骨髓间充质干细胞抑制磷酰胺氮芥诱导的卵巢颗粒细胞凋亡  

The inhition of bone marrow derived mesenchymal stem cells on apoptosis of ovarian granulosa cells induced by phosphoramide mustard

作　　者：付霞霏[1] 何援利[1] 

机构地区：[1]南方医科大学珠江医院妇产科,广州510280

出　　处：《现代妇产科进展》2009年第2期99-102,共4页Progress in Obstetrics and Gynecology

基　　金：广东省科技计划资助项目(No:2007B030501005)

摘　　要：目的:探讨骨髓间充质干细胞(mesenchymal stem cells,MSCs)对磷酰胺氮芥(Phosphoramide Mustard,PM)诱导卵巢颗粒细胞凋亡的影响及可能机制。方法:培养雌性Wistar大鼠卵巢颗粒细胞,加入PM诱导凋亡后,与MSCs共培养(MSCs:颗粒细胞分别为1:1,1:5和1:10),Hochest33342、PI双染和DNA-ladder检测凋亡情况;Western blot检测凋亡相关蛋白Bcl-2和Bax的变化。结果:PM可引起颗粒细胞凋亡。与MSCs共培养后,电泳图无明显的梯状带形,凋亡细胞比例下降,细胞存活率上升,差异有统计学意义。Western blot显示,与MSCs共培养后,颗粒细胞Bcl-2表达水平增高,Bax表达水平无变化。结论:MSCs对PM诱导的卵巢颗粒细胞凋亡有抑制作用,其作用机制可能与上调Bcl-2在颗粒细胞中的表达密切相关,为进一步的体内实验及临床应用提供了实验依据。Objective：To investigate the influence of mesenchymal stem cells （MSCs） on phosphoramide mustard-induced ovarian granulosa cells apoptosis and the mechanism. Methods：Granulosa cells were isolated from female Wistar rats and cultured in vitro in media with phosphoramide mustard. The apoptosis of co-cultured granulosa cells （ the ratio of granulosa ceils to MSCs were 1：1,5：1 and 10：l,respectively. ） and apoptosis was detected via Hochest 33342 and PI double staining, and DNA ladder. The expressions of apoptosis associated proteins Bcl-2 and Bax were assessed through Western blot. Results：The phosphoramide mustard could induce granulosa cell apoptosis. When granulosa cells were co-cuhured with MSCs, ladder disappeared in electrophoretogram, the apoptotic ration decreased and cellular vitality increased. The expression of Bcl-2 in co-cultured granulosa cells was significantly increased compared with that in control group. And there was no significant difference in Bax expression. Conclusion： MSCs can inhibit the apoptosis of granulosa cells induced by phosphoramide mustard with the possible mechanism of up-regulation of Bcl-2 in granulosa cells,which provides the experimental base of in vivo detection and clinic practice.

关 键 词：间充质干细胞 卵巢颗粒细胞 磷酸酰胺氮芥 细胞凋亡 大鼠 Wistar 

分 类 号：R711.32[医药卫生—妇产科学]