Inhibitory effect of circular dumbbell decoy ODNs targeting NF-кB on cytokines expression in mesangial cells

Objective To investigate the inhibitory effects of circular dumbbell decoy oligodeoxynucleotides(ODNs) targeting NF-κB on the transcriptional activity of NF-κB binding its cis-elements and the expression of cytokines(IL-6 and TGF-β_1) in mesangial cells. Methods The inhibitory effect of circular dumbbell decoy ODNs on the transcriptional activity of NF-κB was detected by electrophoretic mobilitory shift assay (EMSA). Mesangial cells extracted from rats were randomly divided into normal control, LPS-stimulated, circular dumbbell decoy-ODNs treated, scrambled-decoy-ODNs treated and cationic liposome treated group. Circular dumbbell decoy ODNs at the doses of 2 mg/L, 4 mg/L, and 8 mg/L were transfected respectively into mesangial cells from rats using cationic liposome vector. Mesangial cell were stimulated with LPS at 6 h after circular dumbbell decoy-ODNs transfection. Then, the transfected supernatants and cells were collected at 8 h, 12 h. Cytokines mRNA expression in mesangial cells was detected by RT-PCR and cytokines protein level by ELISA. Results Circular dumbbell decoy ODNs could effectively inhibit the DNA binding activity of NF-кB to its cis-elements in vitro. Circular dumbbell decoy ODNs at the doses of 4 and 8 mg/L could effectively inhibit the expression levels of IL-6, TGF-β1 mRNA and protein in mesangial cells.Conclusion Circular dumbbell decoy ODNs targeting NF-κB could effectively inhibit the expression of cytokines in mesangial cells through the inhibition of NF-κB activity.