中国农业科学2012，Vol.45Issue(2)：346-352,7.DOI:10.3864/j.issn.0578-1752.2012.02.017
五指山小型猪近交系主动脉内皮细胞体外培养模型的建立和TLR2介导炎症相关因子的表达
Establishment of Aortic Endothelial Cells Model in Vitro from Wuzhishan of Inbred Miniature Pig and Expression of Inflammatorv Cvtokines Mediated by TLR2
摘要
Abstract
[Objective] This study aimed to explore a protocol of isolation, culture and identification of Wuzhishan Inbred Miniature Pig aortic endothelial cells (EC), and to study the regulation of Toll-like receptor 2 (TLR2) expression by LPS in EC with the expression of inflammatory cytokines mediated by TLR2. [Method] Thoracoabdominal aortic were isolated from 3-4 month-old Wuzhishan Inbred Miniature Pig, disassociated with 0.1% collagenase I. Aortic EC were identified with the methods of uptake of fluorescence-labeled acetylated low density lipoproteins (Dil-Ac-LDL) and detecting of CD31 expression in the cells by flow cytometry. After stimulation of aortic derived EC with 1 ugmL"1 lipopolysaccharide (LPS) for 0, 6, 8 and 12 h, the level of TLR2 and TLR4 mRNA was detected by real-time PCR. Furthermore, we detected the influence of lipoteichoic acid (LTA) on IL-6, IL-8 and ICAM-1 expression in aortic EC pretreated for 12 h with 1 ugmL"1 LPS. [ Result] Obtained Wuzhishan Inbred Miniature Pigaortic EC displayed paving stone-like pattern with high vitality. The cultured cells expressed CD31 and Ac-LDL Receptor in membrane, demonstrating that they were indeed EC. Stimulation of aortic derived EC with LPS induced a strong increase in TLR2 mRNA but not in TLR4 mRNA. LTA potentiated the increase of the inflammatory cytokines IL-6, IL-8 and ICAM-1 expression in EC pretreated with LPS, but not in EC without LTA. [Conclusion]The present study successfully isolated and identified Wuzhishan Inbred Miniature Pig aortic EC in vitro culture system. LPS stimuli induced a high TLR2 expression in EC, which mediated the expression of inflammatory cyokines.关键词
五指山小型猪近交系/主动脉内皮细胞/TLR2Key words
Wuzhishan Inbred Miniature Pig/ aortic endothelial cells/ TLR2引用本文复制引用
王攀,陶晓莉,沈祖楠,李和刚,潘杰,牟玉莲,李奎..五指山小型猪近交系主动脉内皮细胞体外培养模型的建立和TLR2介导炎症相关因子的表达[J].中国农业科学,2012,45(2):346-352,7.基金项目
国家自然科学基金项目 (30871775) （30871775）
