摘要
目的:基于斑马鱼模型与网络药理学技术研究苦参碱治疗炎症性肠病(IBD)的作用机制。方法:以2,4,6-三硝基苯磺酸(TNBS)建立斑马鱼IBD模型,使用中性红染色、阿尔新蓝染色及中性粒细胞数量变化评估肠道吞噬功能、杯状细胞分泌及中性粒细胞聚集情况,利用相关试剂盒测定苦参碱(40、60 mg·L^(-1))对斑马鱼体内肿瘤坏死因子(TNF)-α与胆囊收缩素(CCK)含量变化;使用网络药理学及分子对接技术预测苦参碱治疗IBD的潜在作用机制;并通过实时荧光定量聚合酶链式反应(Real-time PCR)对相关靶点验证基因表达情况。结果:与模型组比较,苦参碱组能够呈剂量依赖性增加中性红染色面积(P<0.05,P<0.01),改善肠道吞噬功能;能够增加阿尔新蓝染色面积(P<0.05,P<0.01),影响肠道杯状细胞分泌;能够减少中性粒细胞数量(P<0.01)及缓解其聚集情况,显著减少TNF-α含量(P<0.01);苦参高剂量组显著增加CCK含量(P<0.01),苦参低剂量组CCK含量差异无统计学意义。网络药理学分析得到苦参碱治疗IBD潜在靶点28个,蛋白质-蛋白质相互作用(PPI)网络分析得度值排名前5的靶点分别为α7烟碱乙酰胆碱受体(CHRNA7)、多巴胺受体D1(DRD1)、烟碱型乙酰胆碱α4亚型(CHRNA4)、溶质载体家族6成员3(SLC6A3)、代谢型谷氨酸受体5(GRM5);京都基因与基因组百科全书(KEGG)结果显示,苦参碱治疗IBD可能与神经活性配体-受体相互作用(neuroactiveligand-receptorinteraction)、胆碱能突触(cholinergic synapse)、中性粒细胞胞外诱捕网形成(neutrophil extracellular trap formation)相关,且Real-time PCR结果表明苦参碱能够影响相关靶点基因表达水平。结论:苦参碱对炎症性肠病具有一定的治疗作用,能够影响IBD炎症反应,其治疗作用可能与神经活性配体-受体相互作用等通路相关。
Objective:To study the mechanism of matrine in the treatment of inflammatory bowel disease(IBD)based on the zebrafish model and network pharmacology.Method:The IBD model of zebrafish was established using 2,4,6-trinitro-benzenesulfonicacid(TNBS),and the intestinal phagocytic function,goblet cell secretion,and neutrophil aggregation were evaluated using neutral red staining,alcian blue staining,and neutrophil number changes.Changes in tumor necrosis factor(TNF)-αand cholecystokinin(CCK)content in zebrafish were determined by using relevant reagent kits.Network pharmacology and molecular docking techniques were used to predict the potential mechanism of matrine in the treatment of IBD.Gene expression of relevant targets was verified through Real-time polymerase chain reaction(Real-time PCR).Result:Compared with the model group,the matrine administration group can increase the neutral red staining area in a dose-dependent manner and improve intestinal phagocytic function(P<0.05,P<0.01).It can reduce the staining area of alcian blue and affect the secretion of intestinal goblet cells(P<0.01).It can reduce the number of neutrophil granulocytes,relieve its aggregation,significantly reduce TNF-αconten(t P<0.01),and increase the CCK content.Network pharmacology analysis identifies 28 potential targets for matrine in the treatment of IBD.The top five targets by protein-protein interaction(PPI)network analysis are CHRNA7,DRD1,CHRNA4,SLC6A3,and GRM5.The Kyoto encyclopedia of genes and genomes(KEGG)results show that the treatment of IBD with matrine may be related to neuroactive ligand-receptor interaction,cholinergic synapse,and neutrophil extracellular trap formation.Real-time PCR results show that matrine can affect the expression level of related target genes.Conclusion:matrine has a certain therapeutic effect on IBD and can affect the inflammatory response of IBD.Its therapeutic effect may be related to neuroactive ligand-receptor interaction and other pathways.
作者
陈林珍
张雪
陈祺
于雪
戴胜云
马志强
赵崇军
CHEN Linzhen;ZHANG Xue;CHEN Qi;YU Xue;DAI Shengyun;MA Zhiqiang;ZHAO Chongjun(Beijing Key Laboratory of Traditional Chinese Medicine(TCM)Quality Evaluation,Beijing University of Chinese Medicine,Beijing 102488,China;School of TCM,Beijing University of Chinese Medicine,Beijing 100029,China;National Institutes for Food and Drug Control,Beijing 102629,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2023年第22期88-94,共7页
Chinese Journal of Experimental Traditional Medical Formulae
基金
公益性行业科研专项经费项目(201507004)
北京中医药大学新教师启动基金项目(2020-JYB-XJSJJ-009)。
关键词
苦参碱
炎症性肠病
斑马鱼
网络药理学
作用机制
matrine
inflammatory bowel disease
zebrafish
network pharmacology
mechanism
