【摘要】：目的:探討mmu-mi R-24在小鼠子宮內(nèi)膜基質(zhì)細(xì)胞和蛻膜細(xì)胞中的作用。方法:收集妊娠第1、4、5、6天(D1、D4、D5、D6)的小鼠子宮;分離小鼠基質(zhì)細(xì)胞,體外激素人工誘導(dǎo)蛻膜化。實(shí)時(shí)定量PCR(real-time quantitative PCR,q RT-PCR)、原位雜交(in situ hybridization,ISH)以及蛋白印跡(Western blot)檢測(cè)mmu-mi R-24及相應(yīng)因子的表達(dá)。采用脂質(zhì)體2000轉(zhuǎn)染mmumi R-24模擬物(mimics)及抑制物(inhibitor)模型上調(diào)或下調(diào)mmu-mi R-24表達(dá)后,檢測(cè)細(xì)胞凋亡與細(xì)胞周期。結(jié)果:mmumi R-24在D1高表達(dá),且表達(dá)量高于D4(P=0.003)。mmu-mi R-24在D4的腔上皮、腺上皮及少量基質(zhì)細(xì)胞中表達(dá),而在妊娠第5天著床點(diǎn)(D5implantation site,D5IS)及妊娠第6天著床點(diǎn)(D6IS)的蛻膜區(qū)表達(dá),且D5IS的表達(dá)量高于第5天著床旁(D5interimplantation site,D5IIS)的表達(dá)量,但無統(tǒng)計(jì)差異(P=0.094)。在基質(zhì)細(xì)胞中,mimics干擾后,S期的細(xì)胞數(shù)降低(P=0.000),增殖因子PCNA表達(dá)降低,總凋亡細(xì)胞數(shù)目增加(P=0.042),凋亡因子BAX表達(dá)增高;通過inhibitor干擾后,G1期細(xì)胞數(shù)降低(P=0.005),S期與G2期細(xì)胞數(shù)有所升高(P=0.075,P=0.054),PCNA表達(dá)增多,晚期凋亡細(xì)胞數(shù)目減少(P=0.009),BAX表達(dá)降低。在蛻膜細(xì)胞中,mimics干擾后,與對(duì)照組相比,G1期細(xì)胞數(shù)升高(P=0.002),S期細(xì)胞數(shù)降低(P=0.000),PCNA表達(dá)降低,晚期凋亡細(xì)胞數(shù)目增加(P=0.025),BAX表達(dá)增多;inhibitor干擾后,與對(duì)照組相比,S期細(xì)胞數(shù)升高(P=0.026),PCNA表達(dá)增多;晚期凋亡細(xì)胞數(shù)目減少(P=0.006),BAX表達(dá)降低。結(jié)論:在胚胎植入前,mmu-mi R-24的低表達(dá)可促進(jìn)基質(zhì)細(xì)胞的增殖;在胚胎植入期,mmu-mi R-24的高表達(dá)可促進(jìn)蛻膜細(xì)胞凋亡,有利于妊娠的維持。
[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.
【作者單位】： 重慶醫(yī)科大學(xué)公共衛(wèi)生與管理學(xué)院生殖生物研究室;重慶醫(yī)科大學(xué)附屬第一醫(yī)院婦產(chǎn)科;
【基金】：國(guó)家自然科學(xué)基金青年基金資助項(xiàng)目(編號(hào):31501207)
【分類號(hào)】：R714

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