Guo Jun, Xia Jie, Zhang Hong-Wei, Wang Xiao-Yi, Hou Ji-Xue, Chen Xue-Ling, Wu Xiang-Wei
School of Medical Sciences, Shihezi University, Shihezi 832008, Xinjiang Uygur Autonomous Region, China.
Department of General Surgery, The First Affiliated Hospital, School of Medical Sciences, Shihezi University, Shihezi 832008, Xinjiang Uygur Autonomous Region, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2016 Feb;24(1):211-6. doi: 10.7534/j.issn.1009-2137.2016.01.039.
To investigate the effects of intercellular adhesion molecule-1(ICAM-1) on the adherence between mesenchymal stem cells (MSC) and endothelial progenitor cells (EPC).
MSC and EPC were isolated, cultured and expanded from the 6-8 weeks aged C57BL/6 murine bone marrow by in vitro. Immuno-fluorescence was used to detect the expression of ICAM-1 in MSC group, EPC group and co-cultured MSC and EPC group. The mRNA and protein levels of ICAM-1 were detected by RT-PCR and Western blot respectively, then, the ICAM-1 adherence between MSC and EPC was observed by adding different concentration of neutralizing antibody.
The expression of ICAM-1 on surface of MSC and EPC could be detected by cell immunofluorescence method. According to results of the semiquantitative fluorescene detection, the fluorescence strength of MSC+EPC co-cultured group (89.02 ± 24.52) was higher than that of MSC group (31.25 ± 2.95) and EPC group (34.32 ± 5.02), and there was statistical difference between them (P < 0.01), but there was no obvious difference between MSC group and EPC group (P > 0.05). RT-PCR detection showed that the expression levels of ICAM-1 in MSC+EPC co-cultured group were higher than that in MSC group and that in EPC group (P < 0.01), and expression level of ICAM in EPC group was higher than that in MSC group (P < 0.01). Western blot detection showed that the expression level of ICAM-1 protein in MSC+EPC co-cultured group (0.33 ± 0.4) was higher than that in MSC group (0.11 ± 0.01) (P < 0.05) and than that in EPC group (0.19+0.02) (P < 0.05), However, the expression level of ICAM-1 protein in EPC group was higher than that in MSC group (P < 0.05). The test of different concentrations against neutralizing antibody showed that with the increasing of concentration of ICAM-1 neutralizing antibody, the adhesion capability of MSC and EPC was gradually decreasing.
The ICAM-1 can mediate the adherence process between MSC and EPC.
研究细胞间黏附分子-1（ICAM-1）对间充质干细胞（MSC）与内皮祖细胞（EPC）黏附的影响。
体外从6-8周龄C57BL/6小鼠骨髓中分离、培养并扩增MSC和EPC。采用免疫荧光法检测MSC组、EPC组及共培养的MSC与EPC组中ICAM-1的表达。分别用RT-PCR和蛋白质免疫印迹法检测ICAM-1的mRNA和蛋白水平，然后通过加入不同浓度的中和抗体观察MSC与EPC之间ICAM-1介导的黏附情况。
细胞免疫荧光法可检测到MSC和EPC表面ICAM-1的表达。根据半定量荧光检测结果，MSC+EPC共培养组的荧光强度（89.02±24.52）高于MSC组（31.25±2.95）和EPC组（34.32±5.02），差异有统计学意义（P<0.01），但MSC组与EPC组之间无明显差异（P>0.05）。RT-PCR检测显示，MSC+EPC共培养组ICAM-1的表达水平高于MSC组和EPC组（P<0.01），EPC组ICAM的表达水平高于MSC组（P<0.01）。蛋白质免疫印迹法检测显示，MSC+EPC共培养组ICAM-1蛋白表达水平（0.33±0.4）高于MSC组（0.11±0.01）（P<0.05）和EPC组（0.19+0.02）（P<0.05），然而，EPC组ICAM-1蛋白表达水平高于MSC组（P<0.05）。不同浓度中和抗体试验显示，随着ICAM-1中和抗体浓度的增加，MSC与EPC的黏附能力逐渐降低。
ICAM-1可介导MSC与EPC之间的黏附过程。
