ampk inhibitor cc  (MedChemExpress)

Journal: Frontiers in Endocrinology

Article Title: Anoctamin 5 mutation leads to abnormal bone homeostasis of GDD by regulating AMPK-dependent glucose metabolism

doi: 10.3389/fendo.2026.1703491

Figure Lengend Snippet: AMPK activation is responsible for enhanced glucose metabolism in Ano5 Cys360Tyr osteoblast. To examine the effect of AMPK on abnormal glucose metabolism, Ano5 KI/KI mCOB was treated with 5μM CC. (A) Immunoblotting analysis of HK2, p-AMPKα/AMPKα, and ACTB at days 0 and 14 of osteogenic induction; (B) qRT-PCR analysis of Hk2 and Ldha ; (C–E) LDH activity (C) , lactate content (D) , and ATP content (E) in mCOB; (F) Seahorse XF glycolysis rate examination at day 14, and relative quantitative analysis of basal and compensatory glycolysis ability according to proton efflux rate (PER); (G) qRT-PCR (left) and immunoblotting (right) analysis of PGC1α; (H) Immunoblotting analysis of OXPHOS complex and ACTB; (I) Seahorse XF mitochondrial stress examination at days 0 and 14, ATP production ability according to oxygen consumption rate (OCR). Data are presented as mean ± SEM. Statistic significances are determined by one-way ANOVAs with Dunnett’s multiple comparison tests, with *P < 0.05, **P < 0.01.

Article Snippet: For examining the effect of AMPK inhibitor on bone phenotype of GDD, 8-week-old male Ano5 KI/KI mice were given AMPK inhibitor CC (5mg/kg, MCE, HY-13418) by gavage three times a week for four weeks, control group was given NaCl.

Techniques: Activation Assay, Western Blot, Quantitative RT-PCR, Activity Assay, Comparison

Journal: Frontiers in Endocrinology

Article Title: Anoctamin 5 mutation leads to abnormal bone homeostasis of GDD by regulating AMPK-dependent glucose metabolism

doi: 10.3389/fendo.2026.1703491

Figure Lengend Snippet: AMPK inhibitor attenuates osteogenesis of Ano5 KI/KI mCOB. To examine the effect of AMPK on abnormal glucose metabolism, Ano5 KI/KI mCOB was treated with 5μM CC. (A) Cell proliferation analysis by CCK8 detection of mCOB at days 0, 1, 2, 3, and 4 without osteoblast differentiation ( A P < 0.01 representing Ano5 KI/KI vs Ano5 +/+ , b P < 0.05 and B P < 0.01 representing Ano5 KI/KI +Compound C vs Ano5 KI/KI ); (B, C) ALP staining [ (B) , bar=100 μm] and activity analysis (C) at day 7 of osteogenic induction; (D, E) Alizarin Red S staining [ (D) , bar=100 μm] and histogram of the corresponding calcium-binding levels in the mineral nodules (E) at day 21; (F) Immunoblotting analysis of Col1α1, Ocn, and ACTB; (G) qRT-PCR analysis of Ocn , Col1α1 , Opg and Opg/Rankl . Data are presented as mean ± SEM. Statistic significances are determined by one-way ANOVAs with Dunnett’s multiple comparison tests, with *P < 0.05, **P < 0.01.

Article Snippet: For examining the effect of AMPK inhibitor on bone phenotype of GDD, 8-week-old male Ano5 KI/KI mice were given AMPK inhibitor CC (5mg/kg, MCE, HY-13418) by gavage three times a week for four weeks, control group was given NaCl.

Techniques: Staining, Activity Assay, Binding Assay, Western Blot, Quantitative RT-PCR, Comparison

Journal: Frontiers in Endocrinology

Article Title: Anoctamin 5 mutation leads to abnormal bone homeostasis of GDD by regulating AMPK-dependent glucose metabolism

doi: 10.3389/fendo.2026.1703491

Figure Lengend Snippet: Ano5 Cys360Tyr mutation interferes with AMPK-dependent mitochondrial function of osteoclast. (A) TEM scan of osteoclast in tibia of 12-week-old male mice, orange arrow indicating nucleus and green arrow indicating mitochondria; (B) Flow cytometry detecting JC-1 monomer (green) and polymer (red) of Ano5 +/+ and Ano5 KI/KI osteoclast at day 5 of osteoclast differentiation; (C) Intracellular ATP level of Ano5 +/+ and Ano5 KI/KI osteoclast at day 5 of M-CSF (30 ng/mL) and RANKL (100 ng/mL) induction; (D) Immunoblotting analysis of OXPHOS complex and ATCB at days0, 3, and 5; (E) Seahorse XF mito stress analysis at day 0 and 5, and histogram of basal respiration ability; (F) Immunoblotting analysis of PGC1β, p-AMPKα/AMPKα, and ACTB; (G, H) In order to examine the relation between AMPK activation and mitochondrial dysregulation of osteoclast, 5 μM AMPK inhibitor Compound C was administered to Ano5 KI/KI BMM. (G, H) Immunoblotting analysis of PGC1β, p-AMPKα/AMPKα (G) , OXPHOS (H) , and ACTB at day 5; (I) Intracellular ATP content analysis at day 5; (J) qRT-PCR analysis of Atp5b , Sdhb , and Cox4 . Data are presented as mean ± SEM. Statistic significances are determined by t-tests (B, C) and one-way ANOVAs with Dunnett’s multiple comparison tests (E, I, J) , with *P < 0.05, **P < 0.01.

Article Snippet: For examining the effect of AMPK inhibitor on bone phenotype of GDD, 8-week-old male Ano5 KI/KI mice were given AMPK inhibitor CC (5mg/kg, MCE, HY-13418) by gavage three times a week for four weeks, control group was given NaCl.

Techniques: Mutagenesis, Flow Cytometry, Polymer, Western Blot, Activation Assay, Quantitative RT-PCR, Comparison

Journal: Frontiers in Endocrinology

Article Title: Anoctamin 5 mutation leads to abnormal bone homeostasis of GDD by regulating AMPK-dependent glucose metabolism

doi: 10.3389/fendo.2026.1703491

Figure Lengend Snippet: Blocking AMPK rescues osteoclastogenesis of Ano5 KI/KI BMM. 5 μM AMPK inhibitor Compound C was administered to Ano5 KI/KI BMM. (A, B) qRT-PCR (A) and immunoblotting (B) analysis of NFATC1, CTSK, and cFOS at days 0 and 5; (C) TRAP (above, bar=100 μm) and Phalloidin (bellow, green, bar=200 μm) staining at days 7 with DAPI labeling cell nucleus; (D) the number of TRAP-positive cells per well; (E) the percentage of nuclei in actin ring-positive osteoclasts to the total number of nuclei; (F) TRAP activity analysis of osteoclast at day 5 of RANKL stimulation normalized to protein content; (G) qRT-PCR analysis of Trap , Dcstamp , and Mmp9 . Data are presented as mean ± SEM. Statistic significances are determined by one-way ANOVAs with Dunnett’s multiple comparison tests, with *P < 0.05, **P < 0.01.

Article Snippet: For examining the effect of AMPK inhibitor on bone phenotype of GDD, 8-week-old male Ano5 KI/KI mice were given AMPK inhibitor CC (5mg/kg, MCE, HY-13418) by gavage three times a week for four weeks, control group was given NaCl.

Techniques: Blocking Assay, Quantitative RT-PCR, Western Blot, Staining, Labeling, Activity Assay, Comparison

Journal: Frontiers in Endocrinology

Article Title: Anoctamin 5 mutation leads to abnormal bone homeostasis of GDD by regulating AMPK-dependent glucose metabolism

doi: 10.3389/fendo.2026.1703491

Figure Lengend Snippet: AMPK inhibitor effectively rescues bone metabolism of GDD. 12-week-old male mice were used to observe bone phenotype. (A) μCT images of vertical plane and trabeculae 3D reconstructions of tibia. (B, C) Quantification analysis of μCT analysis of cortical bone (B) and trabecula bone (C) of tibia; (D) μCT images of vertical plane and trabeculae 3D reconstructions of femur, and quantification analysis of cortical bone thickness; (E) μCT images of vertical plane of mandible and quantification analysis of palatal cortical bone thickness; (F, G) HE staining of tibia (F) and quantification analysis of cortical bone thickness (G) (bar=100 μm); (H) Displacement-load curve and quantification analysis of three-point bending examination. Data are presented as mean ± SEM. Statistic significances are determined by one-way ANOVAs with Dunnett’s multiple comparison tests, with ns: no significance, *P < 0.05, **P < 0.01.

Article Snippet: For examining the effect of AMPK inhibitor on bone phenotype of GDD, 8-week-old male Ano5 KI/KI mice were given AMPK inhibitor CC (5mg/kg, MCE, HY-13418) by gavage three times a week for four weeks, control group was given NaCl.

Techniques: Staining, Comparison

Journal: Frontiers in Endocrinology

Article Title: Anoctamin 5 mutation leads to abnormal bone homeostasis of GDD by regulating AMPK-dependent glucose metabolism

doi: 10.3389/fendo.2026.1703491

Figure Lengend Snippet: AMPK inhibitor regulates osteogenesis and osteoclastogenesis of GDD in vivo . 12-week-old male mice were used to observe bone phenotype. (A) Representative photomicrographs and quantitative immuno-positive area analysis of OCN in tibia (bar=100 μm); (B–D) ELISA analysis of serum level of ALP, PINP (B) , OPG, OPG/RANKL (C) , and CTX (D, E) Representative images of TRAP staining and quantitative analysis of the osteoclast number (Oc.N)/perimeter of bone (B.Pm) in the germinal center of the tibia (bar=100 μm). Data are presented as mean ± SEM. Statistic significances are determined by one-way ANOVAs with Dunnett’s multiple comparison tests, with ns: no significance, *P < 0.05, **P < 0.01.

Article Snippet: For examining the effect of AMPK inhibitor on bone phenotype of GDD, 8-week-old male Ano5 KI/KI mice were given AMPK inhibitor CC (5mg/kg, MCE, HY-13418) by gavage three times a week for four weeks, control group was given NaCl.

Techniques: In Vivo, Enzyme-linked Immunosorbent Assay, Staining, Comparison

Journal: Frontiers in Endocrinology

Article Title: Anoctamin 5 mutation leads to abnormal bone homeostasis of GDD by regulating AMPK-dependent glucose metabolism

doi: 10.3389/fendo.2026.1703491

Figure Lengend Snippet: Schematic illustration of glucose metabolic distemperedness mediated by AMPK activation in GDD. Excessive AMPK activation caused by ANO5 Cys360Tyr mutation stimulated glycolysis in osteoblast and disturbed the mitochondrial homeostasis between osteoblast and osteoclast by enhancing PGC1α and inhibiting PGC1β expression respectively, to augment bone formation and suppress osteoclastogenesis.

Article Snippet: For examining the effect of AMPK inhibitor on bone phenotype of GDD, 8-week-old male Ano5 KI/KI mice were given AMPK inhibitor CC (5mg/kg, MCE, HY-13418) by gavage three times a week for four weeks, control group was given NaCl.

Techniques: Activation Assay, Mutagenesis, Expressing

Journal: Advanced Science

Article Title: Trigonelline Improves Metabolism and Cardiac Function of HFpEF Mice Via Gut Microbiome Alterations‐Mediated AMPK Activation

doi: 10.1002/advs.202513956

Figure Lengend Snippet: AMPK activation was required for the cardioprotective effects of trigonelline in HFpEF mice. A) Representative immunoblot images of total and phosphorylated AMPK in the heart and liver tissues from HFpEF mice receiving trigonelline. B,C) Quantification of cardiac p‐AMPK/AMPK ( n = 6 mice per group). D) Representative immunoblot images of total and phosphorylated AMPK in the heart and liver tissues from HFpEF mice receiving trigonelline with or without AMPK inhibitor. E,F) Quantification of hepatic p‐AMPK/AMPK ratio ( n = 6 mice per group). G,H) SBP and DBP of different experimental groups ( n = 5 mice per group). I) Representative echocardiography‐derived M‐mode tracings (top), pulsed‐wave Doppler (middle), and tissue Doppler (bottom) tracings of mice in the indicated group. J) Percent left ventricular ejection fraction (LVEF%). K) The ratio between mitral E wave and E’ wave (E/E’). L) Running distance during the exercise exhaustion test of mice. M) The ratio between wet and dry lung weight (LW). N) Ratio between heart weight and tibia length (HW/TL) (for LVEF%, E/E’ ratio, running distance, LW wet/LW dry ratio, and HW/TL ratio, n = 6 mice per group). O) Representative images of WGA in transversal sections of the left ventricle of mice of different experimental groups. Scale bar: 50 µm for WGA. P) WGA quantification of cardiomyocyte cross‐sectional area ( n = 5 mice per group). Data are presented as mean ± SEM and analyzed using one‐way ANOVA followed by Tukey's multiple comparisons test. ns, no significant; * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001.

Article Snippet: To test the role of AMPK in the therapeutic effects of trigonelline in vivo, HFpEF mice were pre‐treated with AMPK inhibitor Compound C (CC) (MedChemExpress, HY‐13418, 5 mg kg −1 day −1 ) or vehicle for 1 week.

Techniques: Activation Assay, Western Blot, Derivative Assay

Journal: Advanced Science

Article Title: Trigonelline Improves Metabolism and Cardiac Function of HFpEF Mice Via Gut Microbiome Alterations‐Mediated AMPK Activation

doi: 10.1002/advs.202513956

Figure Lengend Snippet: AMPK inhibition abolished the beneficial effects of trigonelline on metabolic disorders in HFpEF mice. A) Food intake of mice per day of different experimental groups per day ( n = 5). B) Body weight was monitored weekly in each experimental group. C) Representative images of MRI of mice from different experimental groups. Red indicates higher fat content, green represents moderate fat content, and blue corresponds to low fat content. Scale bar: 1 cm. Fat mass D) and lean mass E) ratios of mice in the indicated groups ( n = 5). F) Glucose tolerance tests in the indicated groups ( n = 5). G) Insulin sensitivity tests in the indicated groups ( n = 5). H) Total cholesterol of serum in each group. I) Low‐density lipoprotein cholesterol (LDL‐C) of the serum in each group. J) Representative images of hematoxylin and eosin staining and oil red staining of liver from mice in each treated group, Scale bar = 100 µm. K) Serum ALT in each group. L) Serum AST in each group. M) Serum Cre in each group. N) Serum BUN in each group (for total cholesterol, LDL‐C, Serum ALT, AST, Cre, and BUN, n = 5 mice per group). Data are presented as mean ± SEM and analyzed using one‐way ANOVA followed by Tukey's multiple comparisons test. ns, no significant; * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: To test the role of AMPK in the therapeutic effects of trigonelline in vivo, HFpEF mice were pre‐treated with AMPK inhibitor Compound C (CC) (MedChemExpress, HY‐13418, 5 mg kg −1 day −1 ) or vehicle for 1 week.

Techniques: Inhibition, Staining

Journal: Advanced Science

Article Title: Trigonelline Improves Metabolism and Cardiac Function of HFpEF Mice Via Gut Microbiome Alterations‐Mediated AMPK Activation

doi: 10.1002/advs.202513956

Figure Lengend Snippet: Trigonelline activates AMPK in a gut microbiota‐dependent manner. A–C) Diversity of the gut microbiota in each group, as indicated by the observed species, Shannon, and Chao1 indices. D) PCA score plot analysis based on the relative abundance of OTUs. E) Scheme for the experimental strategy in HFpEF mice treated with trigonelline and antibiotics. F) Representative immunoblot images of total and phosphorylated AMPK in the heart and liver tissues of mice in each group. Quantification of cardiac pAMPK/AMPK ratio in the heart G) and liver H) tissues of mice in each group. ( n = 6 mice per group). I) Representative immunoblot images of total and phosphorylated AMPK in HL‐1 and HepG2 cell lines in each group. Quantification of cardiac pAMPK/AMPK ratio in the HL‐1 J) and HepG2 cell lines K) in each group. ( n = 4 mice per group). Data are presented as mean ± SEM and analyzed using one‐way ANOVA followed by Tukey's multiple comparisons test. ns, no significant; * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001. Abx, antibiotic cocktail.

Article Snippet: To test the role of AMPK in the therapeutic effects of trigonelline in vivo, HFpEF mice were pre‐treated with AMPK inhibitor Compound C (CC) (MedChemExpress, HY‐13418, 5 mg kg −1 day −1 ) or vehicle for 1 week.

Techniques: Western Blot

Journal: Journal of animal science and biotechnology

Article Title: Methionine deficiency inhibited pyroptosis in primary hepatocytes of grass carp (Ctenopharyngodon idella): possibly via activating the ROS-AMPK-autophagy axis.

doi: 10.1186/s40104-024-01069-6

Figure Lengend Snippet: Fig. 7 MD promoted autophagy through the AMPK signaling pathway. Relative protein expression of LKB1, p-AMPK, ULK1, p-TOR (A) and quantification (B) after with or without 1 mmol/L Met treatment for 48 h; relative protein expression of p-AMPK (C) and quantification (D) after with or without 1 mmol/L Met and different concentrations of CC (0, 2, 4, 8, 16 μmol/L) co-treatment for 48 h; relative protein expression of LC3 II and p62 (E) and quantification (F) after with or without 1 mmol/L Met and 16 μmol/L CC co-treatment for 48 h. The results were expressed as mean ± SD of 3 independent observations. *P < 0.05, **P < 0.01, n.s: no significance. Met, methionine; LKB1, liver kinase B1; p-AMPK, phosphorylated-AMP-activated protein kinase; ULK1, Unc-51-like kinase 1; p-TOR, phosphorylated-rapamycin; CC, compound C; p62, sequestosome 1; LC3, microtubule-associated protein 1 light chain 3

Article Snippet: The following reagents were used in the experiments: LPS (L2880, Sigma-Aldrich, St. Louis, Missouri, USA), NLRP3 activator nigericin sodium salt (Nig) (S6653, Selleck Chemicals, Houston, TX, USA), autophagy inhibitor chloroquine (CQ) (S6999, Selleck Chemicals), AMPK inhibitor compound C (CC) (S7840, Selleck Chemicals), ROS scavenger N-acetyl-L-cysteine (NAC) (T0875, TargetMol Chemicals, Boston, MA, USA).

Techniques: Expressing

Journal: Journal of animal science and biotechnology

Article Title: Methionine deficiency inhibited pyroptosis in primary hepatocytes of grass carp (Ctenopharyngodon idella): possibly via activating the ROS-AMPK-autophagy axis.

doi: 10.1186/s40104-024-01069-6

Figure Lengend Snippet: Fig. 8 MD activated the AMPK signaling pathway by increasing ROS. Content of GSH (A); ROS (B) and quantification (C) after with or without 1 mmol/L Met treatment for 48 h; content of ROS (D) and quantification (E) after pre-treatment with NAC (0, 2.5, 5, 7.5 mmol/L) for 1 h then treat with or without 1 mmol/L Met for 48 h; relative protein expression of p-AMPK (F) and quantification (G) after pre-treatment with 5 mmol/L NAC for 1 h then treat with or without 1 mmol/L Met for 48 h. The results were expressed as mean ± SD of 3 or 6 independent observations (WB: n = 3). *P < 0.05, **P < 0.01, ***P < 0.001, n.s: no significance. GSH, glutathione; Met, methionine; Nig, nigericin sodium salt; ROS, reactive oxygen species; NAC, N-acetyl-L-cysteine; p-AMPK, phosphorylated-AMP-activated protein kinase

Article Snippet: The following reagents were used in the experiments: LPS (L2880, Sigma-Aldrich, St. Louis, Missouri, USA), NLRP3 activator nigericin sodium salt (Nig) (S6653, Selleck Chemicals, Houston, TX, USA), autophagy inhibitor chloroquine (CQ) (S6999, Selleck Chemicals), AMPK inhibitor compound C (CC) (S7840, Selleck Chemicals), ROS scavenger N-acetyl-L-cysteine (NAC) (T0875, TargetMol Chemicals, Boston, MA, USA).

Techniques: Expressing

Journal: Journal of animal science and biotechnology

Article Title: Methionine deficiency inhibited pyroptosis in primary hepatocytes of grass carp (Ctenopharyngodon idella): possibly via activating the ROS-AMPK-autophagy axis.

doi: 10.1186/s40104-024-01069-6

Figure Lengend Snippet: Fig. 9 Mechanism of MD induced autophagy but inhibited pyroptosis. MD promoted autophagy to inhibit pyroptosis through the ROS-AMPK signaling pathway of primary hepatocyte in grass carp. ROS, reactive oxygen species; LKB1, liver kinase B1; p-AMPK, phosphorylated-AMP-activated protein kinase; ULK1, Unc-51-like kinase 1; LC3, microtubule-associated protein 1 light chain 3; p62, sequestosome 1; NLRP3, NOD-like receptor thermal protein domain associated protein 3; ASC, apoptosis-associated speck-like protein containing a CARD; CASP-1, cysteinyl aspartate specific proteinase-1; IL-1β, interleukin-1β; GSDME, gasdermin E

Article Snippet: The following reagents were used in the experiments: LPS (L2880, Sigma-Aldrich, St. Louis, Missouri, USA), NLRP3 activator nigericin sodium salt (Nig) (S6653, Selleck Chemicals, Houston, TX, USA), autophagy inhibitor chloroquine (CQ) (S6999, Selleck Chemicals), AMPK inhibitor compound C (CC) (S7840, Selleck Chemicals), ROS scavenger N-acetyl-L-cysteine (NAC) (T0875, TargetMol Chemicals, Boston, MA, USA).

Techniques: