CAJ | 학술논문

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(iNOS)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siRNA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iNOS表达,以及抑制IκB磷酸化和NF-κB激活。而用HO-1 siRNA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。
목적：：관찰근채소대지다당( LPS)유도소서거서세포분비염증개질적영향,병탐토기분자궤제。방법：체외배양소서거서세포RAW 264．7,용불동농도적근채소처리후,가입LPS자격。 Western blot검측혈홍소양합매-1(HO-1)、배양화매-2(COX-2)、유도형일양화담합성매(iNOS)적표체이급p38화IκB적린산화；비색법검측아초산염화초산염(NOx)적함량；ELISA검측PGE2적산생；보고기인분석근채소대NF-κB격활적영향。결과：근채소가현저유도RAW 264．7세포표체HO-1,채용p38억제제SB203580예처리세포후,가억제근채소유도적HO-1표체；동시,근채소능강저포장내핵전록인자Nrf2함량,병증고기재세포핵내수평；채용siRNA간우Nrf2표체후,HO-1표체량강저。차외,근채소야능억제LPS유도RAW264．7세포산생NOx화PGE2,하조COX-2화iNOS표체,이급억제IκB린산화화NF-κB격활。이용HO-1 siRNA전염후,능재일정정도상억제근채소적상술효응。결론：근채소가능통과유도HO-1표체、억제NF-κB적격활이발휘항염증작용。
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 (iNOS),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 prosta-glandin 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 siRNA could inhibit apigenin-induced HO-1 expression. Furthermore,apigenin administration significantly inhibited LPS-induced NOx production and PGE2 secretion, COX-2 and iNOS expression,IκB phosphorylation and NF-κB activation,and transfection of HO-1 siRNA could reverse these actions. Conclusion:Apigenin inhibits LPS-induced inflammatory response through induction of HO-1 and inhibition of NF-κB in macrophages.