Bone marrow mesenchymal stem cell treatment for myocardial ischemia/reperfusion injury: mechanisms of exosomes and factors

doi:10.3969/j.issn.2095-4344.1599

Abstract

Abstract:

BACKGROUND: Myocardial ischemia/reperfusion injury is one of the most common complications in ischemic cardiomyopathy and open heart surgery. The development of bone marrow mesenchymal stem cells provides a new method for clinical prevention and treatment of myocardial ischemia/reperfusion injury.
OBJECTIVE: To review the therapeutic effect and potential mechanisms of bone marrow mesenchymal stem cells in the treatment of myocardial ischemia/reperfusion injury, in order to provide a theoretical basis for the clinical application of bone marrow mesenchymal stem cells.
METHODS: Chinese Journal Full-text Database (CNKI), WanFang, and PubMed were retrieved for articles related to the use of bone marrow mesenchymal stem cells for myocardial ischemia-reperfusion injury published from January 2000 to October 2018. The search terms were “bone marrow mesenchymal stem cells; myocardial ischaemia/reperfusion; research process” in Chinese and “bone marrow mesenchymal stem cells; myocardial ischaemia/reperfusion; cell therapy; clinical trial studies” in English. Old and repetitive viewpoints were excluded, the searched literatures were sorted out, and finally 56 articles were included for further analysis and discussion.
RESULTS AND CONCLUSION: (1) In this paper, we summarize paracrine factors, exosomes miRNA and their effects in the treatment of myocardial ischemia/reperfusion injury with bone marrow mesenchymal stem cells, such as anti-inflammation, anti-apoptosis, anti-fibrosis, repair of myocardium and neovascularization. (2) We also summarize the possible molecular mechanisms of bone marrow mesenchymal stem cells involved in the treatment of myocardial ischemia/reperfusion injury, such as the role of mitochondrial fusion protein 2, regulation of myocardial autophagy, and regulation of AMPK/mTOR signaling pathway. Overall, we attempt to provide a theoretical basis for the clinical application of bone marrow mesenchymal stem cells in the treatment of myocardial ischemia/reperfusion injury.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Bone Marrow, Mesenchymal Stem Cells, Myocardial Ischemia, Reperfusion Injury, Tissue Engineering

CLC Number:

Zhang Changjiang, Liang Guiyou. Bone marrow mesenchymal stem cell treatment for myocardial ischemia/reperfusion injury: mechanisms of exosomes and factors[J]. Chinese Journal of Tissue Engineering Research, 2019, 23(9): 1455-1460.

Figures/Tables 2

2.1 骨髓间充质干细胞概述及生物特性 2.1.1 骨髓间充质干细胞概述骨髓间充质干细胞是一种来源于哺乳动物骨髓基质中的除造血干细胞以外的一类具有保留体外自我复制能力、多向分化潜能的干细胞[17]。在标准培养条件下，可表达细胞表面标记CD105、CD73 和CD90，而不表达细胞表面标记CD45、CD34、CD14或CD11b、CD79、CD19或HLA-DR[18]，其具有自我更新、多向分化潜能、可体外大量扩增等干细胞生物学特性[19-21]。心肌梗死后移植骨髓间充质干细胞能够增加新生血管数量，减小梗死面积，减少瘢痕[22]。骨髓间充质干细胞在一定条件下被诱导分化为成骨细胞、软骨细胞、脂肪细胞、心肌细胞、神经元及肝细胞等[23]。刘煜昊等[24]直接证据表明，人骨髓间充质干细胞能分化为心肌细胞和血管内皮细胞。研究显示，骨髓间充质干细胞具有低免疫原性[25]。骨髓间充质干细胞及其分化细胞表达的细胞外基质分子如黏附分子和整合素参与构成了支持造血的微环境，帮助造血干细胞的黏附和归巢以及细胞间的相互黏附。骨髓间充质干细胞分泌的细胞因子对造血干/祖细胞的自我更新与分化起重要调节作用。 2.1.2 归巢定植特性骨髓间充质干细胞归巢是一种与造血干细胞相关的现象。造血干细胞被认为有能力从血液中迁移到不同器官，并在化学信号导航下返回到骨髓基质中。骨髓间充质干细胞被认为具有相似特征，这有助于它们迁移和移植到缺血心肌中并进行介导修复。实验研究结果显示，骨髓间充质干细胞在体外扩增之后，归巢能力严重受损。将未经过体外培养用绿色荧光蛋白转基因标记的骨髓间充质干细胞，移植到亚致死性辐射小鼠体内，55%-65%标记绿色荧光蛋白的细胞存活于骨髓间质中，4%-7%存活于脾脏中，而经体外培养24 h后用绿色荧光蛋白标记的骨髓间充质干细胞的存活率却下降到10%[26]。传代晚期的骨髓间充质干细胞已失去趋化因子受体CCR1、CCR9、CXCR6、CXCR5、CXCR4的表面表达，也就失去了对相应的趋化反应。因此，需要优化骨髓间充质干细胞的培养条件来维持趋化因子受体的归巢表达[27]。骨髓间充质干细胞在一定诱导条件下可以扩增分化为成骨细胞、软骨细胞、心肌细胞、脂肪细胞等。例如骨髓间充质干细胞在5-氮杂胞苷诱导下分化成心肌样细胞[28]。不同培养条件下骨髓间充质干细胞的分化能力也不相同。骨髓间充质干细胞通过归巢定植于心肌损伤位置，定向分化为心肌细胞，达到治疗心肌缺血再灌注损伤的作用。 2.1.3 旁分泌特性骨髓间充质干细胞发挥改善器官功能的效应依赖于其旁分泌作用。研究表明，骨髓间充质干细胞移植利用旁分泌作用改善心肌梗死后的心脏功能[29]。此外，旁分泌作用还可以解释许多与修复相关的作用机制，包括促进新生血管形成、减小梗死面积和瘢痕形成、改善心肌收缩功能等。有证据表明，骨髓间充质干细胞通过旁分泌相关因子来改善心脏功能，见表1，而不是通过直接分化成心肌细胞。骨髓间充质干细胞外泌体还携带miRNA基因，转录后参与抗凋亡和血管生成蛋白的调控[30-32]，见表2。在这种情况下，外泌体作为一种运输心脏保护分子的载体获得了广泛的关注[33]。骨髓间充质干细胞分泌的外泌体可以通过抗凋亡、抗炎、促血管生成和免疫调节作用来保护心脏和大脑。骨髓间充质干细胞可能通过分泌卵泡抑素样分子1 (FSTL1)从而减少大鼠心肌梗死面积，进而起到心肌保护作用[34]。因此，骨髓间充质干细胞的旁分泌作用是治疗心肌缺血再灌注损伤的又一重要机制。旁分泌特性可能成为骨髓间充质干细胞治疗心肌缺血再灌注损伤的主要论点。"

骨髓间充质干细胞可以通过分泌多种可溶性因子与细胞表面分子及胞外基质组成特异性的微环境发挥治疗作用。研究发现，骨髓间充质干细胞通过产生一些细胞因子发挥其生物学效应，如血管内皮生长因子、干细胞因子、肝细胞生长因子、肿瘤坏死因子α、胰岛素样生长因子1等[35]。这些细胞因子间接的抗凋亡、抗炎、抗纤维化等作用均促进了组织的内源性修复。骨髓间充质干细胞的抗炎、抗凋亡特性是治疗心肌缺血再灌注损伤的基础，可以减少炎症发生及心肌细胞凋亡，缓解心肌损伤发生。许多学者通过研究骨髓间充质干细胞与各种不同免疫细胞的相互作用，以及与同种异体T细胞的相互影响来证明骨髓间充质干细胞的低免疫原性。骨髓间充质干细胞对T细胞应答具有较低的刺激阈，并且不能诱导同种异体T细胞的激活，要充分的激活T细胞，需2个信号途径：①T受体对主要组织相容性复合体(major histocompatibility complex，MHC)分子与抗原递呈细胞表面抗原相结合的识别；②T细胞激活需要一个共刺激的信号，包括激活在T细胞内的CD28和抗原递呈细胞内的CD80或CD86[36]。骨髓间充质干细胞表面虽然能表达较低水平的MHCⅠ类分子，但是其缺乏MHCⅡ类分子和共刺激分子CD80或CD86，CD40的表达就有利于形成骨髓间充质干细胞的低免疫原性。尽管干扰素γ可刺激MHCⅠ类分子和Ⅱ类分子的表达，但仍不能激起可引起免疫反应的骨髓间充质干细胞的免疫原性。自然杀伤细胞是先天免疫系统中重要的效应细胞，这类细胞对缺乏MHCⅠ类分子表达的细胞具有极强的溶解细胞能力。同时，自然杀伤细胞也是重要的造血细胞移植过程的调节物。在体外，骨髓间充质干细胞对自然杀伤细胞具有抑制作用，并能下调其增殖能力、细胞毒活性和细胞因子表达。由于骨髓间充质干细胞表面低表达MHCⅠ类分子，自然杀伤细胞又对其有极强的杀伤作用[37]。Wu等[38]揭示miR-21通过负调控转化生长因子β1来调控骨髓间充质干细胞的免疫调节作用，这为骨髓间充质干细胞的免疫调控作用提供了新的见解。因此，可以利用骨髓间充质干细胞的免疫调控、低免疫性、减少免疫应答、抑制自然杀伤细胞、减少炎症发生达到治疗心肌缺血再灌注损伤的目的。 2.2 骨髓间充质干细胞治疗心肌缺血再灌注损伤作用靶点及作用机制骨髓间充质干细胞治疗心肌缺血再灌注损伤的作用机制知之甚少，结合实验室前期结果以及当前研究进行总结骨髓间充质干细胞治疗心肌缺血再灌注损伤潜在作用机制，见图2。"

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