鲟鱼子酶解产物对酒精损伤肝细胞的保护作用及活性肽虚拟筛选  

Protective Effects of Enzymatic Products from Sturgeon Roe on Alcohol-induced Hepatic Cell Damage and Virtual Screening of Active Peptides

作　　者：张李君 赵甜甜 陈杰琼 钟康荣 关永健 罗睿 林春燕 南海军 张业辉[2] ZHANG Lijun;ZHAO Tiantian;CHEN Jieqiong;ZHONG Kangrong;GUAN Yongjian;LUO Rui;LIN Chunyan;NAN Haijun;ZHANG Yehui(School of Chinese Materia Medica,Guangdong Pharmaceutical University,Guangzhou 510006,China;Sericultural&Agri-food Research Institute,Guangdong Academy of Agriculture Sciences/Key Laboratory of Functional Foods,Ministry of Agriculture and Rural Affairs/Guangdong Key Laboratory of Agricultural Products Processing,Guangzhou 510640,China;Guizhou Qianxun Biological Technology Co.,Ltd.,Qiandongnan 556399,China;Guangdong Bixiang Technological Development Co.,Ltd.,Qingyuan 513023,China)

机构地区：[1]广东药科大学中药学院,广东广州510006 [2]广东省农业科学院蚕业与农产品加工研究所/农业农村部功能食品重点实验室/广东省农产品加工重点实验室,广东广州510640 [3]贵州省千鲟生物科技有限公司,贵州黔东南州556399 [4]广东碧乡科技发展有限公司,广东清远513023

出　　处：《食品工业科技》2024年第19期316-324,共9页Science and Technology of Food Industry

基　　金：国家自然科学基金(31972021,32201980);广东省重大科技专项(2021B0202060001);广东省农业科学院协同创新中心(XT202303)。

摘　　要：本研究旨在探究鲟鱼子酶解产物对酒精损伤肝细胞的保护作用,并深入阐明其作用机理。以鲟鱼子为研究对象,结合酶解效率和体外活性等关键指标,对其酶解工艺进行优化。借助酒精损伤HepG2细胞模型,系统研究鲟鱼子酶解产物对肝细胞的保护作用。运用计算机虚拟筛选技术,探索潜在的生物活性多肽序列。研究结果表明,在碱性蛋白酶(1343 U/g)及胰酶(27 U/g)1:1复配酶解8 h的条件下获得的鲟鱼子酶解产物最优,蛋白回收率为49.30%±0.57%,水解度为46.05%±0.92%,ABTS+自由基清除率为51.45%±0.66%。不同浓度下均表现出显著的乙醇脱氢酶(Alcohol Dehydrogenase,ADH)激活率(P<0.05),在5 mg/mL时为168.64%±1.42%。在此酶解条件下制备的产物,0.3~2 mg/mL时与模型组相比能显著提升酒精损伤HepG2细胞内超氧化物歧化酶(Superoxide Dismutase,SOD)活性、谷胱甘肽(Glutathione,GSH)含量(P<0.05),显著降低丙二醛(Malondialdehyde,MDA)含量(P<0.05)。此外,酶解产物在0.3~2 mg/mL时显著降低丙氨酸氨基转移酶(Alanine Aminotransaminase,ALT)和天门冬氨酸氨基转移酶(Aspartate Aminotransferase,AST)的活性(P<0.05)。虚拟筛选结果显示,鲟鱼子酶解产物中的LPG和FLPR等5条多肽显示潜在的酒精损伤肝细胞保护作用。因此,本研究为进一步发展鲟鱼子等水产原料的精深加工利用提供了基础支持。This study was designed to explore the hepatoprotective effects of enzymatic degradation products derived from sturgeon roe on alcohol-damaged hepatocytes and to elucidate the underlying mechanisms.Utilizing sturgeon roe as the research material,the enzymatic process was optimized by focusing on key parameters such as enzymatic efficiency and in vitro activity.The protective effects on hepatocytes were systematically investigated by using the HepG2 cell model.Advanced computerized virtual screening technology was employed to identify potential bioactive peptides within the sturgeon roe.The findings indicated that the optimum enzymatic product of sturgeon roe was obtained under the conditions of 1:1 compounding of alkaline protease(1343 U/g)and trypsin(27 U/g)for 8 h,yielded a protein recovery of 49.30%±0.57%,a hydrolysis degree of 46.05%±0.92%,and ABTS+free radical scavenging rate of 51.45%±0.66%.Notably,at a concentration of 5 mg/mL,this product significantly enhanced the activity of alcohol dehydrogenase(ADH)by 168.64%±1.42%(P<0.05).The product prepared under this enzymatic condition at 0.3~2 mg/mL significantly elevated superoxide dismutase(SOD)activity,glutathione(GSH)contents,and reduced malondialdehyde(MDA)contents in alcohol-injured HepG2 cells compared to the model group(P<0.05).In addition,the product at 0.3~2 mg/mL also significantly reduced alanine aminotransferase(ALT)and aspartate aminotransferase(AST)activities(P<0.05).Virtual screening identified 5 peptides,including LPG and FLPR,in the sturgeon roe enzymatic degradation product,showed promising hepatoprotective properties against alcohol-induced hepatocyte injury.Consequently,this study lays a solid foundation for the further development and utilization of aquatic resources,such as sturgeon roe,in hepatoprotective applications.

关 键 词：鲟鱼 鲟鱼子 乙醇脱氢酶 抗氧化 生物活性多肽 酒精代谢 

分 类 号：TS254.9[轻工技术与工程—水产品加工及贮藏工程]