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作 者:李艳敏[1] 李翠翠[1] 郑航[1] 田方圆[1] 王台安 田亚东[1] 李转见[1] 康相涛[1] 刘小军[1]
机构地区:[1]河南农业大学牧医工程学院,河南省家禽种质资源创新工程研究中心,河南省家禽育种国际联合实验室,郑州450002
出 处:《畜牧兽医学报》2016年第12期2362-2369,共8页ACTA VETERINARIA ET ZOOTECHNICA SINICA
基 金:河南省国际合作项目(162102410030);地方鸡种质资源保护与利用教育部创新团队发展计划(IRT1236);国家蛋鸡产业技术体系遗传育种岗位专项资金(CARS-41-K04)
摘 要:本研究旨在探讨鸡CYP2C8基因的表达调控规律。采集30周龄卢氏绿壳蛋鸡心、肝、脾、肺、肾、胸肌、腹脂、肌胃、腺胃、卵巢、胰、十二指肠、空肠、回肠14种组织以及从1日龄~30周龄不同发育时期的肝组织,用荧光实时定量PCR分析CYP2C8基因的时空表达规律;用雌激素及其受体拮抗剂对活体和体外培养的鸡胚肝原代细胞进行处理,用荧光实时定量PCR分析CYP2C8基因的表达调控机制。结果表明,鸡CYP2C8基因在各组织中表现出广泛表达的特性,但在脂类代谢旺盛的器官如肝、肾、小肠(十二指肠、空肠、回肠)等器官中表达较高,且在肝中的表达水平随着性成熟而显著升高(P〈0.01);雌激素处理可显著提高鸡肝组织和体外培养的鸡胚胎肝原代细胞中CYP2C8的表达水平(P〈0.05),但雌激素受体ER-α拮抗剂MPP可完全抑制雌激素的作用(P〈0.05)。综上表明,鸡CYP2C8基因在不同组织中广泛表达,而以肝中的表达水平最高,且肝中CYP2C8基因的表达是受雌激素并通过ER-α调控。The objectives of the present study were to investigate the characteristics of expression and regulation of CYP2C8 gene in chicken.A total of 14tissues(including heart,liver,spleen,lung,kidney,pectoral muscle,abdominal fat,gizzard,glandular stomach,ovaries,pancreas,duodenum,jejunum and ileum)were collected from Lushi green-shelled-egg chickens at the age of30 weeks old,and the liver tissues were obtained from the chickens at different development stages(from the age of 1day to 30 weeks old).The samples were used to detect the spatiotemporal expression profiles of CYP2C8 gene using quantitative real-time PCR(qRT-PCR)technology.In addition,the pullets(at the age of 10 weeks old)which were treated with 17β-estradiol,and chicken embryo primary hepatocytes which were treated with either 17β-estradiol alone or combination of 17β-estradiol and estrogen receptor(ER)antagonists were used to analysis the expression regulation mechanism of the CYP2C8 gene.The results showed that CYP2C8 gene was expressed extensively in different tissues of chicken.The relative expression level of CYP2C8 was higher in the main lipid metabolism tissues such as liver,kidney,and small intestine(duodenum,jejunum and ileum)than that in other tissues.The expression level of CYP2C8 gene in liver of hens was significantly increased with the sexual maturity(P〈0.01).Estrogen could significantly up-regulate the expression of CYP2C8 in liver of chicken and hepatocytes(P〈0.05).MPP,as an ER-αspecific antagonist,could completely antagonize the expression of CYP2C8gene(P〈0.05).Therefore we concluded that CYP2C8 gene was expressed extensively in different tissues with higher expression levels in liver,kidney,and small intestine(duodenum,jejunum,and ileum)tissues,and the highest expression level in liver.It was suggested that the effect of estrogen on the expression of CYP2C8 gene was predominantly mediated via ER-αin the chicken liver.
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