大黄鱼鱼鳔低聚肽的制备及其延缓皮肤衰老的研究  

Preparation of swim bladder peptide of Pseudosciaena crocea and its study on delaying skin aging

作　　者：孙彦庆 杜芬 朱宗敏 冯晓梅 尚丽丽 曹婉秀 刘楚怡 SUN Yanqing;DU Fen;ZHU Zongmin;FENG Xiaomei;SHANG Lili;CAO Wanxiu;LIU Chuyi(Qingdao Daiyoujia Biotechnology Co.,Ltd.,Qingdao 266001,China;Marine Biomedical Research Institute of Qingdao,Qingdao 266073,China;School of Medicine and Pharmacy,Ocean University of China,Qingdao 266100,China)

机构地区：[1]青岛黛优佳生物科技有限公司,山东青岛266001 [2]青岛海洋生物医药研究院,山东青岛266073 [3]中国海洋大学医药学院,山东青岛266100

出　　处：《轻工学报》2025年第1期107-119,共13页Journal of Light Industry

基　　金：国家重点研发计划项目(2023YFF1105305);中国工程院院地合作重大咨询研究项目(20220118);中央引导地方科技发展资金项目(YDZX2023090)。

摘　　要：以超氧阴离子自由基清除率为指标,采用单因素试验和响应面试验筛选制备大黄鱼鱼鳔低聚肽(HSBP),并利用人皮肤永生化角质形成细胞(HaCaT细胞)建立氧化损伤模型,检测HSBP对胶原蛋白、透明质酸分泌及抗氧化酶、β-半乳糖苷酶活力的影响,评价其氧化损伤修复活性;建立3D重组人表皮屏障损伤模型,分析HSBP对其组织形态学和皮肤屏障相关蛋白表达的影响,综合评价其延缓皮肤衰老的作用。结果表明:HSBP的最佳制备工艺为酶解时间4 h、酶解温度50℃、pH值7.5、菠萝蛋白酶添加量3400 U/g,在此工艺下,超氧阴离子自由基清除率为65.6%;HSBP的重均分子量为480 Da,无刺激性、无致敏性且无细胞毒性;HSBP能显著促进体外培养的氧化损伤细胞分泌透明质酸、Ⅰ型胶原蛋白和Ⅳ型胶原蛋白,同时提高超氧化物歧化酶和谷胱甘肽过氧化物酶活性;此外,HSBP还能降低丙二醛和β-半乳糖苷酶的含量,显著提高屏障损伤模型丝聚蛋白、兜甲蛋白、谷氨酰胺转移酶Ⅰ和水通道蛋白3的表达量,有效修复屏障损伤的皮肤组织状态,并明显增加活细胞层数。因此,HSBP能有效保护氧化损伤皮肤细胞并修复皮肤屏障损伤,起到延缓皮肤衰老的作用。Using the superoxide anion radical scavenging rate as an indicator,single-factor experiments and response surface methodology were employed to optimize the preparation of peptides from swim bladder of Pseudosciaena crocea(HSBP).The HaCaT cell line,an immortalized human keratinocyte cell line,was employed to establish an oxidative damage model to evaluate the effects of HSBP on collagen and hyaluronic acid secretion,as well as the activities of antioxidant enzymes andβ-galactosidase,to assess its oxidative damage repair activity.The 3D recombinant human epidermal barrier injury model was established to study the effects of HSBP on its tissue morphology and the expression of skin barrier related proteins,and to comprehensively evaluate its delaying aging effect on skin.The results showed that the optimal preparation conditions for HSBP were:enzymatic hydrolysis time of 4 hours,50℃,pH value of 7.5,and a bromelain addition of 3400 U/g.Under these conditions,the superoxide anion radical scavenging rate was 65.6%.The molecular weight of HSBP was 480 Da,with no irritation,sensitization,or cytotoxicity.HSBP could significantly promote the secretion of hyaluronic acid,type I and type IV collagen in oxidation-damaged cells,increase the activities of superoxide dismutase and glutathione peroxidase(GSH-Px),decrease the contents of malondialdehyde andβ-galactosidase,and significantly increase the expressions of LOR,FLG,TGM1 and AQP3 in barrier damaged models,effectively repair the barrier damaged skin tissue,and significantly increase the number of viable cell layers.Therefore,HSBP could effectively protect skin cells from oxidative damage and repair skin barrier injury,thereby delaying skin aging.

关 键 词：大黄鱼鱼鳔 低聚肽 氧化损伤 皮肤屏障损伤 皮肤衰老 

分 类 号：TS254.9[轻工技术与工程—水产品加工及贮藏工程]