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

MA Xuelian; LIAO Xiudong; ZHANG Liyang; LYU Lin; LUO Xugang

doi:10.3969/j.issn.1006-267x.2020.08.042

2020 , Vol. 32 >Issue 8: 3830 - 3842

DOI: https://doi.org/10.3969/j.issn.1006-267x.2020.08.042

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

MA Xuelian ,
LIAO Xiudong ,
ZHANG Liyang ,
LYU Lin ,
LUO Xugang

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1. Mineral Nutrition Research Division, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China;
2. Poultry Mineral Nutrition Laboratory, College of Animal Science and Technology, Yangzhou University, Yangzhou 225000, China

Received date: 2020-03-13

Online published: 2020-08-13

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Abstract

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 protein expression level of FTH1 in hepatocytes (P<0.05). The protein expression level of COX1 in hepatocytes at 24 and 72 h was significantly higher than that at 48 h (P<0.05), and the protein expression level of COX1 in hepatocytes of 1.00 mmol/L Fe level group was significantly higher than that of other Fe level groups (P<0.05). In conclusion, Fe level and incubation time can affect the Fe content, activities and gene expression of SDH, CAT and COX, and the gene expression of FTH1 in primary cultured hepatocytes of broiler embryos. Consider the above indicators comprehensively, the optimal Fe level is 0.50 mmol/L, and the optimal incubation time is 24 h.

Key words： primary cultured hepatocytes; broiler embryos; iron; iron-containing enzyme; gene expression

Cite this article

MA Xuelian , LIAO Xiudong , ZHANG Liyang , LYU Lin , LUO Xugang . 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[J]. Chinese Journal of Animal Nutrition, 2020 , 32(8) : 3830 -3842 . DOI: 10.3969/j.issn.1006-267x.2020.08.042

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