摘要
目的观察百草枯(Paraquat,PQ)诱导人肺间质成纤维细胞HEL299纤维化增殖过程中肺上皮细胞的重要作用及其调节的信号机制。方法应用体外人肺上皮细胞和间质细胞共培养系统,MTT法检测PQ对人肺间质成纤维细胞HEL299和HPAEpiC细胞增殖的影响,ELISA检测共培养体系中转化生长因子-β(TGF-β)变化,Real-time PCR和Western blot检测上皮HPAEpiC细胞的TGF-β信号通路变化对肺间质成纤维细胞HEL299增殖的影响。结果PQ对肺间质成纤维细胞HEL299体外增殖依赖于上皮细胞的存在。PQ促进肺上皮HPAEpiC细胞TGF-β的分泌,进而促进肺间质成纤维细胞增殖;阻断上皮细胞TGF-β信号将明显减少间质HEL299细胞的增殖。结论 PQ促进人肺间质细胞上皮HEL299的增殖依赖微环境中上皮HPAEpiC细胞的TGF-β信号。本研究结果阐述了PQ诱导肺纤维化的一种新机制。
Objective To investigate the paraquat（PQ）-induced proliferation of stroma fibroblast cells and dissect the mechanisms of epithelial cells＇role in the lung microenvironments.Methods HEL299 cell IC50 was determined by MTT assay.TGF-βwas assayed by ELASA.The TGF-βmRNA and protein expressions were also assayed by real-time PCR and Western blot in HPAEpiC-HEL299 cells co-cultured system following paraquat treatment.Results PQ IC50 cutoff in HEL299 cells was 210.3μmol/L.Compared with DMSO control group,PQ could induce the proliferation of lung stroma fibroblast HEL299 cells,which was dependent on the signaling of epithelial HPAEpiC cell TGF-β.However,inhibition of TGF-βsignaling by si RNA or specific antibody abolished PQ-mediated HEL299 cell proliferation.Conclusion PQ＇s promoting the proliferation of human lung stroma cells depends on the signaling of epithelial cell TGF-β.Our conclusion provides a new mechanism for PQ-induced pulmonary fibrosis and blocking epithelia TGF-βsignaling will be a potential therapeutic target for PQ toxicity.
出处
《西安交通大学学报（医学版）》
CAS
CSCD
北大核心
2015年第5期619-622,643,共5页
Journal of Xi’an Jiaotong University（Medical Sciences）
基金
陕西省自然科学基金资助项目(No.2013JM4069)~~
