为了深入探讨生物活性肽对细胞自噬活性的作用并应用于制备露酒,该研究通过倒置显微镜和流式细胞仪观察不同生物活性肽对人源肝细胞(LO2细胞)的影响,随后以生物活性肽和白酒为原料制备露酒,以肽保留率为指标,通过单因素试验和响应面试验对露酒工艺进行优化。结果表明,通过倒置显微镜观察细胞存活情况,最终选取5.00 mg/mL作为检测细胞自噬活性实验的最高作用浓度。玉米肽-1、玉米肽-3在0.63~5.00 mg/mL时有促进细胞自噬作用,小麦肽-1、小麦肽-6在5.00 mg/mL时有抑制作用,其中玉米肽-1对细胞自噬的促进效果最为明显。露酒最佳工艺条件为:基酒度数为48%vol,基酒香型为清香型,基酒温度为室温,生物活性肽类型为玉米肽-1,生物活性肽添加量为15.09 mg/mL,该露酒中肽的保留率高,口感佳,感官评分93分。最后分别用生理盐水、食用酒精、基酒、肽酒灌胃小鼠,处死后观察肝组织病理形态学改变,发现添加玉米肽的露酒在一定程度上减轻了实验小鼠肝损伤程度。
To further investigate the effect of bioactive peptides on autophagy flux, this study used an inverted microscope and flow cytometry to observe the effects of different types of bioactive peptides on LO2 hepatocytes.Then Lujiu was prepared with bioactive peptides and Baijiu as raw materials, the peptide retention rate was used as an indicator, and the technology of Lujiu was optimized by a single factor experiment and response surface experiment according to light transmittance index.Results showed that cell survival was observed through an inverted microscope, and 5.00 mg/mL was finally selected as the highest concentration for the test of autophagy activity.Corn peptide-1 and corn peptide-3 could promote autophagy in the range of 0.63-5.00 mg/mL, while wheat peptide-1 and wheat peptide-6 could inhibit it in the concentration of 5.00 mg/mL, among which corn peptide-1 had the most obvious effect on autophagy.The best technological conditions of Lujiu were that the degree of base liquor was 48%vol, the flavor type of base liquor was fragrant, the temperature of base liquor was room temperature, the type of bioactive peptide was corn peptide-1, and the additional amount of bioactive peptide was 15.09 mg/mL.Under these conditions, the light transmittance of Lujiu was high and the taste was good.The sensory score was 93.Finally, the mice were administered with normal saline, edible alcohol, base liquor, and peptide liquor, respectively.After being killed, the pathological changes of liver tissues were observed.It was found that Lujiu reduced the degree of liver damage in the experimental mice to a certain extent.
[1] THEODORESON M D, AITHAL G P, ALLISON M, et al.Extra-hepatic morbidity and mortality in alcohol-related liver disease:Systematic review and meta-analysis[J].Liver International, 2023, 43(4):763-772.
[2] CHOWDHURY A B, MEHTA K J.Liver biopsy for assessment of chronic liver diseases:A synopsis[J].Clinical and Experimental Medicine, 2023, 23(2):273-285.
[3] OSNA N A, RASINENI K, GANESAN M, et al.Pathogenesis of alcohol-associated liver disease[J].Journal of Clinical and Experimental Hepatology, 2022, 12(6):1492-1513.
[4] GONG X X, AN Q, LE L Q, et al.Prospects of cereal protein-derived bioactive peptides:Sources, bioactivities diversity, and production[J].Critical Reviews in Food Science and Nutrition, 2022, 62(11):2855-2871.
[5] WANG X J, LIU X L, ZHENG X Q, et al.Antagonistic effect of the glycopeptide from zein on acute alcohol-induced liver injury in mice[J].Journal of Functional Foods, 2022, 92:105062.
[6] 于亚莉, 宋雪梅, 关玉, 等.玉米肽Tyr-Phe-Cys-Leu-Thr对酒精性HepG2细胞的保护作用[J].中国食品学报, 2020, 20(5):43-52.
YU Y L, SONG X M, GUAN Y, et al.Protective effect of corn peptide Tyr-Phe-Cys-Leu-Thr on alcoholic HepG2 cells[J].Journal of Chinese Institute of Food Science and Technology, 2020, 20(5):43-52.
[7] 刘效森, 王琳, 蔡转章, 等.大豆低聚肽对HepG2细胞增殖抑制作用的研究[J].营养学报, 2019, 41(3):287-292.
LIU X S, WANG L, CAI Z Z, et al.Inhibition of soybean oligopeptides on proliferation HepG2 cells[J].Acta Nutrimenta Sinica, 2019, 41(3):287-292.
[8] WANG X Q, YU H H, XING R E, et al.Hepatoprotective effect of oyster peptide on alcohol-induced liver disease in mice[J].International Journal of Molecular Sciences, 2022, 23(15):8081-8096.
[9] 曾瑜, 潘兴昌, 张立实, 等.小麦低聚肽对小鼠解酒功能的评价[J].现代预防医学, 2019, 46(7):1255-1259.
ZENG Y, PAN X C, ZHANG L S, et al.Evaluation of wheat oligopeptide on hangover function in mice[J].Modern Preventive Medicine, 2019,46(7):1255-1259.
[10] 王妍凌. 基于细胞自噬的葡萄酒及其酚类物质对肝脏保护的影响研究[D].乌鲁木齐:新疆农业大学, 2023.
WANG Y L.Effects of wine and its phenolic compounds on liver protection based on cellular autophagy[D].Urumqi:Xinjiang Agricultural University, 2023.
[11] 高立敏. 金线莲苷对酒精性脂肪肝和肝纤维化的保护作用及机制探索[D].武汉:华中科技大学, 2022.
GAO L M,.Protective effect and mechanism of kinsenoside on alcoholic fatty liver and liver fibrosis[D].Wuhan:Huazhong University of Science and Technology, 2022.
[12] MANOHAR S M, SHAH P, NAIR A.Flow cytometry:Principles, applications and recent advances[J].Bioanalysis, 2021, 13(3):181-198.
[13] WELSH J A, JONES J C, TANG V A.Fluorescence and light scatter calibration allow comparisons of small particle data in standard units across different flow cytometry platforms and detector settings[J].Cytometry Part A, 2020, 97(6):592-601.
[14] MANOHAR S M, SHAH P, NAIR A.Flow cytometry:Principles, applications and recent advances[J].Bioanalysis, 2021, 13(3):181-198.
[15] REN H, BAKAS N A, VAMOS M, et al.Design, synthesis, and characterization of an orally active dual-specific ULK1/2 autophagy inhibitor that synergizes with the PARP inhibitor olaparib for the treatment of triple-negative breast cancer[J].Journal of Medicinal Chemistry, 2020, 63(23):14609-14625.
[16] 陈国华. 一种风味极佳的木瓜肽酒及其制备方法:中国, CN114106961A[P].2022-03-01.
CHEN G H.A method for preparing papaya peptide wine with excellent flavor:China, CN114106961A[P].2022-03-01.
[17] 曾允灏, 苏文, 郑小莉, 等.大鲵肝肽酒制备及其对小鼠酒后肝脏保护作用研究[J].中国酿造, 2023, 42(2):120-125.
ZENG R H, SU W, ZHENG X L, et al.Preparation of Andrias davidianus liver peptide wine and its protective effect on mouse liver after drinking[J].China Brewing, 2023, 42(2):120-125.
[18] 罗蕊琪, 钟恒, 韩兴林, 等.一种酒体微量成分对细胞自噬通量影响的检测方法:中国, CN116987756A[P].2023-11-03.
LUO R Q, ZHONG H, HAN X L, et al.A method for detecting the effect of trace components of wine body on autophagy flux:China, CN116987756A[P].2023-11-03.
[19] MUELLER L K, BAUMRUCK A C, ZHDANOVA H, et al.Challenges and perspectives in chemical synthesis of highly hydrophobic peptides[J].Frontiers in Bioengineering and Biotechnology, 2020, 8:162.
[20] QAZANFARZADEH Z, KADIVAR M, SHEKARCHIZADEH H, et al.Rye secalin characterisation and use to improve zein-based film performance[J].International Journal of Food Science & Technology, 2021, 56(2):742-752.
[21] 卓俊纳. 三种香型白酒的ICP-OES和GC-QTOF MS分析及判别研究[D].自贡:四川轻化工大学, 2022.
ZHUO J N.Analysis and discrimination research by ICP-OES and GC-QTOF MS of three kinds of flavor Baijiu[D].Zigong:Sichuan University of Science and Engineering, 2022.
[22] JIANG W, CHANG K F, MA S N, et al.Effects of heat treatment on structural and functional properties of velvet antler polypeptides[J].Journal of Food Processing and Preservation, 2021, 45(6):e15490.
[23] 路联, 李海忠.一种养生保健酒的制备方法:中国, CN115433655A[P].2022-12-06.
LU L, LI H Z.The preparation method of healthy wine:China, CN115433655A[P].2022-12-06.
[24] IKARI S, YANG Q, LU S L, et al.Quercetin in tartary buckwheat induces autophagy against protein aggregations[J].Antioxidants, 2021, 10(8):1217.
