Mmu-miR-24对小鼠着床期子宫内膜基质细胞和蜕膜细胞的调节作用  

Effect of Mmu-miR-24 on endometrial stromal and decidual cells during early pregnancy in mice

作　　者：杨洋[1] 吴梦云[1] 李荣[2] 王应雄[1] 刘学庆[1] 

机构地区：[1]重庆医科大学公共卫生与管理学院生殖生物研究室,重庆400016 [2]重庆医科大学附属第一医院妇产科,重庆400016

出　　处：《重庆医科大学学报》2017年第8期959-964,共6页Journal of Chongqing Medical University

基　　金：国家自然科学基金青年基金资助项目(编号:31501207)

摘　　要：目的:探讨mmu-mi R-24在小鼠子宫内膜基质细胞和蜕膜细胞中的作用。方法:收集妊娠第1、4、5、6天(D1、D4、D5、D6)的小鼠子宫;分离小鼠基质细胞,体外激素人工诱导蜕膜化。实时定量PCR(real-time quantitative PCR,q RT-PCR)、原位杂交(in situ hybridization,ISH)以及蛋白印迹(Western blot)检测mmu-mi R-24及相应因子的表达。采用脂质体2000转染mmumi R-24模拟物(mimics)及抑制物(inhibitor)模型上调或下调mmu-mi R-24表达后,检测细胞凋亡与细胞周期。结果:mmumi R-24在D1高表达,且表达量高于D4(P=0.003)。mmu-mi R-24在D4的腔上皮、腺上皮及少量基质细胞中表达,而在妊娠第5天着床点(D5implantation site,D5IS)及妊娠第6天着床点(D6IS)的蜕膜区表达,且D5IS的表达量高于第5天着床旁(D5interimplantation site,D5IIS)的表达量,但无统计差异(P=0.094)。在基质细胞中,mimics干扰后,S期的细胞数降低(P=0.000),增殖因子PCNA表达降低,总凋亡细胞数目增加(P=0.042),凋亡因子BAX表达增高;通过inhibitor干扰后,G1期细胞数降低(P=0.005),S期与G2期细胞数有所升高(P=0.075,P=0.054),PCNA表达增多,晚期凋亡细胞数目减少(P=0.009),BAX表达降低。在蜕膜细胞中,mimics干扰后,与对照组相比,G1期细胞数升高(P=0.002),S期细胞数降低(P=0.000),PCNA表达降低,晚期凋亡细胞数目增加(P=0.025),BAX表达增多;inhibitor干扰后,与对照组相比,S期细胞数升高(P=0.026),PCNA表达增多;晚期凋亡细胞数目减少(P=0.006),BAX表达降低。结论:在胚胎植入前,mmu-mi R-24的低表达可促进基质细胞的增殖;在胚胎植入期,mmu-mi R-24的高表达可促进蜕膜细胞凋亡,有利于妊娠的维持。Objective：To explore the effect of micro RNA-24（mmu-miR-24）in endometrial stromal cells during early pregnancy in mice. Methods：Uterus tissues were collected on pregnancy of day 1,4,5,6（D1,D4,D5,D6）. The expressions of mmu-miR-24 and other factors were detected by real-time quantitative PCR（q RT-PCR）,in situ hybridization,and Western blot. The primary stromal cells were induced by estradiol（E2）and progesterone（P4）for decidulization. Flow cytometry was implemented to detect cell cycle and cell apoptosis. Results：The expression of mmu-miR-24 was detected in uterine luminal,glandular epithelium,and stromal cell on Day 4. The expression level of mmu-miR-24 on D5 IS was higher than the level on D5 IIS. On D5 IS and D6 IS,mmu-miR-24 was located in decidual zone. When mmu-miR-24 mimics being incorporated into stromal cells,the number of stromal cells in the S phase was decreased,the expression of PCNA was decreased,the total apoptotic cell was increased,and the expression of BAX was increased. In contrast,when mmu-miR-24 inhibitor being incorporated into stromal cells,the number of stromal cells in G1 phase and S phase were decreased and increased respectively,and PCNA expression level was increased,the number of non-viable apoptotic cell was decreased,and BAX level was decreased in stormal cells. On the other hand,when incorporating mmu-miR-24 mimics into the decidual cells,the number of decidual cells in the S phase was decreased,and PCNA expression level was decreased,the number of non-viable apoptotic cell was increased,and BAX level was increased in decidual cells. Lastly,when treating decidual cells with mmu-miR-24 inhibitor,the number of decidual cells in the S phase was increased,and PCNA expression level was increased,the number of non-viable apoptotic cell was decreased,and BAX level was decreased in decidual cells. Conclusion：The low expression of mmu-miR-24 in stromal cells may promote the progress of cell proliferation. The high expression of mmu-miR-24 in decidual cells may prom

关 键 词：mmu-miR-24 胚胎植入 蜕膜化 细胞凋亡 细胞增殖 

分 类 号：R714.13[医药卫生—妇产科学]