脂肪酸结合蛋白4对子宫内膜上皮细胞的影响

发布时间：2018-01-14 07:23

本文关键词：脂肪酸结合蛋白4对子宫内膜上皮细胞的影响　出处：《浙江大学》2014年硕士论文　论文类型：学位论文

【摘要】：背景胚胎着床是指胚泡进入子宫腔后与子宫内膜粘附,并逐渐埋入子宫内膜的过程,它是生殖的关键性调控环节,着床失败是早期妊娠丢失的主要原因。胚胎着床受到激素、细胞因子、生长因子、粘附因子及其配基等组成的分子网络的精细调控,但其机制远未阐明。我们前期实验发现人子宫内膜细胞表达脂肪酸结合蛋白4(FABP4,常被称为aP2),利用临床标本,进一步发现分泌期子宫内膜组织表达FABP4较增生期上调,蜕膜组织的FABP4表达较非妊娠期子宫内膜进一步增加；雌、孕激素分别及联合刺激作用子宫内膜细胞,结果显示雌、孕激素分别作用和雌孕激素联合作用,对FABP4的表达均有上调作用。目的明确FABP4是否参与调控子宫内膜细胞的生物学功能,观察FABP4对子宫内膜容受性标志性因子调控作用,应用体外粘附模型验证FABP4是否参与对胚胎粘附和着床的调控；研究和探索FABP4是否参与胚胎着床的过程及其可能的机制。方法 1. Western blot和细胞免疫荧光检测子宫内膜细胞Ishikawa和RL-952细胞FABP4的表达；2、采用siRNA于扰FABP4基因的表达和FABP4特异性抑制剂阻断FABP4功能,利用CCK-8法检测细胞增殖能力,划痕试验检测细胞迁移,Transwell检测细胞侵袭力；3、采用FABP4特异性抑制剂阻断Ishikawa和RL-952细胞FABP4功能,利用实时定量PCR检测子宫内膜容受因子白血病抑制因子(leukemia inhibitory factor,LIF)、整合蛋白beta3(Integrin beta3,Integrin-p3)、紧密连接蛋白4(claudin4)的蛋白水平。结果 (1) Western blot和细胞免疫荧光显示Ishikawa和RL-952细胞表达FABP4;(2).靶向性siRNA干扰Ishikawa和RL-952细胞FABP4的表达和特异性FABP4抑制剂阻断FABP4功能显著抑制子宫内膜细胞增殖、迁移和侵袭力；(3).特异性抑制剂抑制Ishikawa和RL-952细胞FABP4功能显著下调雌、孕激素诱导的子宫内膜容受因子LIF、Integrin-β, claudin4的表达。结论 FABP4表达于人子宫内膜上皮细胞,参与调控子宫内膜上皮细胞的生物学功能,抑制FABP4的表达或其功能可抑制子宫内膜细胞增殖、迁移和侵袭以及子宫内膜容受因子的表达。
[Abstract]:Background Embryo implantation is a process in which the blastocyst adheres to the endometrium and gradually implants into the endometrium. It is the key regulation of reproduction. Implantation failure is the main cause of early pregnancy loss. Embryo implantation is carefully regulated by molecular networks of hormones, cytokines, growth factors, adhesion factors and their ligands. But the mechanism is far from clear. We found that human endometrial cells express fatty acid binding protein 4FABP4, often called aP2P4, using clinical specimens. It was further found that the expression of FABP4 in secretory endometrium was higher than that in proliferative phase, and the expression of FABP4 in decidual tissue was higher than that in non-pregnant endometrium. The effects of estrogen, progesterone and combined stimulation on endometrial cells showed that the expression of FABP4 was up-regulated by estrogen and progesterone combined with estrogen and progesterone. Purpose To determine whether FABP4 is involved in regulating the biological function of endometrial cells and to observe the regulatory effect of FABP4 on endometrial receptivity. The in vitro adhesion model was used to verify whether FABP4 was involved in the regulation of embryo adhesion and implantation. To investigate whether FABP4 is involved in embryo implantation and its possible mechanism. Method 1. Western blot and cellular immunofluorescence were used to detect the expression of FABP4 in Ishikawa and RL-952 cells. 2. SiRNA was used to disturb the expression of FABP4 gene and FABP4 specific inhibitor to block the function of FABP4. The proliferation of cells was detected by CCK-8 assay. Cell migration and cell invasion were detected by scratch test. 3The function of Ishikawa and RL-952 cells was blocked by FABP4 specific inhibitor. The real time quantitative PCR was used to detect the leukemia inhibitory factor (LIF) of endometrial accommodation factor leukemia suppressor. The protein level of integrin beta3(Integrin beta3, integrin-p3, tight junction protein 4, claudin4. 1) Western blot and cellular immunofluorescence showed FABP4 expression in Ishikawa and RL-952 cells. 2). Targeted siRNA interfered with the expression of FABP4 in Ishikawa and RL-952 cells and the specific FABP4 inhibitor blocked the function of FABP4 significantly inhibited the proliferation of endometrial cells. Migration and invasiveness; Inhibition of FABP4 function of Ishikawa and RL-952 cells by specific inhibitors significantly down-regulated estrogen and progesterone-induced endometrial receptor-factor LIF. Expression of Integrin- 尾, claudin4. Conclusion FABP4 is expressed in human endometrial epithelial cells and participates in regulating the biological function of endometrial epithelial cells. Inhibiting the expression or function of FABP4 can inhibit the proliferation of endometrial cells. Migration and invasion and expression of endometrial receptive factors.
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R714.8

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