NF-κB“诱骗”寡核苷酸对小鼠巨噬细胞iNOS表达的影响

Effects of NF-κB “decoy” oligodeoxynucleotides on inducible nitric oxide synthase expression in LPS-induced mouse macrophages

目的:研究NF- κB“诱骗(decoy)”寡核苷酸(ODNs)对脂多糖(LPS)诱导小鼠巨噬细胞株J774.1 细胞诱导型一氧化氮合酶(iNOS)表达的影响。方法:体外培养巨噬细胞,LPS刺激后脂质体转染NF -κB“decoy”ODNs,用硝酸还原酶法测定培养液中NO水平以及iNOS含量,RT PCR观察细胞iNOS的mRNA表达变化。结果:NF- κB“decoy”ODNs可降低巨噬细胞培养体系LPS诱导的NO合成,阻断iNOS- mRNA的表达;对照序列的ODNs和转染剂对NO、iNOS无明显影响。结论:NF -κB“decoy”ODNs可以抑制iNOS的活性,减少NO生成,可能与其特异性竞争抑制活化NF -κB 结合位点有关。

Objective:To study the effect of NF-κB “decoy” oligodeoxynucleotides on nitric oxide synthase expression in LPS-induced mouse macrophages. Methods: Mouse macrophage cell lines J774.1 were cultured with LPS and liposome-mediated oligodeoxynucleotides, and the level of nitric oxide and inducible nitric oxide synthase were measured in the different culture fluids. RNA was extracted from macrophages, and the mRNA expression of macrophages inducible nitric oxide synthase was observed by RT-PCR. Results: NF-κB “decoy” oligodeoxynucleotides decreased the activity of inducible nitric oxide synthase in LPS-induced macrophage, and inhibited the generation of nitric oxide. The level of nitric oxide and inducible nitric oxide synthase did not change in control group. Conclusion: NF-κB “decoy” oligodeoxynucleotides inhibit the expression of inducible nitric oxide synthase in LPS-induced macrophages, reducing the amount of nitric oxide, which is probably due to specific inhibition of the binding sites of activated nuclear factor-kappa B.