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目的探讨TNF-α对小鼠卵母细胞体外成熟的影响。方法收集未成熟卵母细胞,在含不同浓度TNF-α的培养液中进行培养,浓度梯度为0(作为对照),5,10,100,200,400和600ng/ml培养3、9、12h,所有卵母细胞体外培养12h后统计MⅡ期成熟卵母细胞数量。结果当TNF-α浓度≥5ng/ml作用3h,与对照组相比,卵母细胞生发泡破裂(GVBD)比例及成熟率明显降低(分别为89.54%与73.12%及78.43%与50.54%,P<0.05)。将已经发生生发泡破裂的小鼠卵母细胞移到含有不同浓度TNF-α的培养液中培养9h,当TNF-α浓度≥10ng/ml时,小鼠卵母细胞自GVBD发育到MⅡ的比例为74.55%,明显低于对照87.59%(P<0.05)。TNF-α作用12h组,当TNF-α浓度≥5ng/ml时,MⅡ期卵母细胞比例显著低于对照组(P<0.01)。结论TNF-α明显抑制卵母细胞的体外成熟,包括抑制生发泡的破裂及第一极体的排放,并具有一定的剂量依赖性,低剂量组还具有时间依赖性。
Objective To investigate the effect of TNF-α on mouse oocyte maturation in vitro. Methods Immature oocytes were collected and cultured in culture medium containing different concentrations of TNF-α. The concentration gradient was 0 (as control), and all the oocytes were cultured in vitro at 5, 10, 100, 200, 400 and 600ng / ml After 12 hours of cultivation, the number of MⅡ mature oocytes was counted. Results Compared with the control group, the rate of oocytes foaming rupture (GVBD) and maturation rate were significantly decreased (89.54% vs 73.12% and 78.43% vs 50.54%, respectively) when TNF-α concentration was> 5ng / ml for 3h <0.05). The mouse oocytes that had ruptured vesicles were cultured in culture medium containing different concentrations of TNF-α for 9h. The ratio of mouse oocytes developing from GVBD to MⅡ when the concentration of TNF-α≥10ng / ml Was 74.55%, significantly lower than the control 87.59% (P <0.05). In the 12h group, the proportion of MⅡ oocytes was significantly lower than that of the control group (P <0.01) when the concentration of TNF-α≥5ng / ml. Conclusion TNF-α significantly inhibits oocyte maturation in vitro, including inhibition of rupture of primary vesicles and discharge of the first polar body in a dose-dependent manner. The low-dose group also has a time-dependent manner.