摘要
目的运用网络药理学方法预测安石榴苷治疗炎症性肠病(IBD)的作用机制。方法通过数据库获取安石榴苷和IBD的交集基因,并进行PPI、GO功能和KEGG通路富集分析。安石榴苷和靶点通过分子对接验证。C57BL/6J小鼠采用葡聚糖硫酸钠建立IBD模型,给予安石榴苷干预7天,给药过程中观察各组小鼠的体征并计算每天的疾病活动指数(DAI),给药结束后肠道通透性检测;取结肠组织苏木素-伊红染色法观察病理改变,并计算组织学损伤评分;ELISA法检测小鼠结肠组织中肿瘤坏死因子(TNF-α)、白细胞介素-10(IL-10)、髓过氧化物酶(MPO)、趋化因子1(CXCL1)等细胞因子水平。CCK8法测定安石榴苷对caco-2细胞增殖活性的影响。ELISA法检测脂多糖(LPS)诱导的caco-2细胞释放细胞因子水平。结果获得安石榴苷与IBD共同靶点14个,包括TNF、花生四烯酸-5-脂氧合酶(ALOX5)、血管内皮生长因子A(VEGFA)等;KEGG富集分析预测安石榴苷治疗IBD主要作用于花生四烯酸代谢、AGE-RAGE、VEGFA、Ras等信号通路;分子对接显示安石榴苷与TNF表现出较高的对接活性。与模型组比较,安石榴苷组小鼠体质量下降幅度减小(P<0.01);DAI显著降低(P<0.01);结肠黏膜充血、水肿明显减轻,组织学损伤评分显著降低(P<0.01);结肠组织TNF-α、IL-1β、MPO、CXCL1、IL-6、IL-18、IFN-γ水平显著降低(P<0.01);IL-10水平显著升高(P<0.01);20-300μmol·L^(-1)安石榴苷促进caco-2细胞增殖,抑制LPS诱导的TNF-α分泌,上调IL-10水平。结论安石榴苷通过抑制TNF-α等促炎因子分泌,上调抗炎因子IL-10水平,改善IBD小鼠的结肠炎症反应。
Objective To predict the potential mechanism of Punicalagin in the treatment of Inflammatory bowel disease by network pharmacology.Methods The intersection genes of Punicalagin and IBD were obtained from the database,and PPI,GO and KEGG pathways were enriched and analyzed.Punicalagin and the target were verified by molecular docking.C57BL/6J mice were drunk dextran sulfate sodium to establish inflammatory enteritis model,and were given Punicalagin for 7 d of intervention.During the administration,signs of mice in each group were observed,daily disease activity index was calculated;Intestinal permeability test after administration;The colon tissue was stained with hematoxylin eosin to observe the pathological changes and calculate the histological damage score;Detection of tumor necrosis factor(TNF-α),Interleukin-10(IL-10),myeloperoxidase(MPO),chemokine 1(CXCL1)and other cytokines in colon tissue of mice by ELISA.Detection of TNF-α,IL-6,MPO and CXCL1 level in mouse serum by ELISA.CCK8 method was used to determine the effect of Punicalagin on the proliferation activity of caco-2 cells.The levels of cytokines released by caco-2 cells induced by lipopolysaccharide(LPS)were detected by ELISA.Results 14 common targets of Punicalagin and IBD were obtained,including tumor necrosis factor(TNF),arachidonic acid-5-lipoxygenase(ALOX5)and vascular endothelial growth factor A(VEGFA).KEGG enrichment analysis predicted that the treatment of IBD by Punicalagin mainly acted on arachidonic acid signaling pathway,age-rage signaling pathway,VEGR signaling pathway and Ras signaling pathway.Molecular docking showed that Punicalagin had good docking activity with TNF receptor.Compared with the model group,the decreasing range of body mass in Punicalagin group abated(P<0.01);the disease activity index of Punicalagin group decreased significantly(P<0.01);The congestion and edema of colonic mucosa were significantly reduced,and the histological injury score was significantly reduced(P<0.01);The level of TNF-α,IL-1β,MPO,CXCL1,IL-6,IL-18,IFN-γ in colon tissue was significantly decreased(P<0.01);20-300μmol·L^(-1)Punicalagin promoted caco-2 cell proliferation and inhibited TNF-α secretion induced by LPS,up-regulation of IL-10 levels.Conclusion Punicalagin inhibits the secretion of TNF-α and other proinflammatory factors,up-regulation of the level of anti-inflammatory factor IL-10,and improvement of colonic inflammatory response in IBD mice.
作者
毛旭文
古丽若依·帕尔哈提
张雍正
阿米尔·泽布
程路峰
Mao Xuwen;Paerhati Guliruoyi;Zhang Yongzheng;Amir Zeb;Cheng Lufeng(Department of Pharmacy,Xinjiang Medical University,Urumqi 830017,China;Key Laboratory of Active Components of Natural Medicine and Drug Release Technology,Urumqi 830017,China;College of International Education,Xinjiang Medical University,Urumqi 830017,China)
出处
《世界科学技术-中医药现代化》
CSCD
北大核心
2023年第7期2437-2449,共13页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
基金
新疆维吾尔自治区科学技术厅自然科学基金面上项目(2022D01C190):钩藤碱通过芳香烃受体途径促进肠屏障缓解炎症性肠炎机制研究,负责人:毛旭文。
作者简介
通讯作者:程路峰,教授,博士研究生导师,主要研究方向:肠炎治疗药物研发。
