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

Wi Dain|Park Chan Yoon
한국영양학회 (The Korean Nutrition Society)|2024.02|Nutrition Research and Practice|제18권 제1호|pp. 1-18|DOI 10.4162/nrp.2024.18.1.1

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13T3-L1 cells25-dihydroxyvitamin D3adipogenesisadipocytesendoplasmic reticulum stressthapsigargin

초록

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.

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.

참고문헌 (57)

  1. Ouchi N, "Adipokines in inflammation and metabolic disease", Nat Rev Immunol, 11, pp.85~97 (2011)
  2. Choe SS, "Adipose tissue remodeling: its role in energy metabolism and metabolic disorders", Front Endocrinol(Lausanne), 7, pp.30~ (2016)
  3. Bogdanovic E, "Endoplasmic reticulum stress in adipose tissue augments lipolysis", J Cell Mol Med, 19, pp.82~91 (2015)
  4. Cnop M, "Endoplasmic reticulum stress, obesity and diabetes", Trends Mol Med, 18, pp.59~68 (2012)
  5. Ghemrawi R, "Endoplasmic reticulum stress in metabolic disorders", Cells, 7, pp.63~ (2018)
  6. Olzmann JA, "Dynamics and functions of lipid droplets", Nat Rev Mol Cell Biol, 20, pp.137~155 (2019)
  7. Flamment M, "New insights into ER stress-induced insulin resistance", Trends Endocrinol Metab, 23, pp.381~390 (2012)
  8. Kawasaki N, "Obesity-induced endoplasmic reticulum stress causes chronic inflammation in adipose tissue", Sci Rep, 2, pp.799~ (2012)
  9. Ozcan U, "Endoplasmic reticulum stress links obesity, insulin action, and type 2 diabetes", Science, 306, pp.457~461 (2004)
  10. Liang L, "Endoplasmic reticulum stress impairs insulin receptor signaling in the brains of obese rats", PLoS One, 10, pp.e0126384~ (2015)
  11. Jiao P, "FFA-induced adipocyte inflammation and insulin resistance: involvement of ER stress and IKKβ pathways", Obesity(Silver Spring), 19, pp.483~491 (2011)
  12. Longo M, "Pathologic endoplasmic reticulum stress induced by glucotoxic insults inhibits adipocyte differentiation and induces an inflammatory phenotype", Biochim Biophys Acta, 1863, pp.1146~1156 (2016)
  13. Basseri S, "The chemical chaperone 4-phenylbutyrate inhibits adipogenesis by modulating the unfolded protein response", J Lipid Res, 50, pp.2486~2501 (2009)
  14. Wood RJ, "Vitamin D and adipogenesis : new molecular insights", Nutr Rev, 66, pp.40~46 (2008)
  15. Abbas MA, "Physiological functions of vitamin D in adipose tissue", J Steroid Biochem Mol Biol, 165, pp.369~381 (2017)
  16. Wamberg L, "Expression of vitamin D-metabolizing enzymes in human adipose tissue--the effect of obesity and diet-induced weight loss", Int J Obes(Lond), 37, pp.651~657 (2013)
  17. Park CY, "The role of vitamin D in adipose tissue biology : adipocyte differentiation, energy metabolism, and inflammation", J Lipid Atheroscler, 10, pp.130~144 (2021)
  18. Marcotorchino J, "Vitamin D reduces the inflammatory response and restores glucose uptake in adipocytes", Mol Nutr Food Res, 56, pp.1771~1782 (2012)
  19. Sun X, "Calcitriol and calcium regulate cytokine production and adipocyte-macrophage cross-talk", J Nutr Biochem, 19, pp.392~399 (2008)
  20. Sun X, "Calcium and 1, 25-dihydroxyvitamin D3 regulation of adipokine expression", Obesity(Silver Spring), 15, pp.340~348 (2007)
  21. Karkeni E, "Vitamin D limits chemokine expression in adipocytes and macrophage migration in vitro and in male mice", Endocrinology, 156, pp.1782~1793 (2015)
  22. Cheng S, "Adiposity, cardiometabolic risk, and vitamin D status : the Framingham Heart Study", Diabetes, 59, pp.242~248 (2010)
  23. Mai XM, "Cross-sectional and prospective cohort study of serum 25-hydroxyvitamin D level and obesity in adults : the HUNT study", Am J Epidemiol, 175, pp.1029~1036 (2012)
  24. Pereira-Santos M, "Obesity and vitamin D deficiency : a systematic review and meta-analysis", Obes Rev, 16, pp.341~349 (2015)
  25. Roth CL, "Vitamin D deficiency in obese rats exacerbates nonalcoholic fatty liver disease and increases hepatic resistin and Toll-like receptor activation", Hepatology, 55, pp.1103~1111 (2012)
  26. Mezza T, "Vitamin D deficiency : a new risk factor for type 2 diabetes", Ann Nutr Metab, 61, pp.337~348 (2012)
  27. Wen G, "1, 25-hydroxyvitamin D3 decreases endoplasmic reticulum stress-induced inflammatory response in mammary epithelial cells", PLoS One, 15, pp.e0228945~ (2020)
  28. Riek AE, "Vitamin D suppression of endoplasmic reticulum stress promotes an antiatherogenic monocyte", macrophage phenotype in type 2 diabetic patients, 2012, pp.287, 38482~38494
  29. Rayalam S, "Enhanced effects of 1, 25(OH)(2)D(3)plus genistein on adipogenesis and apoptosis in 3T3-L1 adipocytes", Obesity(Silver Spring), 16, pp.539~546 (2008)
  30. Chang E, "Vitamin D decreases adipocyte lipid storage and increases NAD-SIRT1 pathway in 3T3-L1 adipocytes", Nutrition, 32, pp.702~708 (2016)
  31. Żmijewski MA, "Nongenomic activities of vitamin D", Nutrients, 14, pp.5104~ (2022)
  32. Dai N, "Encyclopedia of Biological Chemistry III (Third Edition)" (2021), Elsevier
  33. Sha H, "The IRE1alpha-XBP1 pathway of the unfolded protein response is required for adipogenesis", Cell Metab, 9, pp.556~564 (2009)
  34. Kamiya T, "Effect of endoplasmic reticulum(ER)stress inducer thapsigargin on the expression of extracellular-superoxide dismutase in mouse 3T3-L1 adipocytes", J Clin Biochem Nutr, 52, pp.101~105 (2013)
  35. Han J, "ER stress signalling through eIF2α and CHOP, but not IRE1α, attenuates adipogenesis in mice", Diabetologia, 56, pp.911~924 (2013)
  36. Trump BF, "The role of cytosolic Ca2+ in cell injury, necrosis and apoptosis", Curr Opin Cell Biol, 4, pp.227~232 (1992)
  37. Ntambi JM, "Role of Ca2+ in the early stages of murine adipocyte differentiation as evidenced by calcium mobilizing agents", Differentiation, 60, pp.151~158 (1996)
  38. Deng J, "Lipolysis response to endoplasmic reticulum stress in adipose cells", J Biol Chem, 287, pp.6240~6249 (2012)
  39. Bettaieb A, "Protein tyrosine phosphatase 1B deficiency potentiates PERK", eIF2α signaling in brown adipocytes, 2012, pp.7, e34412~
  40. Park CY, "Dysregulation of lipid droplet protein expression in adipose tissues and association with metabolic risk factors in adult females with obesity and type 2 diabetes", J Nutr, 153, pp.691~702 (2023)
  41. Lim J, "Depot-specific UCP1 expression in human white adipose tissue and its association with obesity-related markers", Int J Obes, 44, pp.697~706 (2020)
  42. Hu X, "1, 25-(OH)2D3 protects pancreatic beta cells against H2O2-induced apoptosis through inhibiting the PERK-ATF4-CHOP pathway", Acta Biochim Biophys Sin(Shanghai), 53, pp.46~53 (2021)
  43. Haas MJ, "Inhibition of endoplasmic reticulum stress and oxidative stress by vitamin D in endothelial cells", Free Radic Biol Med, 99, pp.1~10 (2016)
  44. Rozpędek-Kamińska W, "The PERK-dependent molecular mechanisms as a novel therapeutic target for neurodegenerative diseases", Int J Mol Sci, 21, pp.2108~ (2020)
  45. Wang X, "The unfolded protein response in ischemic heart disease", J Mol Cell Cardiol, 117, pp.19~25 (2018)
  46. Oyadomari S, "Roles of CHOP", GADD153 in endoplasmic reticulum stress, 2004, pp.11, 381~389
  47. Gregor MF, "Thematic review series : adipocyte biology. Adipocyte stress : the endoplasmic reticulum and metabolic disease", J Lipid Res, 48, pp.1905~1914 (2007)
  48. Park CY, "Effects of 1, 25-dihydroxyvitamin D3 on the inflammatory responses of stromal vascular cells and adipocytes from lean and obese mice", Nutrients, 12, pp.364~ (2020)
  49. Ding C, "1, 25-dihydroxyvitamin D3 protects against macrophage-induced activation of NFκB and MAPK signalling and chemokine release in human adipocytes", PLoS One, 8, pp.e61707~ (2013)
  50. Zhang Y, "Vitamin D inhibits monocyte", macrophage proinflammatory cytokine production by targeting MAPK phosphatase-1, 2012, pp.188, 2127~2135
  51. Antoniotti V, "Calreticulin and PDIA3, two markers of endoplasmic reticulum stress, are associated with metabolic alterations and insulin resistance in pediatric obesity: a pilot study", Front Endocrinol (Lausanne), pp.1003919~ (2022)
  52. Farmer SR, "Transcriptional control of adipocyte formation", Cell Metab, 4, pp.263~273 (2006)
  53. Kong J, "Molecular mechanism of 1, 25-dihydroxyvitamin D3 inhibition of adipogenesis in 3T3-L1 cells", Am J Physiol Endocrinol Metab, 290, pp.E916~24 (2006)
  54. Lee H, "Anti-adipogenic effects of 1, 25-dihydroxyvitamin D3 are mediated by the maintenance of the wingless-type MMTV integration site", β-catenin pathway, 2012, pp.30, 1219~1224
  55. Ji S, "Regulation of adipogenesis and key adipogenic gene expression by 1, 25-dihydroxyvitamin D in 3T3-L1 cells", PLoS One, 10, pp.e0126142~ (2015)
  56. Turishcheva E, "The role of endoplasmic reticulum stress in differentiation of cells of mesenchymal origin", Biochemistry(Mosc), 87, pp.916~931 (2022)
  57. Li Y, "Ca2+-dependent redox modulation of SERCA 2b by ERp57", J Cell Biol, 164, pp.35~46 (2004)

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