核因子-κB反义核酸对前炎症细胞因子表达的实验研究

Effect of NF-κB antisense oligodeoxynucleotides on expression of proinflammatory factors

目的 研究核因子(NF) κB反义寡核苷酸对NF κB表达及其下游前炎症细胞因子表达的影响。方法 将不同浓度经硫代磷酸化修饰的反义寡核苷酸(PS AODN)导入培养的RAW 2 6 4 .7细胞中(与脂多糖预孵育) ,设立空白对照组和错义寡核苷酸(MODN)组。采用免疫组织化学SABC法检测细胞中NF κBp65、内皮细胞黏附因子(ICAM )、一氧化氮合酶(iNOS)的表达。结果 AODN组的NF κB p65,ICAM ,iNOS表达较空白对照组及MODN组明显降低(P <0 .0 5 ) ,且在1.0～5 .0 μmol/L范围内AODN的作用随其浓度增加而增加。结论 AODN能有效阻抑NF κB在脂多糖刺激下的表达,从而抑制各前炎症细胞因子的表达。

Objective To study the effect of NF-κB antisense oligodeoxynucleotides (AODN) on expression of NF-κB and proinflammatory factors. Methods The synthesized AODN modified by phosphorothioate at different concentrations were added to the medium of cultured RAW 264.7 cells (preincubation with lipopolysaccharide). Protein expressions of NF-κB p 65 、ICAM、iNOS were detected by SABC immuno-histochemical technique. Results The expressions of NF-κB p 65 、ICAM and iNOS of AODN groups were significantly lower than blank control and MODN groups. From the concentration of 1.0 μM to 5.0 μM , the inhibitory effects increased with the increasing concentration. Conclusion AODN can effectively inhibit the expression of NF-κB protein, thus can depress the expression of proinflammatory factors.