血管紧张素Ⅱ预处理提高骨髓间充质干细胞抗凋亡能力
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国家自然科学基金项目(30973534,8117 3052);南京医科大学科技发展基金(2013NJMU004)


AngiotensinⅡ preconditioning prevents mesenchymal stem cells from apoptosis
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    摘要:

    目的:研究血管紧张素Ⅱ(angiotensinⅡ,AngⅡ)预处理是否能提高骨髓间充质干细胞(mesenchymal stem cells,MSCs)的抗凋亡能力,探讨AngⅡ对MSCs预适应保护作用的可能机制-方法:用AngⅡ预处理大鼠MSCs细胞,采用H2O2或缺氧联合无血清(hypoxia and serum deprivation,Hypoxia/SD)的方法诱导细胞凋亡-用乳酸脱氢酶(lactate dehyarogenase,LDH)检测法-四唑盐比色法(MTT)和Hoechst 33342染色测定细胞活力和凋亡率;用Annexin V-FITC法定量检测细胞凋亡率;对MSCs进行罗丹明-123染色,用流式细胞术检测线粒体跨膜电位-结果:AngⅡ预处理MSCs能够明显减轻H2O2或Hypoxia/SD诱导的细胞损伤,显著提高细胞活力;逆转由Hypoxia/SD引起的线粒体跨膜电位的降低;ERK1/2阻断剂U0126和Akt阻断剂LY294002都能明显减弱AngⅡ对MSCs的预适应保护作用,增加细胞凋亡率-结论:AngⅡ对MSCs具有预适应保护作用,且主要通过Akt-ERK1/2途径发挥其抗凋亡能力-

    Abstract:

    Objective:To examine whether AngⅡ preconditioning improves the anti-apoptotic ability of bone marrow mesenchymal stem cells (MSCs),and if so,what is its possible mechanism. Methods:MSCs cells from rats were pretreated with Ang Ⅱ , and apoptosis of MSCs was induced by H2O2 or hypoxia combined serum deprivation (Hypoxia/SD) method. Cell viability and apoptotic proportion were detected by lactate dehyarogenase (LDH) release, methye thiazolye telrazlium (MTT) assays and Hoechst 33342 staining. Annexin V-FITC assay was performed for quantification of apoptosis rate. The mitochondrial transmembrane potentia (ΔΨm) was detected by a flow cytometer using ΔΨm specific stain Rhodamine 123. Results:AngⅡ preconditioning significantly reduced H2O2- or Hypoxia/SD-induced cell damage,increased cell viability,reversed Hypoxia/SD-induced mitochondrial membrane potential reduction. Both of ERK1/2 inhibitor U0126 and Akt inhibitor LY294002 significantly attenuated the the protective effects of AngⅡ preconditioning on the viability and apoptosis rate of MSCs. Conclusion: AngⅡ preconditioning prevents MSCs from apoptosis,and Akt and ERK1/2 signalling pathway may be implicated in the effect of AngⅡ.

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宋奕辰,胡 亮,周 露,范 越,黄松花,施睿臻,李庆平.血管紧张素Ⅱ预处理提高骨髓间充质干细胞抗凋亡能力[J].南京医科大学学报(自然科学版),2014,(9):1157-1162

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  • 收稿日期:2014-04-23
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  • 在线发布日期: 2014-09-11
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