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: Bioactive Materials

Article Title: Melatonin-incorporated brain extracellular matrix hydrogel enhances NSCs mitochondrial metabolism to promote neuroregeneration via the AMPK-PGC-1α-NRF1/TFAM axis after spinal cord injury

doi: 10.1016/j.bioactmat.2026.04.006

Figure Lengend Snippet: Melatonin activates AMPK signaling and enhances mitochondrial function in vitro. (A) GO enrichment bar plot of differentially expressed genes (DEGs) between Control and Melatonin-treated NSCs. (B) KEGG pathway enrichment bar plot of DEGs between Control and Melatonin groups. (C) Heatmap of selected DEGs associated with neuronal differentiation and mitochondrial function. DEGs were defined as transcripts with FDR <0.05. (D) Representative Western blots showing phosphorylated AMPK (p-AMPK, Thr172) and phosphorylated ACC (p-ACC, Ser79) in Control, Melatonin, Inhibitor, and Melatonin + Inhibitor groups. (E) Densitometric analysis of p-AMPK/total AMPK and p-ACC/GAPDH ratios. (F) RT-qPCR analysis of Ppargc1a and Tfam expression, normalized to GAPDH and presented as fold change relative to the Control group. (G) Representative Western blots of mitochondrial oxidative phosphorylation (OXPHOS) complexes I-V. (H) Densitometric quantification of OXPHOS complexes I-V, normalized to GAPDH (or the corresponding loading control). (I) Representative JC-1 fluorescence images indicating mitochondrial membrane potential (ΔΨm). (J) Quantification of the red/green JC-1 fluorescence ratio from (I). (K) Schematic representation of the proposed melatonin-AMPK-ACC-PGC-1α-NRF1/TFAM signaling axis driving mitochondrial biogenesis in NSCs. All quantitative data (E, F, H, J) are presented as mean ± SD. Statistical significance was assessed using one-way ANOVA followed by Holm–Sidak's multiple comparisons test. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001. K created with BioRender.com .

Article Snippet: For AMPK inhibition experiments, BAY-3827 (HY-112083, MedChemExpress, USA), a selective AMPK inhibitor, was used at a final concentration of 2 μM for 24 h. The mitochondrial membrane potential was measured using the JC-1 Mitochondrial Membrane Potential Assay Kit (C2003S, Beyotime Biotechnology, China).

Techniques: In Vitro, Control, Western Blot, Quantitative RT-PCR, Expressing, Phospho-proteomics, Fluorescence, Membrane

Journal: Bioactive Materials

Article Title: Melatonin-incorporated brain extracellular matrix hydrogel enhances NSCs mitochondrial metabolism to promote neuroregeneration via the AMPK-PGC-1α-NRF1/TFAM axis after spinal cord injury

doi: 10.1016/j.bioactmat.2026.04.006

Figure Lengend Snippet: Molecular validation of neural repair and mechanism activation in spinal cord tissue. Western blot and qPCR analyses of spinal cord tissue lysates from Sham, SCI, BEM, NSCs@BEM, and NSCs@MT/BEM groups. (A) Representative Western blots for the neuronal marker TUJ1 and the glial scar marker GFAP. (B) Representative Western blots for phosphorylated AMPK (p-AMPK), phosphorylated ACC (p-ACC), and their respective total proteins. (C) Representative Western blots for the five oxidative phosphorylation (OXPHOS) complex subunits. (D) Densitometric quantification of TUJ1 and GFAP protein levels. (E) Densitometric quantification of the p-AMPK/total AMPK and p-ACC/total ACC ratios. (F) Densitometric quantification of OXPHOS complex protein levels. (G) Relative mRNA expression of neural markers (TUJ1, GFAP, Olig2) and key mitochondrial biogenesis regulators (Ppargc1a, Tfam) determined by qPCR. Data are presented as mean ± SD. Statistical significance was determined by one-way ANOVA with Holm–Sidak's multiple comparisons test. (∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001).

Article Snippet: For AMPK inhibition experiments, BAY-3827 (HY-112083, MedChemExpress, USA), a selective AMPK inhibitor, was used at a final concentration of 2 μM for 24 h. The mitochondrial membrane potential was measured using the JC-1 Mitochondrial Membrane Potential Assay Kit (C2003S, Beyotime Biotechnology, China).

Techniques: Biomarker Discovery, Activation Assay, Western Blot, Marker, Phospho-proteomics, Expressing

Journal: Neural Regeneration Research

Article Title: Trans-spinal magnetic stimulation upregulates microglial SOCS3 to attenuate neuroinflammation in chronic constriction injury–induced neuropathic pain

doi: 10.4103/NRR.NRR-D-24-00912

Figure Lengend Snippet: rMS-induced upregulation of SOCS3 in microglia is CaMKKβ/AMPK-dependent in vitro . (A) A schematic diagram of the in vitro study. (B) Western blotting data indicating that the p-CaMKKβ protein level in BV-2 cells increased at 5 minutes after a single administration of rMS and was highest at 15 minutes post-rMS ( n = 4). (C) Western blotting data showing that the p-AMPK protein level in BV-2 cells increased significantly at 30 minutes after rMS ( n = 4). (D, E) Representative western blotting results show that STO-609 and compound C alleviated rMS-induced activation of AMPK in vitro ( n = 4). (F–H) Representative western blotting data showing that compound C abrogated rMS-induced upregulation of SOCS3 and inhibition of p38 and STAT3 in vitro ( n = 4). (I) Representative immunofluorescence images showing that rMS inhibited the translocation of NF-κB p65 (green, Alexa Fluor 488) from the cytosol to the nucleus after LPS treatment in vitro , where this effect was reversed by administration of compound C (20 μmol/mL) 0.5 hours before LPS pretreatment ( n = 3). Arrows indicate BV-2 cells. Scale bars: 10 μm. Data are expressed as mean ± SD. # P < 0.05, ## P < 0.01, vs . Control group; * P < 0.05, ** P < 0.01, **** P < 0.001, vs . LPS group; † P < 0.05, †† P < 0.01, vs. LPS + rMS group (one‐way analysis of variance followed by Tukey’s multiple comparisons tests). AMPK: Adenosine 5′-monophosphate-activated protein kinase; CaMKKβ: calmodulin-dependent protein kinase kinase beta; Compound C: an AMPK inhibitor; DAPI: 4′,6-diamidino-2-phenylindole; LPS: lipopolysaccharide; NF-κB: nuclear factor kappa-B; p-AMPK: phosphorylated AMPK(Thr172); p-CaMKKβ: phospho-CaMKKβ(Ser511); rMS: repetitive magnetic stimulation; SOCS3: suppressor of cytokine signaling 3; STAT3: transducer and activator of transcription 3; STO-609: a CaMKKβ inhibitor.

Article Snippet: Intrathecal injections of compound C, an adenosine 5′-monophosphate-activated protein kinase (AMPK) inhibitor (10 μg/10 μL; MedChemExpress, Newark, NJ, USA, Cat# HY-13418A), and SOCS3 siRNA (500 pmol/10 μL; Santa Cruz Biotechnology, Santa Cruz, CA, USA, Cat# SC-270156) were performed as described previously by Mestre et al. (1994).

Techniques: In Vitro, Western Blot, Activation Assay, Inhibition, Immunofluorescence, Translocation Assay, Control

Journal: Neural Regeneration Research

Article Title: Trans-spinal magnetic stimulation upregulates microglial SOCS3 to attenuate neuroinflammation in chronic constriction injury–induced neuropathic pain

doi: 10.4103/NRR.NRR-D-24-00912

Figure Lengend Snippet: The pain-relieving effect and upregulation of SOCS3 induced by focal rTSMS depended on the activation of AMPK. (A) Timeline diagram of CCI modeling, rTSMS treatment, mechanical withdrawal threshold assessment, and experimental analysis in rats. (B) Western blotting data illustrates that the p-AMPK protein level decreased in CCI model rats and increased in CCI + rTSMS + Vehicle rats ( n = 5). (C, D) Intrathecal injection of compound C largely prevented the pain-relieving effects of focal rTMS, when applied once or repeatedly, in rats with CCI neuropathic pain ( n = 8). (E–H) Representative western blotting results showing that compound C alleviated the rTSMS-induced activation of AMPK, increase in SOCS3, and downregulation of p38 and STAT3 ( n = 5). (I, J) Representative immunofluorescence images showing that compound C abrogated the rTSMS-induced upregulation of SOCS3 (green, Alexa Fluor 488) in the dorsal horn of the spinal cord ( n = 3). Dashed box indicates the region of interest in the dorsal horn of the spinal cord. Scale bars: 100 μm. Data are expressed as mean ± SD. # P < 0.05, ## P < 0.01, ### P < 0.001, vs. Sham + Vehicle group; * P < 0.05, ** P < 0.01, **** P < 0.0001, vs . CCI + Vehicle group; † P < 0.05, †† P < 0.01, ††† P < 0.001, †††† P < 0.0001, vs . CCI + rTSMS + Compound C group (C, D: two‐way analysis of variance; B, E–H, J: one‐way analysis of variance followed by Tukey’s multiple comparisons tests). AMPK: Adenosine 5′-monophosphate-activated protein kinase; CCI: chronic constrictive injury; DAPI: 4′,6-diamidino-2-phenylindole; Iba-1: ionized calcium binding adaptor molecule-1; p-AMPK: phosphorylated AMPK(Thr172); p-p38: phospho-p38 mitogen-activated protein kinase; p-STAT3: phospho-STAT3; rTSMS: repetitive trans-spinal magnetic stimulation; SOCS3: suppressor of cytokine signaling 3; STAT3: transducer and activator of transcription 3.

Article Snippet: Intrathecal injections of compound C, an adenosine 5′-monophosphate-activated protein kinase (AMPK) inhibitor (10 μg/10 μL; MedChemExpress, Newark, NJ, USA, Cat# HY-13418A), and SOCS3 siRNA (500 pmol/10 μL; Santa Cruz Biotechnology, Santa Cruz, CA, USA, Cat# SC-270156) were performed as described previously by Mestre et al. (1994).

Techniques: Activation Assay, Western Blot, Injection, Immunofluorescence, Binding Assay

Journal: Journal of Biomedical Research

Article Title: Remote neuromuscular electrical stimulation upregulates MDK to enhance macrophage efferocytosis via LRP1 in wound healing

doi: 10.7555/JBR.38.20240375

Figure Lengend Snippet: Electrical stimulation upregulated MDK through the AMPK-ERK axis. A: rNMES was applied for 3 days (100 Hz, 3 mA). MDK protein levels in the skeletal muscle. * P < 0.05 and ** P < 0.01 vs. the 0 h group. B: Compound C (20 μmol/L) was used 15 min before ES (0.1 mA, 20 Hz, 1 h) to inhibit AMPK. p-AMPK and p-ERK protein levels in C2C12, and MDK protein levels in the supernatant of the cell cultures. C: SCH772984 (2 μmol/L) was used 30 min before ES (0.1 mA, 4 Hz, 1 h) to inhibit ERK. p-ERK protein levels in C2C12 and MDK protein levels in the supernatant of the cell cultures. Protein levels were analyzed by Western blotting. All data are presented as mean ± standard error of the mean. * P < 0.05 and *** P < 0.001 vs. the control group, # P < 0.05 and ### P < 0.001 vs. the ES group (B–C). Statistical analysis was performed using one-way ANOVA followed by Tukey's multiple comparisons test. Abbreviations: MDK, midkine; p-AMPK, phospho-AMP-activated protein kinase; p-ERK, phospho-extracellular signal-regulated kinase; rNMES, remote neuromuscular electrical stimulation; ES, electrical stimulation.

Article Snippet: Compound C (Cat. #HY-13418A, MedChemExpress) was used to inhibit AMPK, and SCH772984 (Cat. #S7101, Selleck Chemicals, Houston, TX, USA) was used to inhibit ERK1/2.

Techniques: Western Blot, Control