L-肉碱对胖头鱥肌肉细胞和草鱼性腺细胞抗氧化功能的调节作用  

Regulation of L-carnitine on antioxidant function in Pimephales promelas muscle cell line and grass carp Ctenopharyngodon idellus ovary cell line

作　　者：郑男 王秋举[1] 马悦[1] 于婷 陈玉珂[1] 刘洪键 张东鸣 ZHENG Nan;WANG Qiu-ju;MA Yue;YU Ting;CHEN Yu-ke;LIU Hong-jian;ZHANG Dong-ming(College of Animal Science and Technology,Jilin Agricultural University,Changchun 130118,China;Fishery Technical Extension Station of Jilin Province,Changchun 130012,China;Tonghua Normal University,Tonghua 134000,China)

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 [3]通化师范学院,吉林通化134000

出　　处：《大连海洋大学学报》2018年第6期736-742,共7页Journal of Dalian Ocean University

基　　金：吉林省科技厅重点研发项目(20180201017NY);国家自然科学基金资助项目(30671621,30972191,31372540);吉林省产业技术研究与开发项目(2011002-1);吉林农业大学科研启动基金资助项目(201704).

摘　　要：为研究L-肉碱对胖头鱥Pimephales promelas肌肉细胞(FHM)和草鱼Ctenopharyngodon idellus性腺细胞(GCO)抗氧化酶功能的调节作用,以两种鲤科鱼细胞FHM和GCO为研究对象,在培养基中添加不同浓度的L-肉碱(0、0. 001、0. 01、0. 1、0. 5、1、5 mmol/L)分别孵育FHM细胞6 h、GCO细胞12 h,采用生物化学法和荧光定量PCR法检测L-肉碱对细胞抗氧化酶活性和基因相对表达量的影响。结果表明:0. 01～5 mmol/L L-肉碱组FHM细胞SOD、CAT活性显著高于对照组(P<0. 05),仅5 mmol/L L-肉碱组GCO细胞SOD、CAT、GPX、γ-GCS活性显著高于对照组(P<0. 05);与对照组相比,L-肉碱组FHM细胞Cu Zn-SOD mRNA相对表达水平无显著变化(P>0. 05),0. 5、1、5 mmol/L L-肉碱组GCO细胞Cu ZnSOD mRNA相对表达水平显著上调(P<0. 05); 0. 01～5 mmol/L L-肉碱组FHM细胞CAT mRNA相对表达水平显著上调(P<0. 05),所有L-肉碱组GCO细胞CAT mRNA的相对表达水平显著上调(P<0. 05); 0. 5mmol/L L-肉碱组GCO细胞GPX mRNA相对表达水平显著上调(P <0. 05);仅0. 1 mmol/L L-肉碱处理FHM细胞GCLC mRNA相对表达量水平显著上调(P<0. 05),L-肉碱添加范围为0. 5～5 mmol/L时,GCO细胞GCLC mRNA的相对表达水平均显著上调(P<0. 05)。研究表明,在培养基中添加L-肉碱能明显上调FHM和GCO细胞的抗氧化酶活性及其相关基因的相对表达水平,在本试验条件下,细胞培养基中推荐L-肉碱的适宜添加浓度为0. 1～1 mmol/L。The activities of Copper,Zinc-superoxide dismutase(CuZn-SOD),catalase(CAT),glutathione peroxidase(GPX) and glutathione ligase synthesis subunit(GCLC) and the gene relative expression of m RNA s of CuZn-SOD,CAT,GPX and GCLC were investigated in fathead minnow Pimephales promelas muscle cell line(FHM)for6h and grass carp Ctenopharyngodon idellus ovary cell line(GCO)for12h in culture media containing L-carnitine at a dose of0,0.001,0.01,0.1,0.5,1and 5mmol/L by q-PCR to evaluate the regulatory function of L-carnitine on the gene relative expression of antioxidant enzymes in FHM and GCO.The results showed that no significant differences in CuZn-SOD mRNA relative expression was found in the FHM between the L-carnitine groups and the control group(P>0.05).There was significant higher regulated CuZn-SOD mRNA relative expression in0.5 and 5mmol/L L-carnitine group than that in the control group in the GCO(P<0.05).Compared to the control group,there was significantly higher activities of SOD and CAT in0.01-5mmol/L L-carnitine group in the FHM(P<0.05).In the 5mmol/L group,however,there were significantly higher activities of SOD,CAT,GPX andγ-GCS in GCO cells than those in the control group(P<0.05).The significant up-regulation in CAT mRNA relative expression were found in GCO between all of the L-carnitine groups and the control groups(P<0.05)and in FHM cells in 0.01-5mmol/L carnitine groups(P<0.05).The GPX mRNA relative expression in GCO cells was shown to be significantly up-regulation in 0.5mmol/L L-carnitine group(P<0.05),with significantly up-regulated GCLC mRNA relative expression in0.1mmol/L L-carnitine group compared to the control group in GCO(P<0.05).The level of GCLC mRNA relative expression was significantly increased when the GCO was treated by0.5,1 and 5mmol/L L-carnitine(P<0.05).The findings indicate that supplementation of L-carnitine up-regulates the mRNA relative expression of antioxidant enzymes in FHM and GCO,and it is suggested that the appropriate concentration of L-carnitine be 0.1-1mmol/L under the

关 键 词：胖头鱥 草鱼 L-肉碱 基因相对表达 鱼细胞 

分 类 号：S963[农业科学—水产养殖]