摘要
本研究旨在探讨细胞间粘附分子-1(ICAM-1)在小鼠间充质干细胞(MSC)体外迁移中的作用及其相关机制。采用transwell法研究小鼠MSC(C3H10T1/2)、转染空载体的小鼠MSC(C3H10T1/2-MIGR1)和转染了ICAM-1的小鼠MSC(C2H10T1/2-ICAM-1)的体外迁移能力,即将各组MSC种植在孔径为8μm的通透膜上面,以胎牛血清作为趋化物质诱导MSC迁移。分别在迁移8 h和12 h后对膜下的MSC进行结晶紫和荧光染料DAPI染色,然后统计各组的MSC的细胞数和迁移率。为了探究调控MSC迁移的机制,丝裂原活化蛋白激酶通路的抑制剂(SB203580,PD98059和JNK inhibitor II)被添加到transwell体系中,并进一步观察MSC迁移能力的改变。结果表明:3种细胞8 h和12 h的transwell体外迁移结果显示,转染了ICAM-1的MSC组迁移的细胞数和迁移率均显著高于未转染组MSC和转染空载体组MSC(P<0.05),未转染组MSC和转染空载体组MSC之间无统计学差异(P>0.05);加入JNK/SAPK通路的抑制剂JNK inhibitor II可以抑制ICAM-1对MSC的促迁移作用,无论是迁移的细胞数还是迁移率均显著降低(P<0.05)。本研究中p38/MAPK通路的抑制剂SB203580和ERK/MAPK通路的抑制剂PD98059对于ICAM-1的促MSC迁移作用无显著影响。结论:ICAM-1可增强小鼠MSC的体外迁移能力,这种促进作用部分依赖于活化JNK/SAPK通路实现。
This study was aimed to investigate the effect of intercellular adhesion molecule-1 （ICAM-1） on the mi- gration in vitro of the murine mesenchymal stem cells （MSC） and its related mechanisms. The migration ability of mu- fine MSC （C3H10T1/2）, ICAM-1 transfected MSC （C3H10T1/2-MIGRI-ICAM-1） and empty vector-transfected MSC （C3H10T 1/2-MIGR1 ） were assayed in vitro by using the transwell system. Briefly, the cells were seeded on the membrane with 8μm aperture and the fetal bovine serum was used as the chemotactic agent to induce MSC migration.The transmigrated cells were stained by crystal purple as well as DAPI for 8 h and 12 h respectively. The absolute cell numbers were counted and the migration rates of MSC were evaluated in each group. To explore the potential mecha- nisms which control the migration of MSC, the specific chemical inhibitors of MAPK pathway （ SB203580, PD98059 and JNK inhibitor Ⅱ） were added to the transwell system and the alteration of the MSC migration ability were evaluated at 12 h. The results showed that the migration ability at 8 h and 12 h of the ICAM-1-transfected MSC increased. Both absolute cell number and migration rate of MSC were significantly up-regulated by ICAM-1. Furthermore, the promoting effect of ICAM-1 on migration was partially suppressed by the inhibition of JNK/SAPK pathway. The transmigrated cell numbers and the migration rate decreased with the addition of JNK inhibitor Ⅱ. However, the ICAM-1 promoting migra- tion of MSC was not suppressed by the inhibitors for ERK/MAPK and p38/MAPK pathway did not work in the present study. It is concluded that ICAM-1 can induce mouse MSC migration in vitro, and the promoting effect is partially de- pendent on the activation of JNK/SAPK pathway.
出处
《中国实验血液学杂志》
CAS
CSCD
北大核心
2014年第2期479-484,共6页
Journal of Experimental Hematology
基金
国家重点基础研究计划(973项目2010CB833600)
国家自然科学基金面上项目(81371945
31070996
31171084)
国家自然科学基金青年项目(81101342)
北京市自然科学基金面上项目(7132133)
关键词
细胞间粘附分子-1
间充质干细胞
体外迁移
intercellular adhesion molecule-1
mouse mesenchymal stem cells
migration in vitro
