Paracrine actions of bone marrow mesenchymal stem cells: angiogenesis, immunoregulation, and inflammatory regulation

doi:10.3969/j.issn.2095-4344.0527

Abstract

Abstract:

BACKGROUND: Although bone marrow mesenchymal stem cells have the inherent ability to differentiate into different types of tissue cells, there is increasing evidence that bone marrow mesenchymal stem cells may regulate the repair process of damaged tissues through a variety of paracrine factors.
OBJECTIVE: To review the progress in the paracrine effect of bone marrow mesenchymal stem cells in orthopedics.
METHODS: A search of Web of Science and PubMed was performed for articles regarding the paracrine effect of bone marrow mesenchymal stem cells in orthopedics. Then, 49 eligible articles were included after systematical summarization and analysis.
RESULTS AND CONCLUSION: Studies have shown that bone marrow mesenchymal stem cells play a vital role in the angiogenesis, immunoregulation, and inflammatory regulation, which is expected to be a new therapy for bone diseases.

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, Paracrine Communication, Tissue Engineering

CLC Number:

Zhang Xin, Xie Jiabing. Paracrine actions of bone marrow mesenchymal stem cells: angiogenesis, immunoregulation, and inflammatory regulation[J]. Chinese Journal of Tissue Engineering Research, 2019, 23(1): 139-143.

Figures/Tables 1

2.1 骨髓间充质干细胞在实验室体外环境下增殖及培养目前在实验室体外环境下对骨髓间充质干细胞进行培养及分离的方法主要有全骨髓贴壁分离法、密度梯度离心法、流式细胞仪分离法和免疫磁珠分离法。后两种方法主要是应用骨髓间充质干细胞表面特殊抗原，对其经行筛选、分离、洗涤等步骤最终起到纯化的作用，但目前仍未找到骨髓间充质干细胞最具特异性的细胞表面标记，且实验成本较高，操作步骤繁琐，技术难度高。密度梯度离心法需要时间较长，且操作相对复杂。全骨髓贴壁分离法则相对简单，对新鲜的骨髓全血进行初步的离心，分离出所需要的底层细胞经行培养，利用骨髓间充质干细胞贴壁生长的特性，通过多次更换培养液，去除悬浮细胞，最终起到纯化的目的。骨髓间充质干细胞的形态以梭形为主，少数呈三角形、星形，细胞传代后快速增殖，呈漩涡状排列，透射电镜下细胞形态不一，多为圆形，核仁清晰，核浆比例大，胞质中含有核糖体、粗面内质网、分泌小泡、线粒体等，细胞表面有微绒毛突起。缝隙连接可加强相邻细胞间的连接，有利于细胞间信息传递[3-4]。 ISCT已提出了一系列的标准来鉴定间充质干细胞：①在标准培养条件下，细胞能够贴壁生长，并呈成纤维样形态。事实上，一些学者认为，骨髓间充质干细胞和成纤维细胞在功能上是相同的[5]；②细胞表面表达一些特异性抗原(标记物)，骨髓间充质干细胞表面有CD73，CD90和CD105表面标志物表达，而缺乏CD11b，CD14，CD19，CD34，CD45，CD79a和HLA-DR表面标志物的表达[6]；③具有向脂肪细胞、成骨细胞、软骨细胞分化的能力。 2.2 骨髓间充质干细胞分泌的活性物质骨髓原始间充质干细胞不仅仅能够为骨髓中的造血干细胞提供物理性附着场所，而且本身还能分泌出多种有利于造血干细胞分化的生长因子，如白细胞介素6、白细胞介素11、白血病抑制因子、巨噬细胞集落刺激因子及干细胞因子等。既往多项研究发现，骨髓间充质干细胞培养上清可能是通过生长因子、细胞因子等旁分泌效应对损伤组织起修复作用，通过动物实验发现，发挥损伤修复作用的物质可能是一种直径在50-100 nm的磷脂膜包裹的小囊泡，后被证实为骨髓间充质干细胞来源的微泡，它是骨髓间充质干细胞在静息或应激状态下产生的异质性膜分泌体系，由脂质双分子膜包裹，内含脂质、蛋白质、mRNA、miRNA等多种生物活性物质。骨髓间充质干细胞是一类低免疫原性细胞，可调控树突状细胞和T细胞的功能，具有免疫调节和诱导免疫耐受的作用[7-9]，可以改善和调节破坏性炎症反应。已知骨髓间充质干细胞通过细胞与细胞间直接相互作用，分泌可溶性因子，抑制T、B细胞的增殖，抑制中性粒细胞产生H2O2，抑制T细胞和NK细胞的细胞毒作用，以及抑制单核细胞的分化和成熟。 2.3 骨髓间充质干细胞分泌活性物质的作用骨髓间充质干细胞有很强的自我扩增能力，同时能够保持多向分化性能，其能多向分化为成骨细胞、软骨细胞、脂肪细胞、心肌细胞、神经元细胞等。在体内，骨髓间充质干细胞受到各种营养机制激活时能够向多种组织类型的细胞分化，当接触到损伤环境时再生潜力提高。组织损伤发生后，内源性骨髓间充质干细胞感知损伤信号移出骨髓，进入外周血循环向损伤组织迁移，与靶组织血管内皮细胞黏附，并穿过胞外基质屏障，然后到达损伤组织进行修复[10-12]。缺氧是导致细胞和组织损伤最常见的原因。早期炎性反应可以激活凝血系统，启动凝血级联反应，同时生理性纤溶途径被抑制，导致微循环障碍，组织缺血缺氧，出现器官功能障碍。缺氧诱导因子1是缺氧应答中起重要作用的转录因子，它通过控制血管生成来调节氧的运输能力，通过调控糖酵解作用来增强机体对低氧的免疫耐受。多项研究表明，将骨髓间充质干细胞暴露于缺氧条件培养，以模仿损伤后的体内缺血性环境，可上调促生存和血管生成因子表达。低氧预处理骨髓间充质干细胞，可提高血管生成因子的含量，有助于减轻损伤和改善功能修复，提高治疗效果。骨髓间充质干细胞在缺氧环境中暴露24 h，能够分泌成纤维细胞生长因子2、肝细胞生长因子、胰岛素样生长因子1，可能是通过NF-κB信号通路所介导的[13-14]。骨髓间充质干细胞分泌的血管内皮生长因子水平进行性增高。低氧可以上调骨髓间充质干细胞表面的SDF-1受体趋化因子受体CXCR4的表达，促进骨髓间充质干细胞的迁移；低氧还促进骨髓间充质干细胞的增殖，发挥再生潜力[15]。骨髓间充质干细胞在体外可以显著抑制T淋巴细胞增殖，通过分泌免疫抑制性细胞因子以及表达IDO等途径构建局部免疫微环境，发挥免疫抑制作用，在γ-干扰素作用下，骨髓间充质干细胞免疫抑制活性增强[16-20]。炎症刺激条件下，骨髓间充质干细胞通过分泌转化生长因子β、肝细胞生长因子等多种蛋白因子[21]，可以抑制细胞或体液免疫中的T淋巴细胞或B淋巴细胞增殖，并能将已被激活的T淋巴细胞或B淋巴细胞诱导失活。因此，骨髓间充质干细胞可以在炎症组织内发挥免疫调节的作用，该作用在组织修复中具有极为广阔的应用前景[22]。肿瘤坏死因子也是一种炎性介质，可以诱导炎症，诱发组织坏死。肿瘤坏死因子α预处理的骨髓间充质干细胞比未经处理的骨髓间充质干细胞具有更优的体外血管生成活性[23]。另一项研究表明，肿瘤坏死因子α预处理通过上调骨形态发生蛋白2蛋白水平增加细胞增殖、动员和成骨分化。骨形态发生蛋白2基因沉默后抑制了骨髓间充质干细胞诱导成骨分化[24]。最近的研究表明，先天免疫激活剂，如脂多糖、Toll样受体激动剂，也可通过旁分泌因子刺激骨髓间充质干细胞增殖活性[25]。炎症介质干扰素、肿瘤坏死因子可通过细胞外基质调节骨髓间充质干细胞归巢和迁移[26]，在体外，骨髓间充质干细胞表现出对各种伤口愈合细胞因子有趋化作用[26-27]。将小鼠和人骨髓间充质干细胞植入到炎症和组织损伤区域[28-30]，可促进伤口愈合、骨组织再生，调节免疫反应[31]。骨髓间充质干细胞可以抑制肥大细胞[32]、T细胞[33]、B细胞和自然杀伤细胞的募集[34-35]，从而减轻损伤。伤口的炎症环境也激发了骨髓间充质干细胞内COX-2的活性，上调前列腺素E2表达[36]，有利于骨组织的再生。前列腺素E2还可以使白细胞介素2、γ-干扰素表达减少，白细胞介素4和白细胞介素10表达增加[37-40]。抗炎细胞因子的表达有利于成纤维细胞基质金属蛋白酶表达上调和胶原蛋白表达下调[41-42]，从而在创伤部位形成密度较小、纤维化的肉芽组织。骨髓间充质干细胞在损伤部位的活性超过炎症反应，其分化为成骨细胞促进骨组织的再生[42-43]。此外，还有大量的证据表明，骨髓间充质干细胞在增殖阶段继续支持组织再生。在此阶段，血管生成是必需的，以确保有足够的营养输送到损伤部位，加快伤口愈合。损伤部位微血管不足可能会抑制伤口愈合和导致慢性伤口未愈合。骨髓间充质干细胞可以分泌出多种细胞因子，包括血管内皮生长因子A、肾上腺髓质素、碱性成纤维细胞生长因子[44]，这些因子可以促进微血管内皮细胞增殖[45-48]，形成一个长期持久的功能性血管网络[49]。骨髓间充质干细胞还可以表达其他的多种细胞生长因子，包括白细胞介素10、肝细胞生长因子。骨髓间充质干细胞分泌活性物质的相关作用见表1。"

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