| 摘要: |
| 目的：研究抵抗素（resistin）基因对TNFα诱导血管内皮细胞ECV304表达细胞间黏附分子1（ICAM1）的影响。方法：应用分子生物学技术，将resistin基因克隆到载体pEGFPC1中构建质粒pEG/Resi，脂质体转染ECV304，转染6 h加入20 μg/L TNFα诱导，用RTPCR方法分别检测转染18、30、42和54h时resistin和ICAM1 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ICAM1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TNFα诱导条件下转染resistin基因组的ICAM1表达水平始终高于未转染组，在resistin基因表达高峰(30和42 h)ICAM1 mRNA和蛋白表达量较未转染组均显著升高(P＜0.05)；且ICAM1 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TNFα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）. |
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| 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 TNFαinduced expression of ICAM1 in ECV304 cells. Methods: Resistin gene was cloned into vector pEGFPC1 to construct pEG/Resi plasmid; the latter was then used to transfect ECV304 cells via liposome.TNFα(20 μg/L) was added to the system 6 h after transfection. RTPCR was used to detect the expression of resistin and ICAM1 mRNA in the cells 18, 30, 42 and 54 h after transfection; meanwhile, the resistin protein levels were determined by Western blotting analysis. ICAM1 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. ICAM1 levels in resistintransfected group were consistently higher than that in the nontransfected group in the presence of TNFα. ICAM1 mRNA and protein expression in the transfectedgroup was significantly higher than that in the nontransfected group during the peak periods (30 and 42 h) of resistin expression (P<0.05); ICAM1 mRNA and protein expression increased with the increase of resistin gene expression. Conclusion: Resistin gene can enhance TNFαinduced expression of ICAM1 in ECV304 cells. |
| Key words: resistin tumor necrosis factor α intercellular adhesion molecule 1 |
