摘要
本研究旨在观察血管紧张素Ⅱ(angiotensinⅡ,AngⅡ)及其受体阻断剂对SD大鼠胸主动脉外膜成纤维细胞亚群迁移及内皮素-1(endothelin-1,ET-1)表达的影响。采用克隆环法进行血管外膜成纤维细胞单克隆培养,并观察其细胞形态;RT-PCR方法进行细胞纯度鉴定;Transwell小室法检测AngⅡ对细胞亚群迁移的影响;荧光定量-PCR方法检测AngⅡ及其拮抗剂对preproET-1 mRNA表达的影响;ELISA法检测AngⅡ及其拮抗剂对ET-1分泌的影响。结果显示,克隆环法获得血管外膜成纤维细胞两个亚群:纺锤形细胞亚群和圆形细胞亚群。RT-PCR检测显示所获得的成纤维细胞亚群是纯细胞系。Transwell小室实验结果表明,两个细胞亚群均具备迁移能力,与自身对照相比,AngⅡ显著促进圆形细胞迁移,对纺锤形细胞迁移无明显影响;AngⅡ(1×10 8～1×10 6mol.L 1)对纺锤形细胞preproET-1 mRNA的表达及ET-1分泌无显著性影响,而呈浓度依赖性显著增加圆形细胞preproET-1mRNA的表达及ET-1的分泌(P<0.05,P<0.01);氯沙坦阻断AngⅡ诱导的圆形细胞preproET-1 mRNA的表达及ET-1的分泌,对纺锤形细胞preproET-1 mRNA的表达及ET-1的分泌无明显影响。以上结果提示,SD大鼠血管外膜成纤维细胞有两个细胞亚群:纺锤形细胞亚群和圆形细胞亚群。AngⅡ显著促进圆形细胞的迁移和ET-1的表达,提示两个细胞亚群的迁移机制可能不同,两种亚群在血管重建和修复过程中可能发挥不同作用。
Abstract： The study is to investigate the effect of angiotensin Ⅱ （AngⅡ） and its receptor blockers on migration and endothelin-1 （ET-1） expression of rat vascular adventitial fibroblast subpopulations. Vascular adventitial fibroblasts were individually expanded by using cloning rings, and the effects of Ang II on the migration of adventitial fibroblast subpopulations were evaluated by Transwell. Fluorescence quantitative-PCR detected the expression of preproET-1 mRNA induced by Ang II, and its receptor antagonists losartan and PD-123319. The concentration of ET-1 was determined by ELISA. It showed that spindle shaped and epithelioid shaped cells were isolated by using cloning rings, named as spindle cells and round cells. RT-PCR showed that fibroblast subpopulations did not have leukocytes, endothelial cells and smooth muscle cells, namely pure cell lines. Compared with respective control cells, two subpopulations had transferring ability. Ang Ⅱ significantly improved round cells migration in a concentration-dependent manner, and had no obvious influence on spindle cells migration. AngⅡ （1×10-8～1×10-6mol·L-1） significantly increased the expression ofpreproET-1 mRNA in round cells （P 〈 0.01）, and had no significant effect on the expression of preproET-1 mRNA in spindle cells. Losartan blocked the expression ofpreproET-1 mRNA induced by Ang II in round cells, and had no significant effect on the expression ofpreproET-1 mRNA in spindle cells. The effects of AngⅡ and ET-1 receptor inhibitors on the release of ET-1 were similar to the expression ofpreproET-1 mRNA. The results indicate that there are two cell subpopulations： round cells and spindle cells in rat vascular adventitial fibroblasts. Ang II significantly improved cells migration, and increased the expression of ET-1 in round cell subpopulation. It suggested that there may be different migratory mechanisms in two cell subpopulations, and the two subpopulations may play a different role in vascular remodeling and reparative
出处
《药学学报》
CAS
CSCD
北大核心
2012年第11期1428-1433,共6页
Acta Pharmaceutica Sinica
基金
河北省自然科学基金资助项目(C2009001037)
