基于cAMP信号通路研究miR-7a2对青春期大鼠雄性生殖系统发育的影响

Effect of miR-7a2 on the development of the male reproductive system in adolescent rats based on the cAMP signaling pathway

目的研究基于环磷酸腺苷(cAMP)信号通路分析微小RNA-7a2(miR-7a2)对青春期大鼠雄性生殖系统发育的影响。方法本研究选取2020年3—10月健康、清洁级Wistar雄性鼠龄3周大鼠30只,随机分为对照组(生理盐水)、miR-7a2上调组(超表达pmcherry-Flag-miR-7a2)、miR-7a2下调组(慢病毒质粒miR-7a2-sh RNA-1),每组各10只。检测三组激素水平,统计体重、睾丸重量、精子畸变、活动度,观察睾丸形态学,检测cAMP信号通路相关蛋白腺苷酸环化酶(AC)、磷酸二酯酶(PDE)、蛋白激酶(PK)。结果miR-7a2上调组miR-7a2表达高于对照组和miR-7a2下调组;miR-7a2下调组miR-7a2表达低于对照组(P<0.05)。miR-7a2上调组雌二醇、卵泡刺激素水平低于miR-7a2下调组;睾酮水平高于miR-7a2下调组(P<0.05)。miR-7a2上调组体重、睾丸重量、精子密度高于miR-7a2下调组,精子畸变率低于miR-7a2下调组(P<0.05)。miR-7a2上调组良好、一般精子活动度高于miR-7a2下调组;不良、死亡精子活动度低于miR-7a2下调组(P<0.05)。miR-7a2上调组AC蛋白表达低于miR-7a2下调组,PK蛋白表达高于miR-7a2下调组(P<0.05)。结论miR-7a2上调能促进青春期大鼠雄性生殖系统发育,其作用机制与cAMP信号通路有关。

Objective To investigate the effect of microRNA-7a2(miR-7a2)on the development of the male reproductive system in adolescent rats based on the cyclic adenosine monophosphate(cAMP)signaling pathway.Methods Thirty healthy,clean Wistar male rats aged three weeks from to March to October 2020 were randomly divided into the control group(saline),the miR-7a2 up-regulation group(overexpression of pmcherry-Flag-miR-7a2),and miR-7a2 down-regulation group(lentiviral plasmid miR-7a2-shRNA-1),with ten rats in each group.Hormone levels were detected.The body weight,testicular weight,sperm aberrations,and motility were measured.The testicular morphology was observed.The cAMP signaling pathway-related proteins adenylate cyclase(AC),phosphodiesterase(PDE),and protein kinase(PK)were measured.Results miR-7a2 expression was higher in the miR-7a2 up-regulation group than in the control and miR-7a2 down-regulation groups.miR-7a2 expression was lower in the miR-7a2 down-regulation group than in the control group(P<0.05).Estradiol and follicle-stimulating hormone levels were lower,and testosterone levels were higher in the miR-7a2 up-regulation group than in the miR-7a2 down-regulation group(P<0.05).Body weight,testicular weight,and sperm density were higher,and sperm aberration rate was lower in the miR-7a2 up-regulation group than in the miR-7a2 down-regulation group(P<0.05).The excellent and general sperm motility in the miR-7a2 up-regulation group was higher than that in the miR-7a2 down-regulation group.The poor and dead sperm motility in the miR-7a2 up-regulation was lower than in the miR-7a2 down-regulation group(P<0.05).AC protein expression was lower,and PK protein expression was higher in the miR-7a2 up-regulation group than in the miR-7a2 down-regulation group(P<0.05).Conclusion The up-regulation of miR-7a2 can promote the development of the male reproductive system in adolescent rats,and the mechanism is related to the cAMP signaling pathway.