杨博雅, 顾嘉宁, 张新党, 常绪路, 张艳敏, 冯世坤, 杨国坤, 孟晓林. 草鱼subfatin分子鉴定及表达特性分析[J]. 南方水产科学, 2024, 20(5): 159-168. DOI: 10.12131/20240096
引用本文: 杨博雅, 顾嘉宁, 张新党, 常绪路, 张艳敏, 冯世坤, 杨国坤, 孟晓林. 草鱼subfatin分子鉴定及表达特性分析[J]. 南方水产科学, 2024, 20(5): 159-168. DOI: 10.12131/20240096
YANG Boya, GU Jianing, ZHANG Xindang, CHANG Xulu, ZHANG Yanmin, FENG Shikun, YANG Guokun, MENG Xiaolin. Molecular identification and expression characteristics analysis of subfatin gene from grass carp[J]. South China Fisheries Science, 2024, 20(5): 159-168. DOI: 10.12131/20240096
Citation: YANG Boya, GU Jianing, ZHANG Xindang, CHANG Xulu, ZHANG Yanmin, FENG Shikun, YANG Guokun, MENG Xiaolin. Molecular identification and expression characteristics analysis of subfatin gene from grass carp[J]. South China Fisheries Science, 2024, 20(5): 159-168. DOI: 10.12131/20240096

草鱼subfatin分子鉴定及表达特性分析

Molecular identification and expression characteristics analysis of subfatin gene from grass carp

  • 摘要: Subfatin脂肪因子在哺乳动物中具有调节脂质代谢、炎症、能量消耗等多种功能,但其在鱼类中的功能研究较少。以草鱼 (Ctenopharyngodon idella) 为研究对象,利用RT-PCR技术克隆得到草鱼subfatin基因序列,通过葡萄糖灌喂、饥饿再投喂、诱导脂质沉积及原代肝细胞培养实验,探究了Subfatin在草鱼糖脂代谢中的生理功能。研究发现,草鱼subfatin基因全长为861 bp,编码286个氨基酸,包含3个O-连接糖基化位点。葡萄糖灌喂草鱼1和3 h后,肝脏和肠道组织中subfatin mRNA的表达量显著上调,葡萄糖处理3 h后,脑和肾脏组织中subfatin mRNA的表达量显著上调。饥饿再投喂实验结果表明,草鱼禁食14 d后,脑、脂肪、肠道和肝脏组织中subfatin mRNA的表达量显著下调,再投喂后其表达量显著上调。诱导脂肪沉积实验结果表明,诱导组的脑、脂肪、肠道和肝脏组织中subfatin基因的表达量均显著上调。葡萄糖、油酸和胰高血糖素分别孵育原代肝细胞均能显著增加subfatin基因的表达量;而胰岛素却会显著降低肝细胞中subfatin基因的表达量。综上所述,Subfatin参与了草鱼的葡萄糖和脂质代谢过程并发挥调节作用,为阐明Subfatin在鱼类中的生理功能提供了一定的理论依据。

     

    Abstract: Subfatin plays an important role in lipid metabolism, inflammation and energy consumption in mammals, but its functions in fish are rarely reported. To investigate the physiological function of subfatin in glucolipid metabolism of grass carp, we took Ctenopharyngodon idella as the research object and cloned the subfatin gene by RT-PCR. The oral glucose tolerance test (OGTT), fast and refeeding, inducing lipid accumulation and primary hepatocyte culture were proceeded. The results show that the grass carp subfatin was 861 bp, encoding 286 amino acids, and three O-linked glycosylation sites were located in the subfatin. The results of OGTT experiment show that after glucose treatment of grass carp for 1 and 3 h, the expression of subfatin mRNA was significantly up-regulated in liver and intestinal tissues, and it was also significantly up-regulated in the brain and kidney tissues after 3 h of glucose treatment. In fast and refeeding experiment, the results show that the expression of subfatin mRNA in brain, fat, intestinal and liver tissues was significantly down-regulated after 14-day starvation treatment, and was significantly up-regulated after re-feeding. In induced lipid accumulation experiment, the results show that subfatin expression in brain, fat, intestinal and liver tissues was significantly up-regulated in the induced group. In primary hepatocytes, subfatin expression level increased significantly by treatment with glucose, oleic acid and glucagon. However, subfatin level decreased significantly by treatment with insulin. In conclusion, this study demonstrates that Subfatin is involved in the process of glucose and lipid metabolism in grass carp and plays a regulatory role, which provides a theoretical basis for elucidating the physiological functions of Subfatin in fish.

     

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