水通道蛋白1对内皮祖细胞迁移功能的影响  

Effect of aquaporin 1 on the migration of endothelial progenitor cells

作　　者：葛海燕 朱惠莉 夏世金 宋元林 白春学 GE Haiyan;ZHU Huili;XIA Shijin;SONG Yuanlin;BAI Chunxue(Department of Pulmonary Medicine, Huadong Hospital, Fudan University, Shanghai 200040, P. R. China;Department of Geriatrics, Huadong Hospital, Fudan University, Shanghai 200040, P. R. China;Department of Pulmonary Medicine, Zhongshan Hospital, Fudan University, Shanghai 200032. P. R. China)

机构地区：[1]复旦大学附属华东医院呼吸科,上海200040 [2]复旦大学附属华东医院上海市老年医学研究所,上海200040 [3]复旦大学附属中山医院呼吸科,上海200032

出　　处：《中国呼吸与危重监护杂志》2018年第3期290-295,共6页Chinese Journal of Respiratory and Critical Care Medicine

基　　金：国家自然科学基金青年项目(81600056)

摘　　要：目的观察水通道蛋白1(AQP1)对内皮祖细胞(EPC)增殖迁移功能的影响。方法体外分离AQP1野生型(WT)(n=6)和敲除型(KO)小鼠(n=6)骨髓细胞并定向培养分化为EPC。应用免疫荧光法检测细胞表面抗原鉴定EPC,通过无标记活细胞动态分析技术、细胞Transwell迁移实验及划痕实验比较AQP1 WT和KO小鼠中EPC的功能差异。结果对小鼠EPC培养观察发现,细胞最初保持悬浮状态,7 d内逐渐黏附贴壁成典型的间充质干细胞,间充质干细胞经内皮细胞专用培养基培养7 d后逐渐分化贴壁,14 d后细胞继续增殖,形态为梭形或多边形,并融合成铺路石样的EPC细胞。细胞表面标志鉴定发现,应用内皮细胞专用培养基培养7 d后细胞CD133及CD31表达阳性,随着培养时间的延长在14 d时CD34及Flk-1表达阳性。免疫荧光法验证结果显示,AQP1仅在AQP1 WT小鼠的EPC中表达阳性。对不同AQP1状态下EPC的功能研究结果显示,培养72 h的AQP1 WT小鼠与KO小鼠EPC细胞增殖数目无显著差异。Transwell检测结果显示,AQP1 KO小鼠其EPC迁移能力较WT小鼠明显减弱。细胞划痕实验检测结果显示,AQP1 KO小鼠EPC的划痕愈合能力也明显低于WT小鼠。结论 EPC最初表现出干细胞特征,随着培养时间延长,逐渐表现出内皮细胞特征。AQP1可影响EPC的迁移能力而非增殖能力。Objective To observe the effects of aquaporin 1（AQP1） on the proliferation and migration of endothelial progenitor-endothelial progenitor cells（EPC）. Methods Bone marrow cells of AQP1 wild-type（WT）（n=6）and knockout-type（KO） mice（n=6） were isolated and differentiated into EPC in vitro. Immunofluorescence was used to detect cell surface antigens to identify EPC. Live cell kinetic imaging and quantification technology, transwell migration assays, as well as scratch test were used to compare the function of EPC between AQP1 WT and KO mice. Results EPC culture showed that cells were initially suspended and gradually adhered to typical mesenchymal stem cells within 7 days.After cultured on special medium for endothelial cells they were adhered and differentiated, and fusiform or polygonal,paving stone-like EPC were observed around 14 days. When cultured by special medium of EPC, CD133 and CD31 were positively detected after 7 days, and CD34 and Flk-1 were positively detected after 14 days. Positive expression of AQP1 was only detected in EPC of AQP1 WT mice. Functional studies of EPC revealed there was no significant difference in the proliferation of EPC between AQP1 WT and KO group mice. Transwell assay showed that EPC migration ability of AQP1 KO mice was significantly weaker than that of WT mice. The scratch healing ability of EPC in AQP1 KO mice was significantly lower than that of WT mice. Conclusions EPC initially shows the characteristics of stem cells and with the prolongation of culture time, EPC gradually shows the characteristics of endothelial cells. AQP1 affects the EPC migration rather than proliferation.

关 键 词：水通道蛋白1 内皮祖细胞 细胞迁移 急性肺损伤 

分 类 号：R563[医药卫生—呼吸系统]