异甘草素对体外肺纤维化模型的作用及其机制  

Effect and Mechanism of Isoliquiritin on Pulmonary Fibrosis Model in Vitro

作　　者：蔡风林 王梅芳[1] 程雪琴[1] 袁乐永 何金娟 胡雯雯 唐以军[1] CAI Fenglin;WANG Meifang;CHEN Xueqin;YUN Leyong;HE Jinjuan;HU Wenwen;TANG Yijun(Department of Respiratory and Critical Care Medicine,Taihe Hospital,Hubei University of Medicine,Shiyan 442000,China;Department of Immunology,Basic Medical College,Hubei University of Medicine,Shiyan 442000,China;Graduate Training Base of Taihe Hospital,Shiyan City,Jinzhou Medical University,Shiyan 442000,China)

机构地区：[1]湖北医药学院附属太和医院呼吸与危重症医学科,十堰442000 [2]湖北医药学院基础医学院免疫教研室,十堰442000 [3]锦州医科大学十堰市太和医院研究生培养基地,十堰442000

出　　处：《医药导报》2022年第2期167-174,共8页Herald of Medicine

基　　金：湖北医药学院自由探索基金创新团队项目(FDFR201801);湖北医药学院研究生科技创新项目(YC2019037);湖北省医学领军人才项目。

摘　　要：目的观察异甘草素(ISL)对转化生长因子β_(1)(TGF-β_(1))诱导人肺泡Ⅱ型上皮A549细胞所构建的体外肺纤维化模型的影响,探讨其作用机制。方法TGF-β_(1)刺激A549细胞发生上皮-间质转化(EMT)构建体外肺纤维化模型;加入ISL干预后,噻唑蓝(MTT)法检测TGF-β_(1)刺激A549细胞后,ISL对细胞增殖活性的影响;细胞划痕实验检测细胞迁移能力;倒置相差显微镜观察细胞形态变化;实时荧光定量PCR(RT-PCR)检测EMT相关E-cadherin、Vimentin、α-SMA、FN及其相关转录因子Snail、Slug、Twist、ZEB1、ZEB2 mRNA的表达水平;蛋白免疫印迹法(Western blotting)检测EMT相关蛋白E-cadherin、N-cadherin、Vimentin、α-SMA、FN及MAPK/Erk信号通路中Erk1/2磷酸化蛋白的表达水平。结果TGF-β_(1)可使A549细胞形态由上皮细胞向成纤维细胞转变,细胞增殖及迁移能力增强(P<0.05或P<0.01);上皮细胞标志物E-cadherin表达降低,间质细胞标志物N-cadherin、Vimentin、α-SMA、FN表达升高;EMT相关转录因子及磷酸化Erk1/2表达升高(P<0.05或P<0.01)。而经ISL处理后,ISL能有效抑制TGF-β_(1)诱导的A549细胞的增殖、迁移及细胞形态变化(P<0.05或P<0.01);提高E-cadherin,降低N-cadherin、Vimentin、α-SMA、FN、EMT相关转录因子及磷酸化Erk1/2的表达水平(P<0.05或P<0.01)。结论ISL通过干预MAPK/Erk信号通路,抑制TGF-β_(1)诱导的A549细胞的EMT过程,维护细胞上皮样形态,减少肺成纤维细胞的产生,有望成为治疗肺纤维化的一种潜在药物。Objective To explore the role and possible mechanism of Isoliquiritigenin(ISL)on the pulmonary fibrosis by observing the effect of ISL on pulmonary fibrosis model induced by transforming growth factorβ_(1)(TGF-β_(1))on human alveolar type II epithelial A549 cells in vitro.Methods TGF-β_(1)was used to stimulate A549 cells to undergo epithelial-mesenchymal transition(EMT)to construct a pulmonary fibrosis model in vitro.After the intervention by adding ISL,MTT method was used to detect effect of ISL on cell proliferating activity after TGF-β_(1)stimulating A549 cells.Cell scratch test was used to detect cell migration ability.Inverted phase contrast microscope was used to observe cell morphology.Real-time PCR(RT-PCR)was used to detect the expression levels of EMT-related mRNA(E-cadherin,Vimentin,α-SMA,and FN)and EMT related transcription factors(Snail,Slug,Twist,ZEB1,and ZEB2).Western blotting was used to detection the expression levels of EMT-related proteins(E-cadherin,N-cadherin,Vimentin,α-SMA,FN)and Erk phosphorylated protein in MAPK/Erk signaling pathway.Results TGF-β_(1)could change the morphology of A549 cells from epithelial cells to fibroblasts,enhance the ability of cell proliferation and migration(P<0.05 or P<0.01),decrease the expression of E-cadherin,increase the expressions of N-cadherin,Vimentin,α-SMA,FN,and increase the expressions of EMT-related transcription factors and phosphorylated Erk(P<0.05 or P<0.01).ISL could effectively inhibit the proliferation,migration and cell morphology changes of A549 cells induced by TGF-β_(1)(P<0.05 or P<0.01).It could also increase the expression levels of E-cadherin,and decrease N-cadherin,Vimentin,α-SMA,FN,EMT-related transcription factors and phosphorylated Erk(P<0.05 or P<0.01).Conclusion ISL could inhibit TGF-β_(1)induced EMT process of A549 cells by interfering the MAPK/Erk signaling pathway.It could maintain the epithelial morphology of the cells,and reduce the production of lung fibroblasts.Thus,it could be used as a potential treatment for pulmo

关 键 词：异甘草素 肺纤维化 上皮-间质转化 MAPK/ERK信号通路 

分 类 号：R965[医药卫生—药理学] R563[医药卫生—药学]