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引用本文:王小凤,申慧婷,孙小玲,于荣贤,刘兵,苏秀榕.金枪鱼(Thunnus sp.)胰腺酶解液对H2O2诱导的胰岛素瘤细胞(INS-1)氧化损伤的保护作用.海洋与湖沼,2019,50(5):1146-1153.
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金枪鱼(Thunnus sp.)胰腺酶解液对H2O2诱导的胰岛素瘤细胞(INS-1)氧化损伤的保护作用
王小凤1,2, 申慧婷1,2, 孙小玲3, 于荣贤3, 刘兵3, 苏秀榕1
1.宁波大学海洋学院 宁波 315211;2.宁波大学食品与药学学院 宁波 315211;3.深圳市荣格保健品有限公司 深圳 518118
摘要:
本文首先通过MALDI-TOF/TOF-MS技术,测定金枪鱼(Thunnus sp.)胰腺酶解液中的多肽组分及其丰度,确定GQPVR、GLPPK和EHER是其中的优势肽;再利用Discovery Studio平台中的反向找靶和分子对接模块,发现这些优势肽均可以与Keap1蛋白结合,推测其可能具有抗氧化活性;最后,利用细胞实验对酶解液的抗氧化活性进行验证,发现其对H2O2氧化损伤的胰岛素瘤细胞(INS-1)具有剂量依赖型保护效果,高剂量和低剂量酶解液处理后细胞活力较模型组显著提高,凋亡率从79.17%分别下降到43.2%(P<0.01)和28.9%(P<0.01);胰岛素的分泌量从3.65mIU/L分别增加到5.75mIU/L(P<0.01)和4.95mIU/L(P<0.01)。Discovery Studio的抗氧化功能预测和细胞学实验相结合的研究方法,为酶解液的功能组分鉴定提供了新的思路。
关键词:  金枪鱼(Thunnus sp.)胰腺  酶解液多肽  分子对接  氧化损伤  胰岛素瘤细胞
DOI:10.11693/hyhz20190300053
分类号:Q955;Q789
基金项目:2014年海洋经济创新发展区域示范项目(2014—2016);2016年海洋经济创新发展区域示范项目(2016—2018)。
IN-SILICO ANALYSIS AND IN VIVO TESTS OF ANTI-OXIDATION EFFECT OF TUNA PANCREAS HYDROLYSATE ON INS-1 CELL
WANG Xiao-Feng1,2, SHEN Hui-Ting1,2, SUN Xiao-Ling3, YU Rong-Xian3, LIU Bing3, SU Xiu-Rong1
1.School of Marine Sciences, Ningbo University, Ningbo 315211, China;2.College of Food and Pharmaceutical Sciences, Ningbo University, Ningbo 315211, China;3.Shenzhen Rongge Health Products Co., Ltd., Shenzhen 518118, China
Abstract:
In this study, the MALDI-TOF/TOF-MS technique was used to determine the polypeptide components and their abundance in the pancreatic enzymatic hydrolysate, and GQPVR, GLPPK, and EHER were identified as the dominant peptides. The reverse lookup in the Discovery Studio platform was used. It was found that these dominant peptides can bind to Keap1 protein, suggesting that it may have antioxidant activity. The antioxidant activity of the enzymatic hydrolysate was verified in cell experiments, and its insulinoma of oxidative damage to H2O2 was found. The cell (INS-1) had a dose-dependent protective effect. The cell viability of the high-dose and low-dose enzymatic treatment groups was significantly higher than that of the model group, and the apoptotic rate decreased from 79.17% to 43.2% and to 28.9% (P<0.01), respectively; insulin secretion increased from 3.65 to 5.75 and to 4.95 mIU/L, respectively (P<0.01). The combination of the Discovery Studio functional predictive analysis and cytology experiments provides new insights into the functional component identification of enzymatic hydrolysates.
Key words:  tuna pancreas  enzyme hydrolysis  molecular docking  oxidative damage  insulinoma cells
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