Inhibitive effect of peritoneal decoy-oligoligomucleotidesnucleotides targeting nuclear factor-?B on expression of inflammatory mediators in macrophages ? cells / 中华创伤杂志

ABSTRACT

Objective To design and synthesize targeting nuclear factor-?B (NF-?B) decoy-oligodeoxynucleotides (ODNs) and measure their anti-inflammatory actions. Methods Peritoneal macrophages ? (pM?) cells extracted from rats were randomly divided into normal control group, lipopolysaccharides (LPS)-stimulated group (Group A), decoy-ODNs treated group (Group B), non-decoy-ODNstreated group (Group C) and cationic liposome treated group (Group D). Supernatants and cells in the control group and the Group A were collected 1,2,6,12,18 and 24 hours after LPS stimulation. By using cationic liposomes in a ratio of 41 in quantity, pM? cells were transfected by decoy-ODNs at concentrations of 2 mg/L,4 mg/L and 8 mg/L respectively and stimulated with LPS 6 hours later. Then, supernatants and cells were collected 8, 12 and 18 hours after transfection. The expression changes and the distribution of p65 were analyzed by immunocytohistochemical method, the protein expressions of tumor necrosis factor ? (TNF?), interleukin-6 (IL-6) and interleukin-10 (IL-10) in the supernatants by enzyme-linked immunosorbent assay (ELISA) and mRNA transcriptions of TNF?,IL-6 and IL-10 by reverse transcriptase polymerase chain reaction (RT-PCR). Results Decoy-ODNs at concentration of 4 mg/L and 8 mg/L could markedly inhibit the expression and transcription of TNF? and IL-6 of pM? cells in a dose-dependent fashion but had a weak inhibitive effect on the expression and transcription of IL-10. Conclusions The targeting NF-?B Decoy-ODNs can suppress the expressions of inflammatory mediators in pM? cells.