华南农业大学学报2017，Vol.38Issue(5)：1-6,6.DOI:10.7671/j.issn.1001-411X.2017.05.001
不同浓度氨基酸对仔猪原代肝细胞尿素循环的影响
Effects of different concentrations of amino acids on urea cycle in primary porcine hepatocytes
摘要
Abstract
[Objective] To investigate the effects of different concentrations of amino acids on urea cycle in primary hepatocytes isolated from piglet,and explore the potential mechanism.[Method] Primary hepatocytes isolated from 5-day old piglet were cultured in medium containing different concentrations of amino acids (AA)(1,2,or 4 times of physiological concentration in serum).After 24 h,the supernatant and cells were collected.Urea and ammonia concentrations in the supernatant,glutaminase(GLS) and glutamine synthetase(GS) activities and ammonia concentration in cells were examined by colorimetry.The mRNA expression of genes involved in urea cycle,including carbamyl phosphate synthetase 1(CPS1),Argininosuccinate synthetase(ASS),Argininosuccinate lyase(ASL),Omithine transcarbamylase (OTC),Arginase (ARG) were detected by qRT-PCR.[Result] Urea and ammonia concentrations in the supernatant,and GLS activity in cells were significantly elevated in 4×AA group.Meanwhile,4×AA remarkably increased the mRNA expression of CPS1,ASS and ASL genes in hepatocytes.NH4C1 at 1.0 and 2.0 mmol·L-1 concentrations significantly promoted urea synthesis in the cell.[Conclusion] High concentration of AA might accelerate ammonia accumulation by GLS and enhance the expression of the urea cycle enzymes (CPS 1,ASS and ASL),which contribute to increased production of urea.关键词
仔猪/原代肝细胞/氨基酸/尿素循环/氨/基因表达/酶活性Key words
piglet/primary hepatocyte/amino acid/urea cycle/ammonia/gene expression/enzyme activity分类
农业科技引用本文复制引用
张梦媛,庄鲁,万小娟,束刚,王丽娜,朱晓彤,高萍,王松波,江青艳..不同浓度氨基酸对仔猪原代肝细胞尿素循环的影响[J].华南农业大学学报,2017,38(5):1-6,6.基金项目
973项目(2013CB127304) （2013CB127304）
