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目的探讨丹参对小鼠冻融卵巢同种异体移植后的血管化作用及可能机制。方法收集C57BL/6j雌鼠和BALB/c雄鼠杂交的F1代4周龄小鼠卵巢,慢速冻融后移植至B6C2F1代8～12周龄雄鼠的肾被膜下,移植后小鼠按给药不同分为生理盐水组和丹参组,分别于移植后1d(24h)、2d(48h)和7d回收移植物,将冻融以及移植后不同时间段的卵巢组织进行HE染色全卵巢卵泡计数,免疫组织化学及RT-PCR方法检测血管相关蛋白及基因的表达。结果移植卵巢组织各时间段全卵巢卵泡计数及卵泡存活率丹参组均高于生理盐水对照组(P<0.05);移植后小鼠血管内皮生长因子(VEGF)的表达有上升趋势,第7天达最高峰,两组之间无显著性差异(P>0.05);移植卵巢组织各时间段细胞凋亡指数,丹参组均低于生理盐水对照组(P<0.05);移植后48h丹参组VEGF188mRNA和VEGF164mRNA的表达量较对照组高,两组之间有显著性差异(P<0.05)。结论小鼠卵巢组织移植后早期阶段丹参能促进卵泡发育,减少卵泡凋亡,可能与提高移植卵巢组织内血管相关基因VEGF(主要是VEGF164mRNA和VEGF188mRNA)的表达而促进移植后早期血管化有关。
Objective To explore the effect of salvia miltiorrhiza on the vascularization after allogeneic transplantation of mice with frozen-thawed ovary and its possible mechanism. Methods The ovaries of F1 generation 4-week-old mice which were crossed with C57BL / 6j female mice and BALB / c male mice were collected and then transplanted into the subrenal capsule of B6C2F1 generation male rats aged 8-12 weeks after slow freezing and thawing. The rats were randomly divided into normal saline group and Salvia miltiorrhiza group. The grafts were harvested at 1d (24h), 2d (48h) and 7d after transplantation. The ovarian follicles were frozen and thawed at different time after transplantation. , Immunohistochemistry and RT-PCR method to detect the expression of vascular related proteins and genes. Results The total ovarian follicle count and follicular viability in transplanted ovarian tissue were higher than those in saline control group (P <0.05). The expression of vascular endothelial growth factor (VEGF) (P> 0.05). The apoptotic index of ovarian tissue in each time period was lower in Salvia miltiorrhiza than that in saline control group (P <0.05). VEGF188 mRNA in Salvia miltiorrhiza group 48 h after transplantation And VEGF164 mRNA were higher than the control group, there was a significant difference between the two groups (P <0.05). Conclusion Salvia miltiorrhiza can promote follicular development and reduce apoptosis of ovarian follicles in the early stage of ovarian transplantation, which may be related to the increase of vascular related gene VEGF (mainly VEGF164 mRNA and VEGF188 mRNA) and the promotion of early vascularization in transplanted ovarian tissue.