glycochenodeoxycholic acid gcdca  (MedChemExpress)

Journal: Frontiers in Immunology

Article Title: GCDCA promotes hepatocellular carcinoma progression through S1PR2/PI3K/AKT-mediated polarization of M2-type macrophages

doi: 10.3389/fimmu.2026.1640450

Figure Lengend Snippet: GCDCA promotes the HCC progression and increases hepatocellular carcinoma cell stemness. (A) Flow chart of animal experiments. (B) Kaplan–Meier plots of DEN-induced primary hepatocellular carcinoma in rats after administration of GCDCA and cholestyramine ( n = 5 in each group). (C) Representative images of liver tumors in different groups of rats (scale bar = 1 cm) and HE-stained images (scale bar = 200 μm). (D, E) Maximum diameters of liver tumors in rats in different groups and the number of tumors for quantitative analysis ( n = 5 in each group). (F, H) Serum AST and ALT activities and TBA levels in different groups of rats ( n = 4 in each group). (I) Representative immunohistochemical staining of KRT19-, EPCAM-, and SOX9-positive cells in the livers of different groups of rats, scale bar = 100 μm ( n = 3 in each group). (J) Representative immunofluorescence staining of co-localized positive cells of KRT19 (red), EPCAM (green), and SOX9 (pink) in liver CSCs of different groups of rats. Cell nuclei were stained with DAPI (blue). Scale bar = 50 μm. * p < 0.05, ** p < 0.01, and *** p < 0.001, n.s., not significant.

Article Snippet: JTE-013 was injected intraperitoneally 2 h before GCDCA administration at a dose of 2 mg/kg body weight in the JTE-013 (TargetMol, China) interference model, starting at week 12 of DEN treatment.

Techniques: Staining, Immunohistochemical staining, Immunofluorescence

Journal: Frontiers in Immunology

Article Title: GCDCA promotes hepatocellular carcinoma progression through S1PR2/PI3K/AKT-mediated polarization of M2-type macrophages

doi: 10.3389/fimmu.2026.1640450

Figure Lengend Snippet: GCDCA does not directly promote stemness in hepatocellular carcinoma cells. (A) Relative viability values of hepatocellular carcinoma cells in different concentrations of GCDCA ( n = 3 in each group). (B) Effect of GCDCA on clonogenic ability of different hepatocellular carcinoma cell lines. (C) Effect of GCDCA on sphere-forming ability of different hepatocellular carcinoma cell lines. Scale bar = 50 µm. Cell clones were counted using ImageJ 1.43. * p < 0.05, ** p < 0.01, and *** p < 0.001, n.s., not significant.

Article Snippet: JTE-013 was injected intraperitoneally 2 h before GCDCA administration at a dose of 2 mg/kg body weight in the JTE-013 (TargetMol, China) interference model, starting at week 12 of DEN treatment.

Techniques: Clone Assay

Journal: Frontiers in Immunology

Article Title: GCDCA promotes hepatocellular carcinoma progression through S1PR2/PI3K/AKT-mediated polarization of M2-type macrophages

doi: 10.3389/fimmu.2026.1640450

Figure Lengend Snippet: GCDCA-induced polarization of M2 macrophages promotes stemness in hepatocellular carcinoma cells. (A) Expression of M2 macrophage marker genes Mrc1 , Cd163 , Arg1 in NR8383 cells after GCDCA treatment (n = 3 in each group). (B) Expression of CD206, CD163, ARG1 proteins in NR8383 cells. (C) Expression of M2 macrophage marker genes MRC1 , ARG1 , and TGF-β in THP-1 cells after GCDCA treatment (n = 3 in each group). (D) Expression of CD206, CD163, ARG1 proteins in THP-1 cells. (E) Co-culture pattern of NR8383 and RH35 cells. (F) Expression of stemness genes Krt19 , Epcam , and Sox9 in RH35 cells after co-culture with GCDCA-treated NR8383 cells (n =3 in each group). (G) Expression of KRT19, EPCAM, and SOX9 proteins in RH35 cells. (H) Co-culture pattern of THP-1 and Huh7. (I) Expression of stemness genes KRT19 , EPCAM , and SOX9 in Huh7 cells after co-culture with GCDCA-treated THP-1 cells (n = 3 in each group). (J) Expression of KRT19, EPCAM, and SOX9 proteins in Huh7 cells. GAPDH was used as an internal reference. * p < 0.05, ** p < 0.01, and *** p < 0.001, n.s., not significant.

Article Snippet: JTE-013 was injected intraperitoneally 2 h before GCDCA administration at a dose of 2 mg/kg body weight in the JTE-013 (TargetMol, China) interference model, starting at week 12 of DEN treatment.

Techniques: Expressing, Marker, Co-Culture Assay

Journal: Frontiers in Immunology

Article Title: GCDCA promotes hepatocellular carcinoma progression through S1PR2/PI3K/AKT-mediated polarization of M2-type macrophages

doi: 10.3389/fimmu.2026.1640450

Figure Lengend Snippet: GCDCA induces M2-type polarization in macrophages through activation of S1PR2 receptors. (A) Single-cell sequencing to analyze bile acid receptor expression in M2 macrophages. (B) Activation of the S1PR2 receptor by GCDCA was detected using the Tango GPCR assay (n = 3 in each group). (C, D) TCGA database analysis of S1PR2 correlation with KRT19 , EPCAM , SOX9 , CD68 , MRC1 , CD163 . (E) Representative immunohistochemical staining of CD68, CD163, and S1PR2-positive cells in the livers of different groups of rats, scale bar = 100 μm (n = 3 in each group). (F) Representative immunofluorescence staining of S1PR2 (red) co-localized with CD163- (green) and CD206-(green) positive cells in the livers of different groups of rats, scale bar = 100 μm. * p < 0.05, ** p < 0.01, and *** p < 0.001, n.s., not significant.

Article Snippet: JTE-013 was injected intraperitoneally 2 h before GCDCA administration at a dose of 2 mg/kg body weight in the JTE-013 (TargetMol, China) interference model, starting at week 12 of DEN treatment.

Techniques: Activation Assay, Single Cell, Sequencing, Expressing, Immunohistochemical staining, Staining, Immunofluorescence

Journal: Frontiers in Immunology

Article Title: GCDCA promotes hepatocellular carcinoma progression through S1PR2/PI3K/AKT-mediated polarization of M2-type macrophages

doi: 10.3389/fimmu.2026.1640450

Figure Lengend Snippet: Pharmacological inhibition of S1PR2 by JTE-013 reverses GCDCA-induced stemness. (A) Flow chart of animal experiments. (B) Kaplan–Meier plots of DEN-induced primary hepatocellular carcinoma in rats after administration of GCDCA and the receptor inhibitor JTE-013 ( n = 5 in each group). (C) Representative images of liver tumors in different groups of rats (scale bar = 1 cm) and HE-stained images (scale bar = 200 μm). (D, E) Analysis of maximum diameter and number of tumors (n = 4 in each group). (F–H) Serum AST and ALT activities and TBA levels in different groups of rats (n = 4 in each group). (I) Representative immunohistochemical staining of KRT19, EPCAM, and SOX9-positive cells in the livers of different groups of rats, scale bar = 200 μm (n = 3 in each group). (J) Representative immunofluorescence staining of KRT19, EPCAM, and SOX9 co-localized positive cells in the livers of different groups of rats, scale bar = 50 μm. * p < 0.05, ** p < 0.01, and *** p < 0.001, n.s., not significant.

Article Snippet: JTE-013 was injected intraperitoneally 2 h before GCDCA administration at a dose of 2 mg/kg body weight in the JTE-013 (TargetMol, China) interference model, starting at week 12 of DEN treatment.

Techniques: Inhibition, Staining, Immunohistochemical staining, Immunofluorescence

Journal: Frontiers in Immunology

Article Title: GCDCA promotes hepatocellular carcinoma progression through S1PR2/PI3K/AKT-mediated polarization of M2-type macrophages

doi: 10.3389/fimmu.2026.1640450

Figure Lengend Snippet: Inhibition of S1PR2 receptor reverses GCDCA-induced polarization of M2 macrophages. (A) Expression of M2 macrophage marker genes Mrc1 , Cd163 , and Arg1 in NR8383 cells after inhibition of the S1PR2 receptor (n = 3 in each group). (B) Expression of CD206, CD163, and ARG1 proteins in NR8383 cells after inhibition of the S1PR2 receptor. (C) Expression of M2 macrophage marker genes MRC1 , ARG1 , and TGF-β in THP-1 cells after inhibition of the S1PR2 receptor (n = 3 in each group). (D) Expression of CD206, CD163, and ARG1 proteins in THP-1 cells after inhibition of the S1PR2 receptor. (E) Expression of stemness genes Krt19 , Epcam , and Sox9 in RH35 cells after co-culture with GCDCA-induced NR8383 cells pretreated with JTE-013 (n = 3 in each group). (F) Expression of KRT19, EPCAM, and SOX9 proteins in RH35 cells after co-culture with GCDCA-induced NR8383 cells pretreated with JTE-013. (G) Expression of stemness genes KRT19 , EPCAM , and SOX9 in Huh7 cells after co-culture with GCDCA-induced THP-1 cells pretreated with JTE-013 (n = 3 in each group). (H) Expression of KRT19, EPCAM, and SOX9 proteins in Huh7 cells after co-culture with GCDCA-induced THP-1 cells pretreated with JTE-013. (I-J) Representative immunofluorescence staining of M2 macrophage markers CD206 (white) and CD163 (white) co-localized with KRT19- (red) and EPCAM- (green) positive cells in the livers of different groups of rats, scale bar = 100 μm. GAPDH is used as an internal reference. * p < 0.05, ** p < 0.01, and *** p < 0.001, n.s., not significant.

Article Snippet: JTE-013 was injected intraperitoneally 2 h before GCDCA administration at a dose of 2 mg/kg body weight in the JTE-013 (TargetMol, China) interference model, starting at week 12 of DEN treatment.

Techniques: Inhibition, Expressing, Marker, Co-Culture Assay, Immunofluorescence, Staining

Journal: Frontiers in Immunology

Article Title: GCDCA promotes hepatocellular carcinoma progression through S1PR2/PI3K/AKT-mediated polarization of M2-type macrophages

doi: 10.3389/fimmu.2026.1640450

Figure Lengend Snippet: GCDCA induces macrophage polarization toward M2 via the S1PR2/PI3K/AKT signaling pathway. (A) Volcano plot of differentially expressed genes (DEGs) between GCDCA-treated and control groups (n = 3 in each group). (B) Histogram of KEGG enrichment results of DEGs between GCDCA-treated and control groups. (C) Bubble plots of GO enrichment results of DEGs between GCDCA-treated and control groups. (D) Western blotting for p-AKT, AKT, p-PI3K, and PI3K protein expression in GCDCA-treated NR8383 cells. (E) Western blotting was used to detect the expression of p-AKT, AKT, p-PI3K, PI3K, CD206, and ARG1 proteins after pretreatment with LY294002 inhibitor.

Article Snippet: JTE-013 was injected intraperitoneally 2 h before GCDCA administration at a dose of 2 mg/kg body weight in the JTE-013 (TargetMol, China) interference model, starting at week 12 of DEN treatment.

Techniques: Control, Western Blot, Expressing

Journal: Journal of Inflammation Research

Article Title: Glycochenodeoxycholic acid induces the release of IL-1beta from L02 cells via the NLRP3/caspase-1/GSDMD pathway to activate LX2 cells

doi: 10.2147/JIR.S532042

Figure Lengend Snippet: GCDCA activates the NLRP3 inflammasome in hepatocytes. ( A ) Dose-dependent effects of GCDCA (25–400 μM, 24 h) on protein levels effects of NLRP3, ASC, pro-caspase-1, caspase-1 P20, pro-IL-1β and GAPDH. ( B ) Time-dependent effects of these proteins in L02 cells treated with 200 μM GCDCA. ( C and D ) Synergistic priming by LPS (1 μg/mL, 4 h pre-treatment): GCDCA (50–400 μM, 24 h) ( C ) or time-dependent (12–72 h) ( D ) enhancement of NLRP3, pro-caspase-1, and pro-IL-1β expression in LPS-primed cells. ( E ) Comparative analysis of NLRP3 inflammasome activationin L02 cells treated with GCDCA (200 μM, 24 h) with/without LPS co-stimulation. ( F ) Essential role of NLRP3 in GCDCA-induced pyroptosis. L02 cells were pretreated with/without LPS (1 μg/mL, 4 h) and MCC950 (10 μM, 2 h) prior to GCDCA (200 μM, 24 h) exposure. Whole cell lysates were examined by Western Blot analysis. Molecular weight markers (kDa) are indicated. n ≥ 3 for each group. (LG200: LPS+200 μM GCDCA; G200: GCDCA 200 μM).

Article Snippet: L02 and LX2 cells were synchronized in serum-free medium for 12 h. L02 cells were exposed to graded GCDCA concentrations (25–400 μM, HY-N2334, MCE) for 24 h or 200 μM GCDCA for 6–72 h to establish dose/time-response profiles.

Techniques: Expressing, Western Blot, Molecular Weight

Journal: Journal of Inflammation Research

Article Title: Glycochenodeoxycholic acid induces the release of IL-1beta from L02 cells via the NLRP3/caspase-1/GSDMD pathway to activate LX2 cells

doi: 10.2147/JIR.S532042

Figure Lengend Snippet: GCDCA induces GSDMD-mediated pyroptosis in hepatocytes. ( A and B ) Time- and dose-dependent LDH release in LPS-primed L02 cells treated with GCDCA: ( A ) Dose-response (50–400 μM GCDCA, 24 h); ( B ) Time-course (200 μM GCDCA, 12–72 h). ( C ) Dose-dependent GSDMD proteolysis in LPS-primed cells treated with GCDCA (50–400 μM, 24 h); ( D ) Time-dependent GSDMD proteolysis (200 μM GCDCA, 12–72 h). GAPDH serves as loading control. ( E ) Validation of GSDMD knockdown efficiency in L02 cells. ( F ) LDH release in lentiviruses-transfected L02 cells treated with GCDCA (200 μM, 24 h). ( G ) Representative flow plots showing Annexin V/PI staining in lentiviruses-transfected L02 cells. ( H ) Flow cytometry quantification of pyroptotic cells (Annexin V⁺/PI⁺). n ≥ 3 for each group. a P < 0.005, b P < 0.05, c P < 0.0001.

Article Snippet: L02 and LX2 cells were synchronized in serum-free medium for 12 h. L02 cells were exposed to graded GCDCA concentrations (25–400 μM, HY-N2334, MCE) for 24 h or 200 μM GCDCA for 6–72 h to establish dose/time-response profiles.

Techniques: Control, Biomarker Discovery, Knockdown, Transfection, Staining, Flow Cytometry

Journal: Journal of Inflammation Research

Article Title: Glycochenodeoxycholic acid induces the release of IL-1beta from L02 cells via the NLRP3/caspase-1/GSDMD pathway to activate LX2 cells

doi: 10.2147/JIR.S532042

Figure Lengend Snippet: GCDCA triggers caspase-1-dependent pyroptosis in hepatocytes. ( A and B ) Caspase-1/4 activity in LPS-primed L02 cells treated with GCDCA. ( C and D ) Flow cytometry quantification of pyroptotic cells (Annexin V⁺/PI⁺) in L02 cells with caspase-1 inhibitor Ac-YVAD-cmk (AC, 50 μM) followed by LPS + GCDCA (200 μM, 24 h). ( E ) LDH release in L02 cells with AC pretreatment. (F)Western blot analysis of pyroptosis-related proteins in L02 cells with AC pretreatment. n ≥ 3 for each group. a P < 0.005, b P < 0.0001.

Article Snippet: L02 and LX2 cells were synchronized in serum-free medium for 12 h. L02 cells were exposed to graded GCDCA concentrations (25–400 μM, HY-N2334, MCE) for 24 h or 200 μM GCDCA for 6–72 h to establish dose/time-response profiles.

Techniques: Activity Assay, Flow Cytometry, Western Blot

Journal: Journal of Inflammation Research

Article Title: Glycochenodeoxycholic acid induces the release of IL-1beta from L02 cells via the NLRP3/caspase-1/GSDMD pathway to activate LX2 cells

doi: 10.2147/JIR.S532042

Figure Lengend Snippet: GCDCA drives IL-1β secretion through GSDMD-dependent activation. ( A ) Dose-dependent IL-1β secretion in LPS-primed L02 cells treated with GCDCA (50–400 μM, 24 h). ( B )Time-dependent IL-1β secretion (200 μM GCDCA, 12–72 h). ( C ) Western blot analysis of GSDMD-NT and GSDMD in L02 cells and mature IL-1β in cell supernatant. ( D ) ELISA quantification of extracellular IL-1β. n ≥ 3 for each group. a P < 0.01.

Article Snippet: L02 and LX2 cells were synchronized in serum-free medium for 12 h. L02 cells were exposed to graded GCDCA concentrations (25–400 μM, HY-N2334, MCE) for 24 h or 200 μM GCDCA for 6–72 h to establish dose/time-response profiles.

Techniques: Activation Assay, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Journal of Inflammation Research

Article Title: Glycochenodeoxycholic acid induces the release of IL-1beta from L02 cells via the NLRP3/caspase-1/GSDMD pathway to activate LX2 cells

doi: 10.2147/JIR.S532042

Figure Lengend Snippet: GCDCA-mediated hepatocyte IL-1β secretion activates HSCs. ( A ) Western blot analysis ofα-SMA and Collagen-I in LX2 cells treated with conditioned medium (CM) from GCDCA/LPS-stimulated L02 hepatocytes. ( B ) The protein levels of α-SMA and Collagen-I in LX2 cells with treatment of IL-1 receptor antagonist (IL-1RA, 200 ng/mL) upon CM exposure. ( C and D ) Representative images and quantification of LX2 migration upon CM exposure detected by wound healing assay. ( E ) CCK-8 assay of LX2 proliferation by CM treatment. n ≥ 3 for each group. a P < 0.05.

Article Snippet: L02 and LX2 cells were synchronized in serum-free medium for 12 h. L02 cells were exposed to graded GCDCA concentrations (25–400 μM, HY-N2334, MCE) for 24 h or 200 μM GCDCA for 6–72 h to establish dose/time-response profiles.

Techniques: Western Blot, Migration, Wound Healing Assay, CCK-8 Assay