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目的:探索葡萄糖对滋养层细胞系HTR8/SVneo细胞Akt2基因甲基化和mRNA水平的影响及早孕期外周血Akt2 mRNA水平与孕前体质指数(BMI)和妊娠期糖尿病(GDM)的关系。方法:(1)构建GDM孕妇子宫内高血糖环境细胞模型,按培养液的葡萄糖浓度分为2.5、5.0、10.0、25.0、40.0 mmol/L 5组;使用实时荧光定量PCR技术检测Akt2 mRNA水平,质谱检测Akt2基因启动子区域的甲基化水平。(2)收集2014年12月至2016年7月在北京大学第一医院分娩的60例孕妇的早孕期外周血,根据其孕前BMI及有无GDM分为4组:超重非GDM组、超重GDM组、肥胖非GDM组和肥胖GDM组。实时荧光定量PCR技术检测4组孕妇早孕期外周血中Akt2 mRNA水平。结果:(1)随培养液中葡萄糖浓度的升高,Akt2 mRNA水平显著升高,呈浓度依赖性(n P均<0.05)。与25.0 mmol/L组相比,5.0 mmol/L组Akt2基因启动子区域的甲基化位点胞嘧啶-磷酸-鸟嘌呤二核苷酸(CpG)10、CpG23、CpG24.5,2.5 mmol/L组Akt2基因启动子区域CpG23、CpG24.5位点,10.0 mmol/L组Akt2基因启动子区域CpG10位点,均出现显著的甲基化水平的改变(n P均<0.05);与5.0 mmol/L组相比,40.0 mmol/L组Akt2基因启动子区域CpG10位点出现显著的甲基化水平的改变(n P<0.05)。(2)与超重非GDM组[1.04(0.90~1.26)]比较,超重GDM组[2.10(1.85~2.28)]和肥胖GDM组[1.68(0.82~2.43)]Akt2 mRNA水平均升高,差异均有统计学意义(n P均0.05)。n 结论:葡萄糖可能通过改变Akt2基因启动子区域的甲基化状态,从而影响Akt2 mRNA水平,且这种变化可能在GDM孕妇的早孕期已经出现,特别是在孕前BMI较低的孕妇中。“,”Objective:To explore the effect of glucose on Akt2 gene methylation and mRNA level in the trophoblast cell line HTR8/SVneo and the association between first-trimester Akt2 mRNA level in peripheral blood and pre-pregnancy body mass index (BMI) and gestational diabetes mellitus (GDM).Methods:The cell model of intrauterine hyperglycemia in pregnant women with GDM was established, and divided into 5 groups based on the different glucose concentrations (2.5, 5.0, 10.0, 25.0, and 40.0 mmol/L). The mRNA levels of Akt2 gene were detected by real-time quantitative PCR, and the methylation levels of Akt2 gene promoter region were detected by mass spectrometry. According to pre-pregnancy BMI and diagnosis of GDM, 60 pregnant women who had given birth at Peking University First Hospital during December 2014 to July 2016 were classified into overweight non-GDM group, overweight GDM group, obese non-GDM group and obese GDM group. Real-time quantitative PCR analyses were used to detect the levels of Akt2 mRNA in first-trimester peripheral blood of the 4 groups.Results:(1) Level of Akt2 mRNA significantly rose with the glucose concentration of medium increased, showing concentration dependency (all n P<0.05). Compared with the 25.0 mmol/L group, the following methylation levels changed significantly (alln P<0.05): cytosine-phosphate-guanine (CpG) 10, CpG23, and CpG24.5 of Akt2 gene promoter region in the 5.0 mmol/L group, CpG23 and CpG24.5 in the 2.5 mmol/L group, and CpG10 in the 10.0 mmol/L group. Compared with the 5.0 mmol/L group, CpG10 in the 40.0 mmol/L group showed methylation change (n P<0.05). (2) Compared with the overweight non-GDM group [1.04(0.90~1.26)], overweight GDM group [2.10(1.85~2.28)] and obese GDM group [1.68(0.82~2.43)] all had higher Akt2 mRNA levels and the differences were statistically significant (alln P0.05).n Conclusions:Glucose might affect Akt2 mRNA level by changing the methylation of Akt2 gene promoter region, and the change might appear in the first trimester of pregnant women with GDM, especially for women with lower pre-pregnancy BMI.