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

发布时间：2018-10-24 09:53

【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。
[Abstract]:Aim: to investigate the role of mmu-mi R-24 in mouse endometrial stromal cells and decidual cells. Methods: the mouse uterus was collected on the 1st day of pregnancy (D _ 1D _ 4, D _ (5) D _ (6), and mouse stromal cells were isolated and induced decidualization by hormone in vitro. The expression of mmu-mi R-24 and its corresponding factors were detected by real-time quantitative PCR (real-time quantitative PCR,q RT-PCR), in situ hybridization (in situ hybridization,ISH) and Western blot (Western blot). The expression of mmumi R-24 was up-regulated or down-regulated by liposome 2000 transfection into mmumi R-24 mimetic (mimics) and inhibitor (inhibitor). Apoptosis and cell cycle were detected. Results: the expression of mmumi R-24 was higher in D1 and higher than that in D4 (P0. 003). Mmu-mi R-24 was expressed in the luminal epithelium, glandular epithelium and a few stromal cells of D4, but in decidua at the implantation site (D5implantation site,D5IS) on the 5th day of pregnancy and the implantation site (D6IS) on the 6th day of pregnancy. The expression of D5IS was higher than that of D5interimplantation site,D5IIS on the 5th day, but there was no statistical difference (P0.094). In stromal cells, the number of S phase cells decreased (P0. 000), the expression of proliferating factor PCNA decreased, the number of total apoptotic cells increased (P0. 042) and the expression of apoptotic factor BAX increased after mimics interference. After the interference of inhibitor, the number of cells in G1 phase decreased (P0. 005), S phase and G2 phase cells increased (P0. 075 + P0. 054), PCNA expression increased, and the number of late apoptotic cells decreased (P0. 009), BAX expression decreased). In decidual cells, after mimics interference, the number of cells in G1 phase increased (P0. 002), S phase cells decreased (Pn0. 000), PCNA expression decreased, late apoptotic cells increased (P0. 025), BAX expression increased), after inhibitor interference, compared with control group, the number of S phase cells increased (Pn0. 026), PCNA expression increased). The number of late apoptotic cells decreased (P < 0. 006), BAX expression). Conclusion: the low expression of mmu-mi R-24 can promote the proliferation of stromal cells before embryo implantation, and the high expression of mmu-mi R-24 can promote the apoptosis of decidual cells during embryo implantation, which is beneficial to the maintenance of pregnancy.
【作者单位】：重庆医科大学公共卫生与管理学院生殖生物研究室;重庆医科大学附属第一医院妇产科;
【基金】：国家自然科学基金青年基金资助项目(编号:31501207)
【分类号】：R714

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