Journal: Oxidative Medicine and Cellular Longevity

Article Title: Sodium Butyrate Ameliorates Oxidative Stress-Induced Intestinal Epithelium Barrier Injury and Mitochondrial Damage through AMPK-Mitophagy Pathway

doi: 10.1155/2022/3745135

Figure Lengend Snippet: Effects of sodium butyrate (NaB) on oxidative stress, intestinal epithelium barrier, and mitophagy of porcine intestinal epithelial cells (IPEC-J2) after inhibiting mitophagy or AMPK. (a–c) Superoxide dismutase (SOD), glutathione reductase (GSH) activity, and malondialdehyde (MDA) content of IPEC-J2 treated with Mdivi-1 or Compound C (CC). (d) Cellular mitochondrial membrane potential quantification by flow cytometry. (e) Cellular reactive oxygen species (ROS) level quantification by flow cytometry. (f) Intestinal epithelial transepithelial resistance (TER) and fluorescein isothiocyanate dextran 4 kDa (FD4) permeability. (g) Protein expression and quantification of tight junction Claudin-1, Occludin, and ZO-1. ∗ indicates a significant difference compared with the control group ( P < 0.05); # indicates a significant difference compared with the H2O2 group ( P < 0.05); & indicates a significant difference compared with the NaB+H2O2 group ( P < 0.05).

Article Snippet: Trypsin (Beyotime Biotechnology, China), phosphate-buffered saline (PBS) (Bozan Biotechnology, China), penicillin-streptomycin (Solabao Biotechnology, China), fetal bovine serum (Gemini, Australia), CCK-8 kit (Beyotime Biotechnology, China), MitoSpyTM Red CMXRos (Biolegend, USA), immunofluorescence fixative (Sevier Biotechnology, China), Triton-X 100 (Sigma-Aldrich, St. Louis, MO, USA), Glycine (Sinopharm Group, China), DAPI (Beyotime Biotechnology, China), Goat Anti-Mouse IgG Dylight 594 (Earthox, USA), Goat Anti-Mouse IgG Dylight 488 (Earthox, USA), mitochondrial division inhibitor (Mdivi-1) (Selleck, USA), AMPK inhibitor (Compound C, CC) (Selleck, USA), Lipofectamine RNAiMAX, and Lipofectamine 2000 were obtained from Invitrogen (Invitrogen, USA).

Techniques: Activity Assay, Membrane, Flow Cytometry, Permeability, Expressing, Control

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Sodium Butyrate Ameliorates Oxidative Stress-Induced Intestinal Epithelium Barrier Injury and Mitochondrial Damage through AMPK-Mitophagy Pathway

doi: 10.1155/2022/3745135

Figure Lengend Snippet: Effects of sodium butyrate (NaB) on mitophagy, oxidative stress, and intestinal epithelium barrier after interference with AMPK α . (a) Expression and quantification of mitophagy proteins PINK1, Parkin, and P62. (b–d) Superoxide dismutase (SOD), glutathione reductase (GSH) activity, and malondialdehyde (MDA) content of porcine intestinal epithelial cells (IPEC-J2). (e) Cellular reactive oxygen species (ROS) level of IPEC-J2. (f) Cellular mitochondrial membrane potential of IPEC-J2. (g) Protein expression and quantification of recombinant NLR family, pyrin domain-containing protein 3 (NLRP3) and Caspase-1. (h) Intestinal epithelial transepithelial resistance (TER) and fluorescein isothiocyanate dextran 4 kDa (FD4) permeability. ∗ indicates a significant difference compared with the control group ( P < 0.05).

Article Snippet: Trypsin (Beyotime Biotechnology, China), phosphate-buffered saline (PBS) (Bozan Biotechnology, China), penicillin-streptomycin (Solabao Biotechnology, China), fetal bovine serum (Gemini, Australia), CCK-8 kit (Beyotime Biotechnology, China), MitoSpyTM Red CMXRos (Biolegend, USA), immunofluorescence fixative (Sevier Biotechnology, China), Triton-X 100 (Sigma-Aldrich, St. Louis, MO, USA), Glycine (Sinopharm Group, China), DAPI (Beyotime Biotechnology, China), Goat Anti-Mouse IgG Dylight 594 (Earthox, USA), Goat Anti-Mouse IgG Dylight 488 (Earthox, USA), mitochondrial division inhibitor (Mdivi-1) (Selleck, USA), AMPK inhibitor (Compound C, CC) (Selleck, USA), Lipofectamine RNAiMAX, and Lipofectamine 2000 were obtained from Invitrogen (Invitrogen, USA).

Techniques: Expressing, Activity Assay, Membrane, Recombinant, Permeability, 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: 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: BioMed Research International

Article Title: Bilobalide Enhances AMPK Activity to Improve Liver Injury and Metabolic Disorders in STZ-Induced Diabetes in Immature Rats via Regulating HMGB1/TLR4/NF- κ B Signaling Pathway

doi: 10.1155/2021/8835408

Figure Lengend Snippet: BB modulated hepatic antioxidant in STZ-induced diabetic rats through AMPK activation and HMGB1/TLR4/NF- κ B signaling pathway. The one-day-old immature rats were randomly allocated to 5 groups: healthy control, diabetes mellitus model (DM), STZ+BB (10 mg/kg), STZ+AMPK inhibitor (compound C, CC), STZ+BB (10 mg/kg)+CC. (a, b) The expressions of AMPK α 1, HMGB1, TLR4, P65, and p-P65 in hepatic tissues were assayed using Western blot analysis. β -Actin is a loading control. (c) The degree of liver damage was identified by H&E staining and Suzuki score. (d–g) Glucose, TG, AST, and ALT levels were determined using commercial kits. ∗∗ P < 0.01 (vs. control); # P < 0.05 (vs. DM); ## P < 0.01 (vs. DM); & P < 0.05 (vs. DM+BB (10 mg/kg)).

Article Snippet: AMPK inhibitor dorsomorphin (compound C, CC) was purchased from Selleck, Inc., USA.

Techniques: Activation Assay, Control, Western Blot, Staining