铁水平和孵育时间对原代培养肉鸡鸡胚肝细胞中铁含量及含铁酶活性、基因表达和蛋白表达的影响  

Effects of Iron Level and Incubation Time on Iron Content and Activity,Gene Expression and Protein Expression of Iron-Containing Enzyme in Primarily Cultured Hepatocytes of Broiler Embryos

作　　者：马雪莲 廖秀冬[1] 张丽阳[1] 吕林[1] 罗绪刚 MA Xuelian;LIAO Xiudong;ZHANG Liyang;LYU Lin;LUO Xugang(Mineral Nutrition Research Division,Institute of Animal Science,Chinese Academy of Agricultural Sciences,Beijing 100193,China;Poultry Mineral Nutrition Laboratory,College of Animal Science and Technology,Yangzhou University,Yangzhou 225000,China)

机构地区：[1]中国农业科学院北京畜牧兽医研究所,矿物元素营养研究室,北京100193 [2]扬州大学动物科技学院,家禽矿物元素营养研究室,扬州225000

出　　处：《动物营养学报》2020年第8期3830-3842,共13页CHINESE JOURNAL OF ANIMAL NUTRITION

基　　金：国家自然科学基金项目(31672440);国家现代农业产业技术体系岗位专家专项经费(CARS-41);中国农业科学院科技创新工程专项经费(ASTIP-IAS08)。

摘　　要：本试验旨在研究铁水平和孵育时间对原代培养肉鸡鸡胚肝细胞中铁含量及含铁酶活性、基因表达和蛋白表达的影响。采用5×3两因子完全随机设计,铁水平分别为0、0.25、0.50、0.75、1.00 mmol/L,孵育时间分别为24、48和72 h,共形成15个组,每组6个重复。结果表明:1)铁水平、孵育时间及两者的互作对肝细胞铁含量有显著影响(P<0.05)。24、48和72 h时,0.25、0.50、0.75、1.00 mmol/L铁水平组肝细胞中铁含量均显著高于0 mmol/L铁水平组(P<0.05)。2)铁水平和孵育时间对肝细胞中胞琥珀酸脱氢酶(SDH)和细胞色素c氧化酶(COX)活性有显著影响(P<0.05)。与0 mmol/L铁水平组相比,0.25、0.50、0.75和1.00 mmol/L铁水平组肝细胞中SDH活性显著降低(P<0.05);24 h肝细胞中SDH活性显著高于48 h(P<0.05),而72 h又显著高于24和48 h(P<0.05)。与0 mmol/L铁水平组相比,0.75和1.00 mmol/L铁水平组肝细胞中COX活性显著提高(P<0.05);24和48 h肝细胞中COX活性显著高于72 h(P<0.05)。3)铁水平、孵育时间及两者的互作对肝细胞中SDH、过氧化氢酶(CAT)、COX7A2L和铁蛋白重链1(FTH1)的mRNA表达水平有显著影响(P<0.05),铁水平及铁水平与孵育时间的互作对肝细胞中COX1的mRNA表达水平有显著影响(P<0.05)。4)铁水平和孵育时间对肝细胞中COX1的蛋白表达水平有显著影响(P<0.05),铁水平及铁水平与孵育时间的互作对肝细胞中FTH1的蛋白表达水平表达有显著影响(P<0.05)。24和72 h肝细胞中COX1的蛋白表达水平显著高于48 h(P<0.05),1.00 mmol/L铁水平组肝细胞中COX1的蛋白表达水平显著高于其他铁水平组(P<0.05)。由此可见,铁水平和孵育时间可影响原代培养肉鸡鸡胚肝细胞中铁含量,SDH、CAT和COX的活性、基因表达,以及FTH1的基因表达。综合考虑以上指标,适宜铁的水平应为0.50 mmol/L,孵育时间为24 h。This experiment was conducted to investigate the effects of iron(Fe)level and incubation time on Fe concentration,Fe-containing enzyme activity,gene expression and protein expression in primary cultured hepatocytes of broiler embryos.A 5×3 two factor completely randomized design was used in the experiment,the Fe levels were 0,0.25,0.50,0.75 and 1.00 mmol/L,respectively,and the incubation times were 12,24 and 48 h,respectively.There were assigned to 15 groups with 6 replicates per group.The result showed as follows:1)Fe level,incubation time and there interaction had significant effects on content of Fe in hepatocytes(P<0.05).At 24,48 and 72 h,the content of Fe in hepatocytes of 0.25,0.50,0.75,1.00 mmol/L Fe level groups was significantly higher than that of 0 mmol/L Fe level group(P<0.05).2)Fe level and incubation time had significant effects on activities of succinate dehydrogenase(SDH)and cytochrome c oxidase(COX)in hepatocytes(P<0.05).Compared with the 0 mmol/L Fe level group,the activity of SDH in hepatocytes of 0.25,0.50,0.75 and 1.00 mmol/L Fe level groups was significantly decreased(P<0.05);the activity of SDH in hepatocytes at 24 h was significantly higher than that at 48 h(P<0.05),and 72 h was significantly higher than 24 and 48 h(P<0.05).Compared with the 0 mmol/L Fe level group,the activity of COX in hepatocytes of 0.75 and 1.00 mmol/L Fe level groups was significantly increased(P<0.05);the activity of COX in hepatocytes at 24 and 48 h was significantly higher than that at 72 h(P<0.05).3)Fe level,incubation time and there interaction had significant effects on mRNA expression levels of SDH,catalase(CAT),COX7A2L,ferritin heavy chain 1(FTH1)in hepatocytes(P<0.05),and Fe level and interaction of Fe level and incubation time had significant effects on mRNA expression level of COX1 in hepatocytes(P<0.05).4)Fe level and incubation time had significant effects on protein expression level of COX1 in hepatocytes(P<0.05),and Fe level and interaction of Fe level and incubation time had significant effects on pro

关 键 词：原代培养肝细胞 肉鸡鸡胚 铁 含铁酶 基因表达 

分 类 号：S831[农业科学—畜牧学]