Inhibitory Effect of Circular Dumbbell Decoy ODNs Targeting NF-KB on TNF-α Expression in Mesangial Cells

Objective To study the inhibitory effects of circular dumbbell decoy oligodeoxynucleotides targeting NF κB on the expression of TNF-α in mesangial cells. Methods According to κB sequence, circular dumbbell decoy-ODNs including 58 bases and 2 copies of κB sequences were designed and synthesized. Mesangial cells of rats were extracted and randomly divided into normal control, LPS-stimulated, circular dumbbell decoy-ODNs treated, scrambled-decoy-ODNs treated and cationic liposome treated groups. With carriers of cationic liposomes, 2 mg/L,4 mg/L, and 8 mg/L decoy ODNs were respectively transfected into mesangial cells of rats. The mesangial cells were stimulated with LPS 6h after circular dumbbell decoy-ODNs transfection. The supernatants and cells were collected at 8, 12 and 18 hours after decoy-ODNs transfection. TNF-α mRNA transcription in the mesangial cells was assessed by RT-PCR and TNF-αprotein expression was determined by ELISA. Results Circular dumbbell decoy ODNs could effectively inhibit the DNA binding activity of NF -κB in vitro. After using decoy ODNs, the expression levels of TNF-α mRNA and protein in the mesangial cells significantly decreased. Conclusions Circular dumbbell decoy ODNs targeting NF-κB could effectively inhibit the expression of TNF-α in the mesangial cells.