团头鲂PGC1β基因克隆及高糖胁迫、葡萄糖和胰岛素负荷对其mRNA表达的影响  

Molecular characterization of PGC1β from Megalobrama amblycephala and responsiveness to dietary carbohydrate levels and glucose and insulin loadings

作　　者：徐超 刘文斌 谢帝芝 丁志荣 Jean-Jacques Yao Adjoumani 石泉 李向飞 XU Chao;LIU Wenbin;XIE Dizhi;DING Zhirong;Jean-Jacques Yao Adjoumani;SHI Quan;LI Xiangfei(College of Marine Sciences,South China Agricultural University,Guangzhou 510642,China;Key Laboratory of Aquatic Nutrition and Feed Science of Jiangsu Province,College of Animal Science and Technology,Nanjing Agricultural University,Nanjing 210095,China)

机构地区：[1]华南农业大学海洋学院,广东广州510642 [2]南京农业大学动物科技学院,江苏省水产动物营养重点实验室,江苏南京210095

出　　处：《中国水产科学》2021年第8期959-967,共9页Journal of Fishery Sciences of China

基　　金：国家自然科学基金项目(31872576,32002361);财政部和农业农村部国家现代农业产业技术体系资助项目(CARS-45-14).

摘　　要：为探讨过氧化物酶体增殖活化受体γ辅激活因子1β(peroxisome proliferator activated receptorγcoactivator 1β,PGC1β)基因在鱼类糖代谢中的作用,本研究采用RACE方法获得了团头鲂PGC1β的cDNA片段序列,利用生物信息学技术对该基因及其所编码蛋白的结构特征进行了分析;采用实时定量PCR技术,检测了高糖(45%)饲喂、葡萄糖及胰岛素负荷对团头鲂肝脏和肌肉中该基因表达的影响。结果显示,团头鲂PGC1βcDNA片段长为1759 bp,其中开放阅读框为1719bp,编码573个氨基酸;同源性分析结果显示,团头鲂PGC1β与其他鲤科鱼类的PGC1β高度同源(氨基酸相似度78%以上),并与草鱼PGC1β的进化关系最近;高糖组鱼肝脏和肌肉中PGC1β的表达量高于对照组,但无显著差异;此外,葡萄糖及胰岛素负荷后,肝脏中PGC1β基因的表达量在2h内显著降低至最小值,随后逐渐升高至基础水平。以上结果提示, PGC1β可能在调节鱼类糖代谢中发挥重要作用。Carbohydrates are an excellent source of energy and carbon for mammalian cells. A constant supply of glucose prevents hypoglycemia and its potentially catastrophic effects on the cells of the nervous system. Unlike for mammals, it is not usually regarded as the panacea for fish. Although the continual ingestion of a carbohydrate-enriched diet or glucose-loading can provide essential metabolic energy, most fish species appear to have impaired glucose tolerance and often display prolonged postprandial hyperglycemia. Recently, several approaches such as metabolomics and transcriptomics have been employed to assess diet-induced metabolic syndromes in fish. Findings from such studies have suggested that the disruption of energy homeostasis in fish is strongly implicated in the development of disturbances in the glucose metabolism. Peroxisome proliferator-activated receptor γ coactivator 1β(PGC1β) is a coactivator of nuclear receptors and other transcription factors that regulates several components of energy metabolism, particularly aspects of hepatic glucose metabolism. To investigate the molecular regulatory mechanisms of glucose metabolism in fish, the full-length cDNA sequence of PGC1β from Megalobrama amblycephala was identified by rapid amplification complementary DNA ends, and the structural features of the gene and its encoded protein were analyzed through bioinformatics methods. Using real-time PCR, the transcriptional response of this gene to dietary carbohydrate levels and glucose and insulin loadings was studied. The result revealed that the obtained cDNA of PGC1β in M. amblycephala was 1759 bp in length, and consisted of a 1719 bp open reading frame encoding 573 amino acids. Additionally, conserved motifs were identified in the amino acid sequence of the PGC1β protein, including the peroxisome proliferator-activated receptor interaction domain;the RNA recognition motif;and the LXXLL, TPPTTPP, and DHDYC motifs. Alignment based on amino acid sequences showed that M. amblycephala PGC1β had a 78.63%-97

关 键 词：团头鲂 过氧化物酶体增殖活化受体γ辅激活因子1β 基因克隆 糖代谢 基因表达 

分 类 号：S961[农业科学—水产养殖]