海湾扇贝抗氧化肽对酒精性肝损伤的保护作用OA北大核心CSTPCD
Protective Effect of Antioxidant Peptides from Argopecten irradians against Alcoholic Liver Injury
目的:探究海湾扇贝肽的抗氧化作用以及对乙醇诱导的酒精性肝损伤小鼠的保护作用及机制.方法:采用两步酶解和膜分离制备低分子质量海湾扇贝肽,通过体外抗氧化实验以及超氧化物歧化酶(superoxide dismutase,SOD)、乙醇脱氢酶(alcohol dehydrogenase,ADH)激活率实验评价其体外抗氧化性以及促进乙醇代谢潜力;通过急性酒精中毒模型小鼠,测定其血清中丙氨酸氨基转移酶(alanine aminotransferase,ALT)、天冬氨酸氨基转移酶(aspartate aminotransferase,AST)活力,肝脏中ADH、乙醛脱氢酶(acetaldehyde dehydrogenase,ALDH)、谷胱甘肽过氧化酶(glutathione peroxidase,GSH-Px)、过氧化氢酶(catalase,CAT)、SOD活力及丙二醛(malondialdehyde,MDA)含量,评价其对小鼠酒精性肝损伤的保护作用及机制.采用液相色谱-串联质谱(liquid chromatography-tandem mass spectrometry,LC-MS/MS)法分析多肽序列,通过PeptideRanker工具和AnOxPePred 1.0工具进行多肽的潜在生物活性预测,并将抗氧化活性最强的肽段与ALDH进行分子对接分析.结果:所制得海湾扇贝活性肽分子质量在200~1 000 Da的肽段占比为70.38%,抗氧化氨基酸占总氨基酸含量的68.75%.体外实验显示,海湾扇贝肽1,1-二苯基-2-三硝基苯肼自由基和羟自由基清除率的半抑制浓度(half maximal inhibitory concentration,IC50)值分别为19.43 mg/mL和4.53 mg/mL,亚铁离子螯合能力的IC50值为2.19 mg/mL;在海湾扇贝酶解液质量浓度为5mg/mL时,SOD和ADH激活率分别为19.83%和18.18%,显示出良好的抗氧化活性和解酒潜力.体内实验结果显示,海湾扇贝肽能显著降低小鼠血清中ALT、AST水平,显著提升ADH、ALDH、GSH-Px、SOD和CAT活力,并显著降低MDA含量.利用PeptideRanker工具和AnOxPePred 1.0工具对鉴定出的44条多肽进行筛选,有6条达到分子对接筛选的标准,分子对接发现其与ALDH均具有强烈的结合能力,其中DQPHFPF和YSTHPHF、ALDH之间分别形成了7、3个氢键,可以通过与ALDH发生分子相互作用激活其活性.结论:本研究为海湾扇贝肽在解酒护肝产品中的应用提供了理论参考.
Objective:To investigate the antioxidant effect of peptides from Argopecten irradians and its protective effect and mechanism against ethanol-induced alcoholic liver injury in mice.Methods:A.irradians peptides were prepared by two-step enzymatic hydrolysis followed by membrane separation.The in vitro antioxidant activity of A.irradians peptides,the activating effects on superoxide dismutase(SOD)and alcohol dehydrogenase(ADH)and the potential to promote alcohol metabolism were evaluated.The protective effect and mechanism against alcoholic liver injury in mice were investigated by measuring the activities of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)in the serum,and the activities of ADH,acetaldehyde dehydrogenase(ALDH),glutathione peroxidase(GSH-Px),catalase(CAT)and SOD and malondialdehyde(MDA)content in the liver of a mouse model of acute alcoholism.Liquid chromatography-tandem mass spectrometry(LC-MS/MS)was used to analyze the peptide sequences.PeptideRanker and AnOxPePred 1.0 tools were used to predict the potential biological activity of peptides and conduct molecular docking analysis of the peptide with the strongest antioxidant activity to ALDH.Results:Peptides with molecular mass between 200 and 1 000 Da constituted approximately 70.38%of the total peptides.Antioxidant amino acids accounted for 68.75%of the total amino acids.In vitro experiments showed that the half maximal inhibitory concentration(IC50)values of A.irradians peptides were 19.43,4.53 and 2.19 mg/mL in 1,1-diphenyl-2-picrylhydrazyl(DPPH)and hydroxyl radical scavenging and Fe2+chelating assays,respectively.At 5 mg/mL,the hydrolysate of A.irradians activated SOD and ADH by 19.83%and 18.18%,respectively,demonstrating good antioxidant activity and anti-alcoholic potential.In vivo experimental results showed that A.irradians peptides significantly reduced the levels of ALT and AST in mouse serum,enhanced the activity of ADH,ALDH,GSH-Px,SOD and CAT,and reduced the content of MDA.Using PeptideRanker and AnOxPePred 1.0 tools,44 identified peptides were screened,and 6 of them met the criteria for molecular docking screening.Molecular docking revealed that they had strong binding capacity to ALDH.Among them,DQPHFPF and YSTHPHF formed 7 and 3 hydrogen bonds with ALDH,respectively,which could be activated through molecular interactions with ALDH.Conclusion:This study provides a theoretical reference for the application of peptides products from A.irradians for curing alcohol and protecting liver.
曹燕峰;常立炀;张修正;陈娜;蔡芳瑗;张志芹;刘海梅;赵芹
鲁东大学食品工程学院,山东烟台 264025烟台南山学院健康学院,山东烟台 265713
轻工业
海湾扇贝活性肽酒精性肝损伤氧化应激分子对接
Argopecten irradiansactive peptidesalcoholic liver injuryoxidative stressmolecular docking
《食品科学》 2024 (019)
87-93 / 7
烟台市科技计划项目(2022XDRH012);鲁东大学-烟台味禧食品科技有限公司校企合作项目(28000103);烟台市重大科技创新工程项目(2023YT06000829)
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