笛鲷鱼鳞源功能多肽对Caco-2细胞氧化应激损伤的保护作用

Protective Effect of Functional Peptides Derived from Crimson Snapper Scales on Oxidative Stress Damage in Caco-2 Cells

本实验利用650μmol/L H_(2)O_(2)构建Caco-2细胞氧化应激损伤模型,探究笛鲷鱼鳞源功能多肽(crimson snapper scale peptides,CSSPs)对Caco-2细胞氧化应激损伤的保护作用。通过测定细胞上清液乳酸脱氢酶(lactate dehydrogenase,LDH)活力和白细胞介素-8(interleukin-8,IL-8)质量浓度、胞内抗氧化酶活力和丙二醛(malondialdehyde,MDA)含量、胞内活性氧(reactive oxygen species,ROS)相对含量、相关凋亡基因(Caspase-3、Caspase-9、Bax和Bcl-2)的相对表达量,从细胞氧化还原状态和细胞凋亡两方面阐明CSSPs对损伤Caco-2细胞的抗氧化作用。结果显示,CSSPs能够通过抗氧化防御系统来减轻H_(2)O_(2)对Caco-2细胞的损伤,主要包括增加胞内抗氧化酶活性以抑制LDH释放、胞内ROS积累、MDA及IL-8的生成。此外,CSSPs抑制氧化应激损伤诱导的细胞凋亡与线粒体介导的细胞凋亡途径有关,其可显著提高线粒体膜电位,下调Caspase-3、Caspase-9和Bax表达,上调Bcl-2表达。结果表明,CSSPs能够维持细胞氧化还原稳态和抑制细胞凋亡,有效减轻Caco-2细胞的损伤程度。

The purpose of this study was to investigate the effect of crimson snapper scale peptides (CSSPs) on H_(2)O_(2)-induced oxidative damage in Caco-2 cells.The levels of lactate dehydrogenase (LDH) and interleukin-8 (IL-8) in the cell supernatant,the activity of intracellular antioxidant enzymes,the release of intracellular reactive oxygen species (ROS)and malondialdehyde (MDA),and the expression of apoptosis-related genes (Caspase-3,Caspase-9,Bax and Bcl-2) were measured.The antioxidant effects of CSSPs in intestinal cells were elucidated from the aspects of cellular redox state and apoptosis.Results showed that CSSPs mitigated H_(2)O_(2) -induced oxidative damage in Caco-2 cells through the antioxidant defense system.This effect involved increased activities of antioxidant enzymes and consequent inhibition of LDH release,intracellular ROS accumulation,and MDA and IL-8 production.Furthermore,CSSPs inhibited oxidative stress-induced apoptosis by regulating the mitochondrial apoptotic pathway,which significantly increased mitochondrial membrane potential,down-regulated the mRNA expression of Caspase-3,Caspase-9 and Bax,and up-regulated the mRNA expression of Bcl-2.In conclusion,CSSPs can effectively reduce the degree of cell damage by maintaining redox homeostasis and inhibiting apoptosis.