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 and Veterinary Medicine, 2022, 49(6): 2347-2361.

References

[1] 卢丽红.恩诺沙星对小鼠肝微粒体细胞色素P450酶系的影响[D].雅安:四川农业大学, 2013. LU L H.Study of the effects of enrofloxacin on mice hepatic CYP450[D].Ya'an:Sichuan Agricultural University, 2013.(in Chinese)
[2] 王想.两种氟喹诺酮类药物在猪排泄物中降解规律的研究[D].贵阳:贵州大学, 2019. WANG X.Investigation on degradation patterns of two fluoroquinolones in pig excreta[D].Guiyang:Guizhou University, 2019.(in Chinese)
[3] LICATE A, RANDAZZO C, MORREALE I, et al.Fluoroquinolone-induced liver injury:Three new cases and a review of the literature[J].European Journal of Clinical Pharmacology, 2012, 68(5):525-532.
[4] SPODNIEWSKA A, BARSKI D, GIZEJEWSKA A.Effect of enrofloxacin and chlorpyrifos on the levels of vitamins A and E in Wistar rats[J].Environmental Toxicology and Pharmacology, 2015, 40(2):587-591.
[5] LIU B, CUI Y, BROWN P B, et al.Cytotoxic effects and apoptosis induction of enrofloxacin in hepatic cell line of grass carp[J].Fish & Shellfish Immunology, 2015, 47(2):639-644.
[6] LI Y, MOU Y, THUNDERS M, et al.Effects of enrofloxacin on antioxidant system, microsomal enzymatic activity, and proteomics in porcine liver[J].Journal of Veterinary Pharmacology and Therapeutics, 2018, 41(4):562-571.
[7] 董漓波, 叶启薇, 孙永学, 等.恩诺沙星对SD系大白鼠的急性及亚急性毒性研究[J].华南农业大学学报, 1995, 16(4):1-4. DONG L B, YE Q W, SUN Y X, et al.Studies on acute and subacute toxicities of enrofloxacin in rats[J].Journal of South China Agricultural University, 1995, 16(4):1-4.(in Chinese)
[8] 栾学斌, 王荻, 刘红柏.恩诺沙星诱导施氏鲟药物性肝损伤模型的建立及复方中草药保肝作用研究[J].大连海洋大学学报, 2017, 32(1):13-19. LUAN X B, WANG D, LIU H B.A model of drug-induced liver injury caused by enrofloxacin and protective effects of compound Chinese herbal medicines on livers in Amur sturgeon Acipenser schrenckii[J].Journal of Dalian Ocean University, 2017, 32(1):13-19.(in Chinese)
[9] 李紫元, 莫蓝, 肖洁怡, 等.恩诺沙星诱导鸡急性肝损伤模型的建立[J].动物医学进展, 2019, 40(11):73-76. LI Z Y, MO L, XIAO J Y, et al.Establishment of enrofloxactin induced acute liver injury model in chickens[J].Progress in Veterinary Medicine, 2019, 40(11):73-76.(in Chinese)
[10] 梁丽.以肝细胞线粒体为靶点的中药保肝成分筛选方法建立及应用[D].昆明:云南中医学院, 2018. LIANG L.Development and application of screening method of hepatoprotective composition in traditional Chinese medicines with hepatocyte mitochondria as target[D].Kunming:Yunnan University of Chinese Medicine, 2018.(in Chinese)
[11] 王小敏, 陈乔, 郭丽丽, 等.连翘果、连翘叶乙醇提取物的抑菌活性及成分分析[J].食品工业科技, 2019, 40(6):89-94. WANG X M, CHEN Q, GUO L L, et al.Antimicrobial activities and component analysis of ethanol extracts from Forsythia suspensa fruit and Forsythia suspensa leaves[J].Science and Technology of Food Industry, 2019, 40(6):89-94.(in Chinese)
[12] 张天锡, 史磊, 刘雯, 等.连翘化学成分、药理活性现代研究[J].辽宁中医药大学学报, 2016, 18(12):222-224. ZHANG T X, SHI L, LIU W, et al.Research progress on chemical constituents and pharmacological effects of Forsythia[J].Journal of Liaoning University of Traditional Chinese Medicine, 2016, 18(12):222-224.(in Chinese)
[13] 赵晨栋, 王萌, 张昊, 等.连翘提取物对对乙酰氨基酚诱导小鼠肝损伤的保护作用[J].中国畜牧兽医, 2021, 48(10):3845-3854. ZHAO C D, WANG M, ZHANG H, et al.Protective effect of Forsythia suspensa extract on acetaminophen-induced liver injury in mice[J].China Animal Husbandry & Veterinary Medicine, 2021, 48(10):3845-3854.(in Chinese)
[14] 梅雪, 周安琴, 李静, 等.连翘叶的化学成分、药理学与毒理学研究概况[J].中国药房, 2015, 26(22):3143-3146. MEI X, ZHOU A Q, LI J, et al.General situation of research on chemical constituents, pharmacology and toxicology of Forsythia leaves[J].China Pharmacy, 2015, 26(22):3143-3146.(in Chinese)
[15] 王进明, 范圣此, 李安平, 等.连翘不同部位中连翘苷和连翘酯苷A的含量分析及其入药探讨[J].中国现代中药, 2013, 15(7):556-559. WANG J M, FAN S C, LI A P, et al.Analysis of the contents of forsythin and forsythiaside A in different parts of Forsythia suspensa and its medicinal use discussion[J].Modern Chinese Medicine, 2013, 15(7):556-559.(in Chinese)
[16] 叶华, 吴臻, 李发荣.连翘叶入药的问题及思考[J].临床合理用药杂志, 2011, 4(27):74-75. YE H, WU Z, LI F R.Problems and thoughts of Folium forsythia used for medical purpose[J].Chinese Journal of Clinical Rational Drug Use, 2011, 4(27):74-75.(in Chinese)
[17] 王晓燕, 常断玲.连翘叶和连翘果提取物指纹图谱比较研究[J].齐鲁药事, 2011, 30(10):574-575. WANG X Y, CHANG D L.Study on the fingerprints of extracts from Forsythia suspensa leaves and fruit[J].Qilu Pharmaceutical Affairs, 2011, 30(10):574-575.(in Chinese)
[18] 潘婷婷.连翘叶的研究进展[J].价值工程, 2017, 36(6):118-120. PAN T T.Advances in Forsythia suspensa leaves[J].Value Engineering, 2017, 36(6):118-120.(in Chinese)
[19] 白美美.连翘叶茶保肝作用研究[D].太原:山西大学, 2018. BAI M M.Study on hepatoprotective effect of Forsythia suspensa leaves tea[D].Taiyuan:Shanxi University, 2018.(in Chinese)
[20] 刘静.连翘叶茶抗氧化抗衰老及保肝作用的实验研究[D].西安:陕西师范大学, 2004. LIU J.The experimental study on anti-oxidative, anti-senile effect and the effect of protecting liver of Forsythia suspensa leaves tea[D].Xi'an:Shaanxi Normal University, 2004.(in Chinese)
[21] 朱淑云.连翘叶茶提取物抗氧化活性的研究[D].西安:陕西师范大学, 2003. ZHU S Y.The anti-oxidative effect of extracts from Forsythia suspensa leaves tea[D].Xi'an:Shaanxi Normal University, 2003.(in Chinese)
[22] HORAVATHOVA K, VACHALKOVA A, NOVOTNY L.Flavonoids as chemoprotective agents in civilization diseases[J].Neoplasma, 2001, 48(6):435-441.
[23] 程晓华, 熊玉卿.五环三萜皂苷的药理作用研究进展[J].中草药, 2007, 5:792-795. CHENG X H, XIONG Y Q.Advances in studies on pharmacological activities of pentacyclic triterpenoid saponins[J].Chinese Traditional and Herbal Drugs, 2007, 5:792-795.(in Chinese)
[24] KANG S W S, HAYDAR G, TANIANE C, at al.AMPK activation prevents and reverses drug-induced mitochondrial and hepatocyte injury by promoting mitochondrial fusion and function[J].PLoS One, 2016, 11(10):e0165638.
[25] SUN J, WEN Y, ZHOU Y, et al.p53 attenuates acetaminophen-induced hepatotoxicity by regulating drug-metabolizing enzymes and transporter expression[J].Cell Death & Disease, 2018, 9(5):536-548.
[26] 张纳博, 王茜, 张一昕, 等.解毒护肝方经PPARs信号通路激活抗炎系统治疗药物性肝损伤的作用研究[J].中药新药与临床药理, 2019, 30(11):1322-1328. ZHANG N B, WANG Q, ZHANG Y X, et al.Treatment of drug- induced liver injury by Jiedu Hugan dcoction via anti-inflammatory system activated by PPARs signaling pathway[J].Traditional Chinese Drug Research & Clinical Pharmacology, 2019, 30(11):1322-1328.(in Chinese)
[27] 饶敏腊, 张明萌, 石本艳, 等.AMPK的生物学功能及其激活机制研究进展[J].广东医科大学学报, 2018, 36(6):603-608. RAO M L, ZHANG M M, SHI B Y, et al.Biological function and activation mechanism of AMPK:A review[J].Journal of Guangdong Medical University, 2018, 36(6):603-608.(in Chinese)
[28] NAGAPPAN A, KIM J, JUNG D, et al.Cryptotanshinone from the Salvia miltiorrhiza Bunge attenuates ethanol-induced liver injury by activation of AMPK/SIRT1 and Nrf2 signaling pathways[J].International Journal of Molecular Sciences, 2019, 21(1):265-284.
[29] LU Q, SHU Y, WANG L, et al.The protective effect of Veronica ciliata Fisch.Extracts on relieving oxidative stress-induced liver injury via activating AMPK/p62/Nrf2 pathway[J].Journal of Ethnopharmacology, 2021, 270:113775.
[30] SHU G, QIU Y, HAO J, et al.γ-Oryzanol alleviates acetaminophen-induced liver injury:Roles of modulating AMPK/GSK3β/Nrf2 and NF-κB signaling pathways[J].Food & Function, 2019, 10(10):6858-6872.
[31] MEHRZADI S, FATEMI I, MALAYERI A R, et al.Ellagic acid mitigates sodium arsenite-induced renal and hepatic toxicity in male Wistar rats[J].Pharmacological Reports, 2018, 70(4):712-719.
[32] HUANG B P, LIN C H, CHEN H M, et al.AMPK activation inhibits expression of proinflammatory mediators through downregulation of PI3K/p38 MAPK and NF-kappa B signaling in murine macrophages[J].DNA and Cell Biology, 2015, 34(2):133-141.
[33] 李晓钰.α-亚麻酸植物甾醇酯基于AMPK调节线粒体功能及氧化应激改善非酒精性脂肪性肝病的作用和分子机制[D].太原:山西医科大学, 2021. LI X Y.Plant sterol eater of α-linolenic acid prevents non-alcoholic fatty liver disease through improving mitochondrial dysfunction and oxidative stress via AMPK signaling[D].Taiyuan:Shanxi Medical University, 2021.(in Chinese)
[34] CAI Y, MARTENS G, HINKE S, et al.Increased oxygen radical formation and mitochondrial dysfunction mediate beta cell apoptosis under conditions of AMP-activated protein kinase stimulation[J].Free Radical Biology & Medicine, 2007, 42(1):64-78.
[35] ZHU H P, FORETZ M, XIEZ L, et al.PRKAA1/AMPK alpha 1 is required for autophagy-dependent mitochondrial clearance during erythrocyte maturation[J].Autophagy, 2014, 10(9):1522-1534.
[36] WANG Q, ZHANG M, TORRES G, et al.Metformin suppresses diabetes-accelerated atherosclerosis via the inhibition of Drp1-mediated mitochondrial fission[J].Diabetes, 2017, 66(1):193-205.
[37] 王春丽.菊花提取物通过AMPK信号通路缓解对乙酰氨基酚诱导的大鼠肝脏损伤[D].开封:河南大学, 2020. WANG C L.Chrysanthemum extract alleviates acetaminophen-induced hepatotoxicity via AMPK pathway in rats[D].Kaifeng:Henan University, 2020.(in Chinese)
[38] INOUE M, SATO E, NISHIKAWA M, et al.Mitochondrial generation of reactive oxygen species and its role in aerobic life[J].Current Medicinal Chemistry, 2003, 10(23):2495-2505.
[39] RAHAL A, KUMAR A, SINGH V, et al.Oxidative stress, prooxidants, and antioxidants:The interplay[J].BioMed Research International, 2014, 53(1):1264-1283.
[40] HE S, GUO Y, ZHAO J, et al.Ferulic acid protects against heat stress-induced intestinal epithelial barrier dysfunction in IEC-6 cells via the PI3K/Akt-mediated Nrf2/HO-1 signaling pathway[J].International Journal of Hyperthermia, 2019, 35(1):112-121.
[41] LI Y M, WANG H B, ZHENG J G, et al.Dimethyl sulfoxide inhibits zymosan-induced intestinal inflammation and barrier dysfunction[J].World Journal of Gastroenterology, 2015, 21(38):10853-10865.
[42] YOU L, PENG H, LIU J, et al.Catalpol protects ARPE-19 cells against oxidative stress via activation of the Keap1/Nrf2/ARE pathway[J].Cells, 2021, 10(10):2635-2655.
[43] KRITHIKA R, MOHANKUMAR R, VERMA R, et al.Isolation, characterization and antioxidative effect of phyllanthin against CCl₄-induced toxicity in HepG2 cell line[J].Chemico-Biological Interactions, 2009, 181(3):351-358.
[44] 侯改霞, 杨建雄.连翘叶提取物对实验小鼠的降脂保肝作用研究[J].河南大学学报(自然科学版), 2010, 40(5):504-506. HOU G X, YANG J X.Effects of extracts from Forsythia suspense leaves on modulating blood lipids and protecting liver of hyperlipidemic mice[J].Journal of Henan University (Natural Science), 2010, 40(5):504-506.(in Chinese)

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

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