5-Hydroxy-4-methoxycanthin-6-one alleviates dextran sodium sulfate-induced colitis in rats via regulation of metabolic profiling and suppression of NF-κB/p65 signaling pathway
5-Hydroxy-4-methoxycanthin-6-one alleviates dextran sodium sulfate-induced colitis in rats via regulation of metabolic profiling and suppression of NF-κB/p65 signaling pathway
复制标题
5-羟基-4-甲氧基canthin-6-one 通过调节代谢谱和抑制 NF-κB/p65 信号通路减轻大鼠右旋糖酐钠诱导的结肠炎
DOI：
10.1016/j.phymed.2020.153438
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发表时间：
2021-01-07
期刊：
影响因子：
7.9
通讯作者：
Lin, Chaozhan
中科院分区：
文献类型：
--
作者：
Liu, Fangle;Yao, Yufeng;Lin, Chaozhan
背景资料：目的：通过非靶向代谢组学研究，阐明苦木中的主要活性成分5-羟基-4-甲氧基坎黄酮-6-酮（PQ-A）在结肠炎治疗中的作用和机制。观察各组大鼠的体重、疾病活动指数（DAI）、结肠长度、结肠组织病理学评分及生化指标（MDA、SOD）。进行了基于UPLC-Q-TOF-MS/MS方法的代谢组学分析，以探索PQ-A在结肠炎治疗中的潜在机制。测定血清中炎性细胞因子（TNF-α、IL-1 β、IL-6和IL-10）浓度及其在结肠中的蛋白水平。半定量结肠中的CD 3和NF-κ B/p65免疫组织化学。Western blotting法检测IKK-NF-kappa B/p65信号通路中相关蛋白的表达，RT-PCR法检测IKK-NF-kappa B/p65信号通路中相关mRNA的表达。使用DS 2.5软件预测PQ-A与NF-κ B/p65之间潜在的分子相互作用。结果：PQ-A可明显预防结肠炎大鼠体重减轻和结肠缩短，并降低DAI和组织病理学评分。PQ-A可降低UC大鼠血清MDA水平，升高SOD水平。代谢组学结果显示，49种差异代谢物可作为DSS诱导的结肠炎的生物标志物，表明结肠炎的路径机制涉及8条代谢途径的干扰，包括α-亚麻酸和亚油酸代谢、鞘脂代谢、视黄醇代谢、胆汁酸代谢、PQ-A通过调节α-亚麻酸和亚油酸代谢，鞘脂代谢，和视黄醇代谢，有效地提示了PQ-A与NF-κ B B/p65炎症信号相关的潜在药理机制。分子对接结果预测PQ-A和NF-κ B/p65之间的高亲和力相互作用，涉及5个氨基酸残基的氢键相互作用，表明NF-κ B/p65是靶点。PQ-A降低结肠炎大鼠血清中TNF-α、IL-1 β和IL-6浓度及其结肠组织中的蛋白水平。PQ-A治疗后，结肠组织中CD 3、MYD 88、p-I κ B α、NF-κ B/p65和p-NF-κ B/p65表达水平降低，而IKK β和I κ B α表达水平升高。PQ-A可显著抑制NF-κ B B/p65的核转位。结论：从血清非靶向代谢组学的角度对PQ-A治疗结肠炎的可能作用机制进行了综述。PQ-A治疗可以通过抑制NF-κ B/p65信号通路来保护大鼠免受DSS诱导的结肠炎。
Background: 5-Hydroxy-4-methoxycanthin-6-one (PQ-A) is the main active compound in Ramulus et Folium Picrasmae, a Chinese herbal medicine commonly used in colitis treatment.Purpose: To clarify PQ-A's role and mechanism in colitis treatment based on a non-targeted metabolomics study.Methods: Rats with ulcerative colitis (UC) established with 4% dextran sulfate sodium (DSS) were orally treated with PQ-A. Body weight, disease activity index (DAI), colon length, biochemical parameters (MDA and SOD), and histopathological score in colon tissue were measured. A UPLC-Q-TOF-MS/MS approach-based metabolomics analysis was conducted to explore the underlying mechanisms of PQ-A in colitis treatment. Inflammatory cytokines (TNF-alpha, IL-1 beta, IL-6, and IL-10) concentrations in serum and their protein levels in the colon were determined. CD3 and NF-kappa B/p65 immunohistochemistry in the colon was semi-quantified. The related protein or mRNA in IKK-NF-kappa B/p65 signaling pathway was measured by Western blotting or RT-PCR, respectively. Potential molecular interactions between PQ-A and NF-kappa B/p65 was predicted using DS 2.5 software.Results: PQ-A significantly prevented body weight loss and colonic shortening in colitic rats, and reduced the DAI and histopathologic score as well. PQ-A decreased MDA levels in the UC rat serum and increased those of SOD. Metabolomics results revealed forty-nine differential metabolites as biomarkers of DSS-induced colitis, demonstrating that the path-mechanism of colitis involved the perturbation of eight metabolic pathways, including alpha-linolenic acid and linoleic acid metabolism, sphingolipid metabolism, retinol metabolism, bile acid metabolism, et al. Thirty-six biomarkers were especially reversed to normal-like levels by PQ-A via regulation of alpha-linolenic acid and linoleic acid metabolism, sphingolipid metabolism, and retinol metabolism, which effectively hinted the potential pharmacological mechanism of PQ-A related to NF-kappa B/p65 inflammatory signaling. Molecular docking results predicted high affinity interaction between PQ-A and NF-kappa B/p65, involving hydrogen-bond interactions at five amino acid residues, suggesting NF-kappa B/p65 as a target. PQ-A decreased TNF-alpha, IL-1 beta, and IL-6 concentrations in serum and their protein levels in colon tissue in colitic rats. CD3, MYD88, p-I kappa B alpha, NF-kappa B/p65, and p-NF-kappa B/p65 expression levels decreased, whereas those of IKK beta and I kappa B alpha increased in colitic tissue following PQ-A treatment. PQ-A strongly inhibited nuclear translocation of NF-kappa B/p65.Conclusions: We provide an overview of PQ-A's possible mechanism of action in colitis treatment based on serum non-targeted metabolomics. PQ-A treatment can protect rats against DSS-induced colitis by suppressing the NF-kappa B/p65 signaling pathway.