埃克替尼抑制肺泡上皮细胞上皮间充质转化进程改善博来霉素诱导的肺纤维化  

Icotinib Improves Bleomycin-Induced Pulmonary Fibrosis by Inhibiting Epithelial-Mesenchymal Transition of Alveolar Epithelial Cells

作　　者：许文奇 肖雷[1] 徐炜 严静静 顾文强 李先伟[3] XU Wenqi;XIAO Lei;XU Wei;YAN Jingjing;GU Wenqiang;LI Xianwei(Dongcheng District,the First Affiliated Hospital of Anhui Medical University,Hefei 230000,China;Department of Urology,The Second Affiliated Hospital of Wannan Medical College,Wuhu 241000,China;Department of pharmacology,Wannan Medical College,Wuhu 241002,China)

机构地区：[1]安徽医科大学第一附属医院东城院区,合肥230000 [2]皖南医学院第二附属医院泌尿外科,安徽芜湖241000 [3]皖南医学院药理学教研室,安徽芜湖241002

出　　处：《中国药学杂志》2024年第12期1120-1128,共9页Chinese Pharmaceutical Journal

基　　金：安徽省高校自然科学研究重大项目资助(KJ2021ZD0106);安徽省卫生健康委科研重点项目资助(AHWJ2021a033);皖南医学院教育基金会项目资助(HXKT2022028)。

摘　　要：目的探究埃克替尼(icotinib,ICO)的抗肺纤维化(pulmonary fibrosis,PF)作用及其潜在的分子机制。方法C57BL/6小鼠随机分为假手术(Sham)组、PF组、ICO 30 mg·kg^(-1)剂量组及ICO 60 mg·kg^(-1)剂量组,每组n=8。单次气管注射博来霉素(BLM,3 mg·kg^(-1))建立小鼠PF模型。HE及Masson染色观察肺组织病理变化及胶原沉积情况。免疫组化检测肺组织Ⅰ型胶原(collagenⅠ)的表达。体外培养肺泡上皮细胞,实验设对照(control)组、表皮生长因子(epidermal growth factor,EGF)组(100 ng·mL^(-1))及EGF联合ICO(0.1、1、10 mmol·L^(-1))3个剂量组。免疫荧光法检测细胞E-钙黏蛋白(E-cadherin)和α-平滑肌肌动蛋白(α-SMA)的表达及核因子-κB(NF-κB)p65核转移情况。Western blots检测肺组织和(或)细胞collagenⅠ、E-cadherin、α-SMA、Vimentin、磷酸化表皮生长因子受体(phosphorylation epidermal growth factor receptor,p-EGFR)、磷酸化核因子κB抑制物α(p-IκBα)、磷酸化NF-κB p65(p-NF-κB p65)的蛋白水平及NF-κB p65核转移情况。结果动物实验表明,ICO抑制了BLM诱导的胶原沉积,减少了Ⅰ型胶原的表达,减轻了博来霉素诱导的上皮-间充质转化(epithelial-mesenchymal transition,EMT)(E-cadherin表达增加,Vimentin和α-SMA表达减少),降低了EGFR,IκBα和NF-κB p65的磷酸化水平并抑制了NF-κB p65的核转移。细胞实验发现,EGF可激活肺泡上皮细胞EMT和EGFR/NF-κB信号通路,而ICO可逆转这一作用。结论ICO可能通过抑制EGFR/NF-κB信号通路的活化,逆转了EGF诱导的EMT和博来霉素诱导的小鼠肺纤维化。OBJECTIVE This study aimed to investigate the role and potential mechanisms of Icotinib(ICO)on pulmonary fibrosis.METHODS C57BL/6 mice were randomly divided into Sham group,PF group,ICO 30 mg·kg^(-1)group and ICO 60 mg·kg^(-1)group,8 rats in each group.A mouse model of PF was induced by intratracheal injection of bleomycin(3 mg·kg^(-1)).Hematoxylin-eosin staining and Masson trichrome staining for lung tissues were performed to observe the pathological alterations and collagen deposition.Immunohistochemical detection of lung tissue collagen typeⅠ(collagenⅠ)expression.In vitro,the lung epithelial cells were divided into control group,epidermal growth factor(EGF)group,EGF combined with ICO(0.1,1,10 mmol·L^(-1))groups.The protein expression of E-cadherin,α-SMA and nuclear transfer of NF-κB p65 were detected by immunofluorescence.The protein levels of CollagenⅠ,E-cadherin,α-SMA,Vimentin,phosphorylation epidermal growth factor receptor(p-EGFR),p-IκBα,p-NF-κB p65 and nuclear NF-κB p65 were detected by Western blot analysis in lung tissue and(or)cells.RESULTS The results demonstrated that ICO inhibited bleomycin-induced collagen deposition,reduced typeⅠcollagen expression,alleviated bleomycin-induced EMT(increased E-cadherin expression and decreased Vimentin andα-SMA expression),and decreased phosphorylation of EGFR,IκBα,NF-κB p65 and nuclear translocation of NF-κB p65 in vivo.Furthermore,incubation of lung epithelial cells with EGF activated EMT and EGFR/NF-κB signaling pathway,and these effects were reversed by ICO In vitro.CONCLUSION In conclusion,ICO attenuates EGF-induced EMT in lung epithelial cells and bleomycin-induced pulmonary fibrosis in mice by downregulating EGFR/NF-κB pathway.

关 键 词：埃克替尼 肺纤维化 上皮-间充质转化 表皮生长因子受体/核因子-κB信号通路 

分 类 号：R965.2[医药卫生—药理学]