鹅乙酰辅酶A酰基转移酶2基因的克隆及其在鹅肥肝形成过程中的表达变化

doi:10.11843/j.issn.0366-6964.2016.04.009

畜牧兽医学报 ›› 2016, Vol. 47 ›› Issue (4): 700-708.doi: 10.11843/j.issn.0366-6964.2016.04.009

鹅乙酰辅酶A酰基转移酶2基因的克隆及其在鹅肥肝形成过程中的表达变化

王倩倩，杨彪，夏丽丽，孙晓先，耿拓宇，龚道清^*

（扬州大学动物科学与技术学院，扬州 225009）

收稿日期:2015-08-24 出版日期:2016-04-23 发布日期:2016-04-23
通讯作者: 龚道清，博士，教授，博士生导师， E-mail：yzgong@163.com
作者简介:王倩倩（1988-），女，山东栖霞人，硕士生，主要从事家禽分子营养研究，E-mail：xiaoxiaoqian_qian@126.com
基金资助:
国家自然科学基金项目（31372298；31472086）；江苏省农业自主创新项目（ＣＸ（１２）２０３３）

Cloning of Goose Acetyl-Coenzyme A Acyltransferase 2 and Its Expression Pattern in the Development of Fatty Liver

WANG Qian-qian，YANG Biao，XIA Li-li，SUN Xiao-xian，GENG Tuo-yu，GONG Dao-qing^*

(College of Animal Science and Technology，Yangzhou University，Yangzhou 225009，China）

Received:2015-08-24 Online:2016-04-23 Published:2016-04-23

摘要/Abstract

摘要：

为了探讨ACAA2基因与鹅（Anser anser）肝脂肪代谢的关系，本试验选取35只朗德鹅分为填饲组（15只）和对照组（20只），填饲组包括填饲7、14和19天3个阶段。运用RT-PCR方法克隆出朗德鹅ACAA2基因的完整编码序列并进行生物信息学分析，采用实时定量PCR技术测定了朗德鹅填饲不同阶段该基因在肝中的表达水平，并用葡萄糖、脂肪酸和胰岛素分别处理鹅原代肝细胞，观察这些因子对基因表达的影响。朗德鹅ACAA2基因完整CDS区长1 194 bp，编码 397个氨基酸；各阶段填饲组肝中该基因的表达量显著高于对照组（P<0.05），但随填饲时间的延长，表达水平呈下降趋势。另外，在培养的鹅原代肝细胞中，相较于对照组，0.5 mmol•L^-1的油酸能使ACAA2的表达量显著上调，而0.25 mmol•L^-1的棕榈酸和不同浓度的胰岛素能显著下调ACAA2的表达（P<0.05）。结果表明，ACAA2基因的表达与鹅肥肝的形成密切相关，为深入研究ACAA2基因在鹅肥肝形成过程中的作用奠定了基础。

Abstract:

To explore the relationship between ACAA2 gene and liver fatty metabolism in goose，35 Landes geese were divided into overfeeding group(15) and control group(20)，the overfeeding group included 3 periods：overfeeding 7，14 and 19 days.The full coding sequence (CDS) of ACAA2 was cloned by RT-PCR and analyzed，its expression levels at different stages of overfeeding were determined in both normal and fatty liver by qRT-PCR in Landes geese.In addition，goose primary hepatocytes were treated with glucose，fatty acids or insulin to determine how its expression was regulated during fatty liver formation.Data indicated that the complete CDS of goose ACAA2 was 1 194 bp and encoded 397 amino acids.The ACAA2 expression level in the livers of the overfed geese was significantly higher (P<0.05) than the control at every stage，and presented a declining trend with the extension of overfeeding time.Moreover，compared to the control，the expression of ACAA2 gene was significantly increased in the goose primary hepatocytes treated with 0.5 mmol•L^-1 oleate，but was decreased in the primary hepatocytes treated with 0.25 mmol•L^-1 palmitate and insulin with different concentrations (P<0.05).The results suggeste that ACAA2 expression is closely related to the development of goose fatty liver，which will provide a basis for further study the role of ACAA2 gene in the formation of goose fatty liver.

中图分类号:

S835
S813.3

王倩倩，杨彪，夏丽丽，孙晓先，耿拓宇，龚道清. 鹅乙酰辅酶A酰基转移酶2基因的克隆及其在鹅肥肝形成过程中的表达变化[J]. 畜牧兽医学报, 2016, 47(4): 700-708.

WANG Qian-qian，YANG Biao，XIA Li-li，SUN Xiao-xian，GENG Tuo-yu，GONG Dao-qing. Cloning of Goose Acetyl-Coenzyme A Acyltransferase 2 and Its Expression Pattern in the Development of Fatty Liver[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(4): 700-708.

0
/ / 推荐

导出引用管理器 EndNote|Reference Manager|ProCite|BibTeX|RefWorks

链接本文: https://www.xmsyxb.com/CN/10.11843/j.issn.0366-6964.2016.04.009

https://www.xmsyxb.com/CN/Y2016/V47/I4/700

参考文献

［1］卢素芳，何瑞国.鹅肥肝饲料技术的研究近况［J］.饲料广角，2003(3)：34-36.
LU S F，HE R G.The research status of goose fat liver feed technology［J］.Feed Angle，2003(3)：34-36.(in Chinese)
［2］周敏，潘健存，王士长.鹅肥肝的营养作用及其生产技术［J］.广西农学报，2004(5)：40-43.
ZHOU M，PAN J C，WANG S C.The utrition function and production technology of geese fatty liver［J］.Chinese Journal of Guangxi Agriculture，2004(5)：40-43.(in Chinese)
［3］MOUROT J，GUY G，LAGARRIGUE S，et al.Role of hepatic lipogenesis in the susceptibility to fatty liver in the goose (Anser anser)［J］.Com Biochem Physiol B：Biochem Mol Biol，2000，126(1)：81-87.
［4］ZHU L H，MENG H，DUAN X J，et al.Gene expression profile in the liver tissue of goose after overfeeding［J］.Poult Sci，2011，90(1)：107-117.
［5］邵丹，王来娣，张蕊，等.鹅肥肝形成相关基因的研究进展［J］.中国畜牧兽医，2012，39(9)：42-46.
SHAO D，WANG L D，ZHANG R，et al.The research progress of related genes in goose fatty liver formation［J］. China Animal Husbandry & Veterinary Medicine，2012，39(9)：42-46.(in Chinese)
［6］WANDERS R J，VREKEN P，FERDINANDUSSE S，et al.Peroxisomal fatty acid alpha-and beta-oxidation in humans：enzymology，peroxisomal metabolite transporters and peroxisomal diseases［J］.Biochem Soc Trans，2001，29 (Pt 2)：250-267.
［7］EASTON S，BARTLETT K，POURFARZAM M.Mammalian mitochondrial β-oxidation［J］.Biochem，1996，320：345-357.
［8］李文娟．鸡肉品质相关脂肪代谢功能基因的筛选及营养调控研究［D］．北京：中国农业科学院，2008.
LI W J.The genes select of lipid metabolism about chicken quality and its ntrition research［D］.Beijing：Chinese Academy of Agricultural Sciences，2008.(in Chinese)
［9］DE BOER V C，VAN SCHOTHORST E M，DIHAL A A.Chronic quercetin exposure affects fatty acid catabolism in rat lung.［J］.Cell Mol Life Sci，2006，63(23)：2847-2858.
［10］SIDRAH A，ATTYA B，ASMA G.In-silico approaches to sequence and structure based scrutiny of nonsynonymous SNPs and synteny of ACAA2 for its implicated role in metabolomics［J］ Proceedings IWBBIO，2014：7-9.
［11］何桦，潘志雄，刘贺贺，等.鹅DHRS7基因克隆、序列分析及其在鹅肥肝形成过程中的表达特点［J］.畜牧兽医学报，2012，43(9)：1385-1391.
HE H，PAN Z X，LIU H H，et al.Cloning of goose DHRS7 gene，sequence analysis and its expression characteristic in the development of fatty liver［J］.Acta Veterinaria et Zootechinca Sinica，2012，43(9)：1385-1391.(in Chinese)
［12］潘志雄，王继文，唐慧，等.鹅Perilipin基因部分片段的克隆、不同品种及填饲对组织mRNA表达水平的影响［J］.畜牧兽医学报，2010，41(8)：939-943.
DAN Z X，WANG J W，TANG H，et al.Cloning of goose Perilipin gene，tissues expression and the effect of overfeeding on its mRNA level of mRNA in tissues［J］.Acta Veterinaria et Zootechinca Sinica，2010，41(8)：939-943.(in Chinese)
［13］潘志雄.油酸诱导鹅肝细胞脂肪变性对细胞内脂质代谢平衡相关基因表达的影响［D］.雅安：四川农业大学，2009.
PAN Z X.Effects of oleic oil on expression of gene related to lipid metabolic homeostasis in goose primary hepatocytes［D］.Ya’an：Sichuan Agricultural University，2009.(in Chinese)
［14］张兴元，宋向芹，张芳，等.介绍一种简易高效的大鼠原代肝细胞分离方法［J］.滨州医学院学报，2011，34(1)：29-32.
ZHANG X Y，SONG X Q，ZHANG F，et al.The introduce of a simple and effective separation method for rat primary hepatocytes［J］.Chinese Journal of Binzhou Medical University，2011，34(1)：29-32.(in Chinese)
［15］WAN C L，KATE L，DAVID T.Isolation and culture of adult mouse hepatocytes［J］.Methods Mol Biol，2010，633：185-196.
［16］HU T，FOXWORTHY P，SIESKY A，et al.Hepatic peroxisomal fatty acid beta-oxidation is regulated by liver X receptor alpha［J］.Endocrinology，2005，146(12)：5380-5387.
［17］DOI M，KONDO Y，TSUTSUMI K.Lipoprotein lipase activator NO-1886 (ibrolipim) accelerates the mRNA expression of fatty acid oxidation-related enzymes in rat liver［J］.Metabolism，2003，52(12)：1547-1550.
［18］HAN C C，WANG J W，XU H Y，et al.Effect of overfeeding on plasma parametes and mRNA expression of genes associated with hepatic lipogenesis in geese［J］.Asian-Australia J Anim Sci，2008，21(4)：590.
［19］王宝维，舒常平，葛文华，等.填饲期肥肝鹅脂肪沉积、血脂成分和脂类代谢酶的变化规律［J］.中国农业科学，2014，47(8)：1600-1610.
WANG B W，SHU C P，GE W H，et al.The change rule of fat deposition，lipid composition and lipid metabolism enzymes in goose fatty liver［J］.Scientia Agricultura Sinica，2014，47(8)：1600-1610.(in Chinese)
［20］GENG T，HU W，BROADWATER M H，et al.Fatty acids differentially regulate insulin resistance through endoplasm reticulum stress-mediated induction of tribbles homologue 3：a potential link between dietary fat composition and the pathophysiological outcomes of obesity［J］.Diabetologia，2013，56(9)：2078-2087.

[1]	陈兴勇，谢珊珊，周丽，姜润深，耿照玉. 皖西白鹅换羽前后基因表达谱差异分析[J]. 畜牧兽医学报, 2013, 44(7): 1030-1036.
[2]	何桦, 潘志雄, 刘贺贺, 李乐, 夏露, 胡深强, 鲁凯, 董霞, 李亮, 王继文. 鹅DHRS7基因克隆、序列分析及其在鹅肥肝形成过程中的表达特点[J]. 畜牧兽医学报, 2012, 43(9): 1385-1391.

鹅乙酰辅酶A酰基转移酶2基因的克隆及其在鹅肥肝形成过程中的表达变化

Cloning of Goose Acetyl-Coenzyme A Acyltransferase 2 and Its Expression Pattern in the Development of Fatty Liver

RichHTML

PDF

可视化

被引次数

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 2

编辑推荐

Metrics

本文评价