人血红素加氧酶1过表达大鼠骨髓间充质干细胞对缺血缺氧环境损伤的耐受能力

发布时间：2019-03-16 18:42

【摘要】：背景:骨髓间充质干细胞移植后在宿主恶劣的缺血缺氧微环境中生存率过低,限制了其治疗急性心肌梗死的疗效。目的:探讨人血红素加氧酶1基因修饰的大鼠骨髓间充质干细胞对缺血缺氧环境损伤的耐受能力。方法:人血红素加氧酶1重组腺病毒转染大鼠骨髓间充质干细胞,Western blot法确定人血红素加氧酶1蛋白表达的最佳时间;以缺氧无血清条件模拟体内缺血缺氧微环境培养人血红素加氧酶1基因修饰的大鼠骨髓间充质干细胞,采用CCK-8试剂盒和锥虫蓝染色法检测缺血缺氧培养12,24,48,72 h的细胞存活率,流式细胞术检测缺血缺氧培养24 h的细胞凋亡情况。结果与结论:①在转染后第4天人血红素加氧酶1蛋白表达量最多;②锥虫蓝染色法和CCK-8法检测人血红素加氧酶1组缺血缺氧培养12,24,48,72 h的细胞生存率均高于对照组(P0.05);③流式细胞术检测人血红素加氧酶1组缺血缺氧培养24 h的生存率高于对照组(P0.05);④结果表明,人血红素加氧酶1修饰的大鼠骨髓间充质干细胞对缺血缺氧环境损伤的耐受能力明显增强。
[Abstract]:Background: the survival rate of bone marrow mesenchymal stem cells (MSCs) after transplantation in poor ischemic and anoxic microenvironment of the host is too low, which limits the efficacy of MSCs in the treatment of acute myocardial infarction (AMI). Aim: to investigate the tolerance of rat bone marrow mesenchymal stem cells (BMSCs) modified with human heme oxygenase-1 gene to hypoxic-ischemic injury. Methods: human heme oxygenase 1 recombinant adenovirus was transfected into rat bone marrow mesenchymal stem cells (BMSCs) by, Western blot assay to determine the optimal time for the expression of human heme oxygenase 1 protein. Human heme oxygenase 1 gene modified rat bone marrow mesenchymal stem cells (BMSCs) were cultured under hypoxia-free condition and simulated in vivo. The survival rate of MSCs was detected by CCK-8 kit and trypanosome blue staining at 12,24,48and 72 h after ischemia and hypoxia, and the survival rate of MSCs cultured with heme oxygenase-1 gene was determined by trypanosome blue staining method. Flow cytometry (FCM) was used to detect the apoptosis of the cells cultured under ischemia and hypoxia for 24 h. Results and conclusion: 1 on the 4th day after transfection, the expression of human heme oxygenase-1 protein was the highest; 2Trypanosome blue staining and CCK-8 assay were used to detect the cell survival rate of 12, 24, 48, 72 h culture in heme oxygenase-1 group (P0.05), and the cell survival rate was significantly higher than that in control group (P0.05). 3The 24-hour survival rate of heme oxygenase-1 group was significantly higher than that of control group (P0.05), and the survival rate was significantly higher than that of control group (P0.05). 4. The results showed that human heme oxygenase 1-modified rat bone marrow mesenchymal stem cells (BMSCs) significantly enhanced their tolerance to ischemia and hypoxia injury in rat bone marrow mesenchymal stem cells (BMSCs).
【作者单位】：广东省中医院珠海医院;解放军第305医院;郴州市第一人民医院;
【基金】：国家自然科学基金(30971235) 军队“十二五”医学科研基金(2013XL010)~~
【分类号】：R542.22

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