目的:观察经气管移植内皮祖细胞(EPCs)在烟雾暴露所致慢性阻塞性肺疾病模型小鼠中的分布及分化。方法:体外分离小鼠骨髓单个核细胞于EGM-2MV培养基中培养并鉴定。24只C57BL/6J小鼠随机分为正常对照组、COPD+PBS干预组及COPD+EPCs干预组;香烟烟雾暴露90d建立慢性阻塞性肺疾病(COPD)小鼠模型;烟雾暴露结束后,COPD+PBS干预组及COPD+EPCs干预组分别经气管注入30μLPBS和30μL细胞悬液(含1×105个CM-DiI标记EPCs)。移植后观察30d处死小鼠,通过荧光显微镜观察移植细胞在肺内的分布及通过免疫荧光检测广谱细胞角蛋白的表达水平。结果:EPCs移植后30d可见EPCs分布于肺血管及气道,部分细胞表达上皮特异标志广谱细胞角蛋白。结论:EPCs移植后可定植COPD模型小鼠肺血管及气道,且可能转化为支气管及肺泡上皮细胞。
Objective:To observe the distribution and transformation of endothelial progenitor cells (EPCs) in mice model of COPD after intratracheal EPCs transplantation.Methods:Isolated mononuclear cells from C57BL/6J mice bone marrow were cultured in EGM-2MV medium and identified as EPCs.Twenty-four male C57BL/6J mice were randomly divided into normal control group, COPD+PBS group and COPD+EPCs group (each n=8).All groups except the control group which was sham-smoked were exposed pas-sively to cigarette smoke (CS) for continuous 90 days).After CS exposure finished, mice in COPD+PBS group and COPD+EPCs group were treated with 30 μL PBS and 30 μL cell suspension (containing 105 EPCs) respectively.Then we detected the distribution and the expression of Pan-cytokeratin of EPCs in lung tissues 30 days after transplantation.Results:After transplantation, we detected EPCs in lung vascular system and air way.With the staining of Pan-cytokeratin, we observed some EPCs expressed Pan-cytokeratin which was a specific marker of lung epithelial cells.Conclusion:EPCs could be located in lung vessels and the air ways and transformed into bronchial and alveolar epithelial cells in mice with COPD after intratracheal transplantation.