鲑鱼皮明胶水解物的抗氧化活性及其对细胞氧化损伤的保护作用
详细信息Antioxidant activity of gelatin hydrolysates derived from salmon (Oncorhynchus keta) skin and their cytoprotective effect on oxidative injury of rat hepatocytes
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摘要: 采用碱性蛋白酶、木瓜蛋白酶和中性蛋白酶水解鲑鱼皮明胶,得到水解度为4.7%到13.5%的7个明胶水解物,分别评价明胶水解物对自由基的清除能力。明胶水解物(0.5、1、2mg/mL)预先作用BRL大鼠肝细胞2h后,通过H2O2(5mmol/L)诱导氧化损伤,分别测定细胞存活率、乳酸脱氢酶(LDH)、丙二醛(MDA)和谷胱甘肽(GSH)等细胞内抗氧化产物的含量变化。结果表明,明胶水解物的抗氧化活性随着水解度的增大呈增强趋势。明胶水解物对细胞具有保护作用,可显著提高细胞存活率,降低LDH渗出量和MDA生成量,且存在一定的剂量关系,但细胞中GSH含量变化不显著;7个明胶水解物对H2O2诱导肝细胞损伤保护时,细胞存活率与水解物的体外抗氧化活性大小显著正相关,而LDH渗出量和MDA生成量与水解物的体外抗氧化活性大小负相关。Abstract: Seven chum salmon skin gelatin hydrolysates with degree of hydrolysis (DH) 4. 7%～13. 5% were prepared by three commercial proteases, Alcalase, papain or Neutrase, respectively. The antioxidant activity of gelatin hydrolysates was evaluated by the ability of scavenging free radicals. The results demonstrated that the antioxidant activity of the gelatin hydrolysates was gradually increased with the increase of degree of hydrolysis in a dose-dependent manner. Rat hepatocyte BRL cells were preincubated with gelatin hydrolysates (0.5, 1, 2mg/mL) and then were oxidative injuried by H2O2 (5mmol/L) . The cell viability, leakage of lactate dehydrogenase (LDH) , content of malondialdehyde (MDA) and GSH were determined by commercial kits. The results indicated that the cell viability was remarkably improved while the leakage of LDH and MDA formation were reduced significantly with the content of GSH almost unchanged. The statistical analysis of the results showed that seven gelatin hydrolysates played a protective role in H2O2-induced oxidative injury, and the cell viability was positively correlated with antioxidant activity of gelatin hydrolysates, while leakage of LDH and the content of MDA formed were negatively correlated with antioxidant activity of gelatin hydrolysates.
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