基质金属蛋白酶14在矽肺中的表达及其对上皮间充质转化的调控作用  

Expression of matrix metalloproteinase 14 in silicosis and its regulation of epithelial-mesenchymal transition

作　　者：丁琼桦 马子怡 钱蕊 陈叙汐 王礼群 陈晴 肖月 姚于勤 许云屹 DING Qiong-hua;MA Zi-yi;QIAN Rui;CHEN Xu-xi;WANG Li-qun;CHEN Qing;XIAO Yue;YAO Yu-qin;XU Yun-yi(Sichuan Provincial Key Laboratory of Molecular Toxicology,West China School of Public Health/West China Fourth Hospital,Sichuan University,Chengdu,Sichuan 610041,China)

机构地区：[1]四川大学华西公共卫生学院/华西第四医院,分子毒理四川省教育厅重点实验室,四川成都610041

出　　处：《现代预防医学》2025年第4期629-635,共7页Modern Preventive Medicine

基　　金：国家自然科学基金(82373548,U22A20359,U23A20495);四川省科学技术厅基金(2023NSFSC1731,2024NSFSC1258);四川大学与达州市战略合作项目(2022CDDZ-14)。

摘　　要：目的通过建立小鼠矽肺模型,探讨潜在致病分子MMP14在二氧化硅致肺损伤中的作用。方法SPF级雄性C57小鼠18只随机分为对照组和矽肺模型组组;人体标本数量16例,来自四川大学华西第四医院尘肺队列;采用单细胞测序技术分析差异表达基因,筛选潜在致病分子,采用非暴露式气管滴注法构建小鼠矽肺模型,通过苏木精-伊红染色和Masson染色评估小鼠肺组织炎症和纤维化程度。采用实时荧光定量聚合酶链式反应、蛋白免疫印迹和免疫组织化学等技术验证小鼠矽肺模型肺组织及正常对照肺组织中MMP14及N-cadherin的表达。通过小干扰RNA敲减上皮细胞16HBE中的MMP14,验证N-cadherin表达,并采用细胞划痕实验验证敲减MMP14对细胞迁移的调控。结果单细胞测序结果显示MMP14为潜在致病分子,与正常对照组相比,小鼠矽肺模型组肺组织中MMP14及N-cadherin表达分别增高3.78和2.42倍(P<0.001)。在SiO2刺激的16HBE细胞中敲减MMP14,可下调N-cadherin的表达约30%(P<0.001),同时抑制16HBE细胞迁移,细胞迁移面积降低50%(P<0.001)。结论MMP14激活N-cadherin表达,调控上皮细胞EMT过程,促进二氧化硅诱导的肺纤维化发生。Objective To investigate the role of the potential pathogenic molecule MMP14 in silicosis.Methods 18 SPF male C57 mice were randomly divided into control and silicosis group,and 16 human specimens from the pneumoconiosis cohort of the Fourth Hospital of West China,Sichuan University.Single-cell sequencing analysis was used to identify differentially expressed genes.A silicosis mouse model was induced by silica dioxide using a non-exposure tracheal instillation method.Hematoxylin-eosin staining and Masson staining were employed to assess the degree of inflammation and fibrosis in mouse lung tissues.Real-time quantitative polymerase chain reaction,western blot,and immunohistochemistry were used to verify the expression of MMP14 and N-cadherin in the lung tissues.Knockdown of MMP14 in epithelial cells(16HBE)was achieved by small interfering RNA,to further verify the expression of N-cadherin.Cell wound healing assays were used to assess the cell migration after MMP14 knockdown.Results MMP14 being selected as a potential pathogenic molecule by single-cell sequencing analysis.Compared with the normal control group,the expression of MMP14 and N-cadherin in silicosis lung tissues was significantly increased 3.78 and 2.42 times,respectively(P<0.001).Knockdown of MMP14 in 16HBE cells stimulated by SiO2 led to the downregulation of N-cadherin by about 30%(P<0.001)and inhibition of cell migration,which area was reduced by 50%(P<0.001).Conclusion MMP14 activates the expression of N-cadherin,regulates the epithelial-mesenchymal transition(EMT)process,and promotes silica-induced pulmonary fibrosis.

关 键 词：矽肺 肺纤维化 基质金属蛋白酶14 上皮间充质转化 神经钙黏蛋白 

分 类 号：R135.2[医药卫生—劳动卫生]