辛伐他汀对大鼠平滑肌祖细胞和内皮祖细胞增殖的影响  

Different effects of simvastatin on proliferation of rat smooth muscle progenitor cells versus endothelial progenitor cells

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

机构地区：[1]第三军医大学新桥医院全军心血管研究所,重庆市400037

出　　处：《中华老年医学杂志》2008年第9期702-705,共4页Chinese Journal of Geriatrics

基　　金：国家自然科学基金资助项目（30270568和30470729）

摘　　要：目的观察辛伐他汀对平滑肌祖细胞（SPC）和内皮祖细胞（EPC）增殖的影响，筛选新一代包被洗脱支架药物。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，加入不同浓度辛伐他汀（0．01～10．00μmol／L）培养6～48h后，平滑肌肌动脉蛋白免疫荧光染色鉴定骨髓源性SPC，激光共聚焦显微镜鉴定异硫氰酸荧光素结合的植物凝集素（FITC-UEA-I）和DiI结合的乙酰化低密度脂蛋白（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。结果辛伐他汀显著抑制骨髓源SPC增殖，0．01μmol／L辛伐他汀作用24h，SPC数量减少了（5．8±3．1）％（对照组与0．01μmol／L辛伐他汀组分别为4070±184与3833±126，P〈0．05）。辛伐他汀可促进EPC增殖，其促进作用随辛伐他汀浓度增加及作用时间延长而增加，1．00μmol／L辛伐他汀作用24hEPC数量增加（2．0±0．1）倍（对照组与1μmol／L辛伐他汀组分别为1249±146与3762±138，P〈0．01）。结论辛伐他汀抑制SPC增殖，促进EPC增殖，局部应用有抑制再狭窄和促进损伤血管再内皮化可能。Objective To investigate the different influences of simvastatin on proliferation of rat smooth muscle progenitor cells（SPCs） versus endothelial progenitor cells （EPCs） and identify the compounds that differentially inhibit SPCs and EPCs proliferation for clinical usefulness. Methods Total mononuclear cells （MNCs） were isolated from bone marrow of rats by Ficoll density gradient centrifugation, and then the cells were plated on fibronectin-coated culture dishes. SPCs outgrew from the culture of MNCs in the presence of platelet-derived growth factor-BB and basic fibroblast growth factor, whereas EPCs were obtained in the presence of vascular endothelial growth factor. SPCs were identified as adherent cells positive for α-smooth muscle actin （α-SMA） by indirect immunofluorescent staining. EPCs were characterized as adherent cells double positive for DiLDL-uptake and lectin binding by direct fluorescent staining. SPCs and EPCs were stimulated by simvastatin （0.01--10.00 μmol/L） or vehicle control for the respective time points （6 h, 12 h, 24 h and 48 h）. SPCs and EPCs proliferation were assayed with ^3 H-TdR incorporation and manual counting respectively. Results Simvastatin obviously inhibited SPCs proliferation. At the concentration of 0.01 μmol/L for 12 h, simvastatin significantly reduced the number of SPCs by （5.8 ± 3.1）% compared with control group （P〈0.05）. Simvastatin significantly stimulated EPCs proliferation, which was dose- and timedependent and reached maximum at 1μmol/L after 24 hours （2.0±0.1 fold increase, P〈0. 01）. Conclusions Simvastatin displays different effects on SPCs （inhibited） and EPCs （promoted） proliferation. Local application of simvastatin may inhibit arterial restenosis and promote reendothelialization of injured vessels.

关 键 词：辛伐他汀 肌细胞 平滑肌 内皮细胞 细胞增殖 

分 类 号：R686[医药卫生—骨科学]