基于NF-κB/Twist 1通路二甲双胍对GDM大鼠子代胎鼠Ⅱ型肺泡上皮细胞EMT的作用机制  

The effect of metformin on type Ⅱ alveolar epithelial cell EMT in gestational diabetic mellitus rats based on NF-κB/Twist 1 pathway

作　　者：段艳芳 刘风伟 秦业强 赵金梅 何文慧 姚晓玲 DUAN Yanfang;LIU Fengwei;QIN Yeqiang;ZHAO Jinmei;HE Wenhui;YAO Xiaoling(Department of Obstetrics,Cangzhou People's Hospital,Cangzhou 061000,Hebei,China)

机构地区：[1]沧州市人民医院产科,河北沧州061000

出　　处：《西部医学》2022年第10期1432-1436,1441,共6页Medical Journal of West China

基　　金：河北省2020年度医学科学研究课题(20200291)。

摘　　要：目的探讨二甲双胍(Met)对妊娠糖尿病(GDM)大鼠子代胎鼠Ⅱ型肺泡上皮细胞上皮间质转化(EMT)的作用及其机制。方法将SD孕鼠随机分为正常组(Normal组)和GDM组。GDM组孕鼠采用高脂饮食联合小剂量链脲佐菌素(STZ)建立GDM模型,测定空腹血糖水平(FBG)。无菌条件下取胎鼠肺组织,分离肺泡上皮细胞,不同浓度Met处理细胞48 h,MTT法检测细胞存活率。将肺泡上皮细胞随机分为对照组(Control组)、GDM组和Met组,Met组细胞用80μmol/L Met处理48 h,Control组和GDM组细胞不做处理。Transwell实验检测细胞侵袭能力,qRT-PCR检测细胞中上皮钙粘蛋白(E-cad)、神经钙粘蛋白(N-cad)和波形蛋白(Vimentin)mRNA相对表达量,蛋白印迹法检测磷酸化核因子-κB(NF-κB)和Twist1蛋白相对表达量。结果GDM组孕鼠的FBG水平明显高于Normal组(P<0.05);细胞存活率随Met浓度的增加而降低,且具有剂量依赖性(P<0.05);与Control组比较,GDM组侵袭细胞数、N-cad、Vimentin mRNA相对表达量以及p-NF-κB p65和Twist1蛋白相对表达量升高,E-cad mRNA相对表达量降低(P<0.05);与GDM组比较,Met组侵袭细胞数、N-cad、Vimentin mRNA相对表达量及p-NF-κB p65和Twist1蛋白相对表达量降低,E-cad mRNA相对表达量升高(P<0.05)。结论Met可抑制GDM大鼠子代胎鼠Ⅱ型肺泡上皮细胞EMT过程,其可能是通过抑制NF-κB/Twist1信号通路发挥作用。Objective To investigate the effect of metformin(Met)on the epithelial-interstitial transformation(EMT)of type Ⅱ alveolar epithelial cells in gestational diabetes mellitus(GDM)rats and its mechanism.Methods SD pregnant rats were randomly divided into Normal group and GDM group.Pregnant rats in GDM group were given high-fat diet combined with low dose streptozotocin(STZ)to establish GDM model,and fasting blood glucose level(FBG)was measured.Lung tissues of fetal rats were collected under aseptic conditions,and alveolar epithelial cells were isolated.Cells were treated with different concentrations of Met for 48 h,and cell survival rate was detected by MTT method.Alveolar epithelial cells were randomly divided into Control group,GDM group and Met group.Met group was treated with 80μmol/L Met for 48 h,and Control group and GDM group were not treated.Transwell assay were used to detect cell invasion.qRT-PCR was used to detect mRNA relative expression levels of epithelial cadherin(E-cad),neurocadherin(N-cad)and Vimentin.The relative expression levels of phosphorylated nuclear factor-κB(NF-κB)and Twist1 protein were detected by Western blot.Results The FBG level in GDM group was significantly higher than that in Normal group(P<0.05).The cell survival rate decreased with the increase of Met concentration in a dose-dependent manner(P<0.05).Compared with the Control group,the number of invasive cells,the mRNA relative expression levels of N-cad and Vimentin,the protein relative expression levels of p-NF-κB p65 and Twist1 in GDM group were increased,while the mRNA relative expression levels of E-cad were decreased(P<0.05).Compared with GDM group,invasive cell number,mRNA relative expression levels of N-cad and Vimentin,protein relative expression levels of p-NF-κB p65 and Twist1 in Met group were decreased,while the mRNA relative expression levels of E-cad were increased(P<0.05).Conclusion Met can inhibit the EMT process of type Ⅱ alveolar epithelial cells in GDM rats,possibly through inhibition of NF-κB/Twist1 s

关 键 词：妊娠期糖尿病 肺泡上皮细胞 上皮间质转化 二甲双胍 NF-κB/Twist1信号通路 

分 类 号：R714.256[医药卫生—妇产科学]