小鼠卵巢玻璃化冻存过程中FSH干预对VEGF及VEGFR-2表达的影响

摘要

Objective To investigate the best protection way of FSH, which interposed in different stage of vitrification process via observing the expression of VEGF and VEGFR -2. Methods:4 -week -old C57BL/ 6J mice were divided into fresh control group( FCG) , vitrification control group( VCG) and the vitrification intervention group by 0. 3IU · mL FSH ( VIG). VIG included the group of FSH interposed in the overall process of vitrification group(OG - FSH) , FSH interposed in the before process of vitrification group(BG -FSH) and FSH interposed in the after process of vitrification group( AG - FSH) . The percentage of normal follicles were observed through morphology and , the expression of VEGF, VEGFR -2 in different cells of ovary were observed by immunohistochemical technique. The effect on the expression of VEGF, VEGFR - 2 protein about FSH intervention in different stage of vitrification was observed and analyzed by western -blot. Results Normal follicles ,VEGF and VEGFR -2. in OG - FSH were significantly higher than those in other groups (P < 0. 05 ) . Conclusion OG - FSH is in favour of improving the percentage of normal follicles and the expression of VEGF, VEGFR -2 protein.%目的 通过在玻璃化冻存不同时段给予小鼠卵巢组织重组人促卵泡刺激素(rFSH)干预,观察rFSH对VEGF以及VEGFR-2表达的影响,探索卵巢玻璃化冻存过程中最佳的FSH保护途径.方法 将4周龄C57BL/6J小鼠卵巢分为新鲜对照组(FCG)、玻璃化冻存对照组(VCG)、0.3IU·mL-1 FSH干预玻璃化冻存组,包括全程添加FSH组(OG-FSH)、冻存前添加FSH组(BG-FSH)以及冻存后添加FSH组(AG-FSH).通过形态学技术观察正常卵泡百分比;采用免疫组织化学技术观察卵巢不同细胞VEGF以及VEGFR-2的表达;通过Western-blot方法观察并分析不同玻璃化冻存时段的FSH干预对卵巢VEGF以及VEGFR-2蛋白表达量的影响.结果 正常卵泡百分比、VEGF及VEGFR免疫组织化学和Western-blot检测结果组间比较差异均有统计学意义.其中,OG-FSH组各项检测指标均高于其他各组(P＜0.05),且BG-FSH、AG-FSH组与FCG相似,高于VCG组(P＜0.05).结论 玻璃化冻存全程添加FSH干预有利于提高正常卵泡数,以及VEGF、VEGFR-2蛋白的表达.