258和PI染色观察凋亡细胞形态变化；琼脂糖DNA凝胶电泳法观察凋亡细胞核小体DNA的断裂现象。并通过SuperArray实时定量PCR细胞周期基因芯片测定FTY720对细胞周期调节蛋白基因表达谱的影响。结果加入FTY720培养6h后，流式细胞术检测发现，GMC出现典型细胞凋亡的亚二倍体峰；12h后Hoechst33258和PI荧光染色观察发现亮蓝色的凋亡小体，且开始出现典型细胞凋亡形态改变；24h后DNA琼脂糖凝胶电泳可见凋亡梯度的出现。随FTY720作用时间延长，细胞凋亡明显增加，不同时间组间细胞凋亡率差异有统计学意义（P〈0．01）。SuperArray实时定量PCR细胞周期基因芯片发现FTY720分别显著上调系膜细胞Dnajc2、LOC688900、RGD1562436_predicted基因表达，分别达41．6、38和16倍。结论FTY720在体外可呈时间依赖诱导正常大鼠GMC凋亡，其机制与影响细胞周期调节蛋白及凋亡相关基因的表达有关。

英文摘要：

Objective To observe the effects of a new immunosuppressive agent, FTY720, on rat glomerular mesangial cell （GMC） apoptosis and on gene expression profiles of cell cycle regulatory proteins. Methods Rat GMCs were cultured with 20 μmol/L FTY720 for 6 h, 12 h, 24 h and 48 h, and then were evaluated for proliferation through M3T method, and for apoptosis by flow cytomctry and fluorescence stainig with Hoechst33258 and PI, and DNA fragmentation analysis. The gene expression profile of cell cycle regulatory proteins was characterized in rat GMCs before and after FTY720 treatment by SuperArray real-time PCR microarray analysis. Results After incubation with FTY720 for 6 h, apoptotic sub-G1 peak was identified in GMC through flow eytometry. After incubation with FTY720 for 12 h, not only apoptosis bodies of GMC were observed by fluorescence staining with Hoechst33258 and PI, but also typical morphological changes of apoptosis were found in GMC. After incubation with FTY720 for 24 h, typical DNA ladder pattern was identified. The percentage of FTY720-induced GMC apoptosis gradually increased with the extension of incubation time. SuperArray real-time PCR microarray analysis revealed that FTY720 could respectively up-regulate the expression of Dnaje2, LOC688900 and RGD1562436_predicted genes to 41.6, 38 and 16 folds. Conclusion FTY720 can induce GMC apoptosis in a time-dependent manner, probably through influencing gene expression of cell cycle regulatory proteins.