子宫内膜异位症患者卵泡液阻碍小鼠卵母细胞体外成熟及内异方药物血清的干预作用

目的 探讨子宫内膜异位症(EM)患者卵泡液阻碍小鼠卵母细胞发育的机制及中药内异方药物血清对此的干预作用.方法 临床获取EM不孕患者、输卵管因素导致不孕患者(对照组)卵泡液各10例样本,对小鼠行中药内异方灌胃获得药物血清,分别制备4组培养基:A组(空白对照)、B组(对照患者卵泡液)、C组(EM患者卵泡液)、D组(EM患者卵泡液十内异方药物血清).将小鼠生发泡(GV)期卵母细胞分别放入各组培养基中进行体外成熟(IVM)培养.观察并统计各组卵母细胞的成熟情况,通过荧光染色对细胞内活性氧(ROS)的水平进行分析比较.结果 C组卵母细胞的成熟率(42.5％)相较A组(81.7％)、B组(56.3％)、D组(51.0％)低(P＜0.05).C组卵母细胞平均荧光强度(0.056 8±0.025 1)高于A组(0.014 8±0.005 1,P＜0.05)和B组(0.037 1±0.010 2,P＜0.05);D组卵母细胞平均荧光强度(0.050 4±0.007 0)低于C组,但差异无统计学意义.结论 EM患者卵泡液能够阻碍小鼠卵母细胞成熟,其机制可能与EM卵泡液产生过多ROS而增强了卵母细胞内的氧化应激有关.内异方药物血清可改善EM患者卵泡液对小鼠卵母细胞发育的阻碍作用.

Effect of Neiyi Recipe-medicated serum on in vitro maturation of mouse oocytes blocked by follicular fluid from endometriosis patients

Objective: To investigate the Block mechanism of follicular fluid from endometriosis patients on mouse oocyte maturation and Effect of Neiyi recipe drug serum on these interfered oocytes. Methods: 20 cases of follicular fluid samples from infertility patients with and without(control group) endometriosis half and half and intragastric administration to mice aimed to obtain Neiyi recipe drug serum. The mouse GV oocytes were placed in culture medium which 4 groups were A group (blank group), B group (control-FF), group C (EM-FF), D group (EM-FF+ Neiyi recipe drug serum) for in vitro maturation. After culturing, maturity rates of groups were calculated and reactive oxygen species concentrations in the cells were analyzed by fluorescence staining and laser scanning confocal microscope. Results: the maturity rate of group C (42.5%) in was lower than that in group B (56.3%) and group D (51.0%) (P<0.05). The average fluorescence intensity of group C (0.0568±0.0251) was significantly higher than that of group A (0.0051±0.0148) (P<0.01) and group B (0.0371±0.0102) (P<0.05),and higher than that of group D ,but no Statistical difference was observed. Conclusion: EM-FF may interfere the maturation of mouse oocytes, which may be induced by enhancing oxidative stress in oocytes. Neiyi recipe drug serum can improve the damage of EM-FF on mouse oocytes development.