Nutrition Research and Practice

The Korean Nutrition Society (한국영양학회)
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1,25-dihydroxyvitamin D3 affects thapsigargin-induced endoplasmic reticulum stress in 3T3-L1 adipocytes

  • Dain Wi (Department of Food & Nutrition, College of Health Science, The University of Suwon) ;
  • Chan Yoon Park (Department of Food & Nutrition, College of Health Science, The University of Suwon)
  • Received : 2023.09.14
  • Accepted : 2023.12.06
  • Published : 2024.02.01
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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.

Keywords

Acknowledgement

The paper was supported by the research grant of the University of Suwon in 2020 and the National Research Foundation of Korea (NRF) grant funded by the Korea government Ministry of Science and ICT (grant number: 2021R1G1A1093362).

References

  1. Ouchi N, Parker JL, Lugus JJ, Walsh K. Adipokines in inflammation and metabolic disease. Nat Rev Immunol 2011;11:85-97.
  2. Choe SS, Huh JY, Hwang IJ, Kim JI, Kim JB. Adipose tissue remodeling: its role in energy metabolism and metabolic disorders. Front Endocrinol (Lausanne) 2016;7:30.
  3. Bogdanovic E, Kraus N, Patsouris D, Diao L, Wang V, Abdullahi A, Jeschke MG. Endoplasmic reticulum stress in adipose tissue augments lipolysis. J Cell Mol Med 2015;19:82-91.
  4. Cnop M, Foufelle F, Velloso LA. Endoplasmic reticulum stress, obesity and diabetes. Trends Mol Med 2012;18:59-68.
  5. Ghemrawi R, Battaglia-Hsu SF, Arnold C. Endoplasmic reticulum stress in metabolic disorders. Cells 2018;7:63.
  6. Olzmann JA, Carvalho P. Dynamics and functions of lipid droplets. Nat Rev Mol Cell Biol 2019;20:137-55.
  7. Flamment M, Hajduch E, Ferre P, Foufelle F. New insights into ER stress-induced insulin resistance. Trends Endocrinol Metab 2012;23:381-90.
  8. Kawasaki N, Asada R, Saito A, Kanemoto S, Imaizumi K. Obesity-induced endoplasmic reticulum stress causes chronic inflammation in adipose tissue. Sci Rep 2012;2:799.
  9. Ozcan U, Cao Q, Yilmaz E, Lee AH, Iwakoshi NN, Ozdelen E, Tuncman G, Gorgun C, Glimcher LH, Hotamisligil GS. Endoplasmic reticulum stress links obesity, insulin action, and type 2 diabetes. Science 2004;306:457-61.
  10. Liang L, Chen J, Zhan L, Lu X, Sun X, Sui H, Zheng L, Xiang H, Zhang F. Endoplasmic reticulum stress impairs insulin receptor signaling in the brains of obese rats. PLoS One 2015;10:e0126384.
  11. Jiao P, Ma J, Feng B, Zhang H, Diehl JA, Chin YE, Yan W, Xu H. FFA-induced adipocyte inflammation and insulin resistance: involvement of ER stress and IKKβ pathways. Obesity (Silver Spring) 2011;19:483-91.
  12. Longo M, Spinelli R, D'Esposito V, Zatterale F, Fiory F, Nigro C, Raciti GA, Miele C, Formisano P, Beguinot F, et al. Pathologic endoplasmic reticulum stress induced by glucotoxic insults inhibits adipocyte differentiation and induces an inflammatory phenotype. Biochim Biophys Acta 2016;1863:1146-56.
  13. Basseri S, Lhotak S, Sharma AM, Austin RC. The chemical chaperone 4-phenylbutyrate inhibits adipogenesis by modulating the unfolded protein response. J Lipid Res 2009;50:2486-501.
  14. Wood RJ. Vitamin D and adipogenesis: new molecular insights. Nutr Rev 2008;66:40-6.
  15. Abbas MA. Physiological functions of vitamin D in adipose tissue. J Steroid Biochem Mol Biol 2017;165:369-81.
  16. Wamberg L, Christiansen T, Paulsen SK, Fisker S, Rask P, Rejnmark L, Richelsen B, Pedersen SB. Expression of vitamin D-metabolizing enzymes in human adipose tissue -- the effect of obesity and dietinduced weight loss. Int J Obes (Lond) 2013;37:651-7.
  17. Park CY, Han SN. The role of vitamin D in adipose tissue biology: adipocyte differentiation, energy metabolism, and inflammation. J Lipid Atheroscler 2021;10:130-44.
  18. Marcotorchino J, Gouranton E, Romier B, Tourniaire F, Astier J, Malezet C, Amiot MJ, Landrier JF. Vitamin D reduces the inflammatory response and restores glucose uptake in adipocytes. Mol Nutr Food Res 2012;56:1771-82.
  19. Sun X, Zemel MB. Calcitriol and calcium regulate cytokine production and adipocyte-macrophage crosstalk. J Nutr Biochem 2008;19:392-9.
  20. Sun X, Zemel MB. Calcium and 1,25-dihydroxyvitamin D3 regulation of adipokine expression. Obesity (Silver Spring) 2007;15:340-8.
  21. Karkeni E, Marcotorchino J, Tourniaire F, Astier J, Peiretti F, Darmon P, Landrier JF. Vitamin D limits chemokine expression in adipocytes and macrophage migration in vitro and in male mice. Endocrinology 2015;156:1782-93.
  22. Cheng S, Massaro JM, Fox CS, Larson MG, Keyes MJ, McCabe EL, Robins SJ, O'Donnell CJ, Hoffmann U, Jacques PF, et al. Adiposity, cardiometabolic risk, and vitamin D status: the Framingham Heart Study. Diabetes 2010;59:242-8.
  23. Mai XM, Chen Y, Camargo CA Jr, Langhammer A. Cross-sectional and prospective cohort study of serum 25-hydroxyvitamin D level and obesity in adults: the HUNT study. Am J Epidemiol 2012;175:1029-36.
  24. Pereira-Santos M, Costa PR, Assis AM, Santos CA, Santos DB. Obesity and vitamin D deficiency: a systematic review and meta-analysis. Obes Rev 2015;16:341-9.
  25. Roth CL, Elfers CT, Figlewicz DP, Melhorn SJ, Morton GJ, Hoofnagle A, Yeh MM, Nelson JE, Kowdley KV. Vitamin D deficiency in obese rats exacerbates nonalcoholic fatty liver disease and increases hepatic resistin and Toll-like receptor activation. Hepatology 2012;55:1103-11.
  26. Mezza T, Muscogiuri G, Sorice GP, Prioletta A, Salomone E, Pontecorvi A, Giaccari A. Vitamin D deficiency: a new risk factor for type 2 diabetes? Ann Nutr Metab 2012;61:337-48.
  27. Wen G, Eder K, Ringseis R. 1,25-hydroxyvitamin D3 decreases endoplasmic reticulum stress-induced inflammatory response in mammary epithelial cells. PLoS One 2020;15:e0228945.
  28. Riek AE, Oh J, Sprague JE, Timpson A, de las Fuentes L, Bernal-Mizrachi L, Schechtman KB, BernalMizrachi C. Vitamin D suppression of endoplasmic reticulum stress promotes an antiatherogenic monocyte/macrophage phenotype in type 2 diabetic patients. J Biol Chem 2012;287:38482-94.
  29. Rayalam S, Della-Fera MA, Ambati S, Yang JY, Park HJ, Baile CA. Enhanced effects of 1,25(OH)(2)D(3) plus genistein on adipogenesis and apoptosis in 3T3-L1 adipocytes. Obesity (Silver Spring) 2008;16:539-46.
  30. Chang E, Kim Y. Vitamin D decreases adipocyte lipid storage and increases NAD-SIRT1 pathway in 3T3-L1 adipocytes. Nutrition 2016;32:702-8.
  31. Zmijewski MA. Nongenomic activities of vitamin D. Nutrients 2022;14:5104.
  32. Dai N, Groenendyk J, Michalak M. Binding proteins | Ca2+ binding/buffering proteins: ER luminal proteins. In: Jez J, editor. Encyclopedia of Biological Chemistry III (Third Edition). Oxford: Elsevier; 2021. p. 534-46.
  33. Sha H, He Y, Chen H, Wang C, Zenno A, Shi H, Yang X, Zhang X, Qi L. The IRE1alpha-XBP1 pathway of the unfolded protein response is required for adipogenesis. Cell Metab 2009;9:556-64.
  34. Kamiya T, Hara H, Adachi 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 2013;52:101-5.
  35. Han J, Murthy R, Wood B, Song B, Wang S, Sun B, Malhi H, Kaufman RJ. ER stress signalling through eIF2α and CHOP, but not IRE1α, attenuates adipogenesis in mice. Diabetologia 2013;56:911-24.
  36. Trump BF, Berezesky IK. The role of cytosolic Ca2+ in cell injury, necrosis and apoptosis. Curr Opin Cell Biol 1992;4:227-32.
  37. Ntambi JM, Takova T. Role of Ca2+ in the early stages of murine adipocyte differentiation as evidenced by calcium mobilizing agents. Differentiation 1996;60:151-8.
  38. Deng J, Liu S, Zou L, Xu C, Geng B, Xu G. Lipolysis response to endoplasmic reticulum stress in adipose cells. J Biol Chem 2012;287:6240-9.
  39. Bettaieb A, Matsuo K, Matsuo I, Wang S, Melhem R, Koromilas AE, Haj FG. Protein tyrosine phosphatase 1B deficiency potentiates PERK/eIF2α signaling in brown adipocytes. PLoS One 2012;7:e34412.
  40. Park CY, Kim D, Seo MK, Kim J, Choe H, Kim JH, Hong JP, Lee YJ, Heo Y, Kim HJ, et al. 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 2023;153:691-702.
  41. Lim J, Park HS, Kim J, Jang YJ, Kim JH, Lee Y, Heo Y. Depot-specific UCP1 expression in human white adipose tissue and its association with obesity-related markers. Int J Obes 2020;44:697-706.
  42. Hu X, Hu C, Liu J, Wu Z, Duan T, Cao Z. 1,25-(OH)2D3 protects pancreatic beta cells against H2O2-induced apoptosis through inhibiting the PERK-ATF4-CHOP pathway. Acta Biochim Biophys Sin (Shanghai) 2021;53:46-53.
  43. Haas MJ, Jafri M, Wehmeier KR, Onstead-Haas LM, Mooradian AD. Inhibition of endoplasmic reticulum stress and oxidative stress by vitamin D in endothelial cells. Free Radic Biol Med 2016;99:1-10.
  44. Rozpedek-Kaminska W, Siwecka N, Wawrzynkiewicz A, Wojtczak R, Pytel D, Diehl JA, Majsterek I. The PERK-dependent molecular mechanisms as a novel therapeutic target for neurodegenerative diseases. Int J Mol Sci 2020;21:2108.
  45. Wang X, Xu L, Gillette TG, Jiang X, Wang ZV. The unfolded protein response in ischemic heart disease. J Mol Cell Cardiol 2018;117:19-25.
  46. Oyadomari S, Mori M. Roles of CHOP/GADD153 in endoplasmic reticulum stress. Cell Death Differ 2004;11:381-9. 
  47. Gregor MF, Hotamisligil GS. Thematic review series: adipocyte biology. Adipocyte stress: the endoplasmic reticulum and metabolic disease. J Lipid Res 2007;48:1905-14.
  48. Park CY, Kim TY, Yoo JS, Seo Y, Pae M, Han SN. Effects of 1,25-dihydroxyvitamin D3 on the inflammatory responses of stromal vascular cells and adipocytes from lean and obese mice. Nutrients 2020;12:364.
  49. Ding C, Wilding JP, Bing C. 1,25-dihydroxyvitamin D3 protects against macrophage-induced activation of NFκB and MAPK signalling and chemokine release in human adipocytes. PLoS One 2013;8:e61707.
  50. Zhang Y, Leung DY, Richers BN, Liu Y, Remigio LK, Riches DW, Goleva E. Vitamin D inhibits monocyte/macrophage proinflammatory cytokine production by targeting MAPK phosphatase-1. J Immunol 2012;188:2127-35.
  51. Antoniotti V, Bellone S, Goncalves Correia FP, Peri C, Tini S, Ricotti R, Mancioppi V, Gagliardi M, Spadaccini D, Caputo M, et al. 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) 2022;13:1003919.
  52. Farmer SR. Transcriptional control of adipocyte formation. Cell Metab 2006;4:263-73.
  53. Kong J, Li YC. Molecular mechanism of 1,25-dihydroxyvitamin D3 inhibition of adipogenesis in 3T3-L1 cells. Am J Physiol Endocrinol Metab 2006;290:E916-24.
  54. Lee H, Bae S, Yoon Y. Anti-adipogenic effects of 1,25-dihydroxyvitamin D3 are mediated by the maintenance of the wingless-type MMTV integration site/β-catenin pathway. Int J Mol Med 2012;30:1219-24.
  55. Ji S, Doumit ME, Hill RA. Regulation of adipogenesis and key adipogenic gene expression by 1, 25-dihydroxyvitamin D in 3T3-L1 cells. PLoS One 2015;10:e0126142.
  56. Turishcheva E, Vildanova M, Onishchenko G, Smirnova E. The role of endoplasmic reticulum stress in differentiation of cells of mesenchymal origin. Biochemistry (Mosc) 2022;87:916-31.
  57. Li Y, Camacho P. Ca2+-dependent redox modulation of SERCA 2b by ERp57. J Cell Biol 2004;164:35-46.