顺铂对人卵巢黄素化颗粒细胞的毒性及凋亡的影响

目的研究顺铂(cisplatin,CDDP)对人卵巢黄素化颗粒细胞的毒性及凋亡的影响。方法收集体外受精—胚胎移植(IVF-ET)时的人黄素化颗粒细胞进行体外培养,采用MTT法检测不同浓度的CDDP(0、0.5、1、2.5、5mg.L-1)对人黄素化颗粒细胞生长的抑制作用;采用Hochest33258荧光染色检测细胞凋亡,流式细胞仪(flowcytometry,FCM)检测bcl-2和bax基因蛋白的表达。结果CDDP浓度≥1mg.L-1可抑制颗粒细胞的生长;1、2.5、5mg.L-1CDPP处理人黄素化颗粒细胞24h后,荧光染色可观察到明显的核浓缩、核碎裂形态。FCM检测bcl-2蛋白表达逐渐减少(P<0.05),各组间比较差别有统计学意义;bax蛋白表达逐渐增加(P<0.05),各组间比较差别有统计学意义。结论CD-DP可抑制人黄素化颗粒细胞生长,促进凋亡,其作用机制可能是通过上调bax和下调bcl-2基因表达实现的。

Effect of cisplatin on toxicity and apoptosis of human luteinized granulose cells

Aim To investigate the effect of cisplatin(CDDP) on toxicity and apoptosis of human luteinized granulose cells.Method Human luteinized granulose cells were prepared from oocyte retrieval of fertilization procedure in vitro and treated with various concentrations of cisplatin(0,0.5,1.0,2.5,5.0 mg·L-1).The inhibitory rate of cells was examined by MTT assay.The change of nucleolus stained with Hochest 33258 was observed under fluorescent microscope,and bcl-2 and bax protein expressions were detected by flow cytometry(FCM).Results The results showed that CDDP inhibited the growth of ovarian granulose cells at the concentration of no less than 1 mg·L-1,in which Hochest 33258 staining demonstrated cell apoptosis.The expression of bax protein increased as compared with that of the control(P<0.05).However,the expression of bcl-2 protein decreased as compared with that of the control(P<0.05).Conclusion CDDP inhibited the growth of human luteinized granulose cells and promoted the cell apoptosis,which possibly resulted from the decreased expression of bcl-2 and increased expression of bax.