E-钙黏蛋白对血管内皮祖细胞和骨髓间充质干细胞之间的黏附作用研究  

Study on the adhesion of E-cadherin between endothelial progenitor cells and bone marrow mesenchymal stem cells

作　　者：夏杰 唐紫萌 吴向未[3] Xia Jie;Tang Zimeng;Wu Xiangwei(Department of General Surgery,Ankang Central Hospital,Ankang,Shaanxi 725000,China;Department of laboratory medicine,Ankang Hospital of Traditional Chinese Medicine,Ankang,Shaanxi 725000,China;School of Medicine,Shihezi University,Shihezi,Xinjiang 832000,China)

机构地区：[1]陕西省安康市中心医院普外科,安康725000 [2]陕西省安康市中医医院检验科,安康725000 [3]新疆石河子大学医学院,石河子832000

出　　处：《中华细胞与干细胞杂志（电子版）》2021年第6期343-350,共8页Chinese Journal of Cell and Stem Cell（Electronic Edition）

基　　金：国家自然科学基金(81760570);安康市科学技术研究发展指导计划项目(AK2020-SFZC-14)。

摘　　要：目的初步探讨E-钙黏蛋白(E-cad)在内皮祖细胞(EPCs)和间充质干细胞(MSCs)之间的黏附作用。方法体外C57BL/6J小鼠骨髓腔分离培养EPCs和MSCs,使用Mito Tracker Red标记EPCs,DAPI标记MSCs;采用免疫荧光的方法观察E-cad在细胞内的表达情况;细胞黏附实验观察EPCs与MSCs体外黏附现象;使用E-cad siRNA干扰EPCs和MSCs中E-cad的表达,RT-qPCR和Western blot检测siRNA转染效率;转染成功后在成血管的凝胶上进一步观察二者黏附现象。多组间比较采用单因素方差分析,组间两两比较使用LSD-t检验。结果(1)与MSCs、EPCs比较,体外共培养MSCs和EPCs上清液中E-cad表达(378.26±34.47、564.72±41.58比1087.28±101.92)升高,差异均具有统计学意义(P<0.05);(2)与Control-siRNA、Mock、Negative相比,E-cad siRNA转染MSCs 48 h后E-cad mRNA表达(0.97±0.07、0.93±0.06、1.00±0.03比0.30±0.05)、E-cad蛋白表达(252.19±11.62、223.70±13.66、257.50±12.08比31.74±4.08)均降低,差异有统计学意义(P<0.05);与Control-siRNA、Mock、Negative比较,E-cad siRNA转染EPCs 48 h后E-cad mRNA表达(0.93±0.07、0.88±0.08、1.00±0.02比0.36±0.07)、E-cad蛋白表达(429.46±31.87、409.13±26.97、436.80±34.69比54.03±15.05)均降低,差异有统计学意义(P<0.05);(3)E-cad siRNA成功转染MSCs和EPCs后,二者黏附现象受到抑制;(4)沉默EPCs和MSCs的E-cad后,二者在成血管凝胶上的黏附及成管现象均受到抑制。结论E-cad介导EPCs和MSCs之间的黏附。Objective To explore the adhesion of E-cadherin(E-cad)between endothelial progenitor cells(EPCs)and mesenchymal stem cells(MSCs).Methods EPCs and MSCs were isolated and cultured from the bone marrow cavity of C57BL/6Jmice.Use Mito Tracker Red to label EPCs and DAPI to label MSCs.Immunofluorescence was used to examine the E-cad expression of the two cell lines.Cell adhesion assay was used to observe the adhesion of EPCs and MSCs in vitro.E-cad expression in EPCs and MSCs was silenced using E-cad siRNA,and siRNA transfection efficiency was determined by RT qPCR and Western blot.After successful transfection,the adhesion of EPCs and MSCs on the angiogenic Matrigel was observed.One way ANOVA was used for comparison between multiple groups,and LSD-t test was used for pairwise comparison between groups.Results(1)Compared with MSCs or EPCs,the expression of E-cad in the supernatant of the co-cultured of MSCs and EPCs in vitro(378.26±34.47,564.72±41.58 vs 1087.28±101.92)was increased,the difference was statistically significant(P<0.05).(2)Compared with ControlsiRNA,Mock and Negative,the expression of E-cad mRNA(0.97±0.07,0.93±0.06,1.00±0.03 vs 0.30±0.05)and E-cad protein(252.19±11.62,223.70±13.66,257.50±12.08 vs 31.74±4.08)of MSCs transfected with siRNA for 48 hours were decreased,the difference was statistically significant(P<0.05);Compared with Control-siRNA,Mock and Negative,the expression of E-cad mRNA(0.93±0.07、0.88±0.08、1.00±0.02 vs 0.36±0.07)and E-cad protein(429.46±31.87、409.13±26.97、436.80±34.69 vs 54.03±15.05)of EPCs transfected with siRNA for 48 hours were decreased,the difference was statistically significant(P<0.05).(3)The adhesion of MSCs and EPCs were significantly inhibited after successful transfection of E-cad siRNA.(4)Adhesion and tube formation were significantly inhibited in the angiogenic Matrigel after E-cad of EPCs and MSCs was silenced.Conclusions E-cad could mediate the adhesion between EPCs and MSCs.

关 键 词：E-钙黏蛋白 血管内皮祖细胞 骨髓间充质干细胞 细胞黏附 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]