目的 比较顺铂、依托泊苷、长春新碱、博来霉素对人卵巢功能的影响.方法 2010年3月至2010年10月在中国人民解放军白求恩国际和平医院科研中心体外培养卵巢癌细胞株PA-1,加入不同浓度的顺铂、依托泊苷、长春新碱、博来霉素,作用48 h后,MTT比色法测定其半数致死量(IC50)；体外受精-胚胎移植患者经阴道取卵获取人卵巢颗粒细胞并进行体外培养,以上述四种药物半数致死量作用于人卵巢颗粒细胞,72 h后MTT比色法测定人卵巢颗粒细胞活性；化学发光法测定颗粒细胞上清液雌二醇(E2)水平；免疫细胞化学法检测颗粒细胞Bcl-2、Bax的表达；流式细胞术测定细胞凋亡率.结果 顺铂、依托泊苷、长春新碱、博来霉素杀伤PA-1细胞株的半数致死量分别为0.55 mg/L、1.67 mg/L、1.8μg/L、1.7 mg/L;除长春新碱外,上述3种药物其IC50均对人卵巢颗粒细胞的生长及分泌E2能力有明显抑制作用,以依托泊苷和博来霉素抑制作用最强,两者比较差异无统计学意义,顺铂次之,长春新碱作用不明显；免疫细胞化学法结果显示,除长春新碱之外,其他3种药物IC50均使人卵巢颗粒细胞中Bcl-2表达下调,Bax表达上调.结论 顺铂、依托泊苷、博来霉素对PA-1细胞株的IC50剂量均能促进人卵巢颗粒细胞凋亡并抑制E2的分泌,以依托泊苷和博来霉素作用最强,顺铂次之,而长春新碱作用不明显,其机制可能是通过下调Bcl-2、上调Bax的表达发挥其促凋亡作用.%Objective To investigate the effect of cisplatin,VP-16,vincristine and bleomycin on the function of human granulosa cell. Methods Ovarian cancer cell line PA-1 was interfered with cisplatin,VP-16,vincristine and bleomycin in different concentration for 48 h. The median lethal dose was measured by MTT. Granulosa cells were obtained from infertile patients undergoing IVF-ET cycles and cultured in vitro. Cultured for 24 h, these cells were interfered with the median lethal dose of four drugs above. After another 72 h, the viability of cells was determined by MTT. The culture medium was collected and the estradiol ( E2) level was detected by chemiluminescence method. Using immunocytochemical technique, the expression of Bcl-2, Bax on granulosa cell was observed. Results It was revealed that the consecutive IC50 of cisplatin,VP-16,vincristine and bleomycin was 0.55 mg/L,1.67 mg/L,l. 8|xg/L,1.7 mg/L. We also demonstrated that the IC50 of cisplatin, VP-16 and bleomycin except vincristine inhibited the growth of human granulose cells and reduced secretion of E2. Immunocytochemical technique revealed the expression of Bcl-2 was down-regulated and Bax was up-regulated. Conclusion It is suggested that cisplatin,VP-16,bleomycin except vincristine can all promote the ap-optosis of human granulosa cell. VP - 16 and bleomycin role to be the strongest, and there was no significant difference between them,cisplatin less, vincristine does not play a significant role.
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