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Resistin基因对TNF-α诱导血管内皮细胞ICAM-1表达的影响
梁明1,栾晓军1,陈小雨1,梁敏2,唐小龙1,3*
0
(1.广东省佛山市第一人民医院内分泌科,佛山 528000;2.广西医科大学第一附属医院内分泌科,南宁 530021;3.暨南大学医学院微生物学与免疫学教研室,广州 510632)
摘要:
目的:研究抵抗素(resistin)基因对TNFα诱导血管内皮细胞ECV304表达细胞间黏附分子1(ICAM1)的影响。方法:应用分子生物学技术,将resistin基因克隆到载体pEGFPC1中构建质粒pEG/Resi,脂质体转染ECV304,转染6 h加入20 μg/L TNFα诱导,用RTPCR方法分别检测转染18、30、42和54h时resistin和ICAM1 mRNA表达;同时用免疫印迹法检测resistin蛋白的表达,并用ELISA方法检测各期ECV304上清ICAM1蛋白表达。 结果:经酶切鉴定和测序证实质粒pEG/Resi构建成功并可在ECV304中表达;相同TNFα诱导条件下转染resistin基因组的ICAM1表达水平始终高于未转染组,在resistin基因表达高峰(30和42 h)ICAM1 mRNA和蛋白表达量较未转染组均显著升高(P<0.05);且ICAM1 mRNA和蛋白表达水平均随resistin基因表达量的增加而升高。 结论:Resistin基因能够增强TNFα诱导ECV304细胞表达ICAM-1。
关键词:  resistin  肿瘤坏死因子α  细胞间黏附分子1
DOI:10.3724/SP.J.1008.2008.00184
投稿时间:2007-06-27
基金项目:广东省医学科研基金(A2005340).
Effects of resistin gene on TNF α induced expression of ICAM 1 in vascular endothelial cells
LIANG Ming1,LUAN Xiao-jun1,CHEN Xiao-yu1,LIANG Min2,TANG Xiao-long1 3*
(1. Department of Endocrinology, First People’s Hospital of Foshan, Foshan 528000, China;2. Department of Endocrinology, The First Affiliated Hospital of Guangxi Medical University, Nanning 530021;3. Department of Microbiology and Immunology, Medical College, Jinan University, Guangzhou 510632)
Abstract:
Objective:To study the effect of resistin gene on TNFαinduced expression of ICAM1 in ECV304 cells. Methods: Resistin gene was cloned into vector pEGFPC1 to construct pEG/Resi plasmid; the latter was then used to transfect ECV304 cells via liposome.TNFα(20 μg/L) was added to the system 6 h after transfection. RTPCR was used to detect the expression of resistin and ICAM1 mRNA in the cells 18, 30, 42 and 54 h after transfection; meanwhile, the resistin protein levels were determined by Western blotting analysis. ICAM1 protein levels in the supernatants of ECV304 cells were determined by ELISA. Results: Restriction enzyme digestion and sequencing demonstrated that the recombinant plasmid was successfully constructed and was expressed in ECV304. ICAM1 levels in resistintransfected group were consistently higher than that in the nontransfected group in the presence of TNFα. ICAM1 mRNA and protein expression in the transfectedgroup was significantly higher than that in the nontransfected group during the peak periods (30 and 42 h) of resistin expression (P<0.05); ICAM1 mRNA and protein expression increased with the increase of resistin gene expression. Conclusion: Resistin gene can enhance TNFαinduced expression of ICAM1 in ECV304 cells.
Key words:  resistin  tumor necrosis factor α  intercellular adhesion molecule 1