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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-1FSH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。
OBJECTIVE: To investigate the effect of rFSH on the expression of VEGF and VEGFR-2 in mouse ovary tissues during different stages of vitrification, and to explore the best way to protect FSH during ovarian vitrification. Methods The ovaries of 4-week-old C57BL / 6J mice were divided into fresh control group (FCG), vitrified control group (VCG) and 0.3IU.mL-1 FSH intervention vitrification group, including FSH group (OG-FSH ), FSH group (BG-FSH) before freezing and FSH group (AG-FSH) after freezing. Morphological techniques were used to observe the percentage of normal follicles. Immunohistochemistry was used to observe the expression of VEGF and VEGFR-2 in different ovarian cells. The expression of VEGF and VEGFR-2 protein in ovarian tissues was observed by Western-blot and analyzed by FSH intervention in different vitrification periods Effect of expression level. Results The percentage of normal follicles, VEGF and VEGFR immunohistochemistry and Western-blot results were significantly different between the two groups were statistically significant. Among them, the detection indexes of OG-FSH group were higher than those of other groups (P <0.05), and the levels of BG-FSH and AG-FSH were similar to that of FCG and higher than that of VCG group (P <0.05). Conclusion FSH intervention during vitrification can improve the number of normal follicles and the expression of VEGF and VEGFR-2.