牛蛙皮胶原蛋白酸提取条件优化及其对细胞生长和粘附性的影响

摘要

应用羟脯氨酸法测定了牛蛙皮胶原蛋白含量,通过正交实验对牛蛙皮胶原蛋白酸法提取条件进行了优化,并结合MTT法测定了胶原蛋白对细胞生长和粘附性的影响.结果显示,牛蛙皮胶原蛋白的含量约为45.1％；温度条件对提取率影响最大,其次依次为酸种、时间和浓度,最优组合为乙酸、1.5 mol/L、37℃、24 h；在低浓度时,牛蛙皮胶原蛋白对正常人类肝细胞的生长和粘附性无显著影响,当浓度升高到一定值时,对细胞生长和粘附性有显著促进作用.%The content of collagen in the skin of Rana catesbeiana was measured by hydroxyproline method, the condition of extraction with acid was optimized by orthogonal experiment, and the effects of collagen on the cell growth and adhesion were determined by MTT method. The results showed that the skin of Rana catesbeiana contained about 45. 1% collagen protein; the factors affected extraction rate from large to small were temperature, type of acids, extraction time and concentration of acids, the optimal condition of extraction with acid was acetic acid, 1. 5 mol/L, 37 ℃ , 24 h. When the concentration of collagen was low, there was no significant effect on cell growth and adhesion, but when concentration of collagen reached a high level, it could promote growth and adhesion of cells cultured in vitro.