γ-分泌酶抑制剂在肺纤维化上皮间质转化中的作用  

The role ofγ-secretase inhibitor in pulmonary fibrosis epithelial-mesenchymal transition

作　　者：韩姣 王华兵[1] 徐玲文 董芳[1] HAN Jiao;WANG Huabing;XU Lingwen;DONG Fang(Department of Critical Care Medicine,Wuhan Third Hospital(Tongren Hospital of Wuhan University),Wuhan 430200,China)

机构地区：[1]武汉市第三医院(武汉大学附属同仁医院)重症医学科,430200

出　　处：《天津医药》2022年第9期917-920,共4页Tianjin Medical Journal

基　　金：武汉市卫健委科研项目(项目编号WX21Z19)。

摘　　要：目的检测体外转化生长因子(TGF)-β1诱导肺纤维化上皮间质转化(EMT)的形成情况并检测Notch信号特异性抑制剂γ-分泌酶抑制剂(DAPT)对其影响及可能机制。方法将A549细胞分成对照组(RPMI 1640完全培养基培养)、TGF-β1组(在含有10μg/L TGF-β1的RPMI 1640培养基中培养)、TGF-β1+DAPT组(在含有10μg/L TGF-β1和2μmol DAPT的RPMI 1640培养基中培养)、DAPT组(在含有2μmol DAPT的RPMI 1640培养基中培养)。通过倒置显微镜观察细胞形态,实时荧光定量逆转录聚合酶链反应检测肺泡上皮细胞特异性蛋白E-钙黏合素以及间质细胞特异性蛋白α-肌动蛋白(α-SMA)mRNA相对表达水平,Western blot检测E-钙黏合素以及α-SMA蛋白表达水平。结果倒置显微镜检查示,对照组细胞呈多边形,铺路石样,细胞间连接紧密;TGF-β1组细胞呈梭行,纺锤状,细胞间连接减少;TGF-β1+DAPT组仅有少量细胞呈梭形,多数细胞形态与对照组相似;DAPT组细胞形态大致同对照组。与对照组相比,TGF-β1组α-SMA蛋白及mRNA表达增加,而E-钙黏合素蛋白及mRNA表达减弱(P<0.05);与TGF-β1组比较,TGF-β1+DAPT组α-SMA蛋白及mRNA表达水平降低,而E-钙黏合素蛋白及mRNA表达水平增加(P<0.05);DAPT组与对照组2指标的蛋白与mRNA相对表达水平差异无统计学意义(P>0.05)。结论TGF-β1可诱导肺上皮间质转化,而Notch信号抑制剂DAPT能够阻断、部分或全部逆转这一过程。Objective To investigate the formation of pulmonary fibrosis epithelial-interstitial transformation(EMT)induced by transforming growth factor TGF-β1 in vitro and to detect the effect of Notch signal specific inhibitor DAPT on this transformation and its possible mechanism.Methods A549 cells were divided into the control group(cultured in RPMI 1640 complete medium),the TGF-β1 group(containing 10μg/L TGF-β1 in RPMI 1640 medium),the TGF-β1+DAPT group(containing 10μg/L TGF-β1 and 2μmol DAPT in RPMI 1640 medium)and the DAPT group(only 2μmol DAPT was cultured in RPMI 1640 medium).The cell morphology was observed by inverted microscope.The mRNA expression of alveolar epithelial cell specific protein levels of E-cadherin and interstitial cell specific proteinα-actin(α-SMA)were detected by quantitative reverse transcription polymerase chain reaction(qRT-PCR),and the expression levels of E cadherin andα-SMA were detected by Western blot assay.Results The cells in the control group were polygonal,irregular and closely linked,while those in the TGF-β1 group were spindle-shaped with reduced intercellular connections.In the TGF-β1+DAPT group,only a few cells showed spindle shape,and most cell morphology was similar to that of the control group.The cell morphology of the DAPT group was similar to that of the control group.Compared with the control group,expression levels ofα-SMA protein and mRNA increased significantly in the TGF-β1 group(P<0.05),while the expression of E-cadherin and its mRNA decreased significantly(P<0.05).Compared with the TGF-β1,the expression ofα-SMA and mRNA decreased significantly in the TGF-β1 and DAPT group,while the expression of E-cadherin and mRNA increased significantly(P<0.05).There were no significant differences in expression levels ofα-SMA and E-cadherin protein and mRNA between the DAPT group and the control group(P>0.05).Conclusion TGF-β1 can induce lung epithelial interstitial transformation,while Notch signal inhibitor DAPT can block,partially or completely reverse this

关 键 词：转化生长因子Β1 肺纤维化 上皮-间充质转化 Γ-分泌酶抑制剂 

分 类 号：R563.9[医药卫生—呼吸系统]