血管内皮细胞YY1条件性敲除对小鼠卵巢功能的影响及其作用机制  

Effect of conditional YY1 knockout in vascular endothelial cells on ovarian function in mice and its mechanism

作　　者：王娜[1] 芦晓红[1] 袁璟 马虹 WANG Na;LU Xiaohong;YUAN Jing;MA Hong(School of Basic Medicine,Ningxia Medical University,Yinchuan 750004,Ningxia Hui Autonomous Region,China;不详)

机构地区：[1]宁夏医科大学基础医学院,宁夏银川750004 [2]宁夏大学西部特色生物资源保护与利用教育部重点实验室,宁夏银川750021

出　　处：《中国生物制品学杂志》2024年第11期1307-1312,共6页Chinese Journal of Biologicals

基　　金：宁夏自然科学基金(2020AAC03144);宁夏医科大学校级科研项目资助(XT202203)。

摘　　要：目的探讨血管内皮细胞YY1条件性敲除对小鼠卵巢功能的影响及其作用机制,以期为血管新生障碍导致不孕症的诊疗提供新靶点及策略。方法采用CreER^(T2)/LoxP系统构建YY1^(flox/flox)(WT型)和Cdh5-CreER^(T2)小鼠,两者进行交配,获得血管内皮细胞YY1诱导性条件性敲除小鼠(Cdh5-CreER^(T2);YY1^(flox/flox),简称YY1^(iΔEC)),PCR法鉴定基因型。通过对YY1^(iΔEC)小鼠注射他莫昔芬诱导Cre酶活性,实现在血管内皮细胞内特异性敲除YY1,敲除1个月后,分离小鼠肺血管内皮细胞,免疫荧光法及Western blot法检测其敲除效果;检测敲除后YY1^(iΔEC)雌鼠的生育力、动情周期、卵巢血管新生情况,并进行卵巢及子宫的大体观察,以WT雌鼠为对照。结果敲除YY11个月后,YY1^(iΔEC)小鼠血管内皮细胞数目下降,且细胞上的YY1被高效特异性敲除。与WT雌鼠比较,YY1^(iΔEC)雌鼠怀孕生产幼崽的能力大幅降低;缺乏规律动情周期,存在自发排卵和黄体生成障碍;卵巢血管新生受到明显抑制(F=1.288,P<0.01);卵巢和子宫体积较小,卵巢颜色苍白,表面黄体缺乏。结论血管内皮细胞YY1条件性敲除可导致小鼠生殖功能障碍,可能是由于血管内皮细胞YY1缺失抑制了卵巢血管新生,导致排卵及黄体生成障碍。Objective To investigate the effect of conditional YY1 knockout in vascular endothelial cells on ovarian function in mice and its mechanism,so as to provide a new target and strategy for the diagnosis and treatment of infertility caused by angiogenesis disorder.Methods YY1^(flox/flox)(WT)and Cdh5-CreER^(T2)mice were constructed by CreER^(T2)/LoxP system,and then mated to obtain the YY1 vascular endothelial conditional knockout mice(Cdh5-CreER^(T2);YY1^(flox/flox),referred to as YY1^(i△EC)).The genotypes were identified by PCR.By administering tamoxifen to YY1^(iΔEC)mice to induce Cre enzyme activity,temporal-specific knockout of YY1 in vascular endothelial cells was achieved.After knockout for one month,the mouse pulmonary vascular endothelial cells were isolated,and the knockout efficiency was verified by immunofluorescence and Western blot.Using WT female mice as control,the YY1^(i△EC)female mice after knockout were detected for the fertility,estrous cycle and ovary angiogenesis,and observed for the ovary and uterus.Results After knockout of YY1 for one month,the number of vascular endothelial cells in YY1^(i△EC)mice decreased,and YY1 in vascular endothelial cells was knocked out efficiently and specifically.Compared with WT female mice,the ability of female YY1^(i△EC)mice to conceive and give birth to cubs was greatly reduced;the female YY1^(iΔEC)mice exhibited disrupted estrous cycles,and had spontaneous ovulation and impaired formation of corpus luteum;the ovarian angiogenesis was significantly inhibited(F=1.288,P<0.01);the ovary and uterus were small in size,and the ovary was pale in color and lacked corpus luteum on the surface.Conclusion Conditional YY1knockout in vascular endothelial cells can cause reproductive dysfunction in mice,which may be due to the inhibition of ovarian angiogenesis by vascular endothelial cell YY1 deficiency,resulting in impaired ovulation and luteinization function.

关 键 词：血管内皮细胞 YY1 卵巢功能 血管新生 CreER^(T2)/LoxP系统 

分 类 号：R319[医药卫生—基础医学]