鸡肝脏中PNPLA3基因表达及调控研究  

Characteristics of Expression and Regulation of PNPLA3 in the Liver of Chicken(Gallus gallus)

作　　者：杨丽玉 李艳敏 郑航 田卫华 王彦彬 田亚东 康相涛 刘小军 李红 YANG Li-Yu;LI Yan-Min;ZHENG Hang;TIAN Wei-Hua;WANG Yan-Bin;TIAN Ya-Dong;KANG Xiang-Tao;LIU Xiao-Jun;LI Hong(College of Animal Science and Veterinary Medicine,Henan Agricultural University/Henan Innovative Engineering Research Center of Poultry GermPlasm Resource/International Joint Research Laboratory for Poultry Breeding of Henan,Zhengzhou 450002,Chin)

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：《农业生物技术学报》2018年第8期1361-1370,共10页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金(No.31372330);河南省国际合作项目(No.162102410030);河南省产学研合作专项(No.162107000066);农业部现代农业产业技术体系项目(No.CARS-40-K04)

摘　　要：PNPLA3(patatin like phospholipase domain-containing 3)也称为脂肪细胞营养素(adiponutrin)或不依赖于钙的磷脂酶A2ε(calcium-independent phospholipase A2ε,i PLA2ε),是PNPLAs家族中的一员,具有水解磷脂和中性脂质的活性以及酯酰基转移酶活性,能够参与脂肪的积累和脂肪动员。本研究旨在探讨鸡(Gallus gallus)肝脏中PNPLA3基因表达调控规律,为进一步阐明PNPLA3基因在鸡肝脏脂质代谢中的生物学功能奠定基础。首先运用生物信息学方法对鸡和其他12种动物的PNPLA3氨基酸序列进行分子进化分析;然后利用q RT-PCR技术,分析PNPLA3基因在不同发育时期卢氏绿壳蛋鸡各组织的时空表达规律,并分别从个体与细胞水平分析雌激素及其受体拮抗剂对PNPLA3表达的调控机制。结果表明,鸡与两栖类、爬行类和鱼类等物种的PNPLA3氨基酸序列相似性较高,在哺乳动物人(Homo sapiens)和小鼠(Mus musculus)等物种中的序列相似性较低;鸡PNPLA3基因在各组织中表现出广泛表达的特性,其中在10周龄的鸡胸肌、腹脂、胰腺中表达水平较高,在30周龄鸡的肝脏、胸肌、腹脂中表达水平较高,且在肝脏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脏组织和体外培养的鸡胚胎肝原代细胞中PNPLA3的表达水平(P<0.05),雌激素受体(estrogen receptor,ER)-α特异性拮抗剂MPP(methyl-piperidino-pyrazole)可完全抑制雌激素的作用(P<0.05)。综上所述,鸡与两栖类和鱼类等物种的PNPLA3氨基酸序列相似性较高,与哺乳动物序列相似性较低;PNPLA3在肝脏组织表达丰度较高,且在性成熟后显著升高;雌激素通过雌激素受体ER-α调控PNPLA3表达。上述发现为深入研究鸡PNPLA3与肝脏脂肪代谢调控机制提供了基础资料。The patatin like phospholipase domain-containing 3 （PNPLA3）, also known as fat cell nutrients or calcium-independent phospholipase A2ε （iPLA2ε）, is a member of the PNPLAs family. PNPLA3 has the activity of hydrolyzing phospholipids and neutral lipids and ester acyltransferase activity, and is able to participate in fat accumulation and fat mobilization. The objective of the present study is to investigate the characteristics of expression and regulation of PNPPLA3 in liver of chicken （Gallus gallus） and to pave the way for further elucidating the biological function of PNPLA3 gene in lipid metabolism of liver in chicken. The amino acid sequences of PNPLA3 in chicken and 12 other different species were retrieved from the genomic data bank using the Basic Local Alignment Search Tool （BLAST） homology search. Bioinformatic methods were used for phylogenetic and syntenic analysis. The qRT-PCR technique was used to analyze the temporal and spatial expression of PNPLA3 gene in different tissues of chicken. The regulatory mechanism of PNPLA3 gene was analyzed through stimulating juvenile hens in vivo, and chicken embryo hepatocytes treated with 17β-estradiol and estrogen receptor antagonists MPP, ICI182780 （fulvestrant） and ICI146474 （tamoxifen） in vitro. The results showed that the amino acid sequence similarity of PNPLA3 among chicken and amphibian and fish species was relatively higher than that among chicken and mammalian species. The PNPLA3 gene was extensively expressed in various tissues tested, in particular, the expression levels were higher in pectoral muscle, abdominal fat and pancreas in 10-week-old chicken, higher in liver, pectoral muscle, abdominal fat in 30-week-old chicken （P〈0.01）. The 17β-estradiol treatment significantly increased the expression level of PNPLA3 in chicken liver tissue and primary hepatocytes （P〈0.05）. The MPP, a specific antagonist of estrogen receptor ER-α, could completely inhibit the effect of 17β-estradiol on PNPLA3 expression （P〈0.05�

关 键 词：鸡 肝脏 PNPLA3基因 表达调控 雌激素 

分 类 号：S831.2[农业科学—畜牧学]