基于网络药理学探讨沙棘治疗酒精性肝损伤的作用机制

邓斌; 陈洁; 李想; 娄可轩; 周立权; 周一苗; 肖作为

doi:10.13386/j.issn1002-0306.2023100057

基于网络药理学探讨沙棘治疗酒精性肝损伤的作用机制

Exploring the Mechanism of Action of Sea Buckthorn in the Treatment of Alcoholic Liver Injury Based on Network Pharmacology

摘要

摘要: 目的：基于网络药理学预测沙棘治疗酒精性肝损伤（Alcohol Liver Injury）的作用机制，并通过建立斑马鱼酒精性肝损伤动物模型来验证沙棘治疗酒精性肝损伤的功效。方法：通过中药系统药理学数据库与分析平台（Traditional Chinese Medicine Systems Pharmacology，TCMSP）、Uniprot数据库收集有效成分及其作用靶标，Venny2.1寻找交集靶点。通过GeneCards及OMIM数据库收集和筛选疾病靶点，STRING v12.0数据库进行PPI网络分析，PDB及PubChem进行蛋白质结构及小分子结构确认。通过Cytoscape（Version 3.9.1）软件网络图构建沙棘治疗酒精性肝损伤关联网络。利用Metascape数据库对共有靶点进行基因本体（Gene Ontology，GO）、京都基因与基因组百科全书（Kyoto Encyclopedia of Gene，KEGG）数据库通路富集分析。通过斑马鱼实验进行功能验证：选取受精后3 d（3 dpf）野生型AB品系斑马鱼，正常组喂养于正常饲养用水，其余各组饲养于2%的无水乙醇溶液中建立酒精性肝损伤模型。沙棘组给予不同浓度的沙棘溶液，28 ℃处理2 d后，确定沙棘对酒精性肝损伤模型斑马鱼的最大耐受浓度（MTC）剂量，并根据MTC结果进行下一步给药，测定样品肝保护功效评价表型实验结果。结果：筛选后得到沙棘活性成分33个，主要包括槲皮素、花葵素、儿茶酸等；治疗酒精性肝损伤的潜在靶点1434个，主要包括ADH1C、CTNNB1、TGFB1等。调控这些核心靶点的信号通路主要富集在Lipid and atherosclerosis、Fluid shear stress and atherosclerosis、PI3K-Akt signaling pathway等多条信号通路中。动物实验结果显示：与模型组相比，沙棘可显著降低酒精性肝损伤模型斑马鱼肝脏不透明值（P<0.01），改善肝脏和卵黄囊肿大（P<0.01），下调AST和ALT的活力值（P<0.001），改善肝细胞核肿大，减少肝组织脂肪空泡样变性的功效。结论：沙棘可改善酒精性肝损伤，其机制可能与改善脂肪酸氧化、细胞代谢和抑制细胞凋亡有关。

Abstract: Objective: To predict the mechanism of sea buckthorn in the treatment of alcoholic liver injury based on network pharmacology, and to verify the efficacy of sea buckthorn in the treatment of alcoholic liver injury by establishing an animal model of alcoholic liver injury in zebrafish. Methods: Through the traditional chinese medicine systems pharmacology (TCMSP) and Uniprot database, the effective components and their targets were collected. Venny2.1 was used to find the intersection targets. GeneCards and OMIM databases were used to collect and screen disease targets. STRING v12.0 database was used for PPI network analysis. PDB and PubChem were used to confirm protein structure and small molecular structure. The correlation network of sea buckthorn in the treatment of alcoholic liver injury was constructed by Cytoscape (Version 3.9.1) software network diagram. Gene ontology (GO) and Kyoto encyclopedia of genes (KEGG) database pathway enrichment analysis were performed on the common targets using the Metascape database. Functional verification was performed by zebrafish experiments: Wild-type AB strain zebrafish 3 days after fertilization (3 dpf) were selected. The normal group was fed with normal feeding water, and the other groups were fed in 2% anhydrous ethanol solution to establish an alcoholic liver injury model. Results: After screening, 33 active components of sea buckthorn were obtained, mainly including quercetin, sunflower, catechin and so on. There were 1434 potential targets for the treatment of alcoholic liver injury, including ADH1C, CTNNB1, TGFB1 and so on. The signaling pathways that regulate these core targets are mainly enriched in multiple signaling pathways such as lipid and atherosclerosis, fluid shear stress and atherosclerosis and PI3K-Akt signaling pathway. The results of animal experiments showed that sea buckthorn had the effect of reducing the opacity value of zebrafish liver in alcoholic liver injury model (P<0.01), improving the phenomenon of liver and yolk cyst enlargement (P<0.01), down-regulating the activity values of AST and ALT (P<0.001), improving the enlargement of liver nucleus and reducing the fatty vacuolar degeneration of liver tissue. Conclusion: Seabuckthorn can improve alcoholic liver injury, and its mechanism may be related to improving fatty acid oxidation, cell metabolism and inhibiting apoptosis.