The carbon monoxide-releasing molecule CORM-2 inhibits the inflammatory response induced by cytokines in Caco-2 cells
The carbon monoxide-releasing molecule CORM-2 inhibits the inflammatory response induced by cytokines in Caco-2 cells
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DOI：
10.1038/sj.bjp.0707184
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发表时间：
2007-04-01
影响因子：
7.3
通讯作者：
Alcaraz, M. J.
中科院分区：
文献类型：
--
作者：
Megias, J.;Busserolles, J.;Alcaraz, M. J.
背景与目的：最近的证据表明，一氧化碳释放分子（CO-RM）具有潜在的抗炎活性。本研究探讨了CORM-2对细胞因子诱导的人结肠上皮细胞Caco-2炎症反应的调控作用。实验方法：将CORM-2与Caco-2共同孵育30 min后，分别用白细胞介素（IL）-1b、肿瘤坏死因子-α和干扰素-γ刺激Caco-2细胞不同时间。通过实时PCR分析基因表达。Western blot和ELISA检测蛋白表达。通过荧光素酶法测定转录因子激活。关键结果：我们已经表明，CORM-2显着降低一氧化氮合酶-2（NOS-2）的mRNA表达和亚硝酸盐的产生，在Caco-2细胞与细胞因子刺激。IL-8、IL-6和金属蛋白酶-7（MMP-7）的mRNA和蛋白也被CORM-2显著降低。时间进程和小干扰RNA研究表明，抑制IL-6在CORM-2调节MMP-7表达中起作用。CORM-2的这些作用可能依赖于核因子-κ B的调节（NF-κ B）、激活蛋白-1、CCAT/增强子结合蛋白和NF-κ B抑制蛋白-α、c-Jun N-末端蛋白激酶1/2、p38和细胞外信号调节激酶1/2的磷酸化形式。CORM-2可以调节许多与肠道炎症和癌症进展相关的基因。这些发现为这类化合物的抗炎特性和潜在应用提供了新的见解。
Background and purpose: Recent evidence indicates that carbon monoxide-releasing molecules ( CO-RMs) exhibit potential anti-inflammatory properties. In the present study, we have investigated whether tricarbonyl dichloro ruthenium( II) dimer ( CORM-2) can control the inflammatory response induced by cytokines in a human colonic epithelial cell line, Caco-2.Experimental approach: Caco-2 cells were preincubated with CORM-2 for 30 minutes and then stimulated with interleukin ( IL)-1b, tumor necrosis factor-alpha and interferon-gamma for different times. Gene expression was analyzed by real-time PCR. Protein expression was investigated by Western blot and ELISA. Transcription factor activation was determined by the luciferase method.Key results: We have shown that CORM-2 significantly decreased the mRNA expression of nitric oxide synthase-2 ( NOS-2) and the production of nitrite, in Caco-2 cells stimulated with cytokines. IL-8, IL-6 and metalloproteinase-7 ( MMP-7) mRNA and protein were also significantly reduced by CORM-2. Time-course and small interfering RNA studies suggest that inhibition of IL-6 plays a role in the regulation of MMP-7 expression by CORM-2. These effects of CORM-2 can be dependent on the modulation of nuclear factor-kappa B ( NF-kappa B), activator protein-1, CCAT/enhancer binding protein and the phosphorylated forms of NF-kappa B inhibitory protein-alpha, c-Jun N-terminal protein kinase 1/2, p38 and extracellular signal-regulated kinase 1/2.Conclusions and Implications: CORM-2 can regulate a number of genes relevant in intestinal inflammation and cancer progression. These findings provide new insights into the anti-inflammatory properties and potential applications of this class of compounds.