摘要
目的:探讨左归丸含药血清对妊娠期糖尿病(GDM)模型胎盘滋养细胞物质转运蛋白及mTOR通路的作用。方法:利用SD大鼠灌胃左归丸,制备左归丸含药血清,利用Bewo细胞系,使用高糖(15mmol/L葡萄糖)环境制备GDM模型滋养细胞,设空白对照组、GDM模型组、左归丸含药血清组、胰岛素治疗组,分别处理24h后提取mRNA,qPCR法检测胎盘物质转运蛋白的mRNA表达量。Western blot法检测细胞物质转运蛋白及mTOR通路蛋白表达量,细胞免疫荧光法检测物质转运蛋白在细胞中的定位和表达。结果:GDM模型组的GLUT1、LAT1、SNAT2 mRNA表达量增高,较对照组差异有统计学意义(P<0.05),左归丸含药血清组和胰岛素治疗组GLUT1、LAT1、SNAT2、LAT4 mRNA表达量较GMD模型组下降,差异有统计学意义(P<0.05)。左归丸含药血清组GLUT1、SNAT2、LAT4 mRNA表达量较对照组显著下降,较胰岛素组下降,差异均有统计学意义(P<0.05)。GDM模型组LAT1、SNAT2、LAT4蛋白表达量增高,较对照组差异有统计学意义(P<0.05),左归丸含药血清组GLUT1、LAT1、LAT4表达量较GDM模型组下降,差异有统计学意义(P<0.05)。胰岛素治疗组LAT1蛋白表达量较GDM模型组下降,差异有统计学意义(P<0.05)。GDM模型组P70S6K、AKT、4EBP1蛋白磷酸化水平较对照组升高,差异有统计学意义(P<0.05),左归丸含药血清组P70S6K、AKT、4EBP1蛋白磷酸化水平较GDM模型组降低,差异有统计学意义(P<0.05)。左归丸含药血清组较胰岛素治疗组,AKT蛋白磷酸化水平下降,差异有统计学意义(P<0.05)。结论:左归丸含药血清减少GDM模型胎盘滋养细胞的物质转运蛋白表达量,其作用机理可能通过下调胎盘滋养细胞mTOR通路蛋白的表达有关。
Objective:To investigate the effect of Zuogui Wan containing serum on nutrient transporter proteins and the mTOR signaling pathway in gestational diabetes mellitus(GDM)model of placental trophoblast cells.Methods:SD rats were orally administered Zuogui Wan to prepare the Zuogui Wan containing serum.Bewo cell line was cultured under high-glucose conditions(15mmol/L glucose)to establish an in-vitro GDM model.Cells were divided into four groups:blank control group,GDM model group,Zuogui Wan containing serum group,and insulin treatment group.After 24 hours of treatment,mRNA was extracted.Quantitative real-time PCR(qPCR)was used to detect the mRNA expression levels of placental nutrient transporters.Western blot was used to measure the expression levels of cellular nutrient transporter proteins and mTOR pathway proteins.Cellular immunofluorescence was employed to detect the localization and expression of nutrient transporter proteins within the cells.Results:In the GDM model group,the mRNA expression levels of GLUT1,LAT1,and SNAT2 were increased,showing statistically significant differences compared to the control group(P<0.05).In the Zuogui Wan medicated serum group and the insulin treatment group,the mRNA expression levels of GLUT1,LAT1,SNAT2,and LAT4 decreased compared to the GDM model group,with statistically significant differences(P<0.05).The mRNA expression levels of GLUT1,SNAT2,and LAT4 in the Zuogui Wan group were significantly lower than those in the control group and also lower than those in the insulin group,with statistically significant differences(P<0.05).In the GDM model group,the protein expression levels of LAT1,SNAT2,and LAT4 were increased,showing significant differences compared to the control group(P<0.05).In the Zuogui Wan medicated serum group,the protein expression levels of GLUT1,LAT1,and LAT4 decreased compared to the GDM model group,with statistically significant differences(P<0.05).In the insulin treatment group,the protein expression level of LAT1 decreased compared to the GDM model group,showing a statistically significant difference(P<0.05).In the GDM model group,the phosphorylation levels of P70S6K,AKT,and 4EBP1 proteins were elevated compared to the control group,with significant differences(P<0.05).In the Zuogui Wan medicated serum group,the phosphorylation levels of P70S6K,AKT,and 4EBP1 proteins decreased compared to the GDM model group,with statistically significant differences(P<0.05).Compared to the insulin treatment group,the phosphorylation level of AKT protein in the Zuogui Wan medicated serum group decreased,showing a statistically significant difference(P<0.05).Conclusion:In vitro,ZGW-containing serum down-regulates the expression of nutrient transporters in GDM-model trophoblast cells,an effect likely mediated by suppression of the mTOR signaling pathway.
作者
刘梦婷
张青
马燕
王永
杨娜娜
宋海波
于迎春
Liu Mengting;Zhang Qing;Ma Yan(Zibo Maternal and Child Health Hospital,Zibo 255000;Department of Gynecology,Zhejiang Hospital,Hangzhou 310030;The Second Affiliated Hospital of Zhengzhou University,Zhenzhou 450003)
出处
《现代妇产科进展》
2025年第12期923-930,共8页
Progress in Obstetrics and Gynecology
基金
山东省中医药科技项目青年项目(No:2021Q051)
山东省自然科学基金青年基金(No:ZR2022QH376)
山东省医药卫生科技项目(No:202105020217)
浙江省自然科学基金青年基金(No:LQ24H270017)。
关键词
妊娠期糖尿病
左归丸
胎盘物质转运蛋白
MTOR信号通路
Gestational diabetes mellitus
Zuogui Wan
Placental nutrient transporters
mTOR signaling pathway
