氨基双吡酮对小鼠卵母细胞成熟的影响

Effect of amrinone on the maturation of mouse oocytes

目的研究氨基双吡酮(氨力农)对体外和体内小鼠卵母细胞成熟的抑制作用,检测磷酸二酯酶(PDE)3抑制剂氨力农能否作为一个潜在的避孕药。方法将不同浓度药物作用于体外培养的不成熟小鼠卵母细胞,以及注射于小鼠体内,观察其对卵母细胞成熟的阻滞作用并找出合适的浓度。卵母细胞成熟后进行卵胞质内单精子注射(ICSI),并进行移植,观察其受精率、胚胎形成情况以及出生后代比较。并监测体内注射组小鼠的不良反应情况。结果 (1)随着浓度的增加,氨力农导致减数分裂阻滞呈剂量依赖性,体外有效浓度为1μmol/L,而在体内为300 mg/kg。(2)其作用是可逆的。药物去除后,减数分裂恢复,小鼠卵母细胞成熟,并呈正常染色体排列和纺锤体组织。(3)与对照组比较,ICSI受精后,卵母细胞表现出正常的形态、受精率、卵裂、囊胚形成。(4)其出生后代表现出相似的体质量和生育能力。(5)在体内,注入一定浓度药物后小鼠表现为不孕,停药后恢复。结论氨力农可以抑制体外和体内小鼠卵母细胞的成熟,并且是可逆的;交配实验证明氨力农可能可作为一种避孕药物。

Objective To examine the inhibitory effect of amrinone on the maturation of mouse oocytes in vitro and in vivo, and whether it is possible to detect the potential use of the inhibitor as a potential contraceptive. Methods The effects of different concentrations of amrinone on immature mouse oocytes were observed. After oocyte maturation, intracytoplasmic sperm injection（ICSI） was carried out, and the fertilization rate, embryo formation and the comparison of the offspring were observed. The adverse reaction of mice in in vivoinjection group was monitored. Results 1） With the increase of the concentration, the ammonia induced meiosis was dose-dependent. The effective concentration was 1 μmol/L in vitro, but was about 300 mg/kg in vivo. 2） Its function was reversible. After the removal of the drug, the meiosis recovered gradually and mouse oocyte matured with normal chromosome alignment and spindle organization. 3） Compared with control group, after fertilization, oocytes showed normal morphology, fertilization rate, cleavage and blastocyst formation. 4） The birth offspring showed similar body weight and fertility. 5） In vivo, a certain concentration of drug injected into mice exhibited infertility till discontinuation. Conclusion In vitro and in vivo mouse oocytes can be inhibited by amrinone and maturation is reversible, and mating experiments have proved that it may be used as a contraceptive drug.