背景:异常的胆汁酸代谢导致妊娠期间胎盘功能的变化。确定内质网蛋白29(ERp29)是否可以通过改变滋养细胞凋亡水平来介导胆汁淤积的妊娠效应。
方法:采用ELISA法检测66例妊娠期肝内胆汁淤积症(ICP)孕妇和74例健康体检者血清中ERp29的含量。皮下注射乙炔雌二醇(E2)在妊娠大鼠中诱导ICP。牛磺胆酸(TCA)用于模拟ICP环境,加入TGF-β1诱导上皮间质转化(EMT)过程。划痕,迁移,和侵袭试验用于检测EMT过程。构建ERp29过表达/敲低载体并转染以验证ERp29在EMT进程中的感化。通过RNA-seq获得下游基因。
结果:与健康孕妇相比,ERp29在ICP妊娠妇女血清中的表达水平明显升高(P<0.001)。ERp29在ICP孕鼠胎盘组织中显著升高,细胞凋亡水平增加。ICP胎盘组织中E-cadherin高表达,N-cadherin低表达,蜗牛1,波形蛋白。TCA诱导HTR-8/SVneo细胞后,EMT被抑制,而ERp29增加。细胞和动物实验表明,ERp29的敲除降低了EMT的抑制作用,ICP进展有所缓解。FOS的过表达挽救了ERp29对细胞EMT的抑制作用。
结论:胎盘滋养细胞中高水平的ERp29降低了FOSmRNA水平,抑制了EMT过程,加重了ICP的发生和发展。
Abnormal bile acid metabolism leading to changes in placental function during pregnancy. To determine whether endoplasmic reticulum protein 29 (ERp29) can mediate the pregnancy effects of cholestasis by altering the level of trophoblast cell apoptosis.
ERp29 in serum of 66 intrahepatic cholestasis of pregnancy (ICP) pregnant women and 74 healthy were detected by ELISA. Subcutaneous injection of ethinyl estradiol (E2) was used to induce ICP in pregnant rats. Taurocholic acid (TCA) was used to simulate the ICP environment, and TGF-β1 was added to induce the epithelial mesenchymal transformation (EMT) process. The scratch, migration, and invasion test were used to detect the EMT process. ERp29 overexpression/knockdown vector were constructed and transfected to verify the role of ERp29 in the EMT process. Downstream gene was obtained through RNA-seq.
Compared with the healthy pregnant women, the expression levels of ERp29 in serum of ICP pregnancy women were significantly increased (P < 0.001). ERp29 in the placenta tissue of the ICP pregnant rats increased significantly, and the level of apoptosis increased. The placental tissues of the ICP had high expression of E-cadherin and low expression of N-cadherin, snail1, vimentin. After HTR-8/SVneo cells were induced by TCA, EMT was inhibited, while the ERp29 increased. Cell and animal experiments showed that, knockdown of ERp29 reduced the inhibition of EMT, the ICP progress was alleviated. Overexpression of FOS salvaged the inhibitory effects of ERp29 on cell EMT.
The high level of ERp29 in placental trophoblast cells reduced FOS mRNA levels, inhibited the EMT process and aggravated the occurrence and development of ICP.