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GW8510 alleviates muscle atrophy and skeletal muscle dysfunction in mice through AMPK/PGC1α signaling

  • Authors:
    • Yutong Chen
    • Zurui Liu
    • Chen Liu
    • Daqian Yang
    • Mengmeng Xiao
    • Zhengqian Li
    • Zhengwei Xie
  • View Affiliations / Copyright

    Affiliations: Peking University International Cancer Institute, School of Basic Medical Sciences, Peking University Health Science Center, Beijing 100191, P.R. China, Beijing Gigaceuticals Tech. Co. Ltd., Beijing 102206, P.R. China
    Copyright: © Chen et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 128
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    Published online on: June 26, 2025
       https://doi.org/10.3892/ijmm.2025.5569
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Abstract

Preventing and restoring muscle loss and function is essential for elderly individuals. GW8510 may accelerate myotube differentiation. The present study aimed to investigate the protective effect of GW8510 (a CDK2 inhibitor) on muscle atrophy. Mouse models of muscle atrophy were induced by denervation, dexamethasone and glycerol. Muscle‑to‑body weight ratio, the cross‑sectional area of muscles, grip strength, fatigue and serum levels of superoxide dismutase and creatine kinase were assessed. In vitro, a dexamethasone‑induced C2C12 myotube atrophy model was used to evaluate mitochondrial function. Reverse transcription‑quantitative PCR, immunoblotting and small interfering RNA transfection were performed to explore the potential molecular mechanisms following treatment with GW8510. GW8510 resulted in a significant increase in the gastrocnemius and soleus muscle ratios in denervation mice (7 and 3%, respectively), alongside an increase in cross‑sectional area. Moreover, GW8510 significantly improved grip strength and superoxide dismutase activity, with similar protective effects in dexamethasone‑ and glycerol‑induced muscle atrophy models. GW8510 decreased reactive oxygen species production, increased mitochondrial DNA copy number, maintained mitochondrial dynamics and enhanced antioxidant activity in C2C12 myotubes. Mechanistically, GW8510 significantly inhibited the expression of atrophy‑associated markers F‑box protein 32 and tripartite motif‑containing 63 while activating AMPK (both P<0.01). The knockdown peroxisome proliferator‑activated receptor‑γ co‑activator‑1α (Pgc1α) negated the effects of GW8510. Overall, GW8510 mitigated muscle atrophy via the activation of the AMPK/PGC1α pathway. GW8510 could serve as a novel therapeutic agent for the prevention of muscle atrophy.
View Figures

Figure 1

GW8510 improves muscle atrophy and
weakness in sciatic nerve denervation mice. Mouse body weight and
muscle/body weight ratio of skeletal muscle. n=5-6/group.
#P<0.05, ####P<0.0001 vs. sham;
*P<0.05, **P<0.01 vs. vehicle. GC,
gastrocnemius; SOL, soleus; TA, tibialis anterior; EDL, extensor
digitorum longus; Quad, quadriceps.

Figure 2

GW8510 improves muscle atrophy in GC
and SOL in denervation mice. (A) Hematoxylin and eosin staining.
Scale bar, 10 μm. (B) Mean CSA distribution of GC muscle.
n=6/group. (C) Maximal grip strength in four limbs and latency to
fall off on day 4 after dissection of the sciatic nerve. (D)
Activity of SOD and creatine kinase. n=5-6/group.
##P<0.01, ####P<0.0001 vs. sham;
*P<0.05, ****P<0.0001 vs. vehicle. GC,
gastrocnemius; SOL, soleus; CSA, cross-sectional area; SOD,
superoxide dismutase.

Figure 3

GW8510 ameliorates DEX-induced
atrophy in C2C12 myotubes in vitro. (A) Viability of C2C12
myoblasts treated with GW8510 for 24 h. n=5/group. (B) Relative
expression of muscle atrophy-associated genes (Fbxo32 and Trim63)
in C2C12 myotubes. n=3/group. (C) Western blot analysis of muscle
atrophy-associated protein (Fbxo32, and Trim63). (D) Hematoxylin
and eosin staining and quantification of diameter of C2C12
myotubes. Scale bar, 50 μm. n=6/group. (E) Relative
expression of muscle atrophy-associated genes (Myog, Fbxo32 and
Trim63). (F) Expression levels of muscle atrophy-related protein
(Myog, Fbxo32 Trim63). n=3/group. (G) SOD and creatine kinase
activity. n=4/group. (H) Relative expression of muscle
fibrosis-related genes (Acta2 and Tgfb) in C2C12 myotubes
stimulated with DEX (10 μM) or vehicle, followed by
treatment with GW8510 (2 μM) or vehicle for 24 h. n=3/group.
#P<0.05, ##P<0.01,
###P<0.001, ####P<0.0001 vs. control;
*P<0.05, **P<0.01,
***P<0.001 vs. DEX. DEX, dexamethasone; Fbxo, F-box
protein; Trim, tripartite motif; SOD, superoxide dismutase.

Figure 4

GW8510 improves mitochondrial
function and biogenesis in C2C12 in vitro. (A) Reactive
oxygen species staining using DCFH-DA in C2C12 myotubes stimulated
with DEX (10 μM) or vehicle, followed by treatment with
GW8510 (2 μM) or vehicle for 24 h. Scale bar, 20 μm.
(B) Relative fluorescence intensity in C2C12 myotubes using ImageJ.
n=5/group. (C) Relative mitochondrial mass using Mitotracker
DeepRed staining. n=8/group. (D) Relative mtDNA copy number in
C2C12 myotubes stimulated with DEX (10 μM) or vehicle,
followed by treatment with GW8510 (2 μM) or vehicle for 24
h. n=4/group. (E) Western blot of mitochondrial fission and
fusion-related protein (Opa1, Mfn1, Drp1 and p-Drp1). (F) Relative
expression of mitochondrial biogenesis-related genes (Tfam, Sirt1,
Nrf1 and Pgc1α) in C2C12 myotubes stimulated with DEX (10
μM) or vehicle, followed by treatment with GW8510 (2
μM) or vehicle for 24 h. n=3/group. #P<0.05,
##P<0.01, ###P<0.001,
####P<0.0001 vs. control; *P<0.05,
**P<0.01, ****P<0.0001 vs. DEX. DEX,
dexamethasone; mtDNA, mitochondrial DNA; Mfn, mitofusin; p-Drp,
phosphorylated dynamin-related protein; Tfam, transcription factor
A, mitochondrial; Sirt1, sirtuin 1; Pgc, peroxisome
proliferator-activated receptor gamma, coactivator.

Figure 5

GW8510 increases NAD+
levels and ATP production and protects against oxidative stress.
(A) Levels of NAD+ and NADH, and the ratio of
NAD+/NADH in C2C12 myotubes. n=4/group. Concentration of
(B) ATP and (C) MDA in C2C12 myotubes stimulated with DEX (10
μM) or vehicle, followed by treatment with GW8510 (2
μM) or vehicle for 24 h. (D) Levels of NAD+ and
NADH, and the ratio of NAD+/NADH in GC tissue.
Concentration of (E) ATP and (F) MDA in GC tissue. (G). Activity of
GSH in GC tissue. n=4/group. (H) Activity of GSH in serum.
n=5/group. #P<0.05, ##P<0.01,
###P<0.001 vs. control/sham; *P<0.05,
**P<0.01, ***P<0.001 vs. DEX/vehicle.
MDA, malondialdehyde; DEX, dexamethasone; GC, gastrocnemius; GSH,
glutathione; prot, protein.

Figure 6

GW8510 exerts a protective effect on
expression of genes associated with muscle atrophy and development.
(A) Principal component analysis score plot on C2C12 myotubes
stimulated with DEX (10 μM) or vehicle, followed by
treatment with GW8510 (2 μM) or vehicle for 24 h. (B)
Overlap between genes down- and upregulated by dexamethasone and
GW8510. (C) Heatmap of muscle atrophy and development-associated
genes, including mitochondrial dynamics- (Fis1, Mfn1, Mfn2 and
Opa1), antioxidation-related genes (Sod2, Sod1 and Cat), myoblast
differentiation-related genes (Myf5, MyF6 and Myof),
mitophagy-related genes (Sqstm1, Pink1, Map1lc3b, Atg12, Atg5 and
Bnip4) and denervation-related genes (Chrna1, Gadd45a, Ncam1 and
Musk). (D) Kyoto Encyclopedia of Genes and Genomes pathway
enrichment analysis for significantly differentially expressed
genes. (E) Validation of relative mRNA expression of muscle
atrophy- and development-associated genes. n=3/group.
##P<0.01 vs. control; **P<0.01 vs. DEX.
DEX, dexamethasone; PC, principal component; Fis, fission,
mitochondrial 1; Mfn, mitofusin; Opa1, OPA1 mitochondrial dynamin
like GTPase; Sod, superoxide dismutase; Cat, catalase; Myf,
myogenic factor; Myof, myoferlin; Gadd45a, growth arrest and DNA
damage inducible alpha; Ncam, neural cell adhesion molecule; Chma,
cholinergic receptor nicotinic α; Pink, PTEN induced kinase; Sqstm,
sequestosome; Map1lc3b, microtubule associated protein 1 light
chain 3 beta; Atg, autophagy related; Bnip, BCL2 interacting
protein; Pgc, peroxisome proliferator-activated receptor γ,
coactivator.

Figure 7

GW8510 activates AMPK signaling
cascade in C2C12 myotubes and sciatic nerve denervation mice. (A)
Western blotting of Mstn, AMPKα and p-AMPKα and quantification of
relative protein levels in C2C12 myotubes stimulated with DEX (10
μM) or vehicle, followed by treatment with GW8510 (2
μM) or vehicle for 24 h. n=3/group. (B) Relative expression
of muscle atrophy-related genes (Myog, Fbxo32 and Trim63) in GC and
SOL tissue. n=3-4/group. (C) Western blot of muscle atrophy- (Myog,
Fbxo32, Trim63) and mitochondrial fission- and fusion-related
protein (Opa1, Mfn1and p-Drp1) and Mstn, AMPK, and p-AMPK in GC
tissue in non-denervated, denervated and denervated mice treated
with GW8510. (D) Relative protein levels. n=3/group.
#P<0.05, ##P<0.01,
###P<0.001, ####P<0.0001 vs.
control/sham; *P<0.05, **P<0.01,
***P<0.001 vs. DEX/vehicle. Mstn, myostatin; p-,
phosphorylated; DEX, dexamethasone; Myog, myogenin; Fbxo, F-box
protein; Trim, tripartite motif; GC, gastrocnemius; SOL, soleus;
Opa1, OPA1 mitochondrial dynamin like GTPase; Mfn, mitofusin; Drp,
Dynamin related protein.

Figure 8

GW8510 alleviates muscle atrophy via
the AMPK/PGC1α signaling cascade. (A) mRNA and (B) protein
expression of Cdk2 in C2C12 myotubes and in GC tissue. (C). Protein
expression of Pgc1α in GC tissue. (D) Western blotting of Pgc1α in
C2C12 myotubes after transfection of siRNAs targeting Pgc1α.
Western blotting of Pgc1α, Fbxo32, Trim63, Mfn1, Drp1, p-Drp1, Myog
and Mstn in C2C12 myotubes transfected with siRNA targeting Pgc1α,
followed by stimulation with (E) DEX (10 μM) or (F) TNFα (20
ng/ml) or vehicle and treatment with GW8510 (2 μM) or
vehicle for 24 h. n=3/group. #P<0.05,
##P<0.01, ###P<0.001 vs. control/sham;
*P<0.05, ***P<0.001,
****P<0.0001 vs. DEX/vehicle. PGC, peroxisome
proliferator-activated receptor γ, coactivator; Cdk, cyclin
dependent kinase; GC, gastrocnemius; si, Small Interfering; Fbxo,
F-box protein; Trim, tripartite motif; Mfn, mitofusin; p-Drp1,
phosphorylated Dynamin related protein 1; Myog, myogenin; Mstn,
myostatin; DEX, dexamethasone; NC, Negative control.

Figure 9

Proposed mechanism by which GW8510
alleviates skeletal muscle atrophy in vitro and in
vivo. GW8510 inhibited muscle atrophy related-genes (Fbxo32 and
Trim63) though activation of AMPK/PGC1α and decreases levels of ROS
and MDA, increased the levels of ATP and GSH, thus protected
against muscle atrophy. CSA, cross-sectional area; CK, creatine
kinase; SOD, superoxide dismutase; ROS, reactive oxygen species;
MDA, malondialdehyde; GSH, glutathione; PGC, peroxisome
proliferator-activated receptor gamma, coactivator; Fbxo, F-box
protein; Trim, tripartite motif. Figure created with BioGDP.com.
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Copy and paste a formatted citation
Spandidos Publications style
Chen Y, Liu Z, Liu C, Yang D, Xiao M, Li Z and Xie Z: GW8510 alleviates muscle atrophy and skeletal muscle dysfunction in mice through AMPK/PGC1&alpha; signaling. Int J Mol Med 56: 128, 2025.
APA
Chen, Y., Liu, Z., Liu, C., Yang, D., Xiao, M., Li, Z., & Xie, Z. (2025). GW8510 alleviates muscle atrophy and skeletal muscle dysfunction in mice through AMPK/PGC1&alpha; signaling. International Journal of Molecular Medicine, 56, 128. https://doi.org/10.3892/ijmm.2025.5569
MLA
Chen, Y., Liu, Z., Liu, C., Yang, D., Xiao, M., Li, Z., Xie, Z."GW8510 alleviates muscle atrophy and skeletal muscle dysfunction in mice through AMPK/PGC1&alpha; signaling". International Journal of Molecular Medicine 56.3 (2025): 128.
Chicago
Chen, Y., Liu, Z., Liu, C., Yang, D., Xiao, M., Li, Z., Xie, Z."GW8510 alleviates muscle atrophy and skeletal muscle dysfunction in mice through AMPK/PGC1&alpha; signaling". International Journal of Molecular Medicine 56, no. 3 (2025): 128. https://doi.org/10.3892/ijmm.2025.5569
Copy and paste a formatted citation
x
Spandidos Publications style
Chen Y, Liu Z, Liu C, Yang D, Xiao M, Li Z and Xie Z: GW8510 alleviates muscle atrophy and skeletal muscle dysfunction in mice through AMPK/PGC1&alpha; signaling. Int J Mol Med 56: 128, 2025.
APA
Chen, Y., Liu, Z., Liu, C., Yang, D., Xiao, M., Li, Z., & Xie, Z. (2025). GW8510 alleviates muscle atrophy and skeletal muscle dysfunction in mice through AMPK/PGC1&alpha; signaling. International Journal of Molecular Medicine, 56, 128. https://doi.org/10.3892/ijmm.2025.5569
MLA
Chen, Y., Liu, Z., Liu, C., Yang, D., Xiao, M., Li, Z., Xie, Z."GW8510 alleviates muscle atrophy and skeletal muscle dysfunction in mice through AMPK/PGC1&alpha; signaling". International Journal of Molecular Medicine 56.3 (2025): 128.
Chicago
Chen, Y., Liu, Z., Liu, C., Yang, D., Xiao, M., Li, Z., Xie, Z."GW8510 alleviates muscle atrophy and skeletal muscle dysfunction in mice through AMPK/PGC1&alpha; signaling". International Journal of Molecular Medicine 56, no. 3 (2025): 128. https://doi.org/10.3892/ijmm.2025.5569
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