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| 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 |
| 李安吉1,张洁2,刘璐茜1,翟东霞1,张丹英1,匡延平2,吕祁峰2,俞超芹1* |
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(1. 第二军医大学长海医院中医系, 上海 200433;
2. 上海交通大学医学院附属上海市第九人民医院辅助生殖科, 上海 200011
*通信作者) |
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| 摘要: |
| 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+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。 |
| 关键词: 子宫内膜异位症 卵泡液 内异方 卵母细胞 |
| DOI：10.16781/j.0258-879x.2016.06.0694 |
| 投稿时间：2016-04-26修订日期：2016-06-02 |
| 基金项目:国家自然科学基金(30930113,81573755). |
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| Effect of Neiyi Recipe-medicated serum on in vitro maturation of mouse oocytes blocked by follicular fluid from endometriosis patients |
| LI An-ji1,ZHANG Jie2,LIU Lu-xi1,ZHAI Dong-xia1,ZHANG Dan-ying1,KUANG Yan-ping2,Lü Qi-feng2,YU Chao-qin1* |
(1. Department of Traditional Chinese Medicine, Changhai Hospital, Second Military Medical University, Shanghai 200433, China;
2. Department of Assisted Reproduction, The 9th People's Hospital of Shanghai, School of Medicine, Shanghai Jiaotong University, Shanghai 200011, China
*Corresponding author) |
| Abstract: |
| Objective To investigate the mechanism by which follicular fluid from endometriosis (EM) patients blocks mouse oocyte maturation and the effect of Neiyi Recipe-medicated serum on the effect of the follicular fluid. Methods A total of 20 follicular fluid samples from infertility patients due to EM (n=10) or fallopian tube factors (control group, n=10) were clinically obtained. Neiyi Recipe-medicated serum was obtained by treating mice with intragastric administration, and were divided into 4 medium groups: Group A (control), group B (control follicular fluid), group C (EM-follicular fluid) and group D (EM-follicular fluid + Neiyi Recipe-medicated serum). The mouse germinal vesicle (GV) oocytes were cultured in the above 4 types of media for in vitro maturation. And the maturity rates of oocytes in different groups were calculated and the concentrations of reactive oxygen species in the cells were analyzed by fluorescence staining and laser scanning confocal microscope. Results The maturity rate of oocytes in group C (42.5%) was significantly lower than those in group A (81.7%), group B (56.3%) and group D (51.0%) (P<0.05). The average fluorescence intensity of group C (0.056 8±0.025 1) was significantly higher than those of group A (0.005 1±0.014 8, P<0.01) and group B (0.037 1±0.010 2, P<0.05), but with no significant difference found when compared with that of group D (0.050 4±0.007 0). Conclusion EM-follicular fluid can block the maturation of mouse oocytes, which may be due to the enhanced oxidative stress in oocytes caused by EM-follicular fluid. Neiyi Recipe-medicated serum can improve the blocking effect of EM-follicular fluid on mouse oocyte development. |
| Key words: endometriosis follicular fluid Neiyi Recipe oocytes |
