MicroRNA-92a-mediated endothelial to mesenchymal transition controls vein graft neointimal lesion formation
MicroRNA-92a-mediated endothelial to mesenchymal transition controls vein graft neointimal lesion formation
复制标题
MicroRNA-92a介导的内皮向间质转变控制静脉移植物新内膜病变的形成
DOI：
10.1016/j.yexcr.2020.112402
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发表时间：
2021-01-01
影响因子：
3.7
通讯作者：
Wu, Qing-Chen
中科院分区：
文献类型：
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作者：
Zhong, Chang-Ming;Li, Song;Wu, Qing-Chen
目的：新内膜增生导致的静脉移植物长期衰竭仍然是冠状动脉搭桥手术中的一个重要问题。内皮到间充质转化（EndMT）有助于静脉移植血管重构。然而，关于microrna介导的EndMT在静脉移植中新内膜形成中的作用的研究很少。我们假设microRNA-92a （miR-92a）可能在决定EndMT对新内膜形成的贡献中发挥重要作用。方法：体外和体内采用qRT-PCR和Western blot检测miR-92a和endmt相关蛋白。采用腺相关病毒6 （AAV6）传递基因疗法在体内抑制新生内膜的形成。HE染色检测移植静脉内膜增生情况，免疫荧光法检测移植静脉中endmt相关蛋白的表达。采用免疫组织化学和荧光素酶检测miR-92a的潜在靶点。结果：发现miR-92a在静脉移植后的新生内膜增生性病变中表达上调。利用培养的人脐静脉内皮细胞（HUVECs），我们发现TGF-p1处理HUVECs显著增加了miR-92a的表达并诱导了EndMT，其特征是内皮特异性标志物（CD31和VE-cadherin）受到抑制，间质特异性标志物（a-SMA和vimentin）增加，而miR-92a表达抑制了培养的HUVECs中的EndMT。此外，AAV6介导的miR-92a抑制基因治疗有效地减少了体内静脉移植物的EndMT和新生内膜形成。我们进一步发现整合素α 5 （ITGA5）是参与这些静脉移植物中新内膜形成发展的潜在靶基因。结论：这些数据表明，新生内膜的形成不仅依赖于血管平滑肌细胞的表型转换，还与EndMT有关，mir -92a介导的EndMT是移植物血管新生内膜形成的重要机制。
Purpose: Long-term failure of vein grafts due to neointimal hyperplasia remains an important problem in coronary artery bypass graft surgery. Endothelial to mesenchymal transition (EndMT) contributes to vein graft vascular remodeling. However, there is little study on microRNA-mediated EndMT contributions to neointimal formation in vein graft. We hypothesized that microRNA-92a (miR-92a) might play an important role in determining EndMT contributions to neointimal formation.Methods: miR-92a and EndMT-related proteins detected by qRT-PCR and Western blot in vitro and in vivo. Adeno-associated virus 6 (AAV6) delivery gene therapy was used to inhibit neointimal formation in vivo. The intimal hyperplasia of vein grafts was measured by HE staining, the expression of EndMT-related protein in vein grafts was measured by immunofluorescence. Immunohistochemistry and luciferase assay were used to detect potential targets of miR-92a.Results: The expression of miR-92a was found to be upregulated in neointimal hyperplasic lesions after vein grafting. Using cultured human umbilical vein endothelial cells (HUVECs), we show that TGF-p1 treatment of HUVECs significantly increased miR-92a expression and induced EndMT, characterized by suppression of endothelial-specific markers (CD31 and VE-cadherin) and an increase in mesenchymal-specific markers (a-SMA and vimentin), while inhibition of miR-92a expression blunted EndMT in cultured HUVECs. Furthermore, AAV6 mediated miR-92a suppression gene therapy effectively resulted in decreased EndMT and less neointimal formation in vein grafts in vivo. We further identified that integrin alpha 5 (ITGA5) is a potential target gene involved in the development of neointima formation in these vein grafts.Conclusion: This data suggests that neointimal formation does not solely rely on vascular smooth muscle cell phenotypic switching but is also related to EndMT, and miR-92a-mediated EndMT is an important mechanism underlying neointimal formation in vein grafts.