1,25-dihydroxyvitamin D3 affects thapsigargin-induced endoplasmic reticulum stress in 3T3-L1 adipocytes.

IF 2 4区 医学 Q3 NUTRITION & DIETETICS Nutrition Research and Practice Pub Date : 2024-02-01 Epub Date: 2023-12-21 DOI:10.4162/nrp.2024.18.1.1
Dain Wi, Chan Yoon Park
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Abstract

Background/objectives: Endoplasmic reticulum (ER) stress in adipose tissue causes an inflammatory response and leads to metabolic diseases. However, the association between vitamin D and adipose ER stress remains poorly understood. In this study, we investigated whether 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) alleviates ER stress in adipocytes.

Materials/methods: 3T3-L1 cells were treated with different concentrations (i.e., 10-100 nM) of 1,25(OH)2D3 after or during differentiation (i.e., on day 0-7, 3-7, or 7). They were then incubated with thapsigargin (TG, 500 nM) for an additional 24 h to induce ER stress. Next, we measured the mRNA and protein levels of genes involved in unfold protein response (UPR) and adipogenesis using real-time polymerase chain reaction and western blotting and quantified the secreted protein levels of pro-inflammatory cytokines. Finally, the mRNA levels of UPR pathway genes were measured in adipocytes transfected with siRNA-targeting Vdr.

Results: Treatment with 1,25(OH)2D3 during various stages of adipocyte differentiation significantly inhibited ER stress induced by TG. In fully differentiated 3T3-L1 adipocytes, 1,25(OH)2D3 treatment suppressed mRNA levels of Ddit3, sXbp1, and Atf4 and decreased the secretion of monocyte chemoattractant protein-1, interleukin-6, and tumor necrosis factor-α. However, downregulation of the mRNA levels of Ddit3, sXbp1, and Atf4 following 1,25(OH)2D3 administration was not observed in Vdr-knockdown adipocytes. In addition, exposure of 3T3-L1 preadipocytes to 1,25(OH)2D3 inhibited transcription of Ddit3, sXbp1, Atf4, Bip, and Atf6 and reduced the p-alpha subunit of translation initiation factor 2 (eIF2α)/eIF2α and p-protein kinase RNA-like ER kinase (PERK)/PERK protein ratios. Furthermore, 1,25(OH)2D3 treatment before adipocyte differentiation reduced adipogenesis and the mRNA levels of adipogenic genes.

Conclusions: Our data suggest that 1,25(OH)2D3 prevents TG-induced ER stress and inflammatory responses in mature adipocytes by downregulating UPR signaling via binding with Vdr. In addition, the inhibition of adipogenesis by vitamin D may contribute to the reduction of ER stress in adipocytes.

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1,25-二羟维生素 D3 会影响硫辛酸诱导的 3T3-L1 脂肪细胞内质网应激。
背景/目的:脂肪组织中的内质网(ER)应激会引起炎症反应并导致代谢性疾病。然而,人们对维生素 D 与脂肪组织内质网应激之间的关系仍然知之甚少。在这项研究中,我们探讨了 1,25-二羟维生素 D3(1,25(OH)2D3)是否能减轻脂肪细胞的 ER 应激。材料/方法:在分化后或分化过程中(即第 0-7、3-7 或 7 天),用不同浓度(即 10-100 nM)的 1,25(OH)2D3 处理 3T3-L1 细胞。然后将它们与硫司加精(TG,500 nM)孵育24小时,以诱导ER应激。接着,我们用实时聚合酶链式反应和免疫印迹法测定了参与折叠蛋白反应(UPR)和脂肪生成的基因的 mRNA 和蛋白水平,并量化了促炎细胞因子的分泌蛋白水平。最后,在转染了靶向 Vdr 的 siRNA 的脂肪细胞中测定了 UPR 通路基因的 mRNA 水平:结果:在脂肪细胞分化的不同阶段使用 1,25(OH)2D3 能显著抑制 TG 诱导的 ER 应激。在完全分化的 3T3-L1 脂肪细胞中,1,25(OH)2D3 可抑制 Ddit3、sXbp1 和 Atf4 的 mRNA 水平,减少单核细胞趋化蛋白-1、白细胞介素-6 和肿瘤坏死因子-α 的分泌。然而,在 Vdr 敲除的脂肪细胞中,1,25(OH)2D3 给药后并未观察到 Ddit3、sXbp1 和 Atf4 mRNA 水平的下调。此外,将 3T3-L1 前脂肪细胞暴露于 1,25(OH)2D3,可抑制 Ddit3、sXbp1、Atf4、Bip 和 Atf6 的转录,并降低翻译起始因子 2 的 p-α 亚基(eIF2α)/eIF2α 和 p 蛋白激酶 RNA 样 ER 激酶(PERK)/PERK 蛋白的比率。此外,在脂肪细胞分化前处理1,25(OH)2D3可减少脂肪生成和脂肪生成基因的mRNA水平:我们的数据表明,1,25(OH)2D3 可通过与 Vdr 结合下调 UPR 信号,从而防止 TG 诱导的成熟脂肪细胞 ER 应激和炎症反应。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Nutrition Research and Practice
Nutrition Research and Practice NUTRITION & DIETETICS-
CiteScore
3.50
自引率
4.20%
发文量
62
审稿时长
6-12 weeks
期刊介绍: Nutrition Research and Practice (NRP) is an official journal, jointly published by the Korean Nutrition Society and the Korean Society of Community Nutrition since 2007. The journal had been published quarterly at the initial stage and has been published bimonthly since 2010. NRP aims to stimulate research and practice across diverse areas of human nutrition. The Journal publishes peer-reviewed original manuscripts on nutrition biochemistry and metabolism, community nutrition, nutrition and disease management, nutritional epidemiology, nutrition education, foodservice management in the following categories: Original Research Articles, Notes, Communications, and Reviews. Reviews will be received by the invitation of the editors only. Statements made and opinions expressed in the manuscripts published in this Journal represent the views of authors and do not necessarily reflect the opinion of the Societies.
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