Effects of resistin gene on TNF-α-induced expression of ICAM-1 in vascular endothelial cells
10.3724/SP.J.1008.2008.00184
- Author:
Ming LIANG
1
Author Information
1. Department of Endocrinology
- Publication Type:Journal Article
- Keywords:
Intercellular adhesion molecule-1;
Resistin;
Tumor necrosis factor-α
- From:
Academic Journal of Second Military Medical University
2010;29(2):184-188
- CountryChina
- Language:Chinese
-
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.
