Human umbilical cord mesenchymal stem cell-derived exosomal microRNA-148a-3p inhibits neointimal hyperplasia by targeting Serpine1
Human umbilical cord mesenchymal stem cell-derived exosomal microRNA-148a-3p inhibits neointimal hyperplasia by targeting Serpine1
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DOI：
10.1016/j.abb.2022.109155
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发表时间：
2022-03-01
影响因子：
3.9
通讯作者：
Wang, Shenming
中科院分区：
文献类型：
--
作者：
Zhang, Xiaoyu;Zhou, Yu;Wang, Shenming
背景：血管内膜增生症(NIH)患者行经皮腔内血管成形术时，再狭窄是不可避免的。人脐带间充质干细胞来源的外切体(hucMSC-Exos)在心血管疾病领域已有研究。然而，hucMSC-Exos对NIH的作用及其机制尚不清楚。我们的目的是研究MSC-Exos是否调节血管平滑肌细胞(VSMC)的功能以抑制NIH并探讨其可能的机制。方法：分离培养人脐血间充质干细胞和小鼠血管平滑肌细胞，用流式细胞仪和免疫荧光法进行鉴定。通过透射电子显微镜、纳米颗粒跟踪分析和Western blotts对HucMSC-Exos进行鉴定。给结扎左颈总动脉的小鼠静脉注射Exosome(Exos)，用苏木精-伊红(H&E)染色和免疫组织化学染色观察Exos对NIH的影响。用细胞计数试剂盒8、划痕、Transwell和Western印迹法检测hucMSC-Exos对VSMCs的影响。基因表达总表中的microRNA测序数据和mRNA测序结果分别用于鉴定hucMSC-Exos中的潜在分子和VSMC中的靶基因。我们通过过表达和敲除实验检测了外源microRNAs和VSMC中的靶基因的调控作用。结果：分离并鉴定了原代hucMSCs、VSMCs和hucMSC-Exos。动脉结扎后给予hucMSCExos可抑制NIH。H&E和免疫组织化学结果显示，hucMSC-Exos减少了颈动脉内膜和中膜面积，减少了内膜/中膜比值，增加了中层收缩表型蛋白SM22a，下调了颈动脉SERPINE1的表达。在体外，VSMCs通过抑制SERPINE1的表达来抑制SM22a的迁移和上调SM22a的表达。MIR-148A-3P富含hucMSC-Exos，并通过靶向其3‘非翻译区抑制SERPINE1。此外，外体miR-148A-3p通过靶向SERPINE1抑制VSMC的表型转换和迁移。结论：在小鼠颈动脉结扎模型中，hucMSC-Exos可抑制NIH，其抑制VSMC表型转换和迁移的作用是通过将miR-148A-3p转导到靶向SERPINE1的VSMC中实现的。
Background: Restenosis is inevitable when patients undergo percutaneous transluminal angioplasty due to neointimal hyperplasia (NIH). Human umbilical cord mesenchymal stem cell-derived exosomes (hucMSC-Exos) have been studied in the field of cardiovascular diseases. However, the effects and mechanisms of hucMSC-Exos on NIH are unclear. We aimed to investigate whether MSC-Exos regulate vascular smooth muscle cell (VSMC) functions to inhibit NIH and explore the underlying mechanisms. Methods: HucMSCs and mouse VSMCs were isolated and characterized by flow cytometry and immunofluorescence. HucMSC-Exos were identified by transmission electron microscopy, nanoparticle tracking analysis and western blots. Exosomes (Exos) were intravenously injected into mice with left common carotid artery ligation, and their effects on NIH were assessed by haematoxylin and eosin (H&E) and immunohistochemistry staining. The effects of hucMSC-Exos on VSMCs were evaluated by Cell Counting Kit-8, scratch wound, Transwell and Western blot assays. MicroRNA sequencing data in the Gene Expression Omnibus and mRNA sequencing results were used to identify potential molecules in hucMSC-Exos and target genes in VSMCs, respectively. We tested the regulatory effect of microRNAs in Exos and target genes in VSMCs using overexpression and knockdown experiments. Results: Primary hucMSCs, VSMCs and hucMSC-Exos were isolated and characterized. Administration of hucMSCExos suppressed NIH after artery ligation. H&E and immunohistochemistry results showed that hucMSC-Exos decreased the intima and media area and intima/media ratio, increased the contractile phenotype protein SM22a in the media layer and downregulated Serpine1 expression in the carotid artery. Exos were ingested by VSMCs, which inhibited migration and upregulated SM22a expression by suppressing Serpine1 expression in vitro. MiR-148a-3p was enriched in hucMSC-Exos and repressed Serpine1 by targeting its 3' untranslated region. Moreover, exosomal miR-148a-3p suppressed VSMC phenotypic switching and migration by targeting Serpine1. Conclusions: We found that hucMSC-Exos inhibited NIH in a mouse carotid artery ligation model and that the inhibitory effects on VSMC phenotypic switching and migration were mediated by delivery of miR-148a-3p to VSMCs to target Serpine1.