基于网络药理学研究连翘叶提取物通过AMPK抗恩诺沙星诱导肝损伤的作用

doi:10.16431/j.cnki.1671-7236.2022.06.035

Abstract

Abstract: 【Objective】 This study was aimed to explore the mechanism of Forsythia suspense leaves extract (FSLE) against enrofloxacin (ENR)-induced liver injury (EILI) by network pharmacology and in vivo experiments.【Method】 The Forsythia suspense leaves were extracted by ethanol-enzymatic method, whose main chemical components were identified by liquid chromatography mass spectrometry (LC-MS).The effective active ingredients and targets of FSLE were analyzed and selected by PubChem, Swiss ADME and Swiss Target Prediction database, while the EILI targets were screened by GeneCards disease database.STRING online analysis and Cytoscape 3.8.0 software were applied to analyze and construct the core targets of FSLE treats EILI.Metascape website was used to perform GO function and KEGG pathway enrichment analysis of the targets, and Cytoscape 3.8.0 software was hired to construct the "component-target-signal pathway" visual network.The protective effects of FSLE were verified in EILI mice in vivo.The expression of AMP-activated protein kinase (AMPK) was tested by Real-time PCR and Western blotting, and the contents of glutathione (GSH), hydrogen peroxide (H₂O₂), malondialdehyde (MDA) and the activity of total superoxide dismutase (SOD) were detected by commercial kits.【Result】 About 20 oral ingredients in FSLE and their related 338 drug targets were analyzed and screened by LC-MS and Network pharmacology, while 913 protein targeting EILI, 124 protein targets which contained 12 core targets, were screened from drug-disease targets intersection.20 signing pathways were obtained by KEGG pathway enrichment analysis.KEGG pathway showed that AMPK signal pathway had a good affinity to EILI.The animal model experiments showed that FSLE could significantly reduce the alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities in the serum of mice (P<0.05), and could significantly increase the expression of gene and protein of AMPK (P<0.05).Compared with the normal control group, FSLE could significantly reduce the contents of MDA and H₂O₂ in liver of EILI mice (P<0.05), and significantly increase the GSH content and SOD activity (P<0.05), which reduced the EILI-induced oxidative stress significantly.【Conclusion】 FSLE could protect the liver against ENR induced injury by inducing the expression of AMPK to reduce the oxidative stress, which was helpful for the development of new drugs targeting liver injury.

Key words: Forsythia suspense leaves extract; enrofloxacin; liver injury; network pharmacology; AMPK

CLC Number:

S853.74

AN Zhixia, PAN Yangyang, FAN Biyue, MA Xiaoyan, YE Dehe, ZHANG Zhijie, QIU Shantong, YAO Yale, WANG Meng. Study on the Effect of Forsythia suspensa Leaf Extract on Enrofloxacin-induced Liver Injury Through AMPK Based on Network Pharmacology[J]. China Animal Husbandry & Veterinary Medicine, 2022, 49(6): 2347-2361.

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Study on the Effect of Forsythia suspensa Leaf Extract on Enrofloxacin-induced Liver Injury Through AMPK Based on Network Pharmacology

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