The effects of 1, 25(OH)2D3 treatment on metabolic reprogramming and maturation in bone marrow-derived dendritic cells from control and diabetic mice. Issue 225 (January 2023)

• Record Type: Journal Article
• Authors: Shin, Ungue; You, Hyeyoung; Lee, Ga Young; Son, YeKyoung; Han, Sung Nim
• Is Part Of: Journal of steroid biochemistry and molecular biology. Issue 225(2022)
• Publication Date: 2023-01
• Subjects: 1, 25(OH)2D3 1, 25-dihydroxyvitamin D3 -- Akt protein kinase B -- BM bone marrow -- BMDCs bone marrow-derived dendritic cells -- CD cluster of differentiation -- cDNA complementary DNA -- CON C57BLKS/J-m+/m+ mice -- db/db C57BLKS/J-db/db mice -- DC medium complete DC culture medium -- DCs dendritic cells -- DDIT4 DNA damage-inducible transcript 4 -- FBS fetal bovine serum -- FCCP carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone -- Gapdh glyceraldehyde-3-phosphate dehydrogenase -- Glut1 glucose transporter 1 -- HIF-1α hypoxia-inducible factor-1 subunit alpha -- Hif1a hypoxia-inducible factor-1 subunit alpha -- HK hexokinase -- Hk2 hexokinase 2 -- IL interleukin -- iNOS inducible nitric oxide synthase -- LPS lipopolysaccharide -- MFI mean fluorescence intensity -- MHC major histocompatibility complex -- mTOR mammalian target of rapamycin -- mTORC1 mammalian target of rapamycin complex 1 -- NO nitric oxide -- OCR oxygen consumption rate -- Oligo oligomycin -- OXPHOS oxidative phosphorylation -- p70S6K 70-kDa ribosomal protein S6 kinase -- PBS phosphate-buffered saline -- Pfkfb3, 6-phosphofructo-2-kinase/fructose-2, 6-biphosphatase 3 -- PI3K phosphatidylinositol 3-kinase -- Pkm2 pyruvate kinase M2 -- rmGM-CSF recombinant murine granulocyte-macrophage colony-stimulating factor -- Rot/AA rotenone/antimycin A mixture -- RT room temperature -- SDs standard deviations -- SEMs standard error of the means -- SRC spare respiratory capacity -- TBS-T Tris-buffered saline with 0.1 % Tween 20 -- TCA cycle tricarboxylic acid cycle -- TLRs Toll-like receptors -- TSC tuberous sclerosis complex -- VDR vitamin D receptor; 1, 25(OH)2D3 -- Diabetes -- Bone marrow-derived dendritic cells -- Metabolic reprogramming -- Aerobic glycolysis -- mTORC1; Steroid hormones -- Periodicals; Biochemistry -- Periodicals; Hormones -- Periodicals; Molecular Biology -- Periodicals; Hormones stéroïdes -- Périodiques; Steroid hormones; Periodicals; 572.579
• DOI: 10.1016/j.jsbmb.2022.106197
• Languages: English
• ISSNs: 0960-0760

Abstract

Abstract: Activated dendritic cells (DCs) undergo significant metabolic reprogramming, which is characterized by an increase in aerobic glycolysis and a concurrent progressive loss of oxidative phosphorylation. The modulation of metabolic reprogramming is believed to be closely related to the function of DCs. Vitamin D has been reported to inhibit the maturation of DCs. DC dysfunction has been reported in diabetic patients, and hyperglycemia is associated with impaired glycolytic metabolism in immune cells. Therefore, vitamin D and diabetes may affect intracellular metabolism, thereby regulating the activity of DCs. We investigated the effect of in vitro treatment of 1, 25-dihydroxyvitamin D3 (1, 25(OH)2 D3 ) on metabolic reprogramming and maturation of bone marrow-derived dendritic cells (BMDCs) from diabetic mouse. Six-week-old male C57BLKS/J-m + /m + mice (CON) and C57BLKS/J-db/db mice (db/db) were fed with a 10% kcal fat diet for seven weeks. BMDCs were generated by culturing bone marrow cells from the mice with rmGM-CSF (20 ng/mL) in the absence or presence of 10 nM 1, 25(OH)2 D3 . The maturation of BMDCs was induced via lipopolysaccharide (LPS, 50 ng/mL) stimulation for 24 h. LPS stimulation induced iNOS protein expression and decreased the mitochondrial respiration, while increased lactate production and the expression of glycolytic pathway-related genes ( Glut1 and Pfkfb3 ) in BMDCs from both CON and db/db groups. In LPS-stimulated mature BMDCs, 1, 25(OH)2 D3 treatment decreased the expression of surface markers related to immunostimulatory functions (MHC class II, CD80, CD86, and CD40) and production of IL-12p70 in both CON and db/db groups. While the mRNA level of the gene related to glucose uptake ( Glut1 ) was increased in both groups, lactate production was decreased by 1, 25(OH)2 D3 treatment. mTORC1 activity was suppressed following 1, 25(OH)2 D3 treatment. Collectively, our findings confirmed that metabolic reprogramming occurred in BMDCs following LPS stimulation. In vitro 1, 25(OH)2 D3 treatment induced tolerogenic phenotypes by reducing the expression of surface markers, as well as cytokine production. However, no significant difference was observed regarding the effects of 1, 25(OH)2 D3 treatment on metabolic conversion and maturation of BMDCs between the control and diabetic mice. Additionally, the decreased aerobic glycolysis induced by the 1, 25(OH)2 D3 treatment appeared to be associated with the diminished maturation of BMDCs, and mTORC1 appears to play a key role in the 1, 25(OH)2 D3 -mediated regulation of glycolysis. Highlights: Metabolic reprogramming occurred in BMDCs following LPS stimulation. Vitamin D treatment induces semi-mature phenotypes of BMDCs. Decreased glycolysis induced by vitamin D treatment appeared to affect BMDCs' maturation. Diabetes did not affect the maturation and metabolic reprogramming of BMDCs. mTORC1 appears to play a key role in the vitamin D -mediated regulation of glycolysis