丹参促进小鼠冻融卵巢移植早期血管内皮生长因子的表达  

Salvia miltiorrhiza promotes vascular endothelial growth factor expression in frozen-thawed mouse ovarian in an early stage after transplantation

作　　者：唐飞[1] 张婵[2] 林海燕[1] 熊成容[1] 王晓娇[1] 吴丹丹[1] 周颖[1,3] 

机构地区：[1]温州医学院附属第一医院生殖医学中心 [2]温州医学院生物学实验教学中心 [3]温州医学院基础医学院组织学胚胎教研室,浙江温州325000

出　　处：《解剖学报》2010年第1期93-99,共7页Acta Anatomica Sinica

基　　金：国家自然科学基金资助项目(30370749);浙江省科技厅资助项目(2003C33001);温州市科技局对外科技合作交流项目资助(H20080061)

摘　　要：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 investigate the angiogenesis effects of Salvia miltiorrhiza in heterotopically grafted frozen-thawed mouse ovaries. Methods The ovaries thawed after cryopreservation were xenografted into the donated kidney capsules of 8- to 12-week adult male mice. The mice were divided into two groups, saline and Salvia miltiorrhiza groups, the mice either in the saline or in Salvia miltiorrhiza groups were administered i.p. daily either saline （0.5ml） or Salvia mihiorrhiza（0.5g） respectively, from the day prior transplantation. The two groups were sacrificed 1 day,2 days and 7 days after transplantation respectively, the grafts from thawed, 1 day, 2 days, 7 days were removed for follicle counting, immunohistochemical studying and detecting of the mRNA expression of vascular endothelial growth factor （VEGF）. Results The number of follicles and survival rates in grafts after transplantation of Salvia mihiorrhiza group were more than that of saline group （ P 〈 0.05 ） ; the expression of VEGF increased after transplantation, peaked on day 7, there was no difference between the two groups （P 〉 0. 05） ; the apoptosis index of Salvia miltiorrhiza group was less than that of saline group （P 〈0. 05 ）, the mRNA expression of VEGF188 and VEGF164 of Salvia miltiorrhiza group was more than that of saline group on 48 hours after transplantation （P 〈 O. 05 ）. Conclusion Salvia miltiorrhiza may provide benefits for follieulogensis and decreasing the apoptosis index of follicles. Nevertheless, a increase in the VEGF188 and VEGF164 isoform in the Salvia miltiorrhiza group may suggest the positive effect of exogenous Salvia miltiorrhiza therapy in the early stage of angiogenesis.

关 键 词：丹参 卵巢 移植 血管化 血管内皮生长因子 免疫组织化学 反转录-聚合酶链反应 小鼠 

分 类 号：R339.2[医药卫生—人体生理学]