辛伐他汀对大鼠血管平滑肌细胞和内皮细胞 p27蛋白表达的影响

摘要

Objective:To observe influence of simvastatin on p 27 protein (cyclin‐dependent kinase inhibitor ) expres‐sions of vascular smooth muscle cells (SMC) and endothelial cells (EC) in rats for screening new generation coating drugs of eluting stents .Methods :Primary aortic SMC and EC of rat were isolated and cultured by methods of adher‐ent and enzymatic digestion respectively .Which were inoculated on fibronectin -coated culture plates .α smooth muscle actin immunofluorescence staining was used to identify SMC ,and von Willebrand factor (vWF) immunofluo‐rescence staining was used to identify EC .SMC and EC were cultured for 24h with different concentrations of simv‐astatin (0.01 ,0.1 ,1 and 10 μmol/L) ,then Western blot was used to measure p27 protein expression .Results:Compared with blank control group ,0.01μmol/L simvastatin had no significant influence on p 27 protein expression of SMC ,but 0.1 ,1 and 10 μmol/L simvastatin significantly raised p27 protein expression of SMC [ (0.53 ± 0.08) vs .(0.86 ± 0.05) ,(1.20 ± 0.05) ,(1.60 ± 0.04)] , P< 0.01 all .Besides ,different concentrations of simvastatin had no significant influence on p27 protein expression of EC , P> 0.05 ,indicating that simvastatin only dose‐de‐pendently promoted p27 protein expression of SMC .Conclusion:Simvastatin dose -dependently promotes p27 pro‐tein expression of vascular smooth muscle cells without affecting p 27 protein expression of endothelial cells .So local application of simvastatin may inhibit restenosis and promote reendothelialization of injured vessels .%目的：观察辛伐他汀对大鼠血管平滑肌细胞（SMC）和内皮细胞（EC） p27蛋白（细胞周期依赖激酶抑制剂）表达的影响，筛选新一代包被洗脱支架药物。方法：原代大鼠主动脉SMC和EC细胞被分离和采用贴壁法和胰酶消化法培养，将其接种在纤维连接素包被培养板，以α平滑肌肌动蛋白免疫荧光染色法鉴定SMC ，血管假性血友病因子（vWF）免疫荧光染色法鉴定EC ，加入不同浓度辛伐他汀（0.01，0.1，1，10μmol／L）培养24 h后，以Western blot检测p27蛋白的表达。结果：与空白对照组比较，0.01μmol／L辛伐他汀对SMC细胞的p27蛋白表达无显著影响，而0.1，1和10μmol／L的辛伐他汀均显著增加SMC细胞的p27蛋白表达[（0.53±0.08）比（0.86±0.05），（1.20±0.05），（1.60±0.04）]， P均＜0.01；但各浓度组辛伐他汀均不影响EC细胞 p27蛋白的表达，表明辛伐他汀浓度依赖性促进SMC细胞p27蛋白表达，而不影响EC细胞p27蛋白的表达（ P＞0.05）。结论：辛伐他汀浓度依赖性地促进血管平滑肌细胞p27蛋白表达，而不影响内皮细胞p27蛋白表达，局部应用有抑制再狭窄和促进损伤血管再内皮化的可能。