L-肉碱对H_2O_2诱导氧化应激FHM细胞的影响  

Effects of L- carnitine on H_2O_2-induced oxidative stress in FHM cells

作　　者：王秋举[1] 鞠雪[1] 罗莎[1] 陈玉柯[1] 董晓庆[1] 刘洪键 张东鸣[1] 

机构地区：[1]吉林农业大学动物科学技术学院,吉林长春130118 [2]吉林省水产技术推广站,吉林长春130012

出　　处：《南京农业大学学报》2016年第3期467-472,共6页Journal of Nanjing Agricultural University

基　　金：国家自然科学基金项目(30671621;30972191;31372540);吉林省产业技术研究与开发项目(2011002-1)

摘　　要：[目的]本文旨在研究L-肉碱对氧化应激状态下胖头鱥肌肉细胞系(fathead minnow muscle cell line,FHM)细胞活力、脂质过氧化及抗氧化功能的影响。[方法]以FHM细胞为研究对象,以H_2O_2为氧化应激因子,采用四甲基偶氮唑盐(MTT)法测定不同浓度L-肉碱(0.001、0.01、0.1、0.5、1.0和5.0 mmol·L^(-1))预处理6 h分别对0.2和0.6 mmol·L^(-1)H_2O_2诱导的细胞氧化应激活力的影响,并采用生物化学方法检测L-肉碱预处理对氧化应激FHM细胞中脂质过氧化程度及抗氧化酶的变化。[结果]0.2和0.6 mmol·L^(-1)H_2O_2刺激FHM细胞1 h,细胞活力分别降低至72%和58%。与H_2O_2组相比,0.001~1.0 mmol·L^(-1)L-肉碱预处理使低氧化水平(0.6 mmol·L^(-1)H_2O_2)的FHM细胞活力显著升高;且0.001、0.5和1.0 mmol·L^(-1)L-肉碱均显著降低不同氧化水平细胞中MDA含量。与0.2 mmol·L^(-1)H_2O_2组相比,0.001、0.01、0.1和1.0 mmol·L^(-1)L-肉碱显著提高细胞T-GSH含量;0.001、0.01和1.0 mmol·L^(-1)L-肉碱显著提高细胞中T-SOD活性;0.5~5.0 mmol·L^(-1)L-肉碱组细胞中CAT活性显著增加;添加0.001~1.0 mmol·L^(-1)L-肉碱显著提高细胞GPx活性,但γ-GCS活性只有0.5 mmol·L^(-1)L-肉碱组显著升高。与0.6 mmol·L^(-1)H_2O_2组相比,0.001~1.0 mmol·L^(-1)L-肉碱显著提高细胞中T-GSH含量,0.01~5.0 mmol·L^(-1)L-肉碱显著提高细胞中CAT活性,0.001 mmol·L^(-1)L-肉碱显著提高细胞T-SOD活性,0.01 mmol·L^(-1)L-肉碱显著提升细胞γ-GCS活性;同时,L-肉碱的处理显著增强细胞GPx活性。[结论]高浓度(0.6 mmol·L^(-1))H_2O_2对FHM细胞的损害较大;0.001~1.0 mmol·L^(-1)L-肉碱可提高FHM细胞活力和抗氧化能力,并对细胞抵抗H_2O_2诱导的不同程度氧化应激有积极作用。[Objectives]This study was designed to investigate the effects of L-carnitine on viability,lipid peroxidation and antioxidant function in fathead minnow muscle cell line（ FHM） under oxidative stress. [Methods]Using FHM as study object and H2O2 as inducer of oxidative stress,the effects of defferent concentrations of L-carnitine（ 0.001,0.01,0.1,0.5,1.0 and 5.0 mmol·L^（-1）） on cell viability following 0.2 and 0.6 mmol·L^（-1）H2O2 challenge were measured by methyl thiazolyl tetrazolium（ MTT） method. The changes of lipid peroxidation and antioxidant enzymes in FHM cells were also detected by biochemical methods. [Results]Treatment with H2O2（ 0.2 and 0.6 mmol·L^（-1）） for 6 h could decrease cell viability by 72% and 58%,respectively. Compared with H2O2 group,pretreatment with L-carnitine at concentrations of 0.001-1.0 mmol·L^（-1）significantly inhibited lower level H2O2（ 0.2 mmol·L^（-1））-induced cell viability loss,and MDA levels were significantly reduced in L-carnitine groups of 0. 001,0. 5 and 1. 0 mmol·L^（-1）（ P〈0.05）.Compared with 0. 2 mmol·L^（-1）H2O2 group,L-carnitine（ 0. 001,0. 01,0. 1 and 1. 0 mmol·L^（-1）） significantly increased intracellular T-GSH levels; the activities of T-SOD were significantly increased in L-carnitine groups of 0.001,0.01 and 1.0 mmol·L^（-1）,0.5-5.0mmol·L^（-1）L-carnitine significantly increased the CAT activities; adding 0.001- 1.0 mmol·L^（-1）L-carnitine significantly increased the GPx activities; however,γ-GCS activity was reached significant level in 0.5 mmol·L^（-1）L-carnitine group. Compared with 0.6 mmol·L^（-1）H2O2 group,0.001- 1. 0 mmol·L^（-1）L-carnitine significantly increased intracellular T-GSH levels; 0. 01- 5. 0 mmol·L^（-1）L-carnitine significantly increased intracellular CAT activities; activities of T-SOD and γ-GCS were significantly increased in 0. 001 and 0. 01mmol·L^（-1）L-carnitine group,respectively. Meanwhile,GPx activities of all the L-carnitine groups were signi

关 键 词：L-肉碱 胖头鱥肌肉细胞系(FHM) H2O2 氧化应激 

分 类 号：S816.701.11[农业科学—饲料科学]