雷帕霉素对内皮祖细胞增殖、黏附及迁移能力影响的实验研究  

Sirolimus inhibits the differentiation,proliferation and migration of endothelial progenitor cells in vitro

作　　者：张坡[1] 黄岚[1] 朱光旭[1] 崔斌[1] 宋明宝[1] 周音频[1] 赵晓晖[1] 尹阳光[1] 

机构地区：[1]第三军医大学新桥医院全军心血管内科中心,重庆400037

出　　处：《中华心血管病杂志》2006年第11期1021-1025,共5页Chinese Journal of Cardiology

基　　金：国家自然科学基金资助项目(30270568;30470729)

摘　　要：目的药物洗脱支架可抑制支架处血管内膜再内皮化,但对骨髓源性内皮修复作用的影响尚不清楚,为此观察了雷帕霉素对内皮祖细胞(endothelial progenitor cells,EPC)增殖、黏附及迁移的影响。方法采用密度梯度离心法从大鼠骨髓获取单个核细胞,将其接种在纤维连接素包被培养板,加入不同浓度雷帕霉素(0.01～100ng/ml),培养12天后,激光共聚焦显微镜鉴定FITC-UEA-Ⅰ和DiI-acLDL双染阳性细胞为正在分化的EPC,并在倒置荧光显微镜下计数。收集培养8天贴壁细胞,分别加入不同浓度雷帕霉素(0.1～200ng/ml)培养12～96h。然后分别采用MTT比色法、改良的Boyden小室和黏附能力测定实验观察EPC的增殖、迁移和黏附能力。结果雷帕霉素显著抑制骨髓单个核细胞分化为EPC,0.1ng/ml浓度雷帕霉素作用12天,EPC减少了(59.3±5.8)%(P<0.01)。雷帕霉素也显著抑制EPC增殖,其抑制作用随雷帕霉素浓度增加及作用时间延长而增加,1ng/ml浓度雷帕霉素作用24h使EPC数量减少(41.5±2.2)%(P<0.01)。雷帕霉素也显著抑制EPC的黏附和迁移能力。结论雷帕霉素可抑制骨髓单个核细胞分化为EPC,并抑制EPC的增殖及迁移能力。Objective To investigate the effect of sirolimus on differentiation, proliferation, adhesion and migration of endothelial progenitor cells （ EPC ） in vitro. Methods （ 1 ） Mononuclear cells （MNC） were isolated from rat bone marrow by Ficoll density gradient centrifugation and cultured on fibronectin-coated culture dishes with or without sirolimus （0. 01-100 ng/ml） for 12 days. （2） After 8 days cultured, attached cells were treated with sirolimus （0. 1-200 ng/ml） or vehicle for various time points （ 12 h, 24 h,48 h and 96 h）. EPC were identified as adherent cells double positive stained for FITC-UEA- Ⅰ and DiI-acLDL under laser confocal immunofluence microscope. EPC proliferation, migration were assayed with MTT assay and modified Boyden chamber assay respectively. Results EPC number differentiated from MNC at 12 days was significantly lower in sirolimus treated cells in a dose-dependent manner than that of vehicle- treated cells. Sirolimus also significantly inhibited the proliferative , migratory and adhesive capacity of EPC in a time and dose dependent manner. Conclusion Present results suggested that sirolimus could inhibit EPC differentiation from MNC and reduce the proliferation, migration and adhesion capacities of EPC.

关 键 词：干细胞 西罗莫司 细胞分化 细胞运动 

分 类 号：R541.4[医药卫生—心血管疾病]