香草乙酮改善葡聚糖硫酸钠诱发溃疡性结肠炎小鼠的炎症反应与NOXs-ROS-p38MAPK信号通路的关系

发布时间：2018-05-06 00:05

本文选题：香草乙酮 + NADPH氧化酶　；参考：《生理学报》2015年01期

【摘要】：本文旨在探讨香草乙酮(apocynin)治疗葡聚糖硫酸钠(dextran sulfate sodium,DSS)诱导的溃疡性结肠炎(ulcerative colitis,UC)小鼠的分子机制。以饮用水配制5%DSS诱发UC动物模型,2%香草乙酮治疗UC小鼠,采样并应用HE染色进行结肠组织病理学评估。采用鲁米诺化学发光方法检测结肠组织活性氧自由基(reactive oxygen species,ROS)生成及DPI抑制后的NADPH消耗率分析NADPH氧化酶(NADPH oxidases,NOXs)活性,Western blot法检测p38MAPK磷酸化水平,Griess方法分析NO,酶联免疫法检测前列腺素E2(prostaglandin E2,PGE2),real time PCR及Western blot法检测i NOS、COX2的表达,酶联免疫吸附实验测定细胞因子TNF-α、IL-6、IFN-γ、IL-1β的水平。体外实验采用分离的结肠组织中性粒细胞,检测其ROS生成、NOXs活性、NO、PGE2的变化,并应用Western blot检测其NOX1、p-p38MAPK的表达。结果显示,香草乙酮治疗后UC小鼠结肠组织NOXs活性及ROS生成被抑制(P0.01),p38MAPK磷酸化水平降低,NO、PGE2及相关细胞因子生成减少(P0.01),UC炎症反应得到缓解。在炎症部位的中性粒细胞中,香草乙酮抑制ROS生成及NOX活性(P0.01),并降低NOX1的表达、p38MAPK的磷酸化及NO、PGE2的生成(P0.01)。因此,香草乙酮可能通过NOXs-ROS-p38MAPK信号通路缓解DSS诱发UC小鼠的炎症反应,中性粒细胞是参与其保护机制的主要炎症细胞。
[Abstract]:The aim of this study was to investigate the molecular mechanism of vanilanone apocynin in the treatment of ulcerative colitis induced by dextran sulfate sodiumn (DSS) in mice. Colonic histopathology was evaluated by HE staining in UC mice treated with 2% vanillanone in drinking water prepared with 5%DSS. Luminol Chemiluminescence Assay was used to detect the production of reactive oxygen in Colon tissue and the NADPH consumption rate after DPI inhibition; the activity of NADPH Oxidase NADPH oxide NOXs; Western blot assay to detect the level of p38MAPK phosphorylation; the enzyme linked immunosorbent assay (Elisa) to detect the level of p38MAPK phosphorylation; the enzyme linked immunosorbent assay (Elisa). Prostaglandin E2(prostaglandin E2P real time PCR and Western blot method were used to detect the expression of iNOS and COX2. Enzyme linked immunosorbent assay (Elisa) was used to determine the level of cytokine TNF- 伪, IL-6, IFN- 纬 and IL-1 尾. In vitro, neutrophils were isolated from colonic tissue to detect the changes of the activity of ROS producing noxs and the expression of NOX1 p-p38 MAPK by Western blot. The results showed that the inhibition of NOXs activity and ROS production in colonic tissue of UC mice after treatment with vanilanone reduced the phosphorylation level of P0.01p38MAPK and the inflammatory response of UCU with the decrease of P0.01MAPK phosphorylation and the decrease of cytokine production. In neutrophils at the inflammatory site, vanillin inhibited the production of ROS and the activity of NOX (P0.01), and decreased the phosphorylation of p38 MAPK and the formation of PGE2 in NOX1. Therefore, Vanilla ethanone may alleviate the inflammatory response of UC mice induced by DSS through NOXs-ROS-p38MAPK signaling pathway, and neutrophils are the main inflammatory cells involved in its protective mechanism.
【作者单位】：河南大学淮河医院检验科;河南大学第一附属医院肾内科;河南大学淮河医院消化内科;河南大学淮河医院普通外科;
【基金】：supported by the National Natural Science Foundation of China(No.U1304802)
【分类号】：R574.62

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