Characterization and Functions of Vascular Adventitial Fibroblast Subpopulations
Characterization and Functions of Vascular Adventitial Fibroblast Subpopulations
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血管外膜成纤维细胞亚群的特征和功能。
DOI：
10.1159/000373939
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发表时间：
2015-01-01
影响因子：
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通讯作者：
Fu, Jun Qiu
中科院分区：
文献类型：
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作者：
An, Sheng Jun;Liu, Pei;Fu, Jun Qiu
背景资料：血管外膜成纤维细胞在血管重塑中起重要作用，并促进血管疾病中新生内膜的形成。然而，很少有人知道外膜成纤维细胞亚群。本研究探讨了分离大鼠胸主动脉外膜成纤维细胞亚群的过程，并表征了血管紧张素II（ANG II）刺激后的特性，血管紧张素II是参与心血管疾病如高血压的关键因素。方法：分离培养大鼠主动脉外膜成纤维细胞。使用克隆环技术单独扩增成纤维细胞亚群。用ANG II（10 nM、100 nM和1 μ M）处理细胞0.5、1、1.5、3、6、12或24 h，并通过MTT测定和Transwell测量ANG II诱导的增殖和迁移。在存在或不存在ANG II受体拮抗剂（100 μ M）、氯沙坦（对于AT 1）和PD-123319（对于AT 2）的情况下，用ANG II（100 nM）处理细胞。RT-PCR法检测PreproET-1 mRNA的表达，ELISA法检测ET-1的表达。免疫印迹法检测I型胶原蛋白。结果如下：在血管外膜中发现了两个主要的成纤维细胞亚群，上皮样细胞和梭形样细胞，虽然ANG II促进这两个亚群的生长，但上皮样细胞的增殖在10 nM至1 μ M的ANG II浓度范围内呈剂量依赖性，而梭形样细胞的增殖在100 nM ANG II刺激后达到峰值; ANG Ⅱ刺激增强上皮样细胞迁移，但不增强梭形细胞迁移; ANG II剂量依赖性地增加前原ET-1和I型胶原的表达，并增加上皮样而非梭形细胞中的ET-1分泌，AT 1受体拮抗剂消除了这种作用，而不与AT 2受体拮抗剂联用。结论：外膜成纤维细胞是异质的，对ANG II刺激具有高敏感性的类上皮细胞亚群可能与血管重塑、修复过程和心血管疾病的病理生理机制有关。版权所有（C）2015 S. Karger AG，巴塞尔
Background: Adventitial fibroblasts have been shown to play an important role in vascular remodeling and contribute to neointimal formation in vascular diseases. However, little is known about adventitial fibroblast subpopulations. This study explored the process of isolating rat thoracic aorta adventitial fibroblast subpopulations and characterized their properties following stimulation with angiotensin II (ANG II), a critical factor involved in cardiovascular diseases such as hypertension. Methods: Adventitial fibroblasts were isolated and cultured from rat aorta. Fibroblast subpopulations were individually expanded using cloning ring techniques. Cells were treated with ANG II (10 nM, 100 nM and 1 mu M) for 0.5, 1, 1.5, 3, 6, 12, or 24 h, and ANG II-induced proliferation and migration were measured by MTT assay and Transwell. Cells were treated with ANG II (100 nM) in the presence or absence of ANG II receptor antagonists (100 mu M), losartan (for AT1) and PD-123319 (for AT2). PreproET-1 mRNA and ET-1 were determined by RT-PCR and ELISA, respectively. Collagen type I was detected by western blotting. Results: Two major fibroblast subpopulations were found in the adventitia, epithelioid-like cells and spindle-like cells; Although ANG II promotes the growth of both subpopulations, epithelioid-like cell proliferation shows dose-dependency on ANG II from 10 nM to 1 mu M, while proliferation of spindle-like cells reaches a peak value following 100 nM ANG II stimulation; ANG II stimulation enhanced epithelioid-like but not spindle-like cell migration; ANG II dose-dependently increased the expression of preproET-1 and collagen type I, and enhanced ET-1 secretion in epithelioid-like but not spindle-like cells, effects abolished by the AT1 receptor antagonist, but not with AT2 receptor antagonist. Conclusion: Adventitial fibroblasts are heterogeneous and epithelioid-like subpopulations with high sensitivity to ANG II stimulation may be implicated in the pathophysiological mechanisms of vascular remodeling, reparative processes and cardiovascular diseases. Copyright (C) 2015 S. Karger AG, Basel