摘要
目的 探讨脂氧素A4(lipoxin A4, LXA4)对Ⅱ型肺泡上皮细胞(alveolar epithelial typeⅡcell, ATⅡ)间质转化(epithelial-mesenchymal transition, EMT)的影响及可能机制。方法 将A549细胞分为对照组、转化生长因子β1(transforming growth factor, TGF-β1)组、空质粒+TGF-β1组、pcDNA3.1-Nrf2+TGF-β1组及LXA4+TGF-β1组。TGF-β1组以5 ng/mL TGF-β1干预A549细胞48 h;空质粒+TGF-β1组和pcDNA3.1-Nrf2+TGF-β1组A549细胞先分别转染空质粒或Nrf2过表达质粒,再以5 ng/mL TGF-β1干预48 h;LXA4+TGF-β1组A549细胞先用100 nmol/L LXA4预处理6 h再给予5 ng/mL TGF-β1干预48 h。光镜观察细胞形态变化,免疫印迹及免疫荧光法检测上皮标志物E钙黏蛋白(E-cadherin)、间质标志物α-平滑肌肌动蛋白(α-smooth muscle actin, α-SMA)表达;免疫印迹法检测核因子红细胞2相关因子2(nuclear factor erythroid 2-related factor 2, Nrf2)蛋白水平。结果 与对照组相比,TGF-β1组光镜下可见部分A549细胞形态从鹅卵石状变成细长不规则状,呈间质性改变,E-cadherin表达减少(P<0.05)、α-SMA表达增多(P<0.01),Nrf2蛋白水平下调(P<0.05)。与TGF-β1组相比,pcDNA3.1-Nrf2+TGF-β1组A549细胞E-cadherin表达增多(P<0.05)、α-SMA表达减少(P<0.01);LXA4+TGF-β1组A549细胞E-cadherin上调、α-SMA下调以及Nrf2蛋白水平增多(P<0.05)。结论 LXA4可抑制A549细胞发生EMT,其机制与上调Nrf2表达相关。
Objective To investigate the effect of lipoxin A4(LXA4)on the epithelial-mesenchymal transition(EMT)of alveolar epithelial typeⅡcell(ATⅡ)and its potential mechanisms.Methods A549 cells were divided into five groups:control group,transforming growth factorβ1(TGF-β1)group,empty vector+TGF-β1 group,pcDNA3.1-Nrf2+TGF-β1 group and LXA4+TGF-β1 group.A549 cells were treated with 5 ng/mL TGF-β1 for 48 h in TGF-β1 group.In empty vector+TGF-β1 group and pcDNA3.1-Nrf2+TGF-β1 group,A549 cells were respectively transfected with empty vector and pcDNA3.1-Nrf2,and then treated with 5 ng/mL TGF-β1 for 48 h.In LXA4+TGF-β1 group,A549 cells were pretreated with 100 nmol/L LXA4 for 6 h,and then stimulated with 5 ng/mL TGF-β1 for 48 h.The morphological changes of A549 cells were observed under light microscope,the expression levels of E-cadherin andα-smooth muscle actin(α-SMA)were examined by immunofluorescence assay and immunoblotting,and the protein level of nuclear factor erythroid 2-related factor 2(Nrf2)was measured by immunoblotting.Results Compared with control group,a part of A549 cells showed a morphological change from a pebble-like shape to an irregular elongated shape in TGF-β1 group.Compared with control group,the expression of E-cadherin was downregulated(P<0.05),α-SMA was upregulated(P<0.01),and the protein level of Nrf2 was decreased in TGF-β1 group(P<0.05).Compared with TGF-β1 group,the E-cadherin expression was upregulated(P<0.05)andα-SMA expression was decreased in pcDNA3.1-Nrf2+TGF-β1 group(P<0.01),and the expression levels of E-cadherin and Nrf2 were increased andα-SMA expression was decreased in LXA4+TGF-β1 group(P<0.05).Conclusion LXA4 can inhibit the EMT of A549 cells by upregulating Nrf2.
作者
王贵佐
杨淑梅
刘璐
WANG Guizuo;YANG Shumei;LIU Lu(Department of Respiratory and Critical Care Medicine,Shaanxi Provincial People′s Hospital,Xi′an 710068,China)
出处
《山西医科大学学报》
CAS
2024年第2期157-163,共7页
Journal of Shanxi Medical University
基金
国家自然科学基金青年项目(82000037)
陕西省重点研发计划重点产业创新链(群)项目(2020ZDLSF01-06)
陕西省人民医院科技人才支持计划项目(2021JY-10)。
