低氧预处理诱导骨髓间充质干细胞Pim-1激酶高表达抑制细胞凋亡

发布时间：2018-03-10 17:49

本文选题：骨髓　切入点：间质干细胞　出处：《中国组织工程研究》2016年14期 　论文类型：期刊论文

【摘要】：背景:骨髓间充质干细胞移植入缺血心肌后存活率低,而低氧有可能增强骨髓间充质干细胞的增殖,促进其存活。目的:探讨Pim-1激酶是否介导缺氧预处理对骨髓间充质干细胞的保护作用及其机制。方法:设置不同低氧处理时间(0,6,12,24 h)处理骨髓间充质干细胞,RT-qP CR及Western blot检测Pim-1及凋亡相关基因的表达,确定最佳低氧处理时间为12 h。将骨髓间充质干细胞分为3组:正常对照组、低氧组、低氧+Pim-1抑制剂组,分别进行Transwell迁移实验、细胞凋亡流式检测、线粒体膜电位检测评估各组骨髓间充质干细胞的迁移能力及抗凋亡能力;建立心肌梗死模型,1周后按分组在大鼠梗死心肌周围多点注射骨髓间充质干细胞,2周后制备心肌冰冻切片行DiI染色统计骨髓间充质干细胞存活数量;心肌梗死模型建立前后、细胞移植4周行超声心动图检查评估大鼠心脏功能。结果与结论:(1)低氧处理12 h时,骨髓间充质干细胞的Pim-1、p-Akt及Bcl-2表达量明显升高,Bax、Caspase-3表达量明显降低。(2)低氧组骨髓间充质干细胞抗凋亡能力明显增强,与正常对照组之间差异有显著性意义(P0.01)。(3)低氧组骨髓间充质干细胞移植1周后存活率明显提高(P0.001);移植4周后,大鼠心功能明显改善(P0.05)。上述获益可被Pim-1抑制剂抑制。(4)结果证实,低氧预处理骨髓间充质干细胞通过激活Akt及上调Pim-1表达抑制细胞凋亡,改善骨髓间充质干细胞对缺血性心脏病的治疗效果。
[Abstract]:Background: the survival rate of bone marrow mesenchymal stem cells after transplantation into ischemic myocardium is low, but hypoxia may enhance the proliferation of bone marrow mesenchymal stem cells. Objective: to investigate the protective effect of Pim-1 kinase mediated hypoxia preconditioning on bone marrow mesenchymal stem cells (BMSCs) and its mechanism. Methods: bone marrow mesenchymal stem cells were treated with RT-qP CR and RT QP CR for 24 h. Western blot was used to detect the expression of Pim-1 and apoptosis-related genes. The optimal hypoxia treatment time was 12 h. Bone marrow mesenchymal stem cells were divided into three groups: normal control group, hypoxic group and hypoxic Pim-1 inhibitor group. Transwell migration assay and apoptosis were detected by flow cytometry. The migration and anti-apoptosis ability of bone marrow mesenchymal stem cells were evaluated by mitochondrial membrane potential assay. One week after myocardial infarction model was established, myocardial frozen sections were prepared after multiple injection of bone marrow mesenchymal stem cells into the infarcted myocardium of rats for 2 weeks. DiI staining was used to calculate the number of bone marrow mesenchymal stem cells survival. Before and after the establishment of myocardial infarction model, the survival rate of bone marrow mesenchymal stem cells was calculated. After 4 weeks of transplantation, echocardiography was performed to evaluate cardiac function in rats. The expression of Pim-1p-Akt and Bcl-2 in bone marrow mesenchymal stem cells (BMSCs) increased significantly. The expression of Caspase-3 in Bax-Caspase-3 decreased significantly. The survival rate of bone marrow mesenchymal stem cells in hypoxic group increased significantly after 1 week of transplantation, and the cardiac function of rats improved significantly after 4 weeks of transplantation. The above benefits could be confirmed by the inhibition of Pim-1 inhibitor (P0. 05), and the difference between the two groups was significant (P 0. 01, P 0. 01, P 0. 01, P 0. 01, P 0. 01, P 0. 01, P 0. 01, P 0. 01, P 0. 01). Hypoxic preconditioning of bone marrow mesenchymal stem cells inhibited apoptosis by activating Akt and up-regulating the expression of Pim-1, thus improving the therapeutic effect of bone marrow mesenchymal stem cells on ischemic heart disease.
【作者单位】：苏州大学附属第一医院心内科;苏州大学附属第一医院心外科;苏州大学心血管病研究所;
【分类号】：R457.7

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