Wendan Y, Changzhu J, Xuhong S, Hongjing C, Hong S, Dongxia Y, Fang X
College of Basic Medicine, Binzhou Medical University, Yantai, Shandong 264003, China.
Emergency Department of Cardiology, The Second People's Hospital of Jining, Shandong 272000, China.
Stem Cells Int. 2016;2016:3196071. doi: 10.1155/2016/3196071. Epub 2016 Jan 6.
In this study a model of simulated vascular injury in vitro was used to study the characterization of bone-marrow-derived mesenchymal stem cells (BMSCs) morphology and to investigate the differentiation and migration of BMSCs in the presence of adventitial fibroblasts. BMSCs from rats were indirectly cocultured with adventitial fibroblasts in a transwell chamber apparatus for 7 days, and clonogenic assays demonstrated that BMSCs could be differentiated into smooth muscle-like cells with this process, including smooth muscle α-actin (α-SMA) expression by immunofluorescence staining. Cell morphology of BMSCs was assessed by inverted microscope, while cell proliferation was assessed by MTT assay. The expressions of TGF-β1, MMP-1, and NF-κB were detected by immunofluorescence staining and Smad3 mRNA was measured by reverse transcription PCR. Migration ability of BMSCs with DAPI-labeled nuclei was measured by laser confocal microscopy. Our results demonstrate that indirect interactions with adventitial fibroblasts can induce proliferation, differentiation, and migration of BMSCs that can actively participate in neointimal formation. Our results indicate that the pathogenesis of vascular remodeling might perform via TGF-β1/Smad3 signal transduction pathways.
在本研究中，体外模拟血管损伤模型用于研究骨髓间充质干细胞（BMSCs）的形态特征，并探讨在外膜成纤维细胞存在的情况下BMSCs的分化和迁移。将大鼠的BMSCs与外膜成纤维细胞在Transwell小室装置中间接共培养7天，集落形成试验表明，在此过程中BMSCs可分化为平滑肌样细胞，包括通过免疫荧光染色检测到平滑肌α-肌动蛋白（α-SMA）表达。通过倒置显微镜评估BMSCs的细胞形态，通过MTT法评估细胞增殖。通过免疫荧光染色检测TGF-β1、MMP-1和NF-κB的表达，通过逆转录PCR检测Smad3 mRNA。通过激光共聚焦显微镜测量带有DAPI标记细胞核的BMSCs的迁移能力。我们的结果表明与外膜成纤维细胞的间接相互作用可诱导BMSCs增殖、分化和迁移，其可积极参与新生内膜形成。我们的结果表明血管重塑的发病机制可能通过TGF-β1/Smad3信号转导途径发挥作用。
