摘要
目的探究三卤甲烷(THMs)通过表观遗传调控增加妊娠期糖尿病孕鼠早产风险的动物实验研究。方法将30只SPF级60 d龄Wistar健康成年未产雌性大鼠与雄性大鼠交配,确定妊娠后,将孕鼠按照随机数字表法分为以下3组,每组各10只。对照组于妊娠第0天通过腹腔注射枸橼酸盐缓冲液+干净自来水,GDM组于妊娠第0天通过腹腔注射链脲霉素(STZ)溶液+干净自来水,GDM+THMs组于妊娠第0天通过腹腔注射STZ溶液+孕后6~10 d THMs水灌胃暴露。采用On-Call Plus血糖试纸计算空腹血糖;采用^(14 )C-2-dexoyglucose计算葡萄糖摄取;采用^(14 )C-葡萄糖计算葡萄糖氧化;通过实时PCR计算IRβ和GLUT2的mRNA相对定量;通过亚硫酸氢盐DNA测序检测CDKN2A和CDKN2B启动子中CpG的甲基化水平;通过蛋白质印迹分析CDKN2A和CDKN2B蛋白质水平;统计不同处理组GDM孕鼠妊娠不良结局。结果与对照组相比,GDM组空腹血糖水平升高,空腹血清胰岛素、^(14 )C-2-脱氧葡萄糖摄取和^(14 )C-葡萄糖氧化水平降低,差异均有统计学意义(P<0.05);与GDM组相比,GDM+THMs组空腹血糖水平升高,空腹血清胰岛素、^(14 )C-2-脱氧葡萄糖摄取和^(14 )C-葡萄糖氧化水平降低,差异均有统计学意义(P<0.05)。与对照组相比,GDM组GLUT2和IRβmRNA水平降低,与GDM组相比,GDM+THMs组GLUT2和IRβmRNA水平降低,差异均有统计学意义(P<0.05)。与对照组相比,GDM组CDKN2A和CDKN2B启动子中CpG的甲基化水平降低,与GDM组相比,GDM+THMs组CDKN2A和CDKN2B启动子中CpG的甲基化水平降低,差异均有统计学意义(P<0.05)。与对照组相比,GDM组CDKN2A和CDKN2B蛋白质水平升高,与GDM组相比,GDM+THMs组CDKN2A和CDKN2B蛋白质水平升高,差异均有统计学意义(P<0.05)。与对照组相比,GDM组早产和低出生体重比例升高,与GDM组相比,GDM+THMs组早产和低出生体重比例升高,差异均有统计学意义(P<0.05)。结论宫内暴露于THMs通过降低CDKN2A和CDKN2B低甲基化,增加C
Objective To explore the animal experimental study that trihalomethanes increase the risk of preterm birth in pregnant mice with gestational diabetes mellitus through epigenetic regulation.Methods A total of 30 SPF 60-day-old Wistar healthy adult sterile female rats were mated with male rats.After confirming pregnancy,the pregnant rats were randomly divided into the following three groups:pregnant rats were randomly divided into three groups,each group 10 rats.The control group received intraperitoneal injection of citrate buffer+clean tap water on the first day of pregnancy,GDM group received intraperitoneal injection on the 0th day of pregnancy injection of streptozotocin(STZ)solution+clean tap water,and GDM+THMs group received intraperitoneal injection of STZ solution+THMs water gavage exposure on day 0 of pregnancy.Calculate fasting blood glucose with On-Call Plus blood glucose test strips;glucose uptake was calculated using ^(14 )C-2-dexoyglucose;glucose oxidation was calculated using ^(14 )C-glucose;relative mRNA quantification of IRβand GLUT2 was calculated by real-time PCR;the methylation levels of CpG in CDKN2A and CDKN2B promoters were detected by bisulfite DNA sequencing;CDKN2A and CDKN2B protein levels were analyzed by Western blotting;the adverse pregnancy outcomes of GDM pregnant mice in different treatment groups were counted.Results Compared with the control group,the fasting blood glucose level was increased,and the fasting serum insulin,^(14 )C-2-deoxyglucose uptake and ^(14 )C-glucose oxidation levels were decreased in the GDM group,and compared with the GDM group,the fasting blood glucose level was increased,the fasting serum insulin,^(14 )C-2-deoxyglucose uptake and ^(14 )C-glucose oxidation levels in the GDM+THMs group were decreased,the differences were statistically significant(P<0.05).Compared with the control group,the GLUT2 and IRβmRNA levels were decreased in the GDM group,and compared with the GDM group,the GLUT2 and IRβmRNA levels in the GDM+THMs group were decreased,the differenc
作者
范可辽
赵璟文
曾军安
李清红
石晓琳
刘振国
FAN Ke-liao;ZHAO Jing-wen;ZENG Jun-an(Department of Pharmacy,Northwest Women's and Children's Hospital,Xi'an Shaanxi 710003,China)
出处
《临床和实验医学杂志》
2022年第23期2472-2476,共5页
Journal of Clinical and Experimental Medicine
基金
陕西省自然科学基础研究计划项目(编号:2022JM-585)。
关键词
大鼠
三卤甲烷
表观遗传
甲基化
妊娠期糖尿病
早产
Rats
Trihalomethanes
Epigenetics
Methylation
Gestational diabetes mellitus
Preterm birth
