3,4-二羟基苯乙酮抑制NF-κB核转位减轻脂多糖诱导巨噬细胞炎症反应

3,4-DHAP inhibits NF-kappa B nuclear translocation and reduces LPS induced inflammatory response in RAW264.7 cell

目的探讨3,4-二羟基苯乙酮(3,4-DHAP)抑制脂多糖(LPS)诱导小鼠巨噬细胞株(RAW264.7)炎症反应的可能机制。方法常规培养的RAW264.7细胞随机分为正常组(A);LPS组(B);3,4-DHAP组(C)、辛伐他汀组(D)。酶联免疫吸附(ELISA)法检测肿瘤坏死因子(TNF-α)、血管细胞黏附分子(VCAM-1)水平;免疫印迹(Western blot)法检测磷酸化IκB(P-IκB)、总IκB、核因子-κBp65(NF-κBp65)蛋白表达;实时定量荧光聚合酶链式反应(q RT-PCR)法检测IκB mRNA表达。结果 3,4-DHAP显著抑制TNF-α、VCAM-1分泌;抑制IκB蛋白磷酸化;细胞核内NF-κBp65蛋白表达明显减少(P<0.05),抑制了NF-κBp65核转位。结论 3,4-DHAP通过抑制NF-κB核转位,有效抑制LPS诱导RAW264.7细胞炎症反应。

Objective To investigate the inhibitory effects and mechanisms of 3,4-dihydroxyacetophe- none （3,4-DHAP） on inflammation induced by lipopolysaccharide （LPS） in murine macrophage cells （RAW264.7）. Methods RAW264.7 cell line was cultured and randomly divided into normal group （A） , LPS group （B）, 3,4- DHAP group （C） and simvastatin group （D） . The levels of tumor necrosis factor-α （TNF-α） and vascular cell adhesion molecule-1 （VCAM-1） were detected by ELISA kits. The expression of phosphorylation of I kappa B （P-1κB） , total I kappa B （IκB） and NF-κBp65 protein were detected by Western-blotting. qRT-PCR method was used to detect the expression of IKB mRNA. Results After 3,4-DHAP treatment, the levels of TNF-α and VCAM-1 in 3,4-DHAP group were obviously decreased, the expression of P-IκB protein and IκBmRNA and protein increased significantly, and the expression of NF-κBp65 protein in the nuclear were significantly decreased （P〈0.05）. Conclusion 3,4-DHAP effectively reduce the inflammatory response induced by LPS through inhibi- tion of NF-κB nuclear translation.