罗非鱼鱼皮肽的体外ACE抑制活性、消化性及分子对接对比Study on in Vitro ACE Inhibitory Activity, Digestibility and Molecular Docking Comparison of Tilapia Fish Skin Peptide
陈佳丽;杨胜涛;萧振邦;千忠吉;
摘要(Abstract):
【目的】分析罗非鱼鱼皮多肽LSGYGP的ACE抑制活性、胃肠道消化稳定性以及与降压药卡托普利在分子对接的对比。【方法】用HHL法测定多肽的IC50值和抑制模式,测定多肽在模拟胃肠道消化中的稳定性,MTT法检测细胞活力,Western blot法检测诱导型一氧化氮合酶(i NOS)、环氧合酶-2(COX-2)、内皮素-1(ET-1)等蛋白表达情况,将多肽、卡托普利分别与ACE进行分子对接对比分析。【结果】LSGYGP的半抑制率(IC50)值为2.57μmol/L,表现为混合非竞争性抑制,4 h内模拟胃肠道消化较为稳定;MTT法验证多肽LSGYGP对HUVEC细胞无毒性作用(P> 0.05),且能明显提高Ang Ⅱ诱导组的HUVEC细胞活力(P <0.01);与对照组相比,随实验组多肽浓度增加,i NOS、COX-2和ET-1的表达明显逐渐减少(P<0.001)。对接结果表明,氢键是LSGYGP与ACE结合结构中的支撑力,而卡托普利通常与ACE的Zn(2+)离子形成紧密结合的相互作用。【结论】消化稳定的罗非鱼鱼皮多肽可明显抑制血管收缩因子和炎症因子的表达,展现了较好ACE抑制活性,LSGYGP与卡托普利的活性差异可能是由于ACE分子对接的作用力不同,这些可作为研发少副作用ACE抑制剂的药理基础。
关键词(KeyWords): 罗非鱼鱼皮多肽;HUVEC;模拟胃肠道消化;分子对接
基金项目(Foundation): 广东省“扬帆起航”紧缺人才引进项目(201433009);; 广东海洋大学“创新强校”科研启动基金项目(2013050204)
作者(Authors): 陈佳丽;杨胜涛;萧振邦;千忠吉;
参考文献(References):
- [1]国家卫生计生委合理用药专家委员会.高血压合理用药指南(第2版)[J].中国医学前沿杂志:电子版, 2017,9(7):28-126.
- [2] MONTEZANO A C, AURELIE N D C, RIOS F J, et al.Angiotensin II and Vascular Injury[J]. Current Hypertension Reports, 2014, 16(6):431.
- [3] SITI H N, KAMISAH Y, KAMSIAH J. The role of oxidative stress, antioxidants and vascular inflammation in cardiovascular disease(a review)[J]. Vascular Pharmacology, 2015, 71:40-56.
- [4] FRAGASSO G, MARANTA F, MONTANARO C, et al.Pathophysiologic therapeutic targets in hypertension:a cardiological point of view[J]. Expert Opinion on Therapeutic Targets, 2012, 16(2):179-193.
- [5] FITZGERALD R J, MURRAY B A, WALSH D J.Hypotensive peptides from milk proteins[J]. Journal of Nutrition, 2004, 34(4):980S.
- [6] HUANG B B, LIN H C, CHANG Y W. Analysis of proteins and potential bioactive peptides from tilapia(Oreochromis spp.)processing co-products using proteomic techniques coupled with BIOPEP database[J].Journal of Functional Foods, 2015, 19:629-640.
- [7] ZHUANG Y L, HOU H, ZHAO X, et al. Effects of collagen and collagen hydrolysate from jellyfish(Rhopilema esculentum)on mice skin photoaging induced by UV irradiation[J]. Journal of Food Science,2010, 74(6):H183-H188.
- [8] CHAROENPHUM N, YOURAVONG W, CHEIRSILP B.Determination of reaction kinetics of hydrolysis of tilapia(Oreochromis niloticu)protein for manipulating production of bioactive peptides with antioxidant activity,angiotensin-I-converting enzyme inhibitory activity and Ca-binding properties[J]. International Journal of Food Science&Technology, 2013, 48(2):419-428.
- [9] LEE J K, JEON J K, BYUN H G. Antihypertensive effect of novel angiotensin I converting enzyme inhibitory peptide from chum salmon(Oncorhynchus keta)skin in spontaneously hypertensive rats[J]. Journal of Functional Foods, 2014, 7:381-389.
- [10] SUN L, ZHANG Y, ZHUANG Y L. Antiphotoaging effect and purification of an antioxidant peptide from tilapia(Oreochromis niloticus)gelatin peptides[J].Journal of Functional Foods, 2013, 5(1):154-162.
- [11] CUSHMAN D W, CHEUNG H S. Spectrophotometric assay and properties of the angiotensin-converting engyme of rabbit lung[J]. Biochemical Pharmacology,1971, 20(7):1637-1648.
- [12] BUSH K, HENRY P R, SLUSARCHYK D S.Muraceins-muramyl peptides produced by Nocardia orientalis as angiotensin-converting enzyme inhibitors. I.Taxonomy, fermentation and biological properties[J].Journal of Antibiotics, 1984, 37(4):330-335.
- [13]李丽,施展,钱春香.酶动力学模块软件求解米氏常数和最大反应速率的实验研究[J].洛阳理工学院学报(自然科学版), 2009, 19(4):1-3.
- [14] SAMARANAYAKA A G P, KITTS D D, LI-CHAN E C Y. Antioxidative and angiotensin-I-converting enzyme inhibitory potential of a pacific hake(Merluccius productus)fish protein hydrolysate subjected to simulated gastrointestinal digestion and Caco-2 cell permeation[J]. Journal of Agricultural and Food Chemistry, 2010, 58(3):1535-1542.
- [15]刘国艳,孙贝贝,张洁,等. AngⅡ诱导人脐静脉内皮细胞建立高血压损伤模型[J].食品科学, 2017, 38(13):174-181.
- [16] HEO S Y, KO S C, KIM C, et al. A heptameric peptide purified from Spirulina sp. gastrointestinal hydrolysate inhibits angiotensin I-converting enzyme and angiotensin II-induced vascular dysfunction in human endothelial cells[J]. International Journal of Molecular Medicine, 2017, 39(3):1072-1082.
- [17]李锐,邹茜,孙玉林,等.克氏原螯虾虾头模拟胃肠道消化产物中ACE抑制肽的分离纯化与鉴定[J].食品与发酵工业, 2018, 45(6):139-146.
- [18] MEHTA P K, GRIENDLING K K. Angiotensin II cell signaling:physiological and pathological effects in the cardiovascular system[J]. American Journal of Physiology-cell Physiology, 2007, 292(1):C82-C97.
- [19] THUANTHONG M, DE G C, SIRINUPONG N, et al.Purification and characterization of angiotensin-converting enzyme-inhibitory peptides from Nile tilapia(Oreochromis niloticus)skin gelatine produced by an enzymatic membrane reactor[J]. Journal of Functional Foods, 2017, 36:243-254.
- [20] TOOPCHAM T, ROYTRAKUL S, YONGSAWATDIGUL J. Characterization and identification of angiotensin I-converting enzyme(ACE)inhibitory peptides derived from tilapia using Virgibacillus halodenitrificans SK1-3-7 proteinases[J]. Journal of Functional Foods, 2015, 14:435-444.
- [21] ZHANG Y, DUAN X, ZHUANG Y. Purification and characterization of novel antioxidant peptides from enzymatic hydrolysates of tilapia(Oreochromis niloticus)skin gelatin[J]. Peptides, 2012, 38(1):13-21.
- [22]操德群,何艳丽,余虹,等.海洋生物ACE抑制肽研究进展[J].核农学报, 2017,31(5):927-937.
- [23] CHEN J, WANG Y, YE R, et al. Comparison of analytical methods to assay inhibitors of angiotensin I-converting enzyme[J]. Food Chemistry, 2013,141(4):3329-3334.
- [24] ITO H Y, HIRATA Y, ADACHI S, et al. Endothelin1 is an autocrine/paracrine factor in the mechanism of angiotensin II-induced hypertrophy in cultured rat cardiomyocytes[J]. Journal of Clinical Investigation,1993, 92(1):398-403.
- [25] GRAGASIN, F. S. Estrogen reduces angiotensin ii-induced nitric oxide synthase and NAD(P)H oxidase expression in endothelial cells[J]. Arteriosclerosis,Thrombosis, and Vascular Biology, 2003, 23(1):38-44.
- [26] POPOVICI RF, ALEXA IF, NOVAC O, et al.Pharmacokinetics study on mesoporous silica-captopril[J]. Digest Journal of Nanomaterials and Biostructures, 2011, 6(4):1619-1630.
- [27] MOSKOWITZ DW. Is ‘somatic’ angiotensin i-converting enzyme a mechanosensor?[J]. Diabetes Technology&Therapeutics, 2002, 4:841-858.
- [28] KOBAYASHI Y, YAMAUCHI T, KATSUDA T, et al.Angiotensin-I converting enzyme(ACE)inhibitory mechanism of tripeptides containing aromatic residues[J]. Journal of Bioscience and Bioengineering,2008, 106(3):310-312.
- [29] WANG X, CHEN H, FU X, et al. A novel antioxidant and ACE inhibitory peptide from rice bran protein:Biochemical characterization and molecular docking study[J]. LWT-Food Science and Technology, 2017, 75:93-99.
- [30] ROHIT A C R, SATHISHA K S, APARNA H S. A variant peptide of buffalo colostrum b-lactoglobulin inhibits angiotensin I-converting enzyme activity[J].European Journal of Medicinal Chemistry, 2012, 53:211-219.
- [31] WU Q, JIA J, YAN H, et al. A novel angiotensin-?converting enzyme(ACE)inhibitory peptide from gastrointestinal protease hydrolysate of silkworm pupa(Bombyx mori)protein:Biochemical characterization and molecular docking study[J]. Peptides, 2015, 68:17-24.