背景:梗阻性黄疸(OJ)是一种具有潜在结局的常见临床疾病,包括肝细胞坏死,胆管增生,显著的胆汁淤积性肝纤维化,and,在严重的情况下,肝功能衰竭。白藜芦醇(RES),一种存在于葡萄和浆果中的多酚,已证明在改善OJ方面有效。然而,其作用的确切机制尚不清楚。
方法:在本研究中,我们采用网络药理学研究RES治疗OJ的潜在分子机制。使用中药系统药理学数据库和分析平台(TCMSP)确定RES的目标,SuperPred,和SwissTargetPrediction数据库。与OJ有关的目标是从DisGeNet收集的,GeneCards,DrugBank,和在线孟德尔人继承(OMIM)数据库,这些目标的交点是使用Venny2.1.0确定的。随后,使用Cytoscape软件构建主动组件-目标网络。使用String数据库和Cytoscape软件生成蛋白质-蛋白质相互作用(PPI)网络。在此之后,使用生物导体平台进行基因本体论(GO)和京都基因和基因组百科全书(KEGG)富集分析。最后,实时定量PCR(qRT-PCR),蛋白质印迹(WB),采用酶联免疫吸附试验(ELISA)评估相关途径中的RNA和蛋白质表达水平。
结果:研究结果揭示了RES的56个潜在目标,通过在线数据库搜索确定了2,742个与OJ相关的目标,在27个目标中重叠。在PPI网络中,mTOR,CYP2C9,CYP1A1,CYP3A4,AHR,ESR1和HSD17B1成为核心靶标。KEGG分析表明,RES治疗OJ的主要途径,特别是那些与脂质代谢有关的,包括亚油酸代谢,花生四烯酸代谢,细胞色素P450,脂质和动脉粥样硬化,酪氨酸代谢,类固醇激素生物合成,以及戊糖和葡糖醛酸的相互转化信号通路。此外,体内实验表明,RES可显着改善OJ大鼠胆总管结扎(CBDL)引起的肝损伤。它降低了血清丙氨酸氨基转移酶(ALT)和天冬氨酸氨基转移酶(AST)水平,降低肝组织MDA水平,谷胱甘肽(GSH)含量增加,和增强的超氧化物歧化酶(SOD)的活性,减轻肝脏损害。代谢组学分析显示,RES在OJ中的治疗作用涉及脂质代谢途径的改变,提示RES治疗OJ的潜在机制。ELISA,qRTPCR,WB分析证实mTOR的表达水平较低,RES组CYP1A1、CYP2C9与模子组比拟,验证他们参与脂质代谢途径。
结论:结论:RES对OJ大鼠肝功能有保护作用。潜在的机制似乎与抗氧化活性和脂质代谢途径的调节有关。
BACKGROUND: Obstructive Jaundice (OJ) is a common clinical condition with potential outcomes, including hepatocyte necrosis, bile duct hyperplasia, significant cholestatic liver fibrosis, and, in severe cases, liver failure. Resveratrol (RES), a polyphenol present in grapes and berries, has demonstrated efficacy in improving OJ. However, the precise mechanism of its action remains unclear.
METHODS: In this study, we employed network pharmacology to investigate the underlying molecular mechanism of RES in the treatment of OJ. The targets of RES were identified using the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP), SuperPred, and SwissTargetPrediction database. The targets related to OJ were gathered from the DisGeNET, GeneCards, DrugBank, and Online Mendelian Inheritance in Man (OMIM) databases, and the intersection of these targets was determined using Venny2.1.0. Subsequently, an active component-target network was constructed using Cytoscape software. The Protein-Protein Interaction (PPI) network was generated using the String database and Cytoscape software. Following this, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were conducted using the Bioconductor platform. Finally, quantitative Real-Time PCR (qRT-PCR), Western Blotting (WB), and Enzyme-Linked Immunosorbent Assay (ELISA) were employed to assess RNA and protein expression levels in related pathways.
RESULTS: The findings revealed a selection of 56 potential targets for RES, and a search through the online database identified 2,742 OJ-related targets with overlapping in 27 targets. In the PPI network, mTOR, CYP2C9, CYP1A1, CYP3A4, AHR, ESR1, and HSD17B1 emerged as core targets. KEGG analyses demonstrated that the primary pathways of RES in treating OJ, particularly those related to lipid metabolism, include linoleic acid metabolism, arachidonic acid metabolism, metabolism of xenobiotics by cytochrome P450, lipid and atherosclerosis, tyrosine metabolism, steroid hormone biosynthesis, and pentose and glucuronate interconversions signaling pathways. Furthermore, in vivo experiments indicated that RES significantly ameliorated liver injury induced by Common Bile Duct Ligation (CBDL) in rats with OJ. It lowered serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels, reduced liver tissue MDA levels, increased glutathione (GSH) content, and enhanced activity of superoxide dismutase (SOD), alleviating liver damage. Metabolomics analysis revealed that the therapeutic effect of RES in OJ involved alterations in lipid metabolic pathways, hinting at the potential mechanism of RES in treating OJ. ELISA, qRTPCR, and WB analyses confirmed lower expression levels of mTOR, CYP1A1, and CYP2C9 in the RES group compared to the model group, validating their involvement in the lipid metabolism pathway.
CONCLUSIONS: In conclusion, RES exhibited a protective effect on liver function in rats with OJ. The underlying mechanism appears to be linked to antioxidant activity and modulation of lipid metabolism pathways.