摘要
背景:研究发现,阿托伐他汀具有心血管保护作用,能显著改善内皮功能,促进内皮祖细胞的动员、迁移和分化。现阶段在细胞体外培养水平上进行阿托伐他汀药物浓度筛选所进行的研究较少。目的:探讨不同浓度阿托伐他汀对大鼠骨髓来源内皮祖细胞体外生长特性的影响。方法:取SD大鼠长骨骨髓,用密度梯度离心法结合差异贴壁法提取单核细胞,用选择性培养液诱导培养内皮祖细胞,采用免疫荧光染色法鉴定细胞表面标志物。将内皮祖细胞分为对照组和4个不同浓度阿托伐他汀组(0.01,0.1,1,10μmol/L)进行培养,光镜下观察和MTT法检测细胞生长增殖情况,流式细胞术检测细胞凋亡率,硝酸还原酶法化学比色法检测培养液中一氧化氮和内皮型一氧化氮合酶水平。结果与结论:(1)对照组和阿托伐他汀组的细胞数量均表现为增长趋势,1μmol/L阿托伐他汀组增长最明显,10μmol/L阿托伐他汀组在7 d后出现下降趋势;(2)1μmol/L阿托伐他汀组凋亡率低于其他各组,10μmol/L阿托伐他汀组凋亡率高于其他各组;(3)0.01,0.1,1.0μmol/L阿托伐他汀组一氧化氮和内皮型一氧化氮合酶水平明显高于空白对照组(P<0.01),10μmol/L阿托伐他汀组一氧化氮和内皮型一氧化氮合酶水平明显低于其他各组(P<0.01);(4)结果表明,阿托伐他汀可通过增加内皮型一氧化氮合酶和一氧化氮的生成,促进内皮祖细胞增殖并减少凋亡,其中1μmol/L浓度的阿托伐他汀最适合内皮祖细胞培养。
BACKGROUND： Atorvastatin has a cardiovascular protective effect that significantly improves endothelial function and promotes the mobilization,migration,and differentiation of endothelial progenitor cells.However,the screening of atorvastatin concentration for in vitro cell culture is not well documented. OBJECTIVE： To investigate the effects of different concentrations of atorvastatin on rat bone marrow-derived EPCs growth characteristics. METHODS： Bone marrow mononuclear cells from Sprague-Dawley rats were induced in selective culture fluid to culture EPCs. Immunofluorescence staining was used to identify cell surface markers. Harvested EPCs were divided into control group and atorvastatin groups with four different concentrations （0.01, 0.1, 1, and 10 μmol/L） for culture. The growth and proliferation of EPCs were observed under light microscope and MTT assay. Flow cytometry was used to detect apoptosis in EPCs. Nitric oxide and endothelial nitric oxide synthase levels in the culture fluid were measured by nitrate reductase method. RESULTS AND CONCLUSION： The number of cells tended to increase in the control and atorvastatin groups, and it was highest in the 1 μmol/L atorvastatin group. The cell number in the 10 μmol/L atorvastatin group began to decrease at 7 days of culture. Among the five groups, the apoptotic rate of cells was lowest in the 1 μmol/L atorvastatin group and highest in the 10 μmol/L atorvastatin group. The levels of nitric oxide and endothelial nitric oxide synthase were significantly higher in the 0.01, 0.1 and 1.0 μmol/L atorvastatin groups compared with the control group （P 〈 0.01）, but lower in the 10 μmol/L atorvastatin group compare with the other groups （P 〈 0.01）. Overall, atorvastatin can promote the proliferation of endothelial progenitor cells and reduce apoptosis by increasing the production of endothelial nitric oxide synthase and nitric oxide, and 1 μmol/L atorvastatin is most suitable for the EPCs culture.
作者
张日霖
陈淑玲
李上海
宁奕明
李庆军
叶小敏
梁伟均
Zhang Ri-lin;Chen Shu-ling;Li Shang-hai;Ning Yi-ming;Li Qing-jun;Ye Xiao-min;Liang Wei-jun(Central People’s Hospital of Zhanjiang, Zhanjiang 524000, Guangdong Province, China;Affiliated Hospital of Guangdong Medical University, Zhanjiang 524000, Guangdong Province, China;Maternal and Child Health Hospital of Wuchuan, Wuchuan 524500, Guangdong Province, China)
出处
《中国组织工程研究》
CAS
北大核心
2018年第13期1976-1980,共5页
Chinese Journal of Tissue Engineering Research
基金
广东省医学科学技术研究基金(A2017494)
广东省科技计划项目(2009B030801337)~~
