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本研究以Neutrase 0.8 L对酪蛋白进行水解,水解后的酪蛋白(HC)分别导入外源性氨基酸苯丙氨酸、组氨酸与脯氨酸进行类蛋白反应。所得的修饰产物(HCPHE、HCHIS和HCPRO)体外的DPPH?清除活性,羟自由基清除能力和还原能力与HC相比最大可分别提高30.51%、45.83%和42.81%(p<0.05)。HUPVC与修饰产物单独预孵育24 h后经300μmol/L H2O2氧化损伤考察修饰产物对HUPVC的保护作用,CCK-8法测定细胞存活率结果表明修饰产物高剂量组的细胞成活率分别高于HC组7.86%、5.21%和10.52%(p<0.05),与茶多酚(TP)给药组无明显差异(p>0.05),并存在剂量依赖关系。与氧化损伤模型组相比,HCPHE、HCHIS和HCPRO可有效降低LDH渗出率,MDA生成量,SOD活力,降低效果强于同浓度的HC组,与此同时可使GSH-Rd,CAT的活力显著增强,然而细胞内GSH的含量变化不明显。综上所述,三种类蛋白反应修饰产物对H2O2诱导的HUPVC损伤具有很好的保护作用,且保护效果高于HC。
In this study, casein was hydrolyzed with Neutrase 0.8 L, and hydrolyzed casein (HC) was introduced into the exogenous amino acids phenylalanine, histidine and proline for protein-like reaction. The obtained modified products (HCPHE, HCHIS and HCPRO) could increase DPPH? Scavenging activity, hydroxyl radical scavenging ability and reducing ability by as much as 30.51%, 45.83% and 42.81% (p <0.05) compared with HC in vitro. HUPVC and modified products were preincubated for 24 h, then the protective effect of modified products on HUPVC was examined by 300 μmol / L H2O2 oxidative damage. The cell survival rate was determined by CCK-8 method. The results showed that the cell survival rate of high-dose modified products were higher than that of HC group 7.86%, 5.21% and 10.52%, respectively (p <0.05). There was no significant difference (p> 0.05) between the two groups and the dose of tea polyphenols (TP). Compared with oxidative damage model group, HCPHE, HCHIS and HCPRO can effectively reduce the exudation rate of LDH, the production of MDA, the activity of SOD, and the reduction effect is stronger than that of the same concentration of HC group. At the same time, the activity of GSH-Rd and CAT Significantly increased, however, the intracellular GSH content did not change significantly. In summary, the three kinds of protein-modified products have a good protective effect against H2O2-induced HUPVC injury, and the protective effect is higher than that of HC.