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目的通过在玻璃化冻存全程给予小鼠卵巢组织卵泡刺激素(FSH)及FSH和黄体生成素(LH)共同干预,观察冻融卵巢组织的形态学改变以及两种激素干预对冻存卵巢组织血管内皮生长因子(VEGF)表达的影响,寻找最佳的提高冻融卵巢组织卵泡存活及VEGF表达的激素干预方式。方法 4周龄C57BL/6J小鼠卵巢组织分为新鲜对照组(CG),玻璃化冻存对照组(VCG),300IU/L FSH全程干预玻璃化冻存组(OG-FSH),以及150IU/L FSH+150IU/L LH全程干预玻璃化冻存组(OG-FSH+LH),每组30个卵巢样本。通过常规组织学、Western blotting技术,观察并分析各组卵巢组织形态结构改变及VEGF蛋白表达量;通过荧光定量PCR技术(Real-time PCR)检测VEGF mRNA表达情况。结果 OG-FSH+LH组正常卵泡百分比最高,且显著高于OG-FSH组(P<0.05);VEGF蛋白表达量在OG-FSH+LH组显著高于OG-FSH组(P<0.05);荧光定量PCR检测结果表现为VEGF mRNA表达量在OG-FSH+LH组最高,其次为OG-FSH组,最低是VCG组(P<0.05)。结论玻璃化冻存全程添加FSH+LH的干预方式较单独FSH干预具有更高正常卵泡百分比和更佳的VEGF蛋白表达。
OBJECTIVE: To observe the morphological changes of frozen-thawed ovarian tissue and the effects of two hormone interventions on frozen-thawed ovarian tissue by giving follicle-stimulating hormone (FSH) and FSH and luteinizing hormone (LH) together during mouse vitrification. To investigate the effect of VEGF on the expression of VEGF and VEGF in ovarian tissues. Methods Four-week-old C57BL / 6J mice were divided into three groups: fresh control group (CG), vitrification control group (VCG), 300IU / L FSH interventional vitrification group (OG-FSH) and 150IU / L FSH + 150IU / L LH intervention vitrification group (OG-FSH + LH), each group of 30 ovarian samples. The morphological changes of ovarian tissue and VEGF protein expression were observed and analyzed by routine histology and Western blotting. The expression of VEGF mRNA was detected by Real-time PCR. Results The percentage of normal follicles in OG-FSH + LH group was significantly higher than that in OG-FSH group (P <0.05). The expression of VEGF protein in OG-FSH + LH group was significantly higher than that in OG-FSH group (P <0.05) Fluorescence quantitative PCR showed that the expression of VEGF mRNA was highest in OG-FSH + LH group, followed by OG-FSH group, and lowest in VCG group (P <0.05). Conclusion The intervention of FSH + LH in vitrification all had higher percentage of normal follicles and better expression of VEGF than FSH alone.