摘要
目的研究miRNA-148b-3p在妊娠期肝内胆汁淤积症(intrahepatic cholestasis of pregnancy,ICP)孕妇胎盘组织中的表达及对滋养细胞葡萄糖转运蛋白-1(glucose transporter 1,GLUT1)的调控,探索ICP孕妇子代糖代谢异常的可能机制。方法收集2017年3月至2018年1月于四川大学华西第二医院产科行剖宫分娩的ICP孕妇30例,正常妊娠孕妇30例,分别收集胎盘组织、母血及脐血,提取胎盘组织miRNA,母血及脐血送检生化指标。采用实时荧光定量PCR验证miR-148b-3p在两组胎盘中的表达差异。通过lipofectaine3000脂质体包裹miR-148b-3p inhibitor/mimics转染人滋养细胞HTR-8,实时荧光定量PCR验证转染后细胞中miR-148b-3p的表达水平;Western blot检测转染后细胞中GLUT1的表达水平。结果 ICP孕妇母血空腹血糖(fasting blood glucose,FPG)及其胎儿脐血胰岛素水平较对照组增高(P<0.05)。ICP组胎盘组织中miR-148b-3p表达量较对照组下降(P<0.05)。转染miR-148b-3p inhibitor后,与转染inhibitor control比较,HTR-8细胞中miR-148b-3p的表达水平下降(P<0.05),GLUT1蛋白的表达量升高(P<0.05);转染miR-148b-3p mimics后,与转染mimics control比较,HTR-8细胞中miR-148b-3p的表达水平上升(P<0.05),GLUT1蛋白的表达量降低(P<0.05)。结论 miR-148b-3p可能通过对胎盘滋养细胞GLUT1表达的负向调控参与了ICP子代糖代谢异常的发生。
Objective To investigate the expression of miRNA-148b-3p and its target gene in the placenta between normal pregnant women and pregnant women with intrahepatic cholestasis of pregnancy(ICP) and to explore the possible mechanism of glucose metabolism of offspring with maternal cholestasis. Methods There were 30 cases of normal pregnant women and 30 cases of pregnant women with ICP recruited in the study, all of whom underwent cesarean delivery from Mar. 2017 to Jan. 2018. Placenta tissues, maternal blood and cord blood were collected in each case. Maternal blood and cord blood were sent for biochemical detection. miRNA of placenta tissues was extracted and qRT-PCR was used to measure the expression of miR-148b-3p in the placenta. Normal HTR-8 cells were transfected with miR-148b-3p inhibitor/mimics wrapped with lipofectaine3000. qRT-PCR was used to measure the expression of miR-148b-3p, and Western blot was used to measure the expression of glucose transporter 1(GLUT1) after transfection. Results Maternal fasting blood glucose(FPG) and its fetal cord blood insulin levels in the ICP group were significantly higher than those of control. The expression of miR-148b-3p in the placenta of ICP group was lower than that of control group(P<0.05). Compared with inhibitor control group, the expression of miR-148b-3p was decreased in HTR-8 cells transfected with miR-148b-3p inhibitor(P<0.05), while the expression of GLUT1 was increased(P<0.05). Compared with mimics control group, the expression of miR-148b-3p was increased in HTR-8 cells transfected with miR-148b-3p mimics(P<0.05), while the expression of GLUT1 was decreased(P<0.05). Conclusion miR-148b-3p might participate in glucose metabolism of offspring with maternal cholestasis through the negative regulation of GLUT1 expression in placental trophoblast cells.
引文
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