| 摘要: |
| 【目的】探讨连翘苷-柚皮素（phillyrin-naringenin,PN）缓解对乙酰氨基酚（acetaminophen,APAP）诱导肝损伤的潜在作用机制。【方法】通过网络药理学筛选PN与药物性肝损伤（drug-induced liver injury, DILI）的交集靶点，将其导入STRING数据库进行蛋白互作网络分析筛选核心靶点，并对核心靶点进行GO和KEGG分析。分子对接验证PN与磷脂酰肌醇3-激酶（phosphatidylinositol 3-kinase,PI3K）、蛋白激酶B（protein kinase B,AKT）、肿瘤蛋白P53（tumor protein 53,P53）和细胞色素P450酶2E1（cytochrome P450 2E1,CYP2E1）的亲和力。将小鼠随机分为空白对照组（normal control,NC）、APAP组（APAP）、乙酰半胱氨酸组（N-acetylcysteine,NAC）、连翘苷-柚皮素低/中/高治疗组（low/medium/high dose PN treatnent,LPN/MPN/HPN），采用试剂盒检测不同处理小鼠谷丙转氨酶（alanine transaminase,ALT）、谷草转氨酶（aspartate aminotransferase,AST）、N-乙酰基-对-苯醌亚胺（N-acetyl-p-benzoquinone imine,NAPQI）、谷胱甘肽（Glutathione,GSH）、超氧化物歧化酶（super oxide dismutase,SOD）和过氧化氢（hydrogen peroxide,H2O2）水平及丙二醛（malonaldehyde,MDA）含量，用显微镜观察小鼠肝组织病变，采用探针法检测小鼠肝脏CYP2E1活性，并用Western blot检测CYP2E1、PI3K、AKT和P53蛋白变化。【结果】①网络药理学筛选PN和DILI作用靶点分别为426和2 576个，PN与DILI的交集靶点为288个，核心靶点为37个，且核心靶点集中在PI3K/AKT通路。分子对接结果表明，PN与PI3K、AKT、P53和CYP2E1蛋白结合较强。②与NC组相比，APAP组小鼠ALT和AST活性显著升高，显微观察发现小鼠肝组织有明显出血点与水肿情况；CYP2E1蛋白表达显著上升，APAP毒性产物NAPQI在肝组织中积累；谷胱甘肽和超氧化物歧化酶水平降低，MDA和H2O2含量增加；PI3K/AKT通路被抑制，P53蛋白表达上调。与APAP组相比，LPN、HPN和MPN组ALT和AST活性显著降低，肝组织水肿有所缓解；CYP2E1蛋白表达水平和NAPQI水平降低，谷胱甘肽和超氧化物歧化酶水平升高，MDA和H2O2含量降低，磷酸化PI3K和AKT蛋白表达水平升高，P53蛋白表达水平降低。【结论】PN可通过激活PI3K/AKT通路来抑制APAP代谢，缓解氧化应激，治疗APAP诱导的DILI。 |
| 关键词: 连翘苷 柚皮素 氧化应激 药物性肝损伤 PI3K/AKT通路 |
| DOI：10.13207/j.jnwafu.2026.03.013 |
| 分类号: |
| 基金项目:国家自然科学基金项目（32160850，32160859） |
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| Effect of phillyrin-naringenin combination against APAP liver injury |
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ZHANG Jianhui, PAN Yangyang, PU Sisi, LIU Huimin, ZHANG Qian, WANG Libin, LI Liang, ZHANG Zhiyong, WANG Meng
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College of Veterinary Medicine,Gansu Agricultural University,Lanzhou,Gansu 730070,China
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| Abstract: |
| 【Objective】This research aims to explore the effects of phillyrin-naringenin (PN) combination on alleviating the liver injury induced by acetaminophen (APAP).【Method】Intersecting targets of PN and drug-induced liver injury (DILI) were screened by network pharmacology method.These targets were imported into the STRING database for protein-protein interaction (PPI) network analysis to identify core targets.Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were performed on the core targets.Molecular docking was conducted to validate the binding affinity of PN with phosphatidylinositol 3-kinase (PI3K),protein kinase B (AKT),tumor protein P53 (P53),and cytochrome P450 2E1 (CYP2E1).The mice were randomly divided into normal control group (NC),APAP group (APAP),N-acetylcysteine group (NAC),low/medium/high dose PN treatment (LPN/MPN/HPN).Serum levels of alanine transaminase (ALT),aspartate aminotransferase (AST),N-acetyl p-benzoquinone imine (NAPQI),glutathione (GSH),superoxide dismutase (SOD),malondialdehyde (MDA),and hydrogen peroxide (H2O2) were measured by commercial assay kits.Liver histopathological changes were evaluated by microscopic observation.CYP2E1 activity in the liver was determined by a fluorescent probe-based method.Protein expression levels of CYP2E1,PI3K,AKT,and p53 were analyzed by Western blot.【Result】① Network pharmacology screening identified 426 potential targets of PN and 2 576 targets associated with DILI,with 288 overlapping targets between PN and DILI.Among these,37 core targets were significantly enriched in the PI3K/AKT signaling pathway.Molecular docking results demonstrated that PN exhibited strong binding affinities to PI3K,AKT,P53,and CYP2E1.② Compared with NC group,the activity of ALT/AST in APAP group was significantly increased,and there were obvious bleeding points and edema in liver tissue under microscopic observation.CYP2E1 protein expression level/enzyme activity increased significantly,and APAP toxic product NAPQI accumulated in liver tissue.The levels of GSH and SOD were decreased,and the contents of MDA and H2O2 were increased.The PI3K/AKT pathway was inhibited and the expression of P53 protein was up-regulated.Compared with APAP group,LPN,HPN and MPN could significantly reduce ALT/AST activity and alleviate liver tissue edema.CYP2E1 protein expression/enzyme activity and NAPQI levels were decreased，the levels of GSH and SOD were increased,and the accumulation of MDA and H2O2 was decreased.The expression level of p-PI3 K and p-AKT protein was increased,and the expression level of P53 protein was decreased.【Conclusion】PN exerts its hepatoprotective effects by activating the PI3K/AKT pathway,thereby inhibiting APAP metabolism (reducing toxic metabolite accumulation) and alleviating oxidative stress,ultimately offering a therapeutic strategy against APAP-induced DILI. |
| Key words: phillyrin-naringenin oxidative stress drug-induced liver injury (DILI) PI3K/AKT pathway |
