国际呼吸杂志
國際呼吸雜誌
국제호흡잡지
INTERNATIONAL JOURNAL OF RESPIRATION
2014年
20期
1544-1550
,共7页
陈海华%周贤龙%施余露%杨炯
陳海華%週賢龍%施餘露%楊炯
진해화%주현룡%시여로%양형
p38丝裂原活化蛋白激酶%c-Jun氨基端激酶%肺泡上皮细胞%上皮-间质转化%转化生长因子β1
p38絲裂原活化蛋白激酶%c-Jun氨基耑激酶%肺泡上皮細胞%上皮-間質轉化%轉化生長因子β1
p38사렬원활화단백격매%c-Jun안기단격매%폐포상피세포%상피-간질전화%전화생장인자β1
p38 MAPK%c-Jun N-terminal kinase%Alveolar epithelial cells%Epithelial to mesenchymal transition%Transforming growth factor-β1
目的 探讨p38丝裂原活化蛋白激酶(p38 MAPK)和c-Jun氨基端激酶(JNK)信号通道在转化生长因子β1 (TGF-β1)诱导的人肺泡上皮细胞上皮-间质转化(epithelial to mesenchymal transition,EMT)中的意义,从而进一步揭示肺纤维化的发病机制.方法 使用TGF-β1诱导A549细胞EMT,分别在蛋白水平(使用p38 MAPK抑制剂SB203580和JNK抑制剂SP600125)和RNA水平(RNA干扰)抑制p38 MAPK和JNK信号通道,检测抑制后A549细胞的p38 MAPK和JNK的蛋白和mRNA,以及间质细胞表型蛋白包括结蛋白、波形蛋白、α-平滑肌肌动蛋白和上皮细胞表型蛋白包括E-钙黏素、紧密连接蛋白-1、水通道蛋白-5的表达.结果 TGF-β1可以诱导A549细胞EMT,表现为间质细胞表型蛋白表达的增加和上皮细胞表型蛋白表达的减少,这一过程p38 MAPK和JNK通道表达也增加,无论蛋白水平抑制或基因沉默p38 MAPK和JNK均可减轻A549细胞的EMT.结论 p38 MAPK和JNK信号通道在TGF-β1诱导的A549细胞EMT过程中起重要作用.
目的 探討p38絲裂原活化蛋白激酶(p38 MAPK)和c-Jun氨基耑激酶(JNK)信號通道在轉化生長因子β1 (TGF-β1)誘導的人肺泡上皮細胞上皮-間質轉化(epithelial to mesenchymal transition,EMT)中的意義,從而進一步揭示肺纖維化的髮病機製.方法 使用TGF-β1誘導A549細胞EMT,分彆在蛋白水平(使用p38 MAPK抑製劑SB203580和JNK抑製劑SP600125)和RNA水平(RNA榦擾)抑製p38 MAPK和JNK信號通道,檢測抑製後A549細胞的p38 MAPK和JNK的蛋白和mRNA,以及間質細胞錶型蛋白包括結蛋白、波形蛋白、α-平滑肌肌動蛋白和上皮細胞錶型蛋白包括E-鈣黏素、緊密連接蛋白-1、水通道蛋白-5的錶達.結果 TGF-β1可以誘導A549細胞EMT,錶現為間質細胞錶型蛋白錶達的增加和上皮細胞錶型蛋白錶達的減少,這一過程p38 MAPK和JNK通道錶達也增加,無論蛋白水平抑製或基因沉默p38 MAPK和JNK均可減輕A549細胞的EMT.結論 p38 MAPK和JNK信號通道在TGF-β1誘導的A549細胞EMT過程中起重要作用.
목적 탐토p38사렬원활화단백격매(p38 MAPK)화c-Jun안기단격매(JNK)신호통도재전화생장인자β1 (TGF-β1)유도적인폐포상피세포상피-간질전화(epithelial to mesenchymal transition,EMT)중적의의,종이진일보게시폐섬유화적발병궤제.방법 사용TGF-β1유도A549세포EMT,분별재단백수평(사용p38 MAPK억제제SB203580화JNK억제제SP600125)화RNA수평(RNA간우)억제p38 MAPK화JNK신호통도,검측억제후A549세포적p38 MAPK화JNK적단백화mRNA,이급간질세포표형단백포괄결단백、파형단백、α-평활기기동단백화상피세포표형단백포괄E-개점소、긴밀련접단백-1、수통도단백-5적표체.결과 TGF-β1가이유도A549세포EMT,표현위간질세포표형단백표체적증가화상피세포표형단백표체적감소,저일과정p38 MAPK화JNK통도표체야증가,무론단백수평억제혹기인침묵p38 MAPK화JNK균가감경A549세포적EMT.결론 p38 MAPK화JNK신호통도재TGF-β1유도적A549세포EMT과정중기중요작용.
Objective To investigate the roles of p38 MAPK and JNK in transforming growth factor-β1 (TGF-β1)-induced human alveolar epithelial to mesenchymal transition (EMT),and to demonstrate the possible molecular mechanism of idiopathic pulmonary fibrosis.Methods A549 cells were treated with TGF-β1 (3 μg/L) for 48 hours to induce EMT.The expressions of mesenchymal phenotypic markers including desmin,α-smooth muscle actin (α-SMA) and vimentin,and expressions of epithelial phenotypic markers including E cadherin,zonula occludens-1 (ZO-1) and aquaporin-5 (AQP-5) were detected by Western blot.The role of p38 MAPK and JNK in TGF-β1-mediated EMT was investigated using gene silencing and inhibitor SB203580 and SP600125.Results The data showed that TGF-β1 induced A549 cells with alveolar epithelial type Ⅱ cell phenotype to undergo EMT.The process of EMT was accompanied by increased expression of the mesenchymal cell markers α-SMA,desmin and vimentin,and downregulation of the epithelial cell markers E-cadherin,ZO-1 and AQP-5.TGF-β1-induced EMT occurred through phosphorylation of p38 MAPK and JNK and was inhibited by inhibitor SB203580 and SP600125 and gene silencing.Conclusions Our data shows that TGF-β1 induces A549 alveolar epithelial cells to undergo EMT partially via p38 MAPK and JNK activation.
