二甲双胍对肺泡上皮细胞氧化损伤的抑制作用及其机制  

Inhibitory effect of metformin on oxidative damage of alveolar epithelial cells and its mechanism

作　　者：李鹏[1] 欧阳运萍 陈涛 赵博 杨小军 LI Peng;OUYANG Yunping;CHEN Tao;ZHAO Bo;YANG Xiaojun(Department of Emergency,Tangshan Union Hospital,Tangshan 063000,China;不详)

机构地区：[1]唐山职业技术学院附属医院急诊科,河北唐山063000 [2]唐山职业技术学院附属医院消化内科 [3]唐山职业技术学院附属医院重症医学科

出　　处：《山东医药》2023年第32期40-44,共5页Shandong Medical Journal

基　　金：河北省医学科学研究课题项目(20231813)。

摘　　要：目的观察二甲双胍对过氧化氢(H2O2)诱导的人肺泡上皮细胞A549氧化损伤的抑制作用,并基于上皮—间质转化(EMT)及核因子κB(NF-κB)信号通路调控探讨相关机制。方法体外培养A549细胞并分为对照组、模型组、二甲双胍组、BAY 11-7082组、抑制剂组和激活剂组。对照组不给予药物干预,模型组、二甲双胍组、BAY 11-7082组、抑制剂组、激活剂组均给予400µmol/L H2O2刺激24 h构建体外急性肺损伤细胞模型;二甲双胍组分为三个浓度亚组,分别给予2.5、5、10 mmol/L的二甲双胍;BAY 11-7082组加入5 mmol/L的NF-κB信号通路抑制剂BAY 11-7082;根据CCK-8法检测细胞活力结果,选择5 mmol/L为二甲双胍最佳作用浓度,抑制剂组、激活剂组造模后给予5 mmol/L二甲双胍,并分别加入5µmol/L的BAY 11-7082和1µmol/L的NF-κB信号通路激活剂Prostratin。各组均干预24 h。倒置显微镜下观察细胞形态;采用ELISA法检测细胞培养液上清中的丙二醛(MDA)和超氧化物岐化酶(SOD);采用细胞划痕试验测算细胞迁移率;采用Western blotting法检测细胞中的EMT相关蛋白及NF-κB信号通路相关蛋白。结果模型组细胞较对照组生长受到抑制、细胞形态不规则、细胞碎片增多,MDA、细胞迁移率、N-钙黏蛋白、波形蛋白、纤维黏连蛋白及p-NF-κB p65蛋白水平升高,SOD、E-钙黏蛋白水平降低(P均<0.05);二甲双胍组和BAY 11-7082组细胞生长抑制作用较模型组减弱、细胞形态正常、细胞碎片少,MDA、细胞迁移率、N-钙黏蛋白、波形蛋白、纤维黏连蛋白及p-NF-κB p65蛋白水平降低,SOD、E-钙黏蛋白水平升高,且抑制剂组较二甲双胍组上述指标变化更显著(P均<0.05);激活剂组上述指标变化趋势与二甲双胍组相反(P均<0.05)。结论二甲双胍对H2O2诱导的A549氧化损伤具有抑制作用,可能与其阻滞NF-κB信号通路、抑制细胞迁移和EMT有关。Objective To observe the inhibitory effect of metformin on hydrogen peroxide(H2O2)-induced oxidative damage in human alveolar epithelial cells A549,and to explore the related mechanism based on epithelial-mesenchymal transition(EMT)and nuclear factor-κb(NF-κB)signaling pathway regulation.Methods A549 cells were cultured in vitro and divided into the control group,model group,metformin group,BAY 11-7082 group,inhibitor group,and activator group.Cells in the control group were not treated,while cells in the model group,metformin group,BAY 11-7082 group,inhibitor group and activator group were stimulated with 400μmol/L H2O2 for 24 h to construct the in vitro models of acute lung injury.Cells in the metformin group were divided into three concentration subgroups,which were given 2.5,5 and 10 mmol/L metformin,respectively.Cells in the BAY 11-7082 group were treated with 5 mmol/L BAY 11-7082.According to the results of cell viability detected by CCK-8,5 mmol/L was selected as the optimal concentration of metformin.After modeling,cells in the inhibitor group and activator group were given 5 mmol/L metformin,and then BAY 11-7082(5μmol/L)and Prostratin(1μmol/L)were added,respectively.All groups were treated for 24 h.Cell morphology was observed under inverted microscope.Malondialdehyde(MDA)and superoxide dismutase(SOD)in the supernatant of cell culture medium were detected by enzyme-linked immunosorbent assay.The cell migration rate was measured by cell Scratch test.Western blotting was used to detect EMT-related proteins and NF-κB signaling pathway-related proteins in cells.Results Compared with the control group,cell growth was inhibited,cell morphology was irregular,cell debris increased,MDA,cell mobility,N-cadherin,vimentin,fibronectin and p-NF-κB p65 protein levels increased,and SOD and E-cadherin levels decreased in the model group(all P<0.05).Compared with the model group,the growth inhibition effect of the metformin group and BAY 11-7082 group was weaker,the cell morphology was normal,the cell debris was le

关 键 词：二甲双胍 急性肺损伤 氧化应激 肺泡上皮细胞 核因子ΚB 细胞迁移 上皮—间质转化 

分 类 号：R961[医药卫生—药理学]