ampk inhibitor compound c  (MedChemExpress)

Journal: International Journal of Molecular Medicine

Article Title: Recombinant myonectin ameliorates sepsis-induced cardiomyopathy by alleviating mitochondrial dysfunction via the AdipoR1/AMPK pathway

doi: 10.3892/ijmm.2026.5900

Figure Lengend Snippet: Protective effect of rMyonectin against LPS-induced apoptosis in cardiomyocytes is mediated by AMPK activation. (A) Western blot analysis and semi-quantification of p-AMPK and AMPK in myocardial tissue. (B) Western blot analysis and semi-quantification of p-AMPK and AMPK in NMCMs. (C) Expression of p-AMPK and AMPK in NMCMs after CC treatment. (D) Intracellular LDH activity in NMCMs. (E) The percentage of apoptotic cells detected using flow cytometry. (F) Apoptosis was assessed using flow cytometry after double labeling with Annexin V-FITC and PI. (G) Western blot analysis and semi-quantification of cleaved caspase-3, caspase-3, Bax and Bcl-2 protein expression in NMCMs. The data are presented as mean±SEM. * P<0.05, ** P<0.01, *** P<0.001, **** P<0.0001. rMyonectin, recombinant myonectin; NMCMs, neonatal mouse cardiomyocytes; AMPK, AMP-activated protein kinase; CC, Compound C; LDH, lactate dehydrogenase; LPS, lipopolysaccharide; PI, propidium iodide; p-, phosphorylated.

Article Snippet: NMCMs were pretreated for 2 h with rMyonectin or with the AMPK inhibitor Compound C (CC; 10 μ M; cat. no. HY-13418A; MedChemExpress) ( , ), followed by stimulation with 10 μ g/ml LPS for 24 h ( ).

Techniques: Activation Assay, Western Blot, Expressing, Activity Assay, Flow Cytometry, Labeling, Recombinant

Journal: International Journal of Molecular Medicine

Article Title: Recombinant myonectin ameliorates sepsis-induced cardiomyopathy by alleviating mitochondrial dysfunction via the AdipoR1/AMPK pathway

doi: 10.3892/ijmm.2026.5900

Figure Lengend Snippet: Protective effect of rMyonectin against LPS-induced mitochondrial dysfunction in cardiomyocytes is mediated by AMPK activation. (A) The ATP content in NMCMs. (B) Relative OCR. (C) Detection of the activities of mitochondrial respiratory chain complexes I and III. (D) Analysis of MMP using JC-1 staining. Scale bar, 50 μ m. (E) Western blot analysis and semi-quantification of PGC-1α, NRF1, TFAM, OPA1, Mfn2, p-Drp1 at Ser616, and Drp1 in NMCMs. The data are presented as mean±SEM. * P<0.05, ** P<0.01, *** P<0.001, **** P<0.0001. rMyonectin, recombinant myonectin; NMCMs, neonatal mouse cardiomyocytes; OCR, oxygen consumption rate; MMP, mitochondrial membrane potential; PGC-1α, peroxisome proliferator-activated receptor γ co-activator-1 α; NRF1, nuclear respiratory factor 1; TFAM, mitochondrial transcription factor A; Mfn2, mitofusin 2; OPA1, optic atrophy 1; Drp1, dynamin-related protein 1; LPS, lipopolysaccharide; p-, phosphorylated.

Article Snippet: NMCMs were pretreated for 2 h with rMyonectin or with the AMPK inhibitor Compound C (CC; 10 μ M; cat. no. HY-13418A; MedChemExpress) ( , ), followed by stimulation with 10 μ g/ml LPS for 24 h ( ).

Techniques: Activation Assay, Staining, Western Blot, Recombinant, Membrane

Journal: International Journal of Molecular Medicine

Article Title: Recombinant myonectin ameliorates sepsis-induced cardiomyopathy by alleviating mitochondrial dysfunction via the AdipoR1/AMPK pathway

doi: 10.3892/ijmm.2026.5900

Figure Lengend Snippet: AdipoR1 knockdown abolishes the protective effect of rMyonectin against LPS-induced apoptosis in cardiomyocytes. (A) Western blot analysis and semi-quantification of AdipoR1 in myocardial tissue. (B) Western blot analysis and semi-quantification of AdipoR1 in NMCMs. (C) Representative western blots showing the expression of AdipoR1, p-AMPK and AMPK in NMCMs following AdipoR1 knockdown. (D) Semi-quantification of AdipoR1, p-AMPK and AMPK protein levels in NMCMs following AdipoR1 knockdown. (E) Intracellular LDH activity in NMCMs. (F) The percentage of apoptotic cells detected using flow cytometry. (G) Apoptosis was assessed using flow cytometry after double labeling with Annexin V-FITC and PI. (H) Western blot analysis and semi-quantification of cleaved caspase-3, caspase-3, Bax, and Bcl-2 protein expression in NMCMs. The data are presented as mean±SEM. * P<0.05, ** P<0.01, *** P<0.001, **** P<0.0001. rMyonectin, recombinant myonectin; NMCMs, neonatal mouse cardiomyocytes; AdipoR1, adiponectin receptor 1; AMPK, AMP-activated protein kinase; LDH, lactate dehydrogenase; si, small interfering RNA; NC, negative control; siAdipoR1, siRNA targeting AdipoR1; LPS, lipopolysaccharide; p-, phosphorylated; PI, propidium iodide.

Article Snippet: NMCMs were pretreated for 2 h with rMyonectin or with the AMPK inhibitor Compound C (CC; 10 μ M; cat. no. HY-13418A; MedChemExpress) ( , ), followed by stimulation with 10 μ g/ml LPS for 24 h ( ).

Techniques: Knockdown, Western Blot, Expressing, Activity Assay, Flow Cytometry, Labeling, Recombinant, Small Interfering RNA, Negative Control

Journal: International Journal of Molecular Medicine

Article Title: Recombinant myonectin ameliorates sepsis-induced cardiomyopathy by alleviating mitochondrial dysfunction via the AdipoR1/AMPK pathway

doi: 10.3892/ijmm.2026.5900

Figure Lengend Snippet: AdipoR1 knockdown abolishes the protective effect of rMyonectin against LPS-induced mitochondrial dysfunction in cardiomyocytes. (A) The ATP content in NMCMs. (B) Relative OCR. (C) Detection of the activities of mitochondrial respiratory chain complexes I and III. (D) Analysis of MMP using JC-1 staining. Scale bar, 50 μ m. (E) Western blot analysis and semi-quantification of PGC-1α, NRF1, TFAM, OPA1, Mfn2, p-Drp1 at Ser616 and Drp1 in NMCMs. The data are presented as mean±SEM. * P<0.05, ** P<0.01, *** P<0.001, **** P<0.0001. rMyonectin, recombinant myonectin; NMCMs, neonatal mouse cardiomyocytes; AdipoR1, adiponectin receptor 1; AMPK, AMP-activated protein kinase; OCR, oxygen consumption rate; MMP, mitochondrial membrane potential; PGC-1α, peroxisome proliferator-activated receptor γ co-activator-1 α; NRF1, nuclear respiratory factor 1; TFAM, mitochondrial transcription factor A; Mfn2, mitofusin 2; OPA1, optic atrophy 1; Drp1, dynamin-related protein 1; si, small interfering RNA; NC, negative control; siAdipoR1, siRNA targeting AdipoR1; LPS, lipopolysaccharide; p-, phosphorylated.

Article Snippet: NMCMs were pretreated for 2 h with rMyonectin or with the AMPK inhibitor Compound C (CC; 10 μ M; cat. no. HY-13418A; MedChemExpress) ( , ), followed by stimulation with 10 μ g/ml LPS for 24 h ( ).

Techniques: Knockdown, Staining, Western Blot, Recombinant, Membrane, Small Interfering RNA, Negative Control

Journal: International Journal of Molecular Medicine

Article Title: Recombinant myonectin ameliorates sepsis-induced cardiomyopathy by alleviating mitochondrial dysfunction via the AdipoR1/AMPK pathway

doi: 10.3892/ijmm.2026.5900

Figure Lengend Snippet: Molecular mechanism by which rMyonectin ameliorates SIC. rMyonectin ameliorates SIC by alleviating mitochondrial dysfunction and inhibiting cardiomyocyte apoptosis via activation of the AdipoR1/AMPK pathway. rMyonectin, recombinant myonectin; SIC, sepsis-induced cardiomyopathy; OMM, outer mitochondrial membranes; IMM, inner mitochondrial membranes; AdipoR1, adiponectin receptor 1; AMPK, AMP-activated protein kinase; PGC-1α, peroxisome proliferator-activated receptor γ co-activator-1 α; NRF1, nuclear respiratory factor 1; TFAM, mitochondrial transcription factor A; Mfn2, mitofusin 2; OPA1, optic atrophy 1; Drp1, dynamin-related protein 1; I, mitochondrial respiratory chain complex I; III, mitochondrial respiratory chain complex III; p-, phosphorylated.

Article Snippet: NMCMs were pretreated for 2 h with rMyonectin or with the AMPK inhibitor Compound C (CC; 10 μ M; cat. no. HY-13418A; MedChemExpress) ( , ), followed by stimulation with 10 μ g/ml LPS for 24 h ( ).

Techniques: Activation Assay, Recombinant

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: Journal of Advanced Research

Article Title: Prophylactic supplementation with biogenic selenium nanoparticles mitigated intestinal barrier oxidative damage through suppressing epithelial-immune crosstalk with gut-on-a-chip

doi: 10.1016/j.jare.2025.04.023

Figure Lengend Snippet: Biogenic SeNPs regulated AMPK/NLRP3/Nrf2 signaling pathway to alleviate intestinal epithelial barrier oxidative damage in gut-on-a-chip. A. Schematic diagram of experimental design. B. pAMPK/NLRP3/Nrf2 immunofluorescent staining (Scale bar: 40 μm). C. Villi-like height. D. LDH activity in the upper channel layer. E. IL-1β levels in the upper channel layer. F. IL-18 levels in the upper channel layer. G. Schematic diagram of the mechanism by which SeNPs exert antioxidant effects against oxidative damage to intestinal epithelial cells in vitro . Data are expressed as mean ± S.E.M. n = 4. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: To investigate the role of the AMPK signaling pathway in the protection of the intestinal epithelial barrier from oxidative stress damage by SeNPs, AMPK activator AICAR (MedChemExpres; Cat# HY-13417) and AMPK inhibitor Dorsomorphin (MedChemExpres; Cat# HY-13418A) were introduced into the gut-on-a-chip, respectively.

Techniques: Staining, Activity Assay, In Vitro

Journal: Journal of Advanced Research

Article Title: Prophylactic supplementation with biogenic selenium nanoparticles mitigated intestinal barrier oxidative damage through suppressing epithelial-immune crosstalk with gut-on-a-chip

doi: 10.1016/j.jare.2025.04.023

Figure Lengend Snippet: Biogenic SeNPs regulated AMPK/NLRP3/Nrf2 signaling pathway to attenuate oxidative stress-induced intestinal barrier dysfunction and mast cell overactivation in mice. A. Immunofluorescence analysis of p-AMPK (red), NLRP3 (yellow) and Nrf2 (green) in jejunum of mice with different treatments (Scale bar: 100 μm). B. Western blot analysis of pAMPK and Nrf2 expression levels in mice jejunum. C. Western blot analysis of NLRP3 and its downstream pyroptosis-related protein expression levels in mice jejunum. Data are expressed as mean ± S.E.M. n = 3. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: To investigate the role of the AMPK signaling pathway in the protection of the intestinal epithelial barrier from oxidative stress damage by SeNPs, AMPK activator AICAR (MedChemExpres; Cat# HY-13417) and AMPK inhibitor Dorsomorphin (MedChemExpres; Cat# HY-13418A) were introduced into the gut-on-a-chip, respectively.

Techniques: Immunofluorescence, Western Blot, Expressing