CAJ | 학술논문

目的：探讨血管紧张素Ⅱ（ Ang Ⅱ）及其受体拮抗剂氯沙坦（ Los）和 PD123319对大鼠胸主动脉血管外膜成纤维细胞亚群增殖和胶原生成的影响。方法通过克隆环法获得血管外膜成纤维细胞单克隆，根据细胞表型克隆了两种细胞亚型并分别进行培养，逆转录聚合酶链反应（ RT-PCR）和免疫荧光染色方法鉴定细胞的纯度；流式细胞术对两细胞亚群的形状和大小进行比较；四甲基偶氮唑盐（ MTT）法和Western印迹法检测Ang Ⅱ及其受体拮抗剂Los、PD123319对细胞亚群的增殖活性和Ⅰ型胶原（ CollagenⅠ）合成的影响。结果克隆环法获得血管外膜成纤维细胞两个亚群，经显微镜和流式细胞仪观察根据细胞表型分别命名为：圆形细胞亚群（RC）和纺锤形细胞亚群（SC）。经 RT-PCR和免疫荧光染色对第Ⅷ因子（vWF）、CD8、MHC、Desmin进行检测，结果显示无内皮细胞、平滑肌细胞细胞标志物的表达，分别为两种纯系细胞。实验发现Ang Ⅱ对 RC亚群的增殖活性及CollagenⅠ的合成能力均有显著影响，既促进RC的增殖又提高I型胶原的合成，但经Los预处理后，AngⅡ的促增殖及胶原合成能力被显著性抑制（P＜0.05），而SC亚群的增殖活性和胶原合成情况无显著性改变。经PD123319预处理后，对RC亚群和SC亚群增殖和胶原的合成均无明显影响。结论 AngⅡ在促进两细胞亚群的增殖和胶原合成的能力方面有差异，说明血管外膜成纤维细胞两种亚群在表型、功能和作用机制上具有显著性的差异，提示两种细胞亚群在病理生理机制及其血管重构和修复过程中扮演着不同的角色。
목적：탐토혈관긴장소Ⅱ（ Ang Ⅱ）급기수체길항제록사탄（ Los）화 PD123319대대서흉주동맥혈관외막성섬유세포아군증식화효원생성적영향。방법통과극륭배법획득혈관외막성섬유세포단극륭，근거세포표형극륭료량충세포아형병분별진행배양，역전록취합매련반응（ RT-PCR）화면역형광염색방법감정세포적순도；류식세포술대량세포아군적형상화대소진행비교；사갑기우담서염（ MTT）법화Western인적법검측Ang Ⅱ급기수체길항제Los、PD123319대세포아군적증식활성화Ⅰ형효원（ CollagenⅠ）합성적영향。결과극륭배법획득혈관외막성섬유세포량개아군，경현미경화류식세포의관찰근거세포표형분별명명위：원형세포아군（RC）화방추형세포아군（SC）。경 RT-PCR화면역형광염색대제Ⅷ인자（vWF）、CD8、MHC、Desmin진행검측，결과현시무내피세포、평활기세포세포표지물적표체，분별위량충순계세포。실험발현Ang Ⅱ대 RC아군적증식활성급CollagenⅠ적합성능력균유현저영향，기촉진RC적증식우제고I형효원적합성，단경Los예처리후，AngⅡ적촉증식급효원합성능력피현저성억제（P＜0.05），이SC아군적증식활성화효원합성정황무현저성개변。경PD123319예처리후，대RC아군화SC아군증식화효원적합성균무명현영향。결론 AngⅡ재촉진량세포아군적증식화효원합성적능력방면유차이，설명혈관외막성섬유세포량충아군재표형、공능화작용궤제상구유현저성적차이，제시량충세포아군재병리생리궤제급기혈관중구화수복과정중분연착불동적각색。
Objective To explore the effect of angiotensin Ⅱand its receptor antagonist Losartan on cell proliferation and collagen synthesis in adventitial fibroblast subpopulations from rat thoracic aorta vascular.Methods Vascular adventitial fibroblasts were individually expanded by using cloning rings ,characterization of isolated cells were determined by reverse transcription-polymerase chain reaction ( RT-PCR) method and immunofluorescence staining.The size of subpopulations were compared by flow cytometry ,cells were treated with AngⅡor AngⅡplus Losartan ,PD123319 to detect cellular proliferation and collagen synthesis by MTT and Western blot.Results RT-PCR showed that fibroblast subpopulations did not have endothelial cells ,smooth muscle cells and leukocytes ,which nameed pure cell lines.Ang Ⅱ had different promotion on cell proliferation and collagen synthesis between two subpopulations ,but this effect could be significantly blocked with Losartan in round cells ,and had no effect in spindle cells.AngⅡalso had no significant effect on cell proliferation and collagen synthesis of two subpopulations with PD 123319.Conclusions AngⅡhas significant difference in adventitial fibroblast subpopulations from rat thoracic aorta vascular.It could promote cell proliferation and collagen synthesis of round cells ,and the effect is achieved by angiotensin Ⅱreceptor 1 (AT1),Ang Ⅱhas no significant effect on spindle cells.The conclusion showes that the two subpopulations of adventitial fibroblast have sig -nificant differences in phenotype ,function and mechanism of action ,which suggestes that the two cell subpopulations play different roles in the pathophysiological mechanisms of vascular remodeling and repair processes.