The anti-inflammatory carbazole, LCY-2-CHO, inhibits lipopolysaccharide-induced inflammatory mediator expression through inhibition of the p38 mitogen-activated protein kinase signaling pathway in macrophages
The anti-inflammatory carbazole, LCY-2-CHO, inhibits lipopolysaccharide-induced inflammatory mediator expression through inhibition of the p38 mitogen-activated protein kinase signaling pathway in macrophages
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DOI：
10.1038/sj.bjp.0705700
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发表时间：
2004-03-01
影响因子：
7.3
通讯作者：
Lin, WW
中科院分区：
文献类型：
--
作者：
Ho, FM;Lai, CC;Lin, WW
1本研究旨在研究合成化合物LCY-2-CHO对小鼠RAW 264.7巨噬细胞中诱导型一氧化氮合酶（iNOS）、考克斯-2和TNF-α表达的抗炎作用。2在1-30 μ M范围内，LCY-2-CHO浓度依赖性地抑制脂多糖（LPS）诱导的一氧化氮（NO）、前列腺素E-2（PGE（2））、和肿瘤坏死因子-α（TNF-α）形成，IC 50值分别为2.3、1和0.8 μ M。伴随着LPS诱导的iNOS、环氧合酶-2（考克斯-2）和pro-TNF-α蛋白的抑制作用也被证实。3逆转录-聚合酶链反应（RT-PCR）和启动子分析表明，iNOS的表达在转录水平上受到抑制（IC 50 = 2.3 μ M），考克斯-2表达的抑制仅部分依赖于基因转录（IC 50 = 7.6 μ M），并且TNF-α的转录未受影响。4转录测定显示AP-1的激活，但不包括NF-κ B的激活，同时被LCY-2-CHO阻断。我们的结果表明，LCY-2-CHO能够干扰转录后调节，改变考克斯-2和TNF-α mRNA的稳定性。5由于考克斯-2和TNF-α mRNA的3 '非翻译区（3' UTR）都含有参与mRNA稳定性的p38促分裂原活化蛋白激酶（MAPK）调节元件，我们评估了LCY-2-CHO对p38 MAPK的影响。我们的数据清楚地表明，LPS介导的p38 MAPK活性的抑制（IC 50 = 1.7 μ M），但不细胞外信号调节激酶（ERK）或c-Jun N-末端激酶（JNK）活性。然而，激酶测定排除了p38 MAPK作用的直接抑制。选择性p38 MAPK抑制剂SB 203580抑制iNOS和考克斯-2的启动子活性，而不是TNF-α的启动子活性。6总之，LCY-2-CHO通过干扰p38 MAPK和AP-1的活化而下调巨噬细胞中炎性iNOS、考克斯-2和TNF-α的基因表达。
1 The present study was undertaken to investigate the anti-inflammatory effects of a synthetic compound, LCY-2-CHO, on the expression of inducible nitric oxide synthase (iNOS), COX-2, and TNF-alpha in murine RAW264.7 macrophages.2 Within 1-30 muM, LCY-2-CHO concentration-dependently inhibited lipopolysaccharide (LPS)-induced nitric oxide (NO), prostaglandin E-2 (PGE(2)), and tumor necrosis factor-alpha (TNF-alpha) formation, with IC50 values of 2.3, 1, and 0.8 muM, respectively. Accompanying inhibition of LPS-induced iNOS, cyclooxygenase-2 (COX-2), and pro-TNF-alpha proteins was observed.3 Reverse transcription-polymerase chain reaction (RT-PCR) and promoter analyses indicated that iNOS expression was inhibited at the transcriptional level (IC50 = 2.3 muM), that inhibition of COX-2 expression only partially depended on gene transcription (IC50 = 7.6 muM), and that TNF-alpha transcription was unaffected.4 Transcriptional assays revealed that activation of AP-1, but not NF-kappaB, was concomitantly blocked by LCY-2-CHO. Our results showed that LCY-2-CHO was capable of interfering with post-transcriptional regulation, altering the stability of COX-2and TNF-alpha mRNAs.5 Since the 3'-untranslated region (3' UTR) of both COX-2 and TNF-alpha mRNA contains a p38 mitogen-activated protein kinase (MAPK)-regulated element involved in mRNA stability, we assessed the effect of LCY-2-CHO on p38 MAPK. Our data clearly indicated an inhibition (IC50 = 1.7 muM) of LPS-mediated p38 MAPK activity, but not of extracellular signal-regulated kinase (ERK) or c-Jun N-terminal kinase (JNK) activity. However, kinase assays ruled out a direct inhibition of p38 MAPK action. The selective p38 MAPK inhibitor, SB203580, inhibited the promoter activities of iNOS and COX-2 rather than that of TNF-alpha.6 In conclusion, LCY-2-CHO downregulates inflammatory iNOS, COX-2, and TNF-alpha gene expression in macrophages through interfering with p38 MAPK and AP-1 activation.