中国食品学报2017，Vol.17Issue(8)：92-100,9.DOI:10.16429/j.1009-7848.2017.08.013
鳕鱼皮胶原蛋白肽制备及其对肝细胞损伤的保护作用
Preparation of Melanogrammus aeglefinus Skin Collagen Peptide and It's Protective Effect on the Injury of Liver Cell
摘要
Abstract
Objective:To investigate the preparation of Melanogrammus acglefinus skin collagen peptide and the protection effect of different concentrations peptide against LPS-induced damage in Chang Liver cells.Methods:Considering liquid ratio,enzyme dosage,reaction time and reaction temperature as factors in the experimental,measured by cells OD value with MTT method,determine the extraction of collagen peptide hydrolysis process and purify through ultrafiltration,anion exchange chromatography,molecular subjected.The protection effect of purified peptides was proved by detecting Chang Liver cells antioxidant enzymes in intracellular and cell supernatants transaminase changes with kit.Using Hoechst 33258 staining and Annexin V-FITC / PI double-labeled flow cytometry,observe the preparation of collagen peptide on Chang Liver cells apoptosis induced.Results:The results show that hydrolase was selected trypsin enzymes,enzymatic hydrolysis temperature was selected 50 ℃,pH was selected 6,enzyme dosage was selected 2 000 U/g,hydrolysis time was selected 8 h.GM2-2-3,got by DEAE-Sepharose Fast Flow ion column,sephadex G25 gel column,significantly reduced LPS-induced damage of Chang Liver cells.SOD,CAT,GSH-PX activity in Chang Liver cells intracellular increased and MDA content decreased in a dose-dependent manner;AST,ALT,ALP,γ-GT,LDH activity in the cell supematant decreased in a dose-dependent manner.Compared with the damage cells,Chang Liver cell with collagen peptide morphology stretched,fewer phenomena appeared that chromatin obviously aggregation and margination,nuclear shrinkage,early apoptotic rate decreased,and early apoptotic cells decreases.关键词
鳕鱼皮/胶原蛋白肽/制备技术/辅肝Key words
Melanogrammus aeglefinus skin/collagen peptide/preparation techniques/auxiliary liver引用本文复制引用
滕芳芳,胡晓斐,刘晨晨,杨最素,黄芳芳,丁国芳..鳕鱼皮胶原蛋白肽制备及其对肝细胞损伤的保护作用[J].中国食品学报,2017,17(8):92-100,9.基金项目
国家自然科学基金面上项目(81273429) （81273429）
浙江省科技厅重大专项(2013C03036,2011C02003,2011C23034) （2013C03036,2011C02003,2011C23034）
舟山市科技计划项目(2012C23023) （2012C23023）
