Journal: Chinese medicine

Article Title: Senkyunolide I suppresses hepatic stellate cell activation and liver fibrosis by reprogramming VDR-dependent fatty acid metabolism.

doi: 10.1186/s13020-025-01133-x

Figure Lengend Snippet: Fig. 1 VDR deficiency exacerbated hepatic fibrosis and steatosis in mice. A mRNA levels of fibrogenic markers (α-Sma, Col1a1, and Timp1). B mRNA levels of lipogenic enzymes (Fasn, Acc-1, and Acly) and fatty acid desaturases (Fads1/2 and Scd1) (n = 5). C Western blot analysis of VDR and TGFβ/ Smad pathway (n = 3), D Serum levels of liver function markers (AST, ALT, ALP, and TBIL) (n = 5). *P < 0.05, **P < 0.01

Article Snippet: Membranes were blocked with 5% BSA and probed overnight at 4 °C with antibodies against: α-SMA (1:2000), COL1 A1 (1:1000), FASN (10624-2-AP, Proteintech, 1:1000), VDR (AF8159, Affinity, 1:1000), p-Smad3 (1:2000), Smad3 (AF6362, Affinity, 1:2000), TGF-β1 (1:3000) and β-actin (BM3873, Boster, 1:2000).

Techniques: Western Blot

Journal: Chinese medicine

Article Title: Senkyunolide I suppresses hepatic stellate cell activation and liver fibrosis by reprogramming VDR-dependent fatty acid metabolism.

doi: 10.1186/s13020-025-01133-x

Figure Lengend Snippet: Fig. 2 VDR activation attenuated TGF-β1-induced fibrosteatotic changes in HSCs. A Western blot analysis of VDR and α-SMA (n = 3). B mRNA levels of fibrogenic markers (α-Sma, Col1a1, and Timp1). C mRNA levels of lipogenic enzymes (Fasn, Acc-1, and Acly) and fatty acid desaturases (Fads1/2 and Scd1) (n = 6). *P < 0.05, **P < 0.01

Article Snippet: Membranes were blocked with 5% BSA and probed overnight at 4 °C with antibodies against: α-SMA (1:2000), COL1 A1 (1:1000), FASN (10624-2-AP, Proteintech, 1:1000), VDR (AF8159, Affinity, 1:1000), p-Smad3 (1:2000), Smad3 (AF6362, Affinity, 1:2000), TGF-β1 (1:3000) and β-actin (BM3873, Boster, 1:2000).

Techniques: Activation Assay, Western Blot

Journal: Chinese medicine

Article Title: Senkyunolide I suppresses hepatic stellate cell activation and liver fibrosis by reprogramming VDR-dependent fatty acid metabolism.

doi: 10.1186/s13020-025-01133-x

Figure Lengend Snippet: Fig. 3 SI inhibited TGF-β1-induced activation of HSCs. A Chemical structure of the SI. B Cell viability of HSCs. C mRNA levels of fibrogenic markers (α-Sma, Col1a1, and Timp1). D Immunofluorescence analysis of α-SMA. scale bar = 20 μm. (n = 6). E Western blot analysis of α-SMA. F Molecular docking results of VDR and SI. G CETSA analysis. (n = 3) *P < 0.05, **P < 0.01

Article Snippet: Membranes were blocked with 5% BSA and probed overnight at 4 °C with antibodies against: α-SMA (1:2000), COL1 A1 (1:1000), FASN (10624-2-AP, Proteintech, 1:1000), VDR (AF8159, Affinity, 1:1000), p-Smad3 (1:2000), Smad3 (AF6362, Affinity, 1:2000), TGF-β1 (1:3000) and β-actin (BM3873, Boster, 1:2000).

Techniques: Activation Assay, Immunofluorescence, Western Blot

Journal: Chinese medicine

Article Title: Senkyunolide I suppresses hepatic stellate cell activation and liver fibrosis by reprogramming VDR-dependent fatty acid metabolism.

doi: 10.1186/s13020-025-01133-x

Figure Lengend Snippet: Fig. 5 SI regulated VDR and fatty acid metabolism to suppress TGFβ/Smad pathway in CCl4-induced mice. A Immunohistochemistry analysis of TGF-β1, p-Smad3, α-SMA, COL1 A1. scale bar = 200 μm. (n = 5). B Western blot analysis of VDR, TGF-β1, and α-SMA. C mRNA levels of Cpt1a. D mRNA levels of lipogenic enzymes (Fasn, Acc-1, and Acly) and fatty acid desaturases (Fads1/2 and Scd1) (n = 6) *P < 0.05, **P < 0.01

Article Snippet: Membranes were blocked with 5% BSA and probed overnight at 4 °C with antibodies against: α-SMA (1:2000), COL1 A1 (1:1000), FASN (10624-2-AP, Proteintech, 1:1000), VDR (AF8159, Affinity, 1:1000), p-Smad3 (1:2000), Smad3 (AF6362, Affinity, 1:2000), TGF-β1 (1:3000) and β-actin (BM3873, Boster, 1:2000).

Techniques: Immunohistochemistry, Western Blot

Journal: Chinese medicine

Article Title: Senkyunolide I suppresses hepatic stellate cell activation and liver fibrosis by reprogramming VDR-dependent fatty acid metabolism.

doi: 10.1186/s13020-025-01133-x

Figure Lengend Snippet: Fig. 7 VDR knockout abolished the SI-induced inhibition of HSCs activation. A Western blot analysis (n = 3). B Immunofluorescence analysis of α-SMA. scale bar = 20 μm. (n = 6). C mRNA levels of fibrogenic markers (α-Sma, Col1a1, and Timp1). (n = 6) *P < 0.05, **P < 0.01 compared with si-NC + TGF-β1; $P < 0.05, $$P < 0.01 compared with si-NC; #P < 0.05, ##P < 0.01 compared with si-VDR + TGF-β1

Article Snippet: Membranes were blocked with 5% BSA and probed overnight at 4 °C with antibodies against: α-SMA (1:2000), COL1 A1 (1:1000), FASN (10624-2-AP, Proteintech, 1:1000), VDR (AF8159, Affinity, 1:1000), p-Smad3 (1:2000), Smad3 (AF6362, Affinity, 1:2000), TGF-β1 (1:3000) and β-actin (BM3873, Boster, 1:2000).

Techniques: Knock-Out, Inhibition, Activation Assay, Western Blot, Immunofluorescence

Journal: Chinese medicine

Article Title: Senkyunolide I suppresses hepatic stellate cell activation and liver fibrosis by reprogramming VDR-dependent fatty acid metabolism.

doi: 10.1186/s13020-025-01133-x

Figure Lengend Snippet: Fig. 8 SI reprogramed fatty acid metabolism through activating VDR in TGFβ1-induced HSCs. A Lipid droplet accumulation. scale bar = 20 μm. B Cpt1a expression. C Western blot analysis of FASN. (n = 3). D mRNA levels of lipogenic enzymes (Fasn, Acc-1, and Acly) and fatty acid desaturases (Fads1/2 and Scd1). (n = 6) *P < 0.05, **P < 0.01. *P < 0.05, **P < 0.01 compared with si-NC + TGF-β1; $P < 0.05, $$P < 0.01 compared with si-NC; #P < 0.05, ##P < 0.01 compared with si-VDR + TGF-β1

Article Snippet: Membranes were blocked with 5% BSA and probed overnight at 4 °C with antibodies against: α-SMA (1:2000), COL1 A1 (1:1000), FASN (10624-2-AP, Proteintech, 1:1000), VDR (AF8159, Affinity, 1:1000), p-Smad3 (1:2000), Smad3 (AF6362, Affinity, 1:2000), TGF-β1 (1:3000) and β-actin (BM3873, Boster, 1:2000).

Techniques: Expressing, Western Blot

Journal: Cell Communication and Signaling : CCS

Article Title: Vitamin D receptor (VDR) on the cell membrane of mouse macrophages participates in the formation of lipopolysaccharide tolerance: mVDR is related to the effect of artesunate to reverse LPS tolerance

doi: 10.1186/s12964-023-01137-w

Figure Lengend Snippet: The detection of mVDR in LPS-tolerant cells. A Co-localization of VDR and DIL in LPS-tolerant cells ( n = 3). mVDR was labeled green with the anti-VDR antibody, the cell membrane was labeled red with DIL ( A1 ), and the fluorescence intensities were calculated ( A2 ). B Co-localization of VDR and CD64 in LPS-tolerant cells ( n = 3). mVDR was labeled green with the anti-VDR antibody, the cell membrane was labeled red with CD64 ( B1 ), and mVDR was labeled green with the anti-VDR antibody, the homologous control antibody was labeled red with mouse IgG ( B2 ). C Membrane VDR level in LPS-tolerant cells ( n = 3). **, P < 0.01; # , P > 0.05. LPS5: 5 ng/mL LPS; LPS100: 100 ng/mL LPS (LPS100); T: LPS tolerance; DIL, 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate. mVDR: membrane VDR

Article Snippet: To confirm the binding of AS to mVDR, cell membrane proteins were extracted, and then mVDR in the extracts was captured to the 96-well plates coated with anti-VDR antibodies (Wuhan Fine Biotech, Wuhan, China).

Techniques: Labeling, Membrane, Fluorescence, Control

Journal: Cell Communication and Signaling : CCS

Article Title: Vitamin D receptor (VDR) on the cell membrane of mouse macrophages participates in the formation of lipopolysaccharide tolerance: mVDR is related to the effect of artesunate to reverse LPS tolerance

doi: 10.1186/s12964-023-01137-w

Figure Lengend Snippet: Anti-VDR antibodies abolish the effect of artesunate. A The effect of seven anti-VDR antibodies from different manufacturers on artesunate (AS)-mediated increase in TNF-α levels in LPS-tolerant cells ( n = 3). Note: a—g represent the anti-VDR antibodies from CST, Boster, Proteintech, Santa Cruz Biotechnology, ABclonal, Abcam, and Bioworld Technology, respectively. B Molecular docking. Human VDR (green) and mouse VDR (red) have similar spatial structures ( B1 ). Histidine 305 and 397 of human VDR (equivalent to histidine 300 and 392 of mouse VDR) are important for VDR binding to AS ( B2 ). C Effect of peptides on the AS-mediated TNF-α increase in LPS-tolerant cells ( n = 3). The peptides are the peptide H397 and its mutated peptide H397D ( C1 ) or the peptide H305 and its mutated peptide H305A ( C2 ). D Illustration of the binding of mVDR and AS by ELISA. Schematic diagram of the binding assay ( D1 ). Effect of anti-VDR on the binding of AS and VDR tracked by FITC-AS ( n = 3). AS with fluorophore Fluorescein 5-isothiocyanate was named FITC-AS (D2). *, P < 0.05; **, P < 0.01; # , P > 0.05. LPS5: 5 ng/mL LPS; LPS100: 100 ng/mL LPS (LPS100); T: LPS tolerance

Article Snippet: To confirm the binding of AS to mVDR, cell membrane proteins were extracted, and then mVDR in the extracts was captured to the 96-well plates coated with anti-VDR antibodies (Wuhan Fine Biotech, Wuhan, China).

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay

Journal: Cell Communication and Signaling : CCS

Article Title: Vitamin D receptor (VDR) on the cell membrane of mouse macrophages participates in the formation of lipopolysaccharide tolerance: mVDR is related to the effect of artesunate to reverse LPS tolerance

doi: 10.1186/s12964-023-01137-w

Figure Lengend Snippet: Artesunate affects the level of membrane, cytoplasmic, and nuclear VDR in LPS-tolerant cells. A The effect of artesunate (AS) on the mVDR level in LPS-tolerant cells without a permeabilization reagent treatment under laser confocal microscopy ( n = 3). mVDR was labeled green with the anti-VDR antibody, the cell membrane was labeled red with vimentin. B The effect of AS on the VDR level in LPS-tolerant cells with a permeabilization reagent treatment under laser confocal microscopy ( n = 3). cVDR was labeled green with the anti-VDR antibody (B1) , and the fluorescence intensities were calculated ( B2 ). C Detection of mVDR ( C1 ), cVDR ( C2 ), and nVDR ( C3 ) levels by Western blotting. D Detection of mVDR ( D1 ), cVDR ( D2 ), and nVDR ( D3 ) levels by ELISA, respectively. *, P < 0.05; **, P < 0.01. LPS5: 5 ng/mL LPS; LPS100: 100 ng/mL LPS (LPS100); T: LPS tolerance

Article Snippet: To confirm the binding of AS to mVDR, cell membrane proteins were extracted, and then mVDR in the extracts was captured to the 96-well plates coated with anti-VDR antibodies (Wuhan Fine Biotech, Wuhan, China).

Techniques: Membrane, Confocal Microscopy, Labeling, Fluorescence, Western Blot, Enzyme-linked Immunosorbent Assay