论文部分内容阅读
促胰液素(secretin)属于一种胃肠肽,已证实它在早期妊娠小鼠子宫内膜基质细胞中有表达。本文旨在进一步研究促胰液素在胚胎着床过程中的功能。通过real-time PCR、ELISA和原位杂交方法检测促胰液素、促胰液素受体、胞质型磷脂酶A2(cytosolic phospholipase A2,c PLA2)和膜相关的前列腺素E合成酶-1(membrane prostaglandin E synthase 1,m PGEs-1)在小鼠妊娠第4到8天子宫中的表达。体外培养妊娠第4天小鼠子宫基质细胞,转染促胰液素表达质粒,使用PKA抑制剂H89处理,24 h收集细胞。Real-time PCR和Western blot检测c PLA2、m PGEs-1和环磷腺苷效应元件结合蛋白(c AMP responsive elementbinding protein,CREB)的表达,ELISA方法分析细胞上清液中前列腺素E2(prostaglandin E2,PGE2)的水平。结果显示促胰液素、c PLA2和m PGEs-1的m RNA在妊娠第5到7天着床点表达量逐渐增加,表达趋势一致。促胰液素在妊娠第6、7、8天血清中的含量显著高于第5天的含量,并且在第6到7天着床点中的含量明显高于第5天非着床点含量。转染促胰液素表达质粒,促进小鼠子宫基质细胞c PLA2、磷酸化c PLA2(p-c PLA2)和m PGEs-1的表达,但对PGE2没有明显的诱导。H89能明显降低促胰液素对CREB、磷酸化CREB(p-CREB)、c PLA2和p-c PLA2的诱导。以上结果表明在小鼠胚胎着床过程中促胰液素在子宫内膜基质细胞中表达量增加,促进p-c PLA2、c PLA2和m PGEs-1的表达,并通过PKA信号通路调节c PLA2/p-c PLA2的水平。
Secretin, a gastrointestinal peptide, has been shown to be expressed in endometrial stromal cells in early pregnancy in mice. This article aims to further study the role of secretin in embryo implantation. Serum levels of secretin, secretin receptor, cytosolic phospholipase A2 (c PLA2) and membrane-associated prostaglandin E synthase-1 (membrane) were measured by real-time PCR, ELISA and in situ hybridization prostaglandin E synthase 1, m PGEs-1) in the uterus on days 4 to 8 of pregnancy in mice. Mouse uterine stromal cells were cultured in vitro on day 4 of gestation, transfected with secretin expression plasmids, and treated with PKA inhibitor H89 for 24 h. Real-time PCR and Western blot were used to detect the expression of PLA2, m PGEs-1 and cAMP responsive element binding protein (CREB). The levels of prostaglandin E2 , PGE2) levels. The results showed that the expression levels of secretin, c PLA2 and m PGEs-1 m RNA increased gradually at the implantation site from the 5th to 7th day of gestation, and the expression trends were consistent. Serum levels of secretin at 6, 7, and 8 days of gestation were significantly higher than those at day 5, and were significantly higher at day 6 to day 7 than at day 5. The expression of c PLA2, p-c PLA2 and m PGEs-1 in murine uterine stroma cells was enhanced by transfection with secretin, but there was no obvious induction of PGE2. H89 significantly decreased the induction of CREB, CREB, c PLA2 and p-c PLA2 by CRE. The above results indicate that during the process of mouse embryo implantation, secretin is increased in endometrial stromal cells and promote the expression of pc PLA2, c PLA2 and m PGEs-1, and regulate c PLA2 / pc PLA2 through PKA signaling pathway s level.