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

作　　者：王倩倩[1] 杨彪[1] 夏丽丽[1] 孙晓先[1] 耿拓宇[1] 龚道清[1] 

机构地区：[1]扬州大学动物科学与技术学院,扬州225009

出　　处：《畜牧兽医学报》2016年第4期700-708,共9页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：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)

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 194bp,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,0.5mmol·L^(-1)的油酸能使ACAA2的表达量显著上调,而0.25mmol·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。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 3periods：overfeeding 7,14 and 19days.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 0.25mmol·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,whichwill provide a basis for further study the role of ACAA2 gene in the formation of goose fatty liver.

关 键 词：鹅 乙酰辅酶A酰基转移酶2 脂肪肝 基因克隆 

分 类 号：S835[农业科学—畜牧学] S813.3[农业科学—畜牧兽医]