中国农业科学2017，Vol.50Issue(21)：4205-4211,7.DOI:10.3864/j.issn.0578-1752.2017.21.015
鸡原代肝细胞培养及叶酸对脂质代谢相关基因表达的影响
Effect of Folic Acid on Lipid Metabolism Associated Gene Expression in Primarily Cultured Chickens Hepatocytes
摘要
Abstract
[Objective] Abdominal fat deposition is a universal phenomenon at present in poultry industry.Different from mammals,90％-95％ of lipid metabolism in poultry occurs in the liver.Previous studies have demonstrated folic acid and IGF2 were involved in animals lipid metabolism.The study was conducted to establish the method for primary culture of chicken hepatocytes and investigate the effects of folic acid on IGF2 and genes expression associated with lipid metabolism in primary chicken hepatocytes,further providing a basis for exploring lipid metabolism of poultry in vitro.[Method] Chicken hepatocytes were isolated by collagenase digestion combined with purification of chicken lymphocyte separation fluid,and evaluated by PAS staining,respectively.After 36 h culture of hepatocytes,cells were treated with different concentration of folic acid for 12 h.Hepatocytes proliferation and injury was detected by MTT method and LDH activity in culture medium respectively.Cells were collected to get total RNA for gene expression analysis by RT-PCR.Later,correlation analysis was carried out between IGF2 and lipid metabolism related genes expression.Regression analysis was performed between folic acid concentration and genes expression.[Result] The results showed that hepatocytes were isolated with high purity.Folic acid didn't affect cell proliferation and cellular LDH activity (P ＞0.05).When compared with the control group (1 mg·L-1 folic acid),10,15 and 20 mg·L~ folic acid significantly reduced IGF2 expression in chicken hepatocytes (P＜0.05),and same phenomenon was observed in FAS and ACC expression.However,folic acid had no effects on CPT-1 and PPARt expression (P＞0.05).In addition,FAS and ACC expression had positive correlation with IGF2 mRNA level in chicken hepatocytes (P＜0.05).There existed linear and quadratic regression between genes expression and folic acid concentration (P＜0.05).[Conclusion] In conclusion,folic acid could reduce gene expression associated with fatty acid synthesis in chicken hepatocytes.In addition,FAS and ACC mRNA level might have positive connection with IGF2 expression.Optimum treatment dose of folic acid was 15 mg·L-1 in this study.关键词
叶酸/IGF2/肝细胞/脂质代谢/鸡Key words
folic acid/IGF2/hepatocytes/lipid metabolism/chicken引用本文复制引用
刘艳利,党燕娜,段玉兰,杨小军..鸡原代肝细胞培养及叶酸对脂质代谢相关基因表达的影响[J].中国农业科学,2017,50(21):4205-4211,7.基金项目
国家重点研发计划(20170502200,2017YFD0500500)、陕西省科技统筹创新工程计划项目(2017TSCXL-NY-04-04,2015KTCQ02-19) （20170502200,2017YFD0500500）
