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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 investigate the role of mmu-mi R-24 in mouse endometrial stromal cells and decidual cells. Methods: The uteri of mice were collected on the 1st, 4th, 5th and 6th day of gestation (D1, D4, D5, D6). Mouse stromal cells were isolated and the hormone in vitro was induced to artificially decidua. The expression of Rg-24 and related factors of mmu-mi was detected by real-time quantitative PCR (q RT-PCR), in situ hybridization (ISH) and Western blot. Apoptosis and cell cycle were detected by up-regulation or down-regulation of mmu-mi R-24 expression using liposome 2000 transfection in mmumi R-24 mimics and inhibitor models. Results: The expression of mmumi R-24 in D1 was higher than that in D4 (P = 0.003). mmu-mi R-24 was expressed in the luminal epithelium, glandular epithelium and a small amount of stromal cells in D4, whereas in decidua at D5 implantation site (D5IS) and implantation site (D6IS) on day 6 of gestation The expression of D5IS was higher than that of D5IIS at day 5, but there was no statistical difference (P = 0.094). In the stromal cells, the number of cells in S phase decreased (P = 0.000), the expression of proliferation factor PCNA decreased, the total number of apoptotic cells increased (P = 0.042) and the apoptosis factor BAX increased after interference with mimics; , The number of cells in G1 phase decreased (P = 0.005), the number of S phase and G2 phase cells increased (P = 0.075, P = 0.054), the expression of PCNA increased, the number of apoptotic cells decreased Decreased expression. In decidual cells, the number of cells in G1 phase increased (P = 0.002), the number of S phase cells decreased (P = 0.000), the expression of PCNA decreased and the number of apoptotic cells increased in late stage (P = 0.025), and BAX expression increased. Compared with the control group, the number of S phase cells increased (P = 0.026), the expression of PCNA increased, the number of apoptotic cells decreased (P = 0.006) . Conclusion: The low expression of mmu-mi R-24 can promote the proliferation of stromal cells before embryo implantation. During embryo implantation, the high expression of mmu-mi R-24 can promote the apoptosis of decidual cells, which is benefit to pregnancy The maintenance.