芹菜素抑制脂多糖诱导小鼠巨噬细胞分泌炎症介质的分子机制  

Molecular mechanism of apigenin on inhibition of LPS-induced inflammatory mediators in murine macrophages

作　　者：吴广[1] 符平[2] 周玉生[1] 周润梅[3] 

机构地区：[1]南华大学附属第二医院药剂科,衡阳421001 [2]南华大学附属第三医院,衡阳421000 [3]南华大学药物药理研究所,衡阳421000

出　　处：《中国免疫学杂志》2015年第6期753-757,共5页Chinese Journal of Immunology

基　　金：湖南省自然科学基金衡阳联合基金(13JJ9009)资助

摘　　要：目的:观察芹菜素对脂多糖(LPS)诱导小鼠巨噬细胞分泌炎症介质的影响,并探讨其分子机制。方法:体外培养小鼠巨噬细胞RAW 264.7,用不同浓度的芹菜素处理后,加入LPS刺激。Western blot检测血红素氧合酶-1(HO-1)、环氧化酶-2(COX-2)、诱导型一氧化氮合成酶(i NOS)的表达以及p38和IκB的磷酸化;比色法检测亚硝酸盐和硝酸盐(NOx)的含量;ELISA检测PGE2的产生;报告基因分析芹菜素对NF-κB激活的影响。结果:芹菜素可显著诱导RAW 264.7细胞表达HO-1,采用p38抑制剂SB203580预处理细胞后,可抑制芹菜素诱导的HO-1表达;同时,芹菜素能降低胞浆内核转录因子Nrf2含量,并增高其在细胞核内水平;采用si RNA干扰Nrf2表达后,HO-1表达量降低。此外,芹菜素也能抑制LPS诱导RAW264.7细胞产生NOx和PGE2,下调COX-2和i NOS表达,以及抑制IκB磷酸化和NF-κB激活。而用HO-1 si RNA转染后,能在一定程度上抑制芹菜素的上述效应。结论:芹菜素可能通过诱导HO-1表达、抑制NF-κB的激活而发挥抗炎症作用。Objective： To investigate the effect and the mechanism of Apigenin on lipopolysaccharides（ LPS）-induced inflammatory mediators production in murine macrophages. Methods： The murine macrophage cell line RAW 264. 7 cells were cultured in vitro,and were treated with different concentration of Apigenin followed by LPS administration. Expression of heme oxygenase-1（ HO-1）,cyclooxygenase-2（ COX-2） and inducible nitric oxide synthase（ i NOS）,phosphorylation of p38 and IκB,nuclear translocation of Nrf2 were detected by Western blot. Production of Nitrite and nitrate（ NOx） was analyzed by colorimetric technique. Secretion of prostaglandin E2（ PGE2） was detected by ELISA. Activation of NF-κB was measured by luciferase assay. Results： Western blot indicated that apigenin could induce RAW 264. 7 cells expression of HO-1,and pretreatment of SB203580,an inhibitor of p38 significantly inhibited apigenin induced HO-1 expression. In addition,Apigenin could also decrease the content of nuclear transcription factor Nrf2 in cytoplasm and increase its level in the nucleus. Silencing of Nrf2 by specific si RNA could inhibit apigenin-induced HO-1expression. Furthermore,apigenin administration significantly inhibited LPS-induced NOx production and PGE2 secretion,COX-2 and i NOS expression,IκB phosphorylation and NF-κB activation,and transfection of HO-1 si RNA could reverse these actions. Conclusion：Apigenin inhibits LPS-induced inflammatory response through induction of HO-1 and inhibition of NF-κB in macrophages.

关 键 词：芹菜素 血红素氧合酶-1 小鼠巨噬细胞 

分 类 号：R967[医药卫生—药理学]