骨髓间充质干细胞对磷酰胺氮芥诱导卵巢颗粒细胞凋亡的保护作用  

Protective effect of bone marrow mesenchymal stem cells on chemotherapy-induced apoptosis of ovarian granulosa cells

作　　者：付霞霏[1] 何援利[1] 

机构地区：[1]广州南方医科大学珠江医院妇产科,510280

出　　处：《解放军医学杂志》2009年第8期967-969,共3页Medical Journal of Chinese People's Liberation Army

基　　金：广东省科技计划资助项目(2007B030501005)

摘　　要：目的探讨骨髓间充质干细胞(MSCs)对磷酰胺氮芥诱导卵巢颗粒细胞凋亡的保护作用及可能机制。方法分离培养雌性Wistar大鼠卵巢颗粒细胞,加入磷酰胺氮芥(30μmol/L)至培养的颗粒细胞中,再与MSCs以不同比例共培养72h。实验分为4组:①颗粒细胞单独培养组(对照组);②共培养组1,MSCs∶颗粒细胞=1∶1;③共培养组2,MSCs∶颗粒细胞=1∶5;④共培养组3,MSCs∶颗粒细胞=1∶10。采用AnnexinⅤ检测凋亡率;采用蛋白质免疫印迹法检测凋亡相关蛋白Bcl-2和Bax的变化,ELISA法检测MSCs培养基中血管内皮生长因子(VEGF)、肝细胞生长因子(HGF)、胰岛素样生长因子-1(IGF-1)的浓度。结果对照组化疗药物引起的卵巢颗粒细胞凋亡率为35.04%±5.75%,而共培养组1、2、3的凋亡率分别为12.80%±2.42%,13.94%±3.86%,22.31%±5.67%,与对照组比较差异均有统计学意义(P<0.05)。蛋白质免疫印迹显示与MSCs共培养后,颗粒细胞Bcl-2表达水平明显增高(P<0.05),Bax表达水平无明显变化(P>0.05)。MSCs能分泌VEGF、HGF和IGF-1。结论MSCs对体外化疗药物诱导的卵巢颗粒细胞的凋亡有保护作用,其作用机制可能是其分泌的细胞因子通过旁分泌途径发挥作用,以及Bcl-2在颗粒细胞中表达上调有关。Objective To investigate the anti-apoptotic effect of bone marrow mesenchymal stem cells （NSCs） on chemotherapyinjured ovarian granulosa cells and its mechanism. Methods Granulosa cells were isolated from female Wistar rats and cultured in vitro in media containing chemotherapeutic agents. The experiment consisted of 4 groups： granulosa cells culture group （control group）, co-culture group 1 with MSCs/GCs ratio of 1：1, co-cuhure group 2 with MSCs/GCs ratio of 1：5 and co-culture group 3 with MSCs/GCs ratio of 1 ： 10. Apoptosis was detected by Annexin V. The expressions of Bcl-2 and Bax were assessed using Western blotting. The concentrations of vascular endothelial growth factor （VEGF）, hepatocyte growth factor （HGF） and insulin-like growth factor-1 （IGF-1） in MSCs media were measured by ELISA. Results The drug could induce apoptosis of granulosa cell, and the apoptotic ratio in control group was 35.04%±5. 75%, and in the other three co, culture groups was 12.80%±2. 42%, 13. 94%±3.86%, 22. 31%±5. 67%, respectively. There was significant difference in apoptotic ratio between control group and 3 co-culture groups. The expression of Bcl-2 in co-cultured granulosa cells increased significantly compared with that in control group （P〈0. 05）, while no significant difference was observed in Bax expression. MSCs secreted a large amount of cytokines including VEGF, HGF and IGF 1. Conclusion MSCs have anti-apoptotic effects on granulosa cells cultured in the mediam containing chemotherapeutic agent. The secretion of cytokines and the up-regulation of Bcl-2 expression in granulosa cells might be the possible mechanism.

关 键 词：间质干细胞 粒层细胞 药物疗法 细胞凋亡 

分 类 号：R711.75[医药卫生—妇产科学] R737.31[医药卫生—临床医学]