细胞周期抑制因子p21活化巨噬细胞促进小鼠肺纤维化发生发展  

The cell cycle inhibitor p21 promote mouse lung fibrosis by activating alveolar macrophages

作　　者：魏旭鹏 王婉玉 李云炫 刘畅 吕晓希 赵燕燕[1] 刘姗姗 WEI Xu-peng;WANG Wan-yu;LI Yun-xuan;LIU Chang;Lü Xiao-xi;ZHAO Yan-yan;LIU Shan-shan(Pharmacy College,Hebei University,Baoding 071000,China;State Key Laboratory of Bioactive Substance and Function of Natural Medicines,Institute of Materia Medica,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100050,China)

机构地区：[1]河北大学药学院,河北保定071000 [2]中国医学科学院、北京协和医学院药物研究所,天然药物活性物质与功能国家重点实验室,北京100050

出　　处：《药学学报》2020年第8期1792-1800,共9页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金资助项目(81803604)。

摘　　要：探讨博来霉素(bleomycin,BLM)损伤所致小鼠肺纤维化发病过程中肺泡巨噬细胞的细胞周期抑制因子p21(cell cycle inhibitor p21,p21)蛋白表达及其对巨噬细胞活化的调节作用。免疫荧光染色法检测肺纤维化组织中肺泡巨噬细胞中CD206的表达;RT-PCR(reverse transcription-polymerase chain reaction)法检测巨噬细胞活化标志蛋白的表达;巨噬细胞/成纤维细胞共培养法检测巨噬细胞对成纤维细胞活化及胶原收缩能力的影响;采用免疫荧光和免疫印迹法检测巨噬细胞中p21蛋白表达的变化;采用p21敲低和过表达病毒感染肺泡巨噬细胞,RT-PCR和巨噬细胞/成纤维细胞共培养法检测改变p21表达对肺泡巨噬细胞促纤维化表型的调节作用。动物实验操作过程依照中国医学科学院、北京协和医学院药物研究所实验动物管理与动物福利委员会的要求执行。结果显示:与对照组比较,模型组肺泡巨噬细胞CD206的表达增加、巨噬细胞活化标志蛋白表达增加、模型组肺泡巨噬细胞促进成纤维细胞的活化并增强其胶原收缩能力。模型组小鼠肺泡巨噬细胞中p21蛋白表达增加。敲低p21可显著抑制巨噬细胞促纤维化表型,而过表达对照小鼠肺泡巨噬细胞中p21则促进其促纤维化表型的转化。以上研究结果表明,肺纤维化小鼠肺泡巨噬细胞中p21表达增加,p21可通过促进巨噬细胞活化参与肺纤维化发病。This study was to determine the expression of the cell cycle inhibitor p21 in alveolar macrophages(AMs)and the role of p21 in activation of AMs in bleomycin(BLM)injury-induced lung fibrosis.The expression of CD206 in AMs was measured by immunofluorescence staining.Reverse transcription-polymerase chain reaction(RT-PCR)assay was used to detect the expression of macrophage activation markers.The coculture assay for macrophage and fibroblast was employed to explore the effect of macrophage on fibroblast activation.Immunofluorescence staining and western blotting assay were adopted to detect the expression of p21 in fibrotic tissues.AMs were treated with p21 knockdown or overexpression virus,RT-PCR and the co-culture system were used to explore the effect of p21 expression on macrophage activation.The Experimental Animal Welfare Ethics Committee of the Institute of Materia Medica,Chinese Academy of Medical Sciences and Peking Union Medical College approved all of the protocols for this research.Our results showed that the expression of CD206 and macrophage activation markers was increased in AMs from fibrotic mice,indicating that AMs from fibrotic mice were associated with a profibrotic phenotype.Moreover,the expression of p21 was upregulated in AMs after BLM treatment.Depletion of p21 suppressed macrophage activation,while overexpression of p21 promoted the profibrotic phenotype of AMs from healthy mice.In summary,BLM injury causes the progressive accumulation of p21 in AMs,which induces the production of a number of profibrotic factors promoting the development of pulmonary fibrosis.

关 键 词：肺纤维化 巨噬细胞 P21 成纤维细胞活化 博来霉素损伤 

分 类 号：R967[医药卫生—药理学]