MicroRNA-146a Decreases High Glucose/Thrombin-Induced Endothelial Inflammation by Inhibiting NAPDH Oxidase 4 Expression
Figure 2
Endothelial inflammation was induced by HG/thrombin through an ROS-dependent pathway. (a) From top to bottom, multiple small dots accumulated into 6 horizontal lines, which represented the expression levels of IL-8, IL-1β, IL-6, IL-10, TNF-, and IL-12p70 proteins in conditioned medium. The rightward shift of the dotted lines represents the relative protein expression levels. Analysis using an inflammatory cytokine kit showed that HG/thrombin stimulation increased IL-8 and IL-6 expression in HAECs, while L-1β, IL-10, TNF-, and IL-12p70 had no discernible effect on its protein expression levels. The images shown represent the results from 3 independent experiments. (b) Stimulation of HAECs with HG/thrombin for 5 h increased caused 4.06-fold and 5.05-fold increases of IL-6 and IL-8 mRNA gene expression levels, respectively. The bars represent the means ± SEM from 3 experiments. , as compared with the control. (c) Stimulation of HAECs with HG/thrombin for 5 h induced an 11.43-fold synergistic increase of THP-1 adhesiveness to HAECs compared with the control. The bars represent the means ± SEM from 4 experiments. , as compared with the control. (d) Stimulation of HAECs with HG/thrombin for 5 h induced significant 8.72-, 11.67-, and 67.21-fold increases of VCAM-1, ICAM-1, and SELE gene expression, respectively. The bars represent the means ± SEM from 3 experiments. , as compared with the control. (e) HAECs were pretreated with 5 mM N-acetylcysteine (NAC) for 1 h and then stimulated with HG/thrombin for 5 h. Real-time PCR showed that NAC pretreatment caused significant decreases in the gene expression of VCAM-1, ICAM-1, and SELE to 51%, 70%, and 49% of that in the HG/thrombin-stimulated HAECs. The bars represent the means ± SEM from 4 experiments. and , as compared with the HG/thrombin.
