中文摘要：

目的观察多球壳菌素（ISP-1）对体外培养的正常大鼠肾小球系膜细胞（GMC）的促凋亡作用，研究ISP-1对细胞周期调节蛋白基因表达谱和凋亡相关蛋白Bax、Bcl-2表达的影响。方法体外培养大鼠GMC，加入100μmol／LISP-1干预，分别于作用6、12、24、48h后以流式细胞术检测GMC凋亡，Hoechst33258／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Super Array Real-Time PCR细胞周期基因芯片测定ISP-1对细胞周期调节蛋白基因表达谱的影响，Western blotting法观察Bax和Bcl-2蛋白的表达。结果ISP-1可显著诱导GMC凋亡，并呈一定的时间依赖性，作用48h后细胞凋亡最显著，Hoechst33258／PI荧光染色法可见亮蓝色的凋亡小体，琼脂糖凝胶电泳可见凋亡梯度的出现；PCR细胞周期基因芯片检测结果发现ISP-1可显著上调GMCRad51、Atm、Brcal、Caspase3、cyclinA2、cyclinC、chekl、cyclinBl、cyclinB2、cyclinD2、cyclinF、Cdc25a和P^27等一些与DNA损伤、凋亡和细胞周期调节蛋白相关的基因的表达；Westernblotting发现ISP-1可显著上调GMCBax蛋白表达，下调Bcl-＋2蛋白表达。结论ISP-1可时间依赖性诱导体外培养的正常大鼠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 myriocin （ISP-1） on improvement ot glomerular messangial cell （GMC） apoptosis and on gene expression profiles of cell cycle regulatory proteins in GMC. Methods Rat GMC was cultured with 100 μmol/L ISP-1 for 6, 12, 24, and 48 h, apoptosis was evaluated through flow cytometry, Hoechst33258/PI fluorescence stainig and DNA fragmentation analysis was carried out under Sepharose electrophoresis to observe the changes of apoptosis and DNA fragmentation. Gene expression profiles of cell cycle regulatory proteins were detected by SuperArray Real-Time PCR microarray analysis. The expression of Bax and Bcl-2 proteins was investigated by Western blotting. Results ISP-1 could significantly induce GMC apoptosis in a time-dependent manner, which was the most significant after 48 h. Apoptosis bodys of GMC were observed by Hoechst/PI fluorescence staining, and a typical ladder pattern was identified in DNA electrophoresis. SuperArray Real-Time PCR microarray analysis revealed that ISP-1 could up-regulate the expression of genes involved in DNA damage, apoptosis and cell cycle regulation, such as Rad51, Atm, Brcal, caspases, cyclinA2, cyclinC, chekl, cyclinB1, cyclinB2, cyclinD2, cyclinF, Cdc25a, and p27 Western blotting showed that ISP-1 could significantly up-regulate Bax protein expression and down-regulate Bcl-2 protein expression, respectively. Conclusion ISP-1 could induce GMC apoptosis in a time-dependent manner, propabably through influencing gene expression of cell cycle regulatory proteins and apoptosis proteins.