文蛤多肽的制备及对非酒精性脂肪肝细胞模型的修复作用  

Preparation of NAFLD Cell Model Repair Polypeptide from Meretrix meretrix

作　　者：赵莎莎 赵玉勤 张亚茹 杨最素 黄芳芳 丁国芳 

机构地区：[1]浙江海洋大学食品与医药学院浙江省海洋生物医用制品工程技术研究中心,浙江舟山316022

出　　处：《中国食品学报》2017年第6期33-39,共7页Journal of Chinese Institute Of Food Science and Technology

基　　金：浙江省科技厅优先主题重大专项(2013C03036);浙江省自然科学基金项目(LY15C200016)

摘　　要：目的:探讨文蛤酶解多肽的制备工艺及对非酒精性脂肪肝(NAFLD)细胞模型的修复作用。方法:用质量浓度15μg/m L的软脂酸诱导正常肝Chang liver细胞建立NAFLD模型,以TG含量为指标,筛选酶解文蛤内脏的最佳酶种,通过正交试验、超滤、G-25葡聚糖凝胶层析、高效液相等技术获得酶解产物。将其作用于NAFLD细胞模型,油红O染色后镜下观察形态,检测细胞中的MDA,GSH-ST,ALT,AST,SOD,γ-GT等含量。结果:软脂酸诱导Chang liver细胞48 h后获得NAFLD细胞模型;最佳酶种为碱性蛋白酶,酶解条件为40℃、pH9.5、料液比1∶2、酶解时间12 h,加酶量1 000 U/g;超滤后得到<5 ku酶解液,经G-25葡聚糖凝胶层析得到4个峰;峰Ⅰ组分的氨基酸序列为Gln-Leu-Asn-Trp-Asp。该多肽能明显降低NAFLD细胞模型内的TG含量,使MDA,ALT,AST,γ-GT的含量下降,SOD,GSH-ST的含量上升,与未用药的模型组细胞相比有统计学意义。结论 :经碱性蛋白酶酶解文蛤得到的多肽,对NAFLD细胞模型具有明显的修复作用。Objective： To study the preparation of Meretrix meretrix polypeptide and the role on repairing for NAFLD model. Methods： NAFLD cell model was established by treating with palmitic acid of 15 μg/m L. The best enzymes were chosen according to the contents of TG. Meretrix meretrix polypeptide can be obtained by rthogonal test, ultra-filtration and HPLC. Cell morphological feature was observed under oil red O staining and the changes of MDA, GSH-ST, ALT,AST, SOD, γ-GT of the cell were detected after treating with Meretrix meretrix polypeptide. Results： NAFLD cell model was established by treating with palmitic acid of 15 μg/m L for 48 h. The best hydrolysis condition on the basis of the index of TG content were： 40 ℃, solid-liquid ration 1∶2, pH 9.5, enzyme dosage 1 000 U/g, time 12 h for Alcalase. G-25 gel chromatography was used to separate the 5 ku peptide and four peak were obtained. The sequence of peak Ⅰ was Gln-Leu-Asn-Trp-Asp. TG, MDA, ALT, AST and γ-GT in Chang liver cells decreased in medicinal groups. SOD,GSH-ST in Chang liver cells increased in medicinal groups. Conclusion： Meretrix meretrix polypeptide had a significant role on repairing NAFLD cell model.

关 键 词：文蛤 多肽 CHANG liver细胞 NAFLD细胞模型 修复 

分 类 号：TQ936.16[化学工程]