磷酰胺氮芥对大鼠卵巢功能损伤的体外实验研究

[摘要] 目的：研究磷酰胺氮芥（Glyciphosphoramide ,PM）引起体外培养大鼠卵巢结构和内分泌功能损伤的最低剂量。方法：实验分为4组，每组12个卵巢，来源于出生8周的Wistar大鼠。实验1、2、3组培养基中加入浓度分别为10μg/mL、20μg/mL、50μg/mL的PM，培养48h后均隔天全量更换不含PM的培养基。正常对照组培养基中不含PM。在PM加入前及加入后2d（48h）、6d时更换的培养基中用放射免疫法测定其雌二醇（E2）浓度。在加入PM6d时取出大鼠卵巢，称湿重，HE染色，显微镜下观察各期卵泡形态并计数总卵泡数量。结果：加入PM后2d及6d的3个实验组的E2浓度均低于正常对照组，差异均有统计学意义（P＜0.05），且3个实验组间两两比较差异也均有统计学意义。加入PM6d，3个实验组分别与正常对照组比较，卵泡数差异均有统计学意义；3个实验组间两两比较，卵泡数量差异亦均有统计学意义。结论：PM浓度为10μg/mL时已能引起体外培养大鼠卵巢结构及功能的损伤。

The effect of Glyciphosphoramide on rat ovarian in vitro

〔Abstract〕Objective: To study the lowest dose level of Glyciphosphoramide(PM) that will cause ovarian damage of female rats in vitro. Method: Ovaries were obtained from wistar rats and incubated with PM at concentrations of 10, 20, and 50 micrograms/mL as experimental groups . Normal control group ovaries were added with culture medium only . Ovaries were got from 8 week wistar rats.To culture the ovarian tissue in 24 hole culture plate，one ovary each hole, for 7 days . Estradiol(E2) were measured by radio-immunity method before and at 2,6 day after given PM . Weighting, HE staining and counting the number of follicles of each ovary at 7 day after culture(6 days after given PM). Result : There showed significant difference at 2 and 6 days after given PM in E2 lever between three experimental groups and control group（P﹤0.05）. There were obvious difference between three experimental groups and control group in follicular numbers at day 7（P﹤0.05）. Conclusion: 10μg/mL PM can cause ovarian damage of female rats in vitro.