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草鱼脂肪肝细胞来源的外泌体的蛋白质组学分析。

Proteomic analysis of exosomes derived from fatty hepatocytes of grass carp.

机构信息

College of Fisheries, Henan Normal University, Xinxiang, People's Republic of China.

College of Animal Science and Technology, Northwest A&F University, Yangling, People's Republic of China.

出版信息

J Fish Biol. 2024 Sep;105(3):886-893. doi: 10.1111/jfb.15840. Epub 2024 Jun 19.

Abstract

Exosomes participate in intercellular communication by carrying proteins, messenger RNA, microRNAs, and non-coding RNA. Fatty liver is a common phenomenon in farmed fish, but there has been little study of fatty hepatocytes-derived exosomes. Here, we successfully isolated exosomes from hepatocytes of grass carp, named Exos (hepatocytes-derived exosomes) and OA-Exos (fatty hepatocytes-derived exosomes), from which 617 differentially expressed proteins were identified using liquid chromatography tandem mass spectrometry. Of these, 320 proteins were promoted and 297 proteins were restrained, which were gathered in biological processes and cellular components (cellular processes, cells, and intracellular structures). The results of kyoto encyclopedia of genes and genomes (KEGG) analysis revealed that the differential expression proteins were gathered in "carbohydrate transport and metabolism", "translation, ribosomal structure and biogenesis", "posttranslational modification, protein turnover, chaperones", and "intracellular trafficking, secretion, and vesicular transport". In addition, five differentially expressed exosomal proteins were further confirmed by parallel reaction monitoring, including 2-phospho-D-glycerate hydrolyase, cytochrome b5, fatty acid-binding protein domain-containing protein, metallothionein, and malate dehydrogenas, which were downregulated. These findings provided evidence that exosomes derived from fatty hepatocytes of grass carp may be biomarkers for the early diagnosis, treatment, and prevention of fatty liver in fishery development.

摘要

外泌体通过携带蛋白质、信使 RNA、microRNAs 和非编码 RNA 参与细胞间通讯。脂肪肝是养殖鱼类中常见的现象,但对脂肪性肝细胞衍生的外泌体研究甚少。在这里,我们成功地从草鱼肝细胞中分离出外泌体,命名为 Exos(肝细胞衍生的外泌体)和 OA-Exos(脂肪性肝细胞衍生的外泌体),并使用液相色谱串联质谱法鉴定了 617 种差异表达蛋白。其中,320 种蛋白被促进,297 种蛋白被抑制,这些蛋白聚集在生物过程和细胞成分(细胞过程、细胞和细胞内结构)中。京都基因与基因组百科全书(KEGG)分析的结果表明,差异表达蛋白聚集在“碳水化合物运输和代谢”、“翻译、核糖体结构和生物发生”、“翻译后修饰、蛋白质周转、伴侣”和“细胞内运输、分泌和小泡运输”。此外,通过平行反应监测进一步证实了 5 种差异表达的外泌体蛋白,包括 2-磷酸-D-甘油酸水解酶、细胞色素 b5、脂肪酸结合蛋白结构域蛋白、金属硫蛋白和苹果酸脱氢酶,它们的表达下调。这些发现为草鱼脂肪性肝细胞衍生的外泌体可能成为渔业发展中脂肪肝早期诊断、治疗和预防的生物标志物提供了证据。

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