Fig. 7

Inhibition of HIF-1α decreases inflammation and oxidative stress in HUVECs via JMJD1A induced by high glucose and hypoxia. a Quantitative gene expression changes of JMJD families under high glucose and hypoxia. Cells were treated with high glucose and hypoxia for 6, 12, 24, and 48 h, qRT-PCR (b) and western blotting (c) showing the expression of JMJD1A. d Cells were treated with or without KC7F2 (10 µM) before exposure to high glucose and hypoxia for 48 h. The total proteins of HIF-1α and JMJD1A were collected and analyzed using western blotting as described under Materials and Methods. The relative densities of HIF-1α and JMJD1A were calculated according to GAPDH. e Cells were treated with high glucose and hypoxia for 24 h. ChIP assay showing HIF-1α bound to the JMJD1A promoter in vitro. The promoter regions of JMJD1A (–2.5 kb to –500 bp) were amplified using the input and immunoprecipitated DNA as templates. g Cells were treated with si-HIF-1α/KC7F2 and JMJD1A overexpression induced by high glucose and hypoxia for 48 h. ELISA and flow cytometry showing the secretion of (g) IL-6, ICAM-1, and h ROS. n = 3; *p < 0.05 and **p < 0.01 vs. DMSO. DMSO-treated cells, DMSO; HIF-1α inhibitors KC7F2 (10 µM)-treated cells, KC7F2 (10 µM); HG, high glucose; HG + Hypoxia, combined stimulus with high glucose and hypoxia; NG, control