FSH干预对小鼠玻璃化冻融移植卵巢血流重建的影响

The Effects of FSH Intervention in Vitrification Freezing on the Revascularization after Mouse Ovarian Transplantation

目的观察小鼠卵巢玻璃化冻融中卵泡刺激素(FSH)干预对移植48h卵巢血流重建的影响及相关机制。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(FSH-VG),采用卵巢异体肾被膜下移植技术,于移植48h通过卵巢形态学观察、荧光血管灌注、EdU检测细胞增殖情况、免疫组织化学及western-blot方法观察并分析血流重建的影响因素。结果小鼠玻璃化冻融卵巢移植48h,与VCG比较,FSH-VG血流灌注较广,且达到卵巢近中央部;FSH-VG组正常卵泡百分比明显高于VCG组(P<0.01),VCG组闭锁卵泡百分比明显高于FSH-VG组(P<0.01),增殖细胞主要为卵巢间质细胞。FSH-VG组CD34、PCNA蛋白表达高于VCG(P<0.01)。结论玻璃化冻融过程中的FSH干预可加速小鼠卵巢冻融后移植的血流重建,CD34及PCNA表达上调。

Objective To investigate the effects of FSH intervention in vitrification freezing ization after mouse ovarian transplantation 48 hours and related mechanisms. Methods on the revaseular- 4 - week - old C57BL/6J mouse ovaries were randomly divided into fresh control group （FCG） and experimental groups,the experimental groups contained vitrification control group （VCG） and FSH intervented vitrification group （ FSH - VG）. The cell proliferation of allotransplantation of ovaries in subrenal capsuler were detected by mor- phology,fluorescence vascular perfusion and EdU methods at 48 hours. Furthermore, the expression of CD34 and PCNA were analyized by immunohistochemistry and western -blot methods. Results After transplanted 48 hours, comparing with VCG group, the distribution of blood flow in FSH - VG group was more wide, nearly reaching the center part of transplanted ovaries. The normal follicles percentages in FSH - VG group were higher than those in VCG （ P 〈 0.01 ）. The percentage of atretic follicles in VCG group were significantly higher than those in FSH -VG group （P 〈0.01 ）. The strom cells consisted mainly of proliferative cells. Up - regulated CD34 ,PCNA in FSH -VG were much more than those in VCG. Conclusion The intervention of FSH in vitrification freezing process can help transplanted mice ovaries to quickly build revascularization, and the expressison of CD34 and PCNA.