鸡源胶原血管保护活性肽的筛选、制备及保护机理研究

Extensive researches have revealed that food-derived bioactive peptides play an important role in diseases prevention, especially in chronic diseases prevention. We previously isolated several collagen peptides from chicken with the property of inhibiting angiotensin converting enzyme (ACE). The results showed that these peptides can protect the blood vessel through absorbing and transporting into blood effectively, which indicate their potential application in cardiovascular diseases prevention. In this project, we attempt to further clarify the blood vessel protection mechanism of the active collagen peptides from chicken. The active peptides with high quality will be obtained by optimizing the preparation conditions and using purification technology such as HPLC. The absorption mechanism of the active peptides will be studied through cell test and animal test, and the function of the absorbed active peptides will be detected. Finally, the effect of the peptides on gene regulation and protein express in cell and animal will be examined by proteomic and transcriptomic methods. This project can provide a foundational support for the utilization of the bioactive peptides in foods and nutraceuticals. It can also improve the comprehensive benefit on agriculture and promote the sustainable development of China's livestock industrial.

研究显示食品来源的活性多肽对疾病预防（特别是慢性疾病）具有非常重要的作用。申请人前期研究从鸡源胶原蛋白中分离得到了几个具有血管紧张素酶(ACE) 抑制功能的小分子活性肽；该肽可被血液和细胞快速吸收，促进血管损伤修复，对血管起保护作用，有望用于高血压等心血管疾病的预防。为了进一步解明鸡源胶原蛋白活性肽在血管保护中的功能和作用机理，本项目拟在前期研究的基础上，通过高效液相色谱等技术优化制备条件，获得高质量具有ACE抑制活性的鸡源胶原蛋白活性肽；并通过细胞实验和动物实验研究其在生物体内的吸收机制，深入了解它的吸收与体内功能之间的关系；利用转录组学和蛋白质组学分析其对细胞和动物基因调控及蛋白表达的影响，从分子水平阐明鸡源胶原活性肽在血管保护的作用机制。本项目的顺利实施不仅能为小分子活性肽在食品、保健品等领域的应用提供重要理论依据，而且对促进农业增收，推动我国畜产业的可持续发展，具有积极的现实意义。

本项目以鸡爪胶原为材料，通过单因素和正交试验优化了胶原多肽蛋白酶解工艺。在此基础上研究了酶解多肽的抗氧化活性，其对DPPH自由基、ABTS自由基、超氧阴离子自由基均有一定的清除效果，且呈浓度依赖关系；IC50各为20.31mg/ml、19.72mg/ml、27.73mg/ml。同时研究了其对酪氨酸酶酶活和B16黑色素瘤细胞黑色素生成的抑制，胶原肽抑制酪氨酸单酚酶的IC50值为0.57mg/mL，抑制二酚酶的IC50值为0.426mg/mL；浓度为200μg/mL时，胶原多肽和熊果苷作用的黑色素瘤细胞黑色素相对含量分别为57.83%和31.53%，鸡胶原多肽有较好的黑色素瘤细胞黑色素生产抑制活性。这为鸡胶原多肽在功能性食品和化妆品等方面的应用提供了基础。