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原代培养虹鳟鱼(Oncorhynchus mykiss)脂肪细胞成脂分化过程中的自噬反应。

The autophagy response during adipogenesis of primary cultured rainbow trout (Oncorhynchus mykiss) adipocytes.

机构信息

Departament de Biologia Cel·lular, Fisiologia i Immunologia, Facultat de Biologia, Universitat de Barcelona, Barcelona 08028, Spain.

Departament de Biologia Cel·lular, Fisiologia i Immunologia, Facultat de Biologia, Universitat de Barcelona, Barcelona 08028, Spain; Université de Pau et des Pays de l'Adour, E2S UPPA, INRAE, UMR1419, Nutrition Métabolisme et Aquaculture, F-64310 Saint-Pée-sur-Nivelle, France.

出版信息

Comp Biochem Physiol B Biochem Mol Biol. 2022 Feb-Mar;258:110700. doi: 10.1016/j.cbpb.2021.110700. Epub 2021 Nov 27.

DOI:10.1016/j.cbpb.2021.110700
PMID:34848371
Abstract

Adipogenesis is a tightly regulated process, and the involvement of autophagy has been recently proposed in mammalian models. In rainbow trout, two well-defined phases describe the development of primary cultured adipocyte cells: proliferation and differentiation. Nevertheless, information on the transcriptional profile at the onset of differentiation and the potential role of autophagy in this process is scarce. In the present study, the cells showed an early and transient induction of several adipogenic transcription factors genes' expression (i.e., cebpa and cebpb) along with the morphological changes (round shape filled with small lipid droplets) typical of the onset of adipogenesis. Then, the expression of various lipid metabolism-related genes involving the synthesis (fas), uptake (fatp1 and cd36), accumulation (plin2) and mobilization (hsl) of lipids, characteristic of the mature adipocyte, increased. In parallel, several autophagy markers (i.e., atg4b, gabarapl1 and lc3b) mirrored the expression of those adipogenic-related genes, suggesting a role of autophagy during in vitro fish adipogenesis. In this regard, the incubation of preadipocytes with lysosomal inhibitors (Bafilomycin A1 or Chloroquine), described to prevent autophagy flux, delayed the process of adipogenesis (i.e., cell remodelling), thus suggesting a possible relationship between autophagy and adipocyte differentiation in trout. Moreover, the disruption of the autophagic flux altered the expression of some key adipogenic genes such as cebpa and pparg. Overall, this study contributes to improve our knowledge on the regulation of rainbow trout adipocyte differentiation, and highlights for the first time in fish the involvement of autophagy on adipogenesis, suggesting a close-fitting connection between both processes.

摘要

脂肪生成是一个受到严格调控的过程,自噬作用的参与最近在哺乳动物模型中被提出。在虹鳟鱼中,两个明确的阶段描述了原代培养脂肪细胞的发育:增殖和分化。然而,关于分化起始时的转录谱以及自噬在这个过程中的潜在作用的信息却很少。在本研究中,细胞表现出几种脂肪生成转录因子基因(如 cebpa 和 cebpb)的早期和短暂诱导表达,同时伴随着脂肪生成起始的形态变化(圆形,充满小脂滴)。然后,涉及脂质合成(fas)、摄取(fatp1 和 cd36)、积累(plin2)和动员(hsl)的各种脂质代谢相关基因的表达增加,这些特征是成熟脂肪细胞的特征。平行地,几种自噬标志物(如 atg4b、gabarapl1 和 lc3b)反映了那些与脂肪生成相关基因的表达,表明自噬在体外鱼脂肪生成中发挥作用。在这方面,用溶酶体抑制剂(巴弗洛霉素 A1 或氯喹)孵育前脂肪细胞,被描述为防止自噬流,延迟脂肪生成过程(即细胞重塑),因此表明自噬和脂肪细胞分化之间可能存在关系在鳜鱼中。此外,自噬流的中断改变了一些关键的脂肪生成基因的表达,如 cebpa 和 pparg。总的来说,这项研究有助于提高我们对虹鳟鱼脂肪细胞分化调控的认识,并首次在鱼类中强调了自噬作用对脂肪生成的参与,表明这两个过程之间存在紧密的联系。

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