Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Eldecalcitol ameliorates diabetic osteoporosis and glucolipid metabolic disorder by promoting Treg cell differentiation through SOCE

doi: 10.1007/s00018-024-05453-3

Figure Lengend Snippet: ED-71 ameliorates glucolipid metabolism disorder in db/db mice. A : Feeding time axis of mice and drug administration. B : Curve and statistical analysis of body weight in mice within 28 days. C : Curve and statistical analysis of blood glucose in mice within 28 days. D : Blood glucose curve and area under curve (AUC) within 120 min after intraperitoneal injection of glucose solution. E : Blood glucose curve and AUC within 120 min after orally of glucose solution. F : Blood glucose curve and AUC within 120 min after intraperitoneal injection of insulin solution. G : Serum insulin curve and AUC within 20 min after intraperitoneal injection of glucose solution. H: Statistical analysis of HOMA-IR, HOMA-IS and HOMA-β in mice. I : H & E staining of pancreas in mice and statistical analysis of islet area. J : Immunofluorescence staining of pancreas in mice. Red: insulin. Green: TUNEL positive. White dashed line: islet region. K : Statistical analysis of BAT, VAT, SCAT and total fat mass weight in mice. L : Biochemical analysis of TC, TG, HDL-C and LDL-C in serum of mice. M : H & E staining, Oil-Red O staining and PAS staining of liver in mice. N : The qRT-PCR detection and statistical analysis of factors related to glucolipid metabolism in mouse liver. Data were shown as mean ± SD of n = 5 mice per group. * P < 0·05; ** P < 0·01; *** P < 0·005 vs. WT + vehicle group; # P < 0·05; ## P < 0·01; ### P < 0·005 vs. db/db + vehicle group

Article Snippet: After 24 h culture, CD4 + T cells were treated with 55 mM glucose solution, 50 μM 2-APB (HY-W009724, MCE) [ ] and 1 nM ED-71 [ ] and cultured in dark for 48 h.

Techniques: Injection, Staining, Immunofluorescence, TUNEL Assay, Quantitative RT-PCR

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Eldecalcitol ameliorates diabetic osteoporosis and glucolipid metabolic disorder by promoting Treg cell differentiation through SOCE

doi: 10.1007/s00018-024-05453-3

Figure Lengend Snippet: ED-71 improves osteoporosis and reduces BMAT in db/db mice. A : 3D images of micro-CT of tibia in mice and the cross-sectional view of the trabecular bone below the growth plate. B : Statistical analysis of BMD, BV/TV, Tb.N, Tb.Th and Tb.sp. C : H & E staining and Masson staining of tibia in mice. D : Relative mRNA expression level of ALP , RUNX2 , OCN and COL1 in tibia of mice. E : Double staining for ALP (brown) and TRAP (red) in tibia of mice and immunostaining of RANKL, RUNX2 and CTSK in tibia of mice. F , G : Statistical analysis of osteogenesis- and osteoclastogenesis-related factor expression levels and osteoclast numbers. H : Number and area of adipocytes in bone marrow. I : The qRT-PCR detection and statistical analysis of adipose-related factors in bone marrow. Data were shown as mean ± SD of n = 5 mice per group. * P < 0·05; ** P < 0·01; *** P < 0·005 vs. WT + vehicle group; # P < 0·05; ## P < 0·01; ### P < 0·005 vs. db/db + vehicle group

Article Snippet: After 24 h culture, CD4 + T cells were treated with 55 mM glucose solution, 50 μM 2-APB (HY-W009724, MCE) [ ] and 1 nM ED-71 [ ] and cultured in dark for 48 h.

Techniques: Micro-CT, Staining, Expressing, Double Staining, Immunostaining, Quantitative RT-PCR

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Eldecalcitol ameliorates diabetic osteoporosis and glucolipid metabolic disorder by promoting Treg cell differentiation through SOCE

doi: 10.1007/s00018-024-05453-3

Figure Lengend Snippet: ED-71 increases Treg cell subsets throughout the body and bone marrow. A : Upregulated and downregulated genes of transcripts. B : Volcano plot of differential gene expression under the diabetic condition. C : GO enrichment analysis of the BP, MF and CC. D : Flow cytometry analysis of CD25 + FOXP3 + T cells gated on CD4-FITC in the spleen lymphocytes and bone marrow. E : Relative mRNA expression level of FOXP3 and TGF-β in the spleen lymphocytes and bone marrow of mice. F : ELISA assay of TGF-β and IL-10 in serum of mice. G : Immunofluorescence staining of tibia in mice. Green: CD4 positive. Red: FOXP3 positive. H : The schematic diagram of CD4 + T cell sorting and induction in mice. I : Flow cytometry analysis of CD25 + FOXP3 + T cells gated on CD4-FITC in the CD4 + T cell and the statistical analyses. J : Relative mRNA expression level of FOXP3 and TGF-β in the CD4 + T cell. K : ELISA assay of TGF-β and IL-10 in the supernatant of cell culture medium. L : The protein expression of FOXP3, TGF-β and β-tubulin detected by western blotting in the CD4 + T cell and the statistical analyses. Data were shown as mean ± SD. D - G , n = 5 mice per group; I - L , representative of n = 3 independent experiments. * P < 0·05; ** P < 0·01; *** P < 0·005 vs. WT + vehicle group or Control group; # P < 0·05; ## P < 0·01; ### P < 0·005 vs. db/db + vehicle group or HG group

Article Snippet: After 24 h culture, CD4 + T cells were treated with 55 mM glucose solution, 50 μM 2-APB (HY-W009724, MCE) [ ] and 1 nM ED-71 [ ] and cultured in dark for 48 h.

Techniques: Gene Expression, Flow Cytometry, Expressing, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Staining, FACS, Cell Culture, Western Blot, Control

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Eldecalcitol ameliorates diabetic osteoporosis and glucolipid metabolic disorder by promoting Treg cell differentiation through SOCE

doi: 10.1007/s00018-024-05453-3

Figure Lengend Snippet: ED-71 improves glucolipid metabolism disorder in db/db mice via Treg subsets. A : Feeding time axis of mice and drug administration. B : Flow cytometry analysis of CD25 + FOXP3 + T cells gated on CD4-FITC in the spleen lymphocytes and bone marrow. C : Curve and statistical analysis of body weight in mice within 28 days. D : Curve and statistical analysis of blood glucose in mice within 28 days. E : Blood glucose curve and AUC within 120 min after intraperitoneal injection of glucose solution. F : Blood glucose curve and AUC within 120 min after orally of glucose solution. G : Blood glucose curve and AUC within 120 min after intraperitoneal injection of insulin solution. H : Serum insulin curve and AUC within 20 min after intraperitoneal injection of glucose solution. I : Statistical analysis of HOMA-IR, HOMA-IS and HOMA-β in mice. J : H & E staining of pancreas in mice and statistical analysis of islet area. K: Immunofluorescence staining of pancreas in mice. Red: insulin. Green: TUNEL positive. White dashed line: islet region. L : Statistical analysis of BAT, VAT, SCAT and total fat mass weight in mice. M : Biochemical analysis of TC, TG, HDL-C and LDL-C in serum of mice. N : H & E staining, Oil-Red O staining and PAS staining of liver in mice. O : The qRT-PCR detection and statistical analysis of factors related to glucolipid metabolism in mouse liver. Data were shown as mean ± SD of n = 5 mice per group. * P < 0·05; ** P < 0·01; *** P < 0·005 vs. db/db + vehicle group; # P < 0·05; ## P < 0·01; ### P < 0·005 vs. db/db + ED-71 group

Article Snippet: After 24 h culture, CD4 + T cells were treated with 55 mM glucose solution, 50 μM 2-APB (HY-W009724, MCE) [ ] and 1 nM ED-71 [ ] and cultured in dark for 48 h.

Techniques: Flow Cytometry, Injection, Staining, Immunofluorescence, TUNEL Assay, Quantitative RT-PCR

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Eldecalcitol ameliorates diabetic osteoporosis and glucolipid metabolic disorder by promoting Treg cell differentiation through SOCE

doi: 10.1007/s00018-024-05453-3

Figure Lengend Snippet: ED-71 improves osteoporosis in db/db mice through Treg subsets. A : 3D images of micro-CT of tibia in mice and the cross-sectional view of the trabecular bone below the growth plate. B : Statistical analysis of BMD, BV/TV, Tb.N, Tb.Th and Tb.sp. C : H & E staining and Masson staining of tibia in mice. D : Relative mRNA expression level of ALP , RUNX2 , OCN and COL1 in tibia of mice. E : Double staining for ALP (brown) and TRAP (red) in tibia of mice and immunostaining of RANKL, RUNX2 and CTSK in tibia of mice. F , G : Statistical analysis of osteogenesis- and osteoclastogenesis-related factor expression levels and osteoclast numbers. H : Number and area of adipocytes in bone marrow. I : The qRT-PCR detection and statistical analysis of adipose-related factors in bone marrow. Data were shown as mean ± SD of n = 5 mice per group. * P < 0·05; ** P < 0·01; *** P < 0·005 vs. db/db + vehicle group; # P < 0·05; ## P < 0·01; ### P < 0·005 vs. db/db + ED-71 group

Article Snippet: After 24 h culture, CD4 + T cells were treated with 55 mM glucose solution, 50 μM 2-APB (HY-W009724, MCE) [ ] and 1 nM ED-71 [ ] and cultured in dark for 48 h.

Techniques: Micro-CT, Staining, Expressing, Double Staining, Immunostaining, Quantitative RT-PCR

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Eldecalcitol ameliorates diabetic osteoporosis and glucolipid metabolic disorder by promoting Treg cell differentiation through SOCE

doi: 10.1007/s00018-024-05453-3

Figure Lengend Snippet: ED-71 activates ORAI1 and STIM1-mediated SOCE in CD4 + T cells. A : KEGG enrichment analysis and the salient pathways. B : GSEA enrichment analysis and the salient pathways. C : Genes associated with the Ca 2+ signaling pathway and statistical analysis. D : Immunofluorescence staining of tibia in mice. Green: STIM1 positive. Red: CD4 positive. E : Immunofluorescence staining of tibia in mice. Green: ORAI1 positive. Red: CD4 positive. F: Relative mRNA expression level of STIM1 and ORAI1 in the spleen lymphocytes and bone marrow of mice. G: Immunofluorescence staining of CD4 + T cell. Green: STIM1 positive. H : Immunofluorescence staining of CD4 + T cell. Green: ORAI1 positive. I : The protein expression of STIM1, ORAI1 and β-tubulin detected by western blotting in the CD4 + T cell and the statistical analyses. J : Relative mRNA expression level of STIM1 and ORAI1 in the CD4 + T cell. K : Flow cytometry analysis of Ca 2+ in the CD4 + T cell. L: Immunofluorescence staining of Ca 2+ in the CD4 + T cell. Data were shown as mean ± SD. D - F , n = 5 mice per group; G - L , representative of n = 3 independent experiments. * P < 0·05; ** P < 0·01; *** P < 0·005 vs. WT + vehicle group or Control group; # P < 0·05; ## P < 0·01; ### P < 0·005 vs. db/db + vehicle group or HG group

Article Snippet: After 24 h culture, CD4 + T cells were treated with 55 mM glucose solution, 50 μM 2-APB (HY-W009724, MCE) [ ] and 1 nM ED-71 [ ] and cultured in dark for 48 h.

Techniques: Immunofluorescence, Staining, Expressing, Western Blot, Flow Cytometry, Control

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Eldecalcitol ameliorates diabetic osteoporosis and glucolipid metabolic disorder by promoting Treg cell differentiation through SOCE

doi: 10.1007/s00018-024-05453-3

Figure Lengend Snippet: ED-71 promotes Treg cell differentiation by regulating SOCE in vitro. A : Flow cytometry analysis of CD25 + FOXP3 + T cells gated on CD4-FITC in the CD4 + T cell. B : Relative mRNA expression level of FOXP3 and TGF-β in the CD4 + T cell. C : ELISA assay of TGF-β and IL-10 in the supernatant of cell culture medium. D : The protein expression of FOXP3, TGF-β and β-tubulin detected by western blotting in the CD4 + T cell and the statistical analyses. E : Putative mechanism for ED-71 in improving diabetic osteoporosis and glucolipid metabolic disorder. Data were shown as mean ± SD. A - D , representative of n = 3 independent experiments. * P < 0·05; ** P < 0·01; *** P < 0·005 vs. HG group; # P < 0·05; ## P < 0·01; ### P < 0·005 vs. HG + ED-71 group

Article Snippet: After 24 h culture, CD4 + T cells were treated with 55 mM glucose solution, 50 μM 2-APB (HY-W009724, MCE) [ ] and 1 nM ED-71 [ ] and cultured in dark for 48 h.

Techniques: Cell Differentiation, In Vitro, Flow Cytometry, Expressing, Enzyme-linked Immunosorbent Assay, Cell Culture, Western Blot