鱼籽多肽降糖及抗氧化活性评价

Evaluation of hypoglycemic and antioxidant activity of fish roe polypeptide

目的:以鱼籽多肽为原料,测定其氨基酸组成,评价其营养价值并对其抗氧化活性进行初步分析,探讨其是否具有降糖及抗氧化活性。方法:参照文献[1]方法测定鱼籽多肽氨基酸含量;通过高糖诱导RIN-m5f胰岛细胞氧化损伤模型,考察鱼籽多肽对氧化损伤胰岛细胞胰岛素分泌的保护作用;体内实验以正常雄性KM小鼠为实验对象,以阿卡波糖为阳性对照,采取鼠尾尖血测定血糖值的方法进行糖耐量实验。采用紫外分光光度法分析鱼籽多肽对2,2'-偶氮(二异丁基脒二盐酸盐)(AAPH)诱发的红细胞溶血的抗氧化活性。结果:鱼籽多肽中氨基酸含量为70.41%,EAA/TAA为39.23%,EAA/NEAA为64.55%,AAS和CS值较高,第一限制性氨基酸为色氨酸;与损伤组相比,鱼籽多肽100μg/mL显著提升高糖损伤RINm5f胰岛β细胞的胰岛素分泌量至(40.0±6.9)pg/mL;鱼籽多肽500mg/kg剂量组可显著降低正常小鼠血糖曲线下面积,与对照组比较差异具有统计学意义。鱼籽多肽对红细胞溶血抑制作用的IC50为1mg/mL。结论:鱼籽多肽中抗氧化活性氨基酸、鲜味氨基酸、药效氨基酸含量丰富,营养价值较高;细胞水平的胰岛素实验筛选到的鱼籽多肽能够改变胰岛细胞胰岛素水平,推测其可能具有降糖活性;动物实验发现其能改善小鼠餐后血糖有效降低血糖曲线下面积,证明其有降糖活性。鱼籽多肽对红细胞溶血的抑制作用阐明其可能通过清除ROS降低细胞氧化应激水平的机制达到降糖效果。

Objective:The amino acid composition of polypeptide from fish roe was determined,and this study was performed to explore whether the polypeptides from fish roes had hypoglycemic activity and antioxidant activity.Methods:The oxidative damage model of RIN-m5f islet cells induced by high glucose was used to investigate the protective effect of the fish roe polypeptide on insulin secretion of oxidatively damaged islet cells.Set male KM normal mice as experimental object in vivo,acarbose as positive control,the experiment of glucose tolerance was performed by the determination of blood glucose levels in mice tailtip.The anti-oxidation activity of the the polypeptides from fish roes on 2,2'-azobis (2-methylpropion amidine dihydrochloride)(AAPH)-induced erythrocyte hemolysis was analyzed by ultraviolet spectrophotometry.Results:The results showed that the amino acid content of fish roe polypeptide was 70.41%,EAA/TAA was 39.23%,EAA/NEAA was 64.55%;AAS and CS were higher, the first limiting amino acid was tryptophan.Compared with the injury group,the fish roe polypeptide 100 μg/mL significantly increased the insulin secretion of high glucose-damaged islet β cells to (40.0±6.9) pg/mL;The 500 mg/kg dose group can significantly reduce the area under the blood glucose curve of normal mice.The difference in the control group was statistically significant.The IC50 of the inhibition of erythrocyte hemolysis by fish roe polypeptide was 1 mg/mL.Conclusion:Antioxidant activity amino acids, delicious amino acids,and pharmacodynamic amino acids were r ich in fi sh roe ploypeptide and had a high nutritional value.The insulin test at the cellular level screened the fish roe polypeptide could change the insulin level of the islet cells,suggesting that it might have hypoglycemic activity;animal experiments found that it could improve the postprandial blood glucose of mice to effectively reduce the area under the blood glucose curve,which proved that it had hypoglycemic activity.The inhibitory effect of the fish roe polypeptide on erythrocyte hemolysis clarified that it might achieve hypoglycemic effect by clearing ROS to reduce the level of cellular oxidative stress.