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: rMyonectin protects cardiac mitochondria in LPS-induced sepsis-induced cardiomyopathy mice by promoting mitochondrial biogenesis and maintaining mitochondrial dynamics. (A) The ATP content in myocardial tissue. (B) Mitochondrial ultrastructure of myocardial tissue was observed by TEM. Magnification, ×3,000, ×7,000 and ×20,000. (C) Western blot analysis and semi-quantification of PGC-1α, NRF1, TFAM, OPA1, Mfn2, p-Drp1 at Ser616 and Drp1 in myocardial tissue. The data are presented as mean±SEM. ** P<0.01, *** P<0.001, **** P<0.0001. rMyonectin, recombinant myonectin; TEM, transmission electron microscopy; 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: The membranes were blocked with 5% skim milk for 1 h at room temperature and then incubated overnight at 4°C with the following primary antibodies: Bax (1:1,000; cat. no. ab32503; Abcam), Bcl-2 (1:1,000; cat. no. A19693; ABclonal Biotech Co., Ltd.), cleaved caspase-3 (1:600; cat. no. WL02117; Wanleibio Co., Ltd.), caspase-3 (1:500; cat. no. T40044 ; Abmart Pharmaceutical Technology Co., Ltd.), PGC-1α (1:2,000; cat. no. ab313559; Abcam), nuclear respiratory factor 1 (NRF1; 1:1,000; cat. no. A3252; ABclonal Biotech Co., Ltd.), mitochondrial transcription factor A (TFAM; 1:1,000; cat. no. 15218; Cell Signaling Technology, Inc.), mitofusin 2 (Mfn2; 1:3,000; cat. no. ab124773; Abcam), optic atrophy 1 (OPA1; 1:1,000; cat. no. ab157457; Abcam), phosphorylated (p)-dynamin-related protein 1 (Drp1; Ser616; 1:1,000; cat. no. ab314755; Abcam), Drp1 (1:4,000; cat. no. ab184247; Abcam), p-AMPK (1:1,000; cat. no. ab133448; Abcam), AMPK (1:1,000; cat. no. ab207442; Abcam), AdipoR1 (1:4,000; cat. no. bs-0610R; Bioss), β-actin (1:8,000; cat. no. P30002 ; Abmart Pharmaceutical Technology Co., Ltd.) and β-tubulin (1:10,000; cat. no. A12289; ABclonal Biotech Co., Ltd.).

Techniques: Western Blot, Recombinant, Transmission Assay, Electron Microscopy

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: rMyonectin protects cardiac mitochondria in CLP-induced sepsis-induced cardiomyopathy mice by promoting mitochondrial biogenesis and maintaining mitochondrial dynamics. (A) The ATP content in myocardial tissue. (B) Mitochondrial ultrastructure of myocardial tissue was observed by TEM. Magnification, ×3,000, ×7,000 and ×20,000. (C) Western blot analysis and semi-quantification of PGC-1α, NRF1, TFAM, OPA1, Mfn2, p-Drp1 at Ser616 and Drp1 in myocardial tissue. The data are presented as mean±SEM. * P<0.05, ** P<0.01, *** P<0.001, **** P<0.0001. rMyonectin, recombinant myonectin; CLP, cecal ligation and puncture; TEM, transmission electron microscopy; 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; p-, phosphorylated.

Article Snippet: The membranes were blocked with 5% skim milk for 1 h at room temperature and then incubated overnight at 4°C with the following primary antibodies: Bax (1:1,000; cat. no. ab32503; Abcam), Bcl-2 (1:1,000; cat. no. A19693; ABclonal Biotech Co., Ltd.), cleaved caspase-3 (1:600; cat. no. WL02117; Wanleibio Co., Ltd.), caspase-3 (1:500; cat. no. T40044 ; Abmart Pharmaceutical Technology Co., Ltd.), PGC-1α (1:2,000; cat. no. ab313559; Abcam), nuclear respiratory factor 1 (NRF1; 1:1,000; cat. no. A3252; ABclonal Biotech Co., Ltd.), mitochondrial transcription factor A (TFAM; 1:1,000; cat. no. 15218; Cell Signaling Technology, Inc.), mitofusin 2 (Mfn2; 1:3,000; cat. no. ab124773; Abcam), optic atrophy 1 (OPA1; 1:1,000; cat. no. ab157457; Abcam), phosphorylated (p)-dynamin-related protein 1 (Drp1; Ser616; 1:1,000; cat. no. ab314755; Abcam), Drp1 (1:4,000; cat. no. ab184247; Abcam), p-AMPK (1:1,000; cat. no. ab133448; Abcam), AMPK (1:1,000; cat. no. ab207442; Abcam), AdipoR1 (1:4,000; cat. no. bs-0610R; Bioss), β-actin (1:8,000; cat. no. P30002 ; Abmart Pharmaceutical Technology Co., Ltd.) and β-tubulin (1:10,000; cat. no. A12289; ABclonal Biotech Co., Ltd.).

Techniques: Western Blot, Recombinant, Ligation, Transmission Assay, Electron Microscopy

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: rMyonectin ameliorates 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; 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: The membranes were blocked with 5% skim milk for 1 h at room temperature and then incubated overnight at 4°C with the following primary antibodies: Bax (1:1,000; cat. no. ab32503; Abcam), Bcl-2 (1:1,000; cat. no. A19693; ABclonal Biotech Co., Ltd.), cleaved caspase-3 (1:600; cat. no. WL02117; Wanleibio Co., Ltd.), caspase-3 (1:500; cat. no. T40044 ; Abmart Pharmaceutical Technology Co., Ltd.), PGC-1α (1:2,000; cat. no. ab313559; Abcam), nuclear respiratory factor 1 (NRF1; 1:1,000; cat. no. A3252; ABclonal Biotech Co., Ltd.), mitochondrial transcription factor A (TFAM; 1:1,000; cat. no. 15218; Cell Signaling Technology, Inc.), mitofusin 2 (Mfn2; 1:3,000; cat. no. ab124773; Abcam), optic atrophy 1 (OPA1; 1:1,000; cat. no. ab157457; Abcam), phosphorylated (p)-dynamin-related protein 1 (Drp1; Ser616; 1:1,000; cat. no. ab314755; Abcam), Drp1 (1:4,000; cat. no. ab184247; Abcam), p-AMPK (1:1,000; cat. no. ab133448; Abcam), AMPK (1:1,000; cat. no. ab207442; Abcam), AdipoR1 (1:4,000; cat. no. bs-0610R; Bioss), β-actin (1:8,000; cat. no. P30002 ; Abmart Pharmaceutical Technology Co., Ltd.) and β-tubulin (1:10,000; cat. no. A12289; ABclonal Biotech Co., Ltd.).

Techniques: 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: 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: The membranes were blocked with 5% skim milk for 1 h at room temperature and then incubated overnight at 4°C with the following primary antibodies: Bax (1:1,000; cat. no. ab32503; Abcam), Bcl-2 (1:1,000; cat. no. A19693; ABclonal Biotech Co., Ltd.), cleaved caspase-3 (1:600; cat. no. WL02117; Wanleibio Co., Ltd.), caspase-3 (1:500; cat. no. T40044 ; Abmart Pharmaceutical Technology Co., Ltd.), PGC-1α (1:2,000; cat. no. ab313559; Abcam), nuclear respiratory factor 1 (NRF1; 1:1,000; cat. no. A3252; ABclonal Biotech Co., Ltd.), mitochondrial transcription factor A (TFAM; 1:1,000; cat. no. 15218; Cell Signaling Technology, Inc.), mitofusin 2 (Mfn2; 1:3,000; cat. no. ab124773; Abcam), optic atrophy 1 (OPA1; 1:1,000; cat. no. ab157457; Abcam), phosphorylated (p)-dynamin-related protein 1 (Drp1; Ser616; 1:1,000; cat. no. ab314755; Abcam), Drp1 (1:4,000; cat. no. ab184247; Abcam), p-AMPK (1:1,000; cat. no. ab133448; Abcam), AMPK (1:1,000; cat. no. ab207442; Abcam), AdipoR1 (1:4,000; cat. no. bs-0610R; Bioss), β-actin (1:8,000; cat. no. P30002 ; Abmart Pharmaceutical Technology Co., Ltd.) and β-tubulin (1:10,000; cat. no. A12289; ABclonal Biotech Co., Ltd.).

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 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: The membranes were blocked with 5% skim milk for 1 h at room temperature and then incubated overnight at 4°C with the following primary antibodies: Bax (1:1,000; cat. no. ab32503; Abcam), Bcl-2 (1:1,000; cat. no. A19693; ABclonal Biotech Co., Ltd.), cleaved caspase-3 (1:600; cat. no. WL02117; Wanleibio Co., Ltd.), caspase-3 (1:500; cat. no. T40044 ; Abmart Pharmaceutical Technology Co., Ltd.), PGC-1α (1:2,000; cat. no. ab313559; Abcam), nuclear respiratory factor 1 (NRF1; 1:1,000; cat. no. A3252; ABclonal Biotech Co., Ltd.), mitochondrial transcription factor A (TFAM; 1:1,000; cat. no. 15218; Cell Signaling Technology, Inc.), mitofusin 2 (Mfn2; 1:3,000; cat. no. ab124773; Abcam), optic atrophy 1 (OPA1; 1:1,000; cat. no. ab157457; Abcam), phosphorylated (p)-dynamin-related protein 1 (Drp1; Ser616; 1:1,000; cat. no. ab314755; Abcam), Drp1 (1:4,000; cat. no. ab184247; Abcam), p-AMPK (1:1,000; cat. no. ab133448; Abcam), AMPK (1:1,000; cat. no. ab207442; Abcam), AdipoR1 (1:4,000; cat. no. bs-0610R; Bioss), β-actin (1:8,000; cat. no. P30002 ; Abmart Pharmaceutical Technology Co., Ltd.) and β-tubulin (1:10,000; cat. no. A12289; ABclonal Biotech Co., Ltd.).

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: The membranes were blocked with 5% skim milk for 1 h at room temperature and then incubated overnight at 4°C with the following primary antibodies: Bax (1:1,000; cat. no. ab32503; Abcam), Bcl-2 (1:1,000; cat. no. A19693; ABclonal Biotech Co., Ltd.), cleaved caspase-3 (1:600; cat. no. WL02117; Wanleibio Co., Ltd.), caspase-3 (1:500; cat. no. T40044 ; Abmart Pharmaceutical Technology Co., Ltd.), PGC-1α (1:2,000; cat. no. ab313559; Abcam), nuclear respiratory factor 1 (NRF1; 1:1,000; cat. no. A3252; ABclonal Biotech Co., Ltd.), mitochondrial transcription factor A (TFAM; 1:1,000; cat. no. 15218; Cell Signaling Technology, Inc.), mitofusin 2 (Mfn2; 1:3,000; cat. no. ab124773; Abcam), optic atrophy 1 (OPA1; 1:1,000; cat. no. ab157457; Abcam), phosphorylated (p)-dynamin-related protein 1 (Drp1; Ser616; 1:1,000; cat. no. ab314755; Abcam), Drp1 (1:4,000; cat. no. ab184247; Abcam), p-AMPK (1:1,000; cat. no. ab133448; Abcam), AMPK (1:1,000; cat. no. ab207442; Abcam), AdipoR1 (1:4,000; cat. no. bs-0610R; Bioss), β-actin (1:8,000; cat. no. P30002 ; Abmart Pharmaceutical Technology Co., Ltd.) and β-tubulin (1:10,000; cat. no. A12289; ABclonal Biotech Co., Ltd.).

Techniques: Activation Assay, Recombinant