{"title":"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。","authors":"Dain Wi, Chan Yoon Park","doi":"10.4162/nrp.2024.18.1.1","DOIUrl":null,"url":null,"abstract":"<p><strong>Background/objectives: </strong>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 D<sub>3</sub> (1,25(OH)<sub>2</sub>D<sub>3</sub>) alleviates ER stress in adipocytes.</p><p><strong>Materials/methods: </strong>3T3-L1 cells were treated with different concentrations (i.e., 10-100 nM) of 1,25(OH)<sub>2</sub>D<sub>3</sub> 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 <i>Vdr</i>.</p><p><strong>Results: </strong>Treatment with 1,25(OH)<sub>2</sub>D<sub>3</sub> during various stages of adipocyte differentiation significantly inhibited ER stress induced by TG. In fully differentiated 3T3-L1 adipocytes, 1,25(OH)<sub>2</sub>D<sub>3</sub> treatment suppressed mRNA levels of <i>Ddit3</i>, <i>sXbp1</i>, and <i>Atf4</i> and decreased the secretion of monocyte chemoattractant protein-1, interleukin-6, and tumor necrosis factor-α. However, downregulation of the mRNA levels of <i>Ddit3</i>, <i>sXbp1</i>, and <i>Atf4</i> following 1,25(OH)<sub>2</sub>D<sub>3</sub> administration was not observed in <i>Vdr</i>-knockdown adipocytes. In addition, exposure of 3T3-L1 preadipocytes to 1,25(OH)<sub>2</sub>D<sub>3</sub> inhibited transcription of <i>Ddit3</i>, <i>sXbp1</i>, <i>Atf4</i>, <i>Bip</i>, and <i>Atf6</i> 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)<sub>2</sub>D<sub>3</sub> treatment before adipocyte differentiation reduced adipogenesis and the mRNA levels of adipogenic genes.</p><p><strong>Conclusions: </strong>Our data suggest that 1,25(OH)<sub>2</sub>D<sub>3</sub> prevents TG-induced ER stress and inflammatory responses in mature adipocytes by downregulating UPR signaling via binding with <i>Vdr</i>. In addition, the inhibition of adipogenesis by vitamin D may contribute to the reduction of ER stress in adipocytes.</p>","PeriodicalId":19232,"journal":{"name":"Nutrition Research and Practice","volume":"18 1","pages":"1-18"},"PeriodicalIF":2.0000,"publicationDate":"2024-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10861344/pdf/","citationCount":"0","resultStr":"{\"title\":\"1,25-dihydroxyvitamin D<sub>3</sub> affects thapsigargin-induced endoplasmic reticulum stress in 3T3-L1 adipocytes.\",\"authors\":\"Dain Wi, Chan Yoon Park\",\"doi\":\"10.4162/nrp.2024.18.1.1\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Background/objectives: </strong>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 D<sub>3</sub> (1,25(OH)<sub>2</sub>D<sub>3</sub>) alleviates ER stress in adipocytes.</p><p><strong>Materials/methods: </strong>3T3-L1 cells were treated with different concentrations (i.e., 10-100 nM) of 1,25(OH)<sub>2</sub>D<sub>3</sub> 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 <i>Vdr</i>.</p><p><strong>Results: </strong>Treatment with 1,25(OH)<sub>2</sub>D<sub>3</sub> during various stages of adipocyte differentiation significantly inhibited ER stress induced by TG. In fully differentiated 3T3-L1 adipocytes, 1,25(OH)<sub>2</sub>D<sub>3</sub> treatment suppressed mRNA levels of <i>Ddit3</i>, <i>sXbp1</i>, and <i>Atf4</i> and decreased the secretion of monocyte chemoattractant protein-1, interleukin-6, and tumor necrosis factor-α. However, downregulation of the mRNA levels of <i>Ddit3</i>, <i>sXbp1</i>, and <i>Atf4</i> following 1,25(OH)<sub>2</sub>D<sub>3</sub> administration was not observed in <i>Vdr</i>-knockdown adipocytes. In addition, exposure of 3T3-L1 preadipocytes to 1,25(OH)<sub>2</sub>D<sub>3</sub> inhibited transcription of <i>Ddit3</i>, <i>sXbp1</i>, <i>Atf4</i>, <i>Bip</i>, and <i>Atf6</i> 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)<sub>2</sub>D<sub>3</sub> treatment before adipocyte differentiation reduced adipogenesis and the mRNA levels of adipogenic genes.</p><p><strong>Conclusions: </strong>Our data suggest that 1,25(OH)<sub>2</sub>D<sub>3</sub> prevents TG-induced ER stress and inflammatory responses in mature adipocytes by downregulating UPR signaling via binding with <i>Vdr</i>. In addition, the inhibition of adipogenesis by vitamin D may contribute to the reduction of ER stress in adipocytes.</p>\",\"PeriodicalId\":19232,\"journal\":{\"name\":\"Nutrition Research and Practice\",\"volume\":\"18 1\",\"pages\":\"1-18\"},\"PeriodicalIF\":2.0000,\"publicationDate\":\"2024-02-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10861344/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Nutrition Research and Practice\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://doi.org/10.4162/nrp.2024.18.1.1\",\"RegionNum\":4,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2023/12/21 0:00:00\",\"PubModel\":\"Epub\",\"JCR\":\"Q3\",\"JCRName\":\"NUTRITION & DIETETICS\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Nutrition Research and Practice","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.4162/nrp.2024.18.1.1","RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2023/12/21 0:00:00","PubModel":"Epub","JCR":"Q3","JCRName":"NUTRITION & DIETETICS","Score":null,"Total":0}
1,25-dihydroxyvitamin D3 affects thapsigargin-induced endoplasmic reticulum stress in 3T3-L1 adipocytes.
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.
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.