不同浓度依达拉奉对低氧环境下c-kit^+心脏干细胞的影响  

Effects of different concentrations of edaravone on c-kit^+ cardiac stem cells under hypoxic conditions

作　　者：文德重[1] 周敏[1] 薛松[1] 黄日太[1] WEN Dezhong ZHOU Min XUE Song HUANG Ritai(Renji Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai 200127, China)

机构地区：[1]上海交通大学医学院附属仁济医院,上海200127

出　　处：《山东医药》2017年第17期10-12,16,共4页Shandong Medical Journal

基　　金：上海市自然科学基金资助项目(134119a5602)

摘　　要：目的观察不同浓度的依达拉奉对于低氧环境下c-kit^+心脏干细胞(CSCs)的影响。方法体外灌注酶解成年小鼠心脏,磁珠分选c-kit^+CSCs,流式细胞仪和免疫荧光染色分别检测分选后干细胞纯度和表型。将ckit^+CSCs培养基中加入不同浓度依达拉奉(0、100、300、500、700、1 000μmol/L),在1%氧条件下分别通过免疫荧光显微镜下DFCH-DA检测细胞活性氧(ROS)生产量、流式细胞仪检测凋亡率、Transwell检测细胞迁移能力、RTPCR检测成纤维细胞生长因子(bFGF)mRNA表达。结果依达拉奉浓度为1 000μmol/L时所产生的ROS量最少,与其他浓度比较有统计学差异(P均<0.05)。依达拉奉浓度为1 000μmol/L时细胞凋亡率最低,与0、100μmol/L浓度比较有统计学差异(P均<0.05)。依达拉奉浓度为700μmol/L时c-kit^+CSCs迁移数最大,与0μmol/L比较有统计学差异(P<0.05)。依达拉奉浓度为700μmol/L时,c-kit^+CSCs bFGF mRNA表达最高(P均<0.05)。结论依达拉奉通过影响细胞的凋亡率、迁移率、ROS生成和bFGF mRNA的表达对c-kit^+CSCs发挥保护作用,且在一定浓度范围内存在剂量依赖性。Objective To observe the effects of different concentrations of edaravone on c-kit＋ cardiac stem cells （CSCs） under hypoxic conditions. Methods The freshly isolated hearts were perfused with collagenase buffer, and the hearts were dissected into single cells, c-kit^ ＋ cells were purified from the cells by immunomagnetic beads. Flow cytometric analysis was used to measure the purity of c-kit^ ＋ cells ; immunofluorescence was used to analyze the phenotype. For further study, c-kit ＋ CSCs＇ were treated with different concentrations of edaravone （0, 100,300,500,700, 1 000 μmol/L）, and then were divided into six groups. The reactive oxygen species （ ROS ） production was measured by DFCH-DA, apoptosis rate was analyzed by flow cytometer, cell migration was measured by 8-mm pore-size Transwell migration chamber, and the expression of basic fibroblast growth factor （bFGF） mRNA was determined by RT-PCR. Results The ROS production in the 1 000 p, mol/L edavarone group was significantly decreased as compared with that of the other groups （all P 〈 0.05 ）. The apoptosis rate in the 1 000 μmol/L edavarone group was the lowest, and significant difference was found as compared with that in the 0 and 100 μmol/L edavarone groups （ all P 〈 0.05 ）. The migration of c-kit^＋ CSCs was the highest when edavarone concentration was 700 μmol/L, and significant difference was found as compared with that of the 0 μmol/L. When the concentration of edavarone was 700 μmol/L, the expression of bFGF mRNA in c-kit^＋ CSCs was the highest （ all P 〈 0.05）. Conclusion Edaravone has the protective effect on c-kit^＋ CSCs by influencing the apoptosis rate, migration rate, ROS production and bFGF mRNA expression with a dose-dependent manner to some extent.

关 键 词：心脏干细胞 依达拉奉 活性氧 细胞凋亡 细胞迁移 成纤维细胞生长因子 

分 类 号：R542.2[医药卫生—心血管疾病]