东海海参胶原蛋白酶解物的制备与抗氧化活性及其对神经细胞损伤的保护作用

Preparation of Acaudina molpadioides Collagen Hydrolysates and Their Antioxidant Activity and Protective Effect on Injured Nerve Cells

目的:研究木瓜蛋白酶酶解东海海参胶原蛋白的最优化条件,测定酶解产物的抗氧化活性及其对神经细胞损伤的保护作用。方法:以水解度为指标,通过响应面法优化东海海参胶原蛋白的酶解条件,确定最佳酶解参数。通过测定酶解产物对超氧自由基、羟基自由基和DPPH自由基的清除能力,研究其抗氧化能力。通过测定SK-N-SH神经细胞的存活率并进行形态观察,研究酶解产物对由H2O2造成的神经细胞损伤的保护作用。结果:木瓜蛋白酶酶解东海海参胶原蛋白的最佳条件为:底物含量34.3 g/g酶,温度43.2℃,pH 7.12,在该条件下东海海参胶原蛋白的水解度为16.73%。东海海参胶原蛋白酶解产物对超氧自由基、羟基自由基和DPPH自由基具有较好的清除作用,且与其浓度呈剂量依赖关系。酶解产物清除上述3种自由基的IC50值分别为34,25和24 mg/mL。胶原蛋白酶解产物对由H2O2造成的神经细胞SK-N-SH的损伤具有较好的保护作用。结论:研究结果为东海海参胶原蛋白的开发和利用奠定了良好基础。

Objective:To investigate optimal enzymatic conditions of Acaudina molpadioides collagen by papain,and to determine the antioxidant activity of collagen hydrolysates and their protective effect on injured nerve cells.Methods: Using the hydrolysis degree as the target,the enzymatic conditions of Acaudina molpadioides collagen by papain were optimized by response surface methodology.The antioxidant ability of collagen hydrolysates was investigated by determining their scavenging ability to O2^-·,·OH and DPPH radicals.The protective effect of collagen hydrolysates on H2O2-injured SK-N-SH nerve cells was studied by determining cell viability and by the morphological observation.Results:The optimal parameters of collagen hydrolysis by papain were as follows:substrate concentration 34.3 g/g,temperature 43.2 ℃ and pH 7.12.Under the optimal conditions,the hydrolysis degree of collagen was 16.73%.The scavenging ability of Acaudina molpadioides collagen hydrolysates to O2^-·,·OH and DPPH radicals was dose-dependent.IC50 of collagen hydrolysates to scavenge the above three free radicals were 34,25 and 24 mg/mL,respectively.Collagen hydrolysates had a strong protective effect on SK-N-SH nerve cells injured by H2O2.Conclusion:This study lays a good foundation for the future development and use of Acaudina molpadioides collagen.