降钙素基因相关肽对卵母细胞体外成熟的影响

Effect of calcitonin gene-related peptide on the maturation of oocyte in vitro

目的：探讨降钙素基因相关肽（calcitoningene．relatedpeptide，CGRP）对小鼠卵母细胞体外成熟的影响。方法：6周龄雌性昆明小鼠腹腔注射孕马血清促性腺激素（pregnantmareserumgonadotropin，PMSG）10U48h后处死，取出卵巢收集卵母细胞以30～40枚／孔的密度接种于培养板内，以不同浓度CGRV（0，10^-10，10^-9，10^-9mol／L）处理24h后检测生发泡破裂（germinalvesiclebreakdown，GVBD）率和第一极体（polarbodyI，PBI）排出率；收集、培养人卵巢颗粒细胞，以不同浓度CGRV（0，10^-10，10^-9，10^-8mol／L）处理24h后测定颗粒细胞内cAMP浓度。结果：外源性CGRP处理小鼠卵母细胞可浓度依赖性地降低GVBD率及PBI排出率；外源性CGRP处理人颗粒细胞，可浓度依赖性地增高细胞内cAMP浓度。结论：CGRP抑制小鼠卵母细胞成熟可能与其增加颗粒细胞内cAMP浓度有关。

Objective： To explore the effect of calcitonin gene-related peptide （CGRP） on murine oocyte maturation. Methods： After injection of pregnant mare serum gonadotropin （PMSG, 10 U, i.p.） for 48 h, 6-week old female Kunming mice were killed, and the cumulus oocyte complexes （COCs） were collected from ovaries and inoculated in the culture plate by 30-40/hole. The COCs were treated with 4 concentrations of CGRP （0, 10-10, 10-9, and 10-8mol/L）, and the germinal vesicle breakdown （GVBD） and polar body I （PBI） were examined. Human granulosa cells were also cultured with CGRP （0, 10 10-9, 10-8 mol/L） and levels of intraceUular cyclic adenosine monophosphate （cAMP） were measured. Results： Exogenous CGRP caused a decrease in GVBD and PBI in COCs, and an increase in cAMP levels in human granulosa cells in a concentration-dependent manner.Conclusion： CGRP can inhibit the oocyte maturation, which may be related to the increased content of cAMP in granulosa cells.