Distinct Anti-Fibrotic Effects of Exosomes Derived from Endothelial Colony-Forming Cells Cultured Under Normoxia and Hypoxia.
Distinct Anti-Fibrotic Effects of Exosomes Derived from Endothelial Colony-Forming Cells Cultured Under Normoxia and Hypoxia.
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常氧和缺氧条件下培养的内皮集落形成细胞衍生的外泌体具有独特的抗纤维化作用。
DOI：
10.12659/msm.911306
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发表时间：
2018-09-05
期刊：
影响因子：
--
通讯作者：
Wang J
中科院分区：
文献类型：
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作者：
Liu W;Zhang H;Mai J;Chen Z;Huang T;Wang S;Chen Y;Wang J
内皮细胞集落形成细胞（ECFCs）的治疗潜力可能在缺血环境中受损。直接注射ECFCs并不是拯救缺血性心脏的有效方法，但这些细胞衍生的外泌体可能是一种有前途的治疗工具。然而，在常氧和缺氧条件下产生的外泌体可能不相同。因此，本研究的目的是研究缺氧处理的ecfc来源的外泌体抗纤维化作用的改变及其潜在机制。从外周血中分离ecfc，并从常氧（non -exo）或缺氧（hyper -exo）处理的ecfc中收集外泌体。体外观察外泌体对心肌成纤维细胞活化的影响。外泌体内的MicroRNAs （miRNAs）被提取并使用下一代RNA测序进行比较。使用双荧光素酶报告基因测定方法验证miR-10b-5p的预测靶mrna。no -exo显著改善体外心肌成纤维细胞活化。这些影响在高外显子处理组减弱。miR-10b-5p在non -exo中富集，而在hyper -exo中不富集。双荧光素酶报告基因检测发现，miR-10b-5p抑制了smad特异性E3泛素蛋白连接酶1 （Smurf1）和组蛋白去乙酰化酶4 (HDAC4) mrna。中性鞘磷脂酶2 （N-SMase2）在缺氧ecfc中的表达降低，这一结果与hypo -exo中miR-10b-5p的变化一致。由于靶向纤维化基因Smurf1和HDAC4的miR-10b-5p的减少，hypexo的抗纤维化作用被取消。
The therapeutic potential of endothelial colony-forming cells (ECFCs) may be impaired in an ischemic environment. Direct injection of ECFCs is not an effective method of rescuing the ischemic heart, but exosomes derived from these cells may be a promising therapeutic tool. However, exosomes produced under normoxia and hypoxia may not be identical. Therefore, the purpose of this study was to investigate alterations in the anti-fibrotic effects of hypoxia-treated ECFC-derived exosomes and the underlying mechanism involved. ECFCs were isolated from peripheral blood and exosomes were collected from ECFCs treated with normoxia (nor-exo) or hypoxia (hyp-exo). Effects of exosomes on cardiac fibroblast activation were evaluated in vitro. MicroRNAs (miRNAs) inside the exosomes were extracted and compared using next-generation RNA sequencing. Predicted target mRNAs of miR-10b-5p were validated using a dual-luciferase reporter gene assay method. Nor-exo significantly ameliorated cardiac fibroblast activation in vitro. These effects were attenuated in the hyp-exo-treated group. miR-10b-5p was enriched in nor-exo but not in hyp-exo. Dual-luciferase reporter gene assay found that both SMAD-specific E3 ubiquitin protein ligase 1 (Smurf1) and histone deacetylase 4 (HDAC4) mRNAs were inhibited by miR-10b-5p. The expression of neutral sphingomyelinase 2 (N-SMase2) was decreased in hypoxia ECFCs, and this result was consistent with the changes in miR-10b-5p in hyp-exo. Due to a reduction of miR-10b-5p, which targets the fibrotic genes Smurf1 and HDAC4, the anti-fibrotic effects of hyp-exo were abolished.