辛伐他汀对大鼠平滑肌祖细胞和内皮祖细胞黏附的不同影响

摘要

Objective: To investigate effects of simvastatin on adhesions of smooth muscle progenitor cells (SPC) and endothelial progenitor cells (EPC) and screen new compounds that could be used for coated stent. Methods: A single mononucicar cell (MNC) was isolated from marrow of rats by density gradient centrifugation. The cell was plated on SPC medium or EPC medium, then cultured on fibroneetin-coated culture dishes. Marrow-derived SPC were identified by smooth muscle actin (α-SMA) immunofluorcsccnt staining. Cells those were double positive for FITC-UEA -I and Dil-acLDL were identified as identified EPC by laser scanning confocal microscope. After cultured for 8d, SPC and EPC were collected respectively and different concentrations of simvastatin (0, 0. 01 , 0. 1 , 1 , 10 μmol/L) were added to them for 24h culture. Adhesive ability of SPC and EPC were measured by adhesion assay. Results: Simvastatin significantly inhibited adhesion of SPC, after 24h -interaction with 0. 01 μmol/L simvastatin, number of adhesive SPC significantly decreased compared with that of control group (0 μmol/L) [ (52 ± 4) vs. (59 ± 5), n=5, P＜0. 05]. On the contrary, simvastatin significantly increased adhesion of EPC, the promotion effect significantly increased along with increase of concentration of simvastatin and it reached maximum effect at the concentration of 1. 0μmol/L, number of adhesive EPC significantly increased compared with that of control group [ (39 ± 5) vs. (12 ± 3) , n=5, P＜0. 01]. Conclusion: Simvastatin inhibits adhesion of smooth muscle progenitor cells and promotes adhesion of endothelial progenitor cells. The double-way regulating effect is depended on concentration. Local application of simvastatin likely to promote re-endothclialization of lesion vessels and inhibit over hyperplasia of new intima.%目的:观察辛伐他汀对平滑肌祖细胞(SPC)和内皮祖细胞(EPC)黏附的影响,筛选新一代用于包被洗脱支架的药物.方法:采用密度梯度离心法获取大鼠骨髓单个核细胞,重悬于SPC培养基或EPC培养基,接种在纤维连接素包被的培养板,以平滑肌肌动蛋白免疫荧光染色鉴定骨髓源性SPC,以激光共聚焦显微镜鉴定的异硫氰酸荧光素-荆豆凝集素-Ⅰ(FITC-UEA-Ⅰ)和Dil标记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.分别收集培养8 d的SPC和EPC,加入不同浓度辛伐他汀(0,0.01,0.1,1,10 μmol/L)培养24 h.采用贴壁法检测SPC和EPC黏附能力.结果:辛伐他汀显著抑制SPC黏附,0.01 μmol/L辛伐他汀作用24h,黏附SPC数量较对照组(0μmol/L)明显减少[ (52±4)个∶(59±5) 个,n=5,P＜0.05].与SPC相反,辛伐他汀显著促进EPC黏附,其促进作用随辛伐他汀浓度升高而明显增加,1.0μmol/L达最大效应,与对照组相比显著增加[(39±5) 个∶(12±3) 个,n=5,P＜0.01].结论:辛伐他汀选择性抑制平滑肌祖细胞黏附,促进内皮祖细胞黏附,其双向调节作用呈浓度依赖性,局部应用有促进损伤血管再内皮化和抑制新内膜过度增生的可能.