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机构地区:[1]沈阳军区总医院干二科,辽宁沈阳110840 [2]沈阳军区总医院全军心血管研究所先心病内科,辽宁沈阳110840 [3]第三军医大学新桥医院全军心血管研究所
出 处:《心血管康复医学杂志》2016年第1期48-51,共4页Chinese Journal of Cardiovascular Rehabilitation Medicine
基 金:国家自然科学基金资助项目(30270568;30470729)~~
摘 要: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24h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。Objective: To observe influence of simvastatin on p27 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, a 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, 0.01/μmol/L simvastatin had no significant influence on p27 protein expression of SMC, but 0.1, i 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 p27 protein expression of endothelial cells. So local application of simvastatin may inhibit restenosis and promote reendothelialization of injured vessels.
关 键 词:平滑肌细胞 内皮细胞 周期素依赖激酶抑制剂P27 辛伐他汀
分 类 号:R543[医药卫生—心血管疾病]
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