miR101调控EZH2抑制胎盘滋养细胞葡萄糖吸收参与妊娠糖尿病机制研究

miR101 regulates EZH2 to inhibit glucose absorption by placental trophoblasts and its mechanism in gestational diabetes

目的研究miR101通过调节组蛋白甲基转移酶同源序列2增强子(EZH2)蛋白对胎盘滋养细胞葡萄糖吸收的影响,以及miR101在妊娠期糖尿病(GDM)中的可能作用机制。方法选取GDM孕妇和正常孕妇各30例分别作为观察组和对照组,qRT-PCR检测胎盘组织miR101和EZH2 mRNA表达,Western blot检测胎盘组织中EZH2蛋白表达。取人绒毛膜滋养层细胞转染miR101模拟物、miR101模拟物对照序列、miR101抑制剂和miR101抑制剂对照序列分别作为M组、M-NC组、I组和I-NC组,设立空白对照B组,每组设立6个平行复孔。qRT-PCR检测转染细胞miR101、EZH2 mRNA表达量,Western blot检测转染细胞EZH2蛋白表达量,葡萄糖测定试剂盒检测转染细胞对葡萄糖的吸收量。结果观察组miR101表达水平高于对照组(t=5.433,P<0.05),EZH2表达水平低于对照组(t=2.178,P<0.05)。细胞转染后:①M组miR101表达水平较B组和M-NC组上升(t=2.133、2.167,P<0.05),I组转染miR101表达水平较B组和I-NC组下降(t=2.918、2.331,P<0.05);②M组EZH2蛋白表达水平较B组和M-NC组下降(t=2.018、2.257,P<0.05),I组转染EZH2蛋白表达水平较B组和I-NC组升高(t=3.271、3.167,P<0.05);③M组葡萄糖消耗量较B组和M-NC组升高(t=3.106、2.115,P<0.05),I组葡萄糖消耗量较B组和I-NC组降低(t=2.002、2.158,P<0.05)。结论GDM患者胎盘组织中miR101表达异常升高,miR101可调控EZH2表达降低胎盘滋养细胞对葡萄糖吸收量,参与GDM病理机制。

Objective To study the effect of miR101 on glucose absorption in placental trophoblast by regulating EZH2 protein,and the possible mechanism of miRNA101 in gestational diabetes mellitus(GDM).Methods The toatal of 30 pregnant women with GDM and 30 normal pregnant women were selected as the observation group and the control group respectively.The expression of miR101 and EZH2 mRNA in placental tissue was detected by qRT-PCR,and the expression of EZH2 protein in placental tissue was detected by Western blot.Human chorionic trophoblast cells were transfected with mir101 mimic,mir101 mimic control sequence,mir101 inhibitor and mir101 inhibitor control sequence as M group,M-NC group,I group and I-NC group respectively.Blank control group B was set up with 6 parallel multiple pores in each group.qRT-PCR was used to detect miR101 and EZH2 mRNA expression.Western blot was used to detect EZH2 protein expression,and glucose detection kit was used to detect glucose uptake.Results The expression level of miR101 in the observation group was significantly higher than that in the control group(t=5.433,P<0.05),and the expression level of EZH2 in the observation group was lower than that in the control group(t=2.178,P<0.05).After cell transfer:①The expression level of miR101 in group M was significantly higher than that in group B and M-NC(t=2.133,2.167,P<0.05).The expression level of miR101 in group M was significantly lower than that in group B and I-NC(t=2.918,2.331,P<0.05).②The expression of EZH2 in group M was significantly lower than that in group B and M-NC(t=2.018,2.257,P<0.05).The expression level of EZH2 in group M was significantly lower than that in group B and I-NC(t=3.271,3.167,P<0.05).③Glucose consumption in group M was lower than that in group B and M-NC(t=3.106,2.115,P<0.05).Glucose consumption in group M was higher than that in group B and I-NC(t=2.002,2.158,P<0.05).Conclusion The expression of miR101 in patients with GDM placental trophoblast abnormally increase.MiR101 could regulate the expression of EZH2 and decrease the absorption of glucose by placental trophoblasts,which was involved in the pathological mechanism of GDM.