论文部分内容阅读
目的通过在玻璃化冻存全程给予小鼠卵巢组织促卵泡刺激素(FSH)、FSH和促黄体生成素(LH)共同干预,观察FSH及FSH+LH干预对冻存卵巢卵泡缝隙连接蛋白(connexin-43,Cx43)表达的影响,从而判断最佳的激素保护方法。方法将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),0.3IU·m L~(-1)FSH全程干预玻璃化冻存组(OG-FSH),以及0.15IU·m L~(-1)FSH+0.15IU·m L~(-1)LH全程干预玻璃化冻存组(OG-FSH+LH)。通过免疫组织化学法、Western blot技术及荧光定量PCR检测,观察并分析各组卵巢组织Cx43蛋白表达量。结果免疫组织化学及Western blot检测结果表明Cx43蛋白表达量在OG-FSH+LH组最高,其次为OG-FSH组、CG组及VCG组(P<0.05),各组间两两比较差异均有统计学意义(P均<0.05);荧光定量PCR检测结果显示Cx43mRNA表达量从高到低依次为OG-FSH+LH组、OG-FSH组、CG组及VCG组(P<0.05),且各组间两两比较差异均有统计学意义(P<0.05)。结论玻璃化冻存全程添加FSH+LH的干预方式更有利于冻融卵巢组织缝隙连接蛋白Cx43蛋白的表达。
OBJECTIVE: To investigate the effect of FSH and FSH + LH on the expression of connexin-2 mRNA in ovarian follicles by the combined intervention of follicle-stimulating hormone (FSH), FSH and luteinizing hormone (LH) 43, Cx43) expression in order to determine the best hormone protection methods. Methods Four-week-old C57BL / 6J mice were divided into four groups: fresh control group (CG), vitrification control group (VCG), 0.3IU · m L -1 FSH intervention group (OG- FSH) and 0.15IU · m L -1 FSH + 0.15IU · m L -1 LH were intervened in vitrification group (OG-FSH + LH). The expression of Cx43 protein in ovarian tissue of each group was observed and analyzed by immunohistochemistry, Western blot and fluorescent quantitative PCR. Results The results of immunohistochemistry and Western blot showed that the expression of Cx43 protein was highest in OG-FSH + LH group, followed by OG-FSH group, CG group and VCG group (P <0.05) (P <0.05). The results of real-time PCR showed that the expression levels of Cx43 mRNA were OG-FSH + LH group, OG-FSH group, CG group and VCG group from high to low (P <0.05) The differences between groups were statistically significant (P <0.05). Conclusion The intervention of FSH + LH during vitrification is more beneficial to the expression of connexin Cx43 in the frozen-thawed ovary tissue.