Li Yun-Xia, Liu Jinzhong, Li Fang
Anorectal Department, The People's Hospital of Huaiyin.Jinan, Jinan, Shandong, 250021, China.
Anorectal Department, Linqu County People's Hospital, Weifang, Shandong, 262699, China.
Cell Biochem Biophys. 2024 Sep;82(3):2747-2757. doi: 10.1007/s12013-024-01391-w. Epub 2024 Jul 8.
As a common inflammatory bowel disease, ulcerative colitis (UC) is featured with inflammation, oxidative damage, and the impairment of intestinal mucosal barrier, which bring threat to patients' quality of live. Hinesol, derived from Atractylodes lancea, is a unique sesquiterpenoid. Our study proposed to survey the effects and mechanism of hinesol in UC. UC mouse model was constructed using dextran sulfate sodium (DSS). Lipopolysaccharide (LPS) was applied for RAW264.7 cells stimulation to construct cell inflammatory model. The changes of disease activity index (DAI), body weight, colon length, and intestinal pathology in mice were analyzed to estimate the severity of colitis. Enzyme-linked immunosorbent assay was applied to check the changes of interleukin (IL)-1β, IL-18, IL-6, and tumor necrosis factor (TNF)-α. The levels of myeloperoxidase (MPO), superoxide dismutase (SOD), glutathione peroxidase (GSH-px), catalase (CAT), and malondialdehyde (MDA) were estimated by corresponding reagent kit. The changes of phosphorylated (p)-NF-κB P65, and p-IκBα, ZO-1, Occludin, Claudin-1, Src, XCL1, CCL2, and CXCL16 protein were examined using western blot. Flow cytometry and cell counting kit-8 assay were utilized for assessment of cell apoptosis and viability. We found that DSS reduced mice body weight, increased DAI, shorten colon length, and led to severe enteric mucosal injury, while hinesol improved the above symptoms induced by DSS. In DSS mice, hinesol raised the levels of ZO-1, Occludin, Claudin-1, SOD, GSH-px, and CAT and decreased the levels of TNF-α, IL-18, IL-1β, IL-6, MPO, and MDA. Additionally, in DSS mice and LPS-stimulated RAW264.7 cells, hinesol inhibited the high expression of Src, XCL1, CCL2, CXCL16, p-NF-κB P65, and p-IκBα. The molecular docking showed that there was a good interaction between hinesol and Src. Moreover, in LPS-stimulated RAW 264.7 cells, Src overexpression partially reversed the inhibition of hinesol on cell apoptosis, pro-inflammatory factors, and oxidative stress. In conclusion, hinesol alleviated DSS-induced colitis, which might have a bearing on the inhibition of Src-mediated NF-κB and chemokine signaling pathway.
作为一种常见的炎症性肠病，溃疡性结肠炎（UC）具有炎症、氧化损伤和肠黏膜屏障受损的特征，这对患者的生活质量构成威胁。苍术素源自白术，是一种独特的倍半萜类化合物。我们的研究旨在探究苍术素在溃疡性结肠炎中的作用及机制。使用葡聚糖硫酸钠（DSS）构建UC小鼠模型。应用脂多糖（LPS）刺激RAW264.7细胞构建细胞炎症模型。分析小鼠疾病活动指数（DAI）、体重、结肠长度和肠道病理学的变化，以评估结肠炎的严重程度。采用酶联免疫吸附测定法检测白细胞介素（IL）-1β、IL-18、IL-6和肿瘤坏死因子（TNF）-α的变化。通过相应试剂盒测定髓过氧化物酶（MPO）、超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GSH-px）、过氧化氢酶（CAT）和丙二醛（MDA）的水平。使用蛋白质印迹法检测磷酸化（p）-NF-κB P65、p-IκBα、紧密连接蛋白1（ZO-1）、闭合蛋白（Occludin）、Claudin-1、Src、趋化因子配体1（XCL1）、趋化因子配体2（CCL2）和趋化因子配体16（CXCL16）蛋白的变化。利用流式细胞术和细胞计数试剂盒-8法评估细胞凋亡和活力。我们发现，DSS降低了小鼠体重，增加了DAI，缩短了结肠长度，并导致严重的肠道黏膜损伤，而苍术素改善了DSS诱导的上述症状。在DSS诱导的小鼠中，苍术素提高了ZO-1、Occludin、Claudin-1、SOD、GSH-px和CAT的水平，降低了TNF-α、IL-18、IL-1β、IL-6、MPO和MDA的水平。此外，在DSS诱导的小鼠和LPS刺激的RAW264.7细胞中，苍术素抑制了Src、XCL1、CCL2、CXCL16、p-NF-κB P65和p-IκBα的高表达。分子对接显示苍术素与Src之间存在良好的相互作用。此外，在LPS刺激的RAW 264.7细胞中，Src过表达部分逆转了苍术素对细胞凋亡、促炎因子和氧化应激的抑制作用。总之，苍术素减轻了DSS诱导的结肠炎，这可能与抑制Src介导的NF-κB和趋化因子信号通路有关。
