Hyperoside protects against poly-GR-mediated neurodegeneration via regulation of mitochondrial fission and oxidative stress in C9orf72-associated ALS

  • Chin Med. 2026 Jun 4;21(1):161. doi: 10.1186/s13020-026-01433-w.
Wen-Chi Hsieh  #  1  2 ,  Chun-Yu Lin  #  1  3 ,  Hsuan-Cheng Wu  1  2 ,  Eddie Feng-Ju Weng  4  5 ,  Shao-Ming Wang  6  7
Affiliations
  • 1. Neuroscience and Brain Disease Center, China Medical University, Taichung, 404328, Taiwan.
  • 2. Graduate Institute of Biomedical Sciences, College of Medicine, China Medical University, Taichung, 404328, Taiwan.
  • 3. School of Medicine, College of Medicine, China Medical University, Taichung, 404328, Taiwan.
  • 4. Neuroscience and Brain Disease Center, China Medical University, Taichung, 404328, Taiwan. [email protected].
  • 5. Graduate Institute of Biomedical Sciences, College of Medicine, China Medical University, Taichung, 404328, Taiwan. [email protected].
  • 6. Neuroscience and Brain Disease Center, China Medical University, Taichung, 404328, Taiwan. [email protected].
  • 7. Graduate Institute of Biomedical Sciences, College of Medicine, China Medical University, Taichung, 404328, Taiwan. [email protected].
  • # Contributed equally.
PMID: 42243993 DOI: 10.1186/s13020-026-01433-w
Abstract

Background: Arginine-rich poly-glycine-arginine (poly-GR), a toxic dipeptide repeat protein generated from C9orf72 hexanucleotide repeat expansion, drives mitochondrial dysfunction, oxidative stress, and neuronal loss in amyotrophic lateral sclerosis (ALS). Hyperoside, a bioactive flavonoid, exhibits antioxidant and cytoprotective properties, but its therapeutic relevance to C9orf72-associated ALS remains unclear.

Purpose: To determine whether hyperoside attenuates poly-GR-induced mitochondrial and oxidative injury and improves neuronal survival in cellular and animal models of C9orf72-ALS.

Methods: A combined in vitro and in vivo experimental study using motor neuron-like cells and an AAV-mediated neonatal mouse model of poly-GR toxicity. NSC34 cells expressing EGFP-GR50 were analyzed for mitochondrial morphology, membrane potential, ROS generation, antioxidant signaling, and Apoptosis using confocal microscopy, CellROX/MitoTracker assays, Western blot analysis, and viability testing. For in vivo assessment, neonatal mice received intracerebroventricular AAV9-EGFP-GR50 followed by intraperitoneal hyperoside (10 mg/kg). Survival, cerebral hemisphere length, and cortical NeuN⁺ neuron numbers were quantified.

Results: Poly-GR expression induced pronounced mitochondrial fragmentation, reduced membrane potential, elevated ROS, and suppressed Nrf2/HO-1/GPx4 signaling, accompanied by increased Drp1 and reduced Opa1 expression. Hyperoside reversed these abnormalities by restoring mitochondrial integrity, normalizing the Drp1/Opa1 balance, enhancing Nrf2 nuclear accumulation, and increasing the expression of HO-1 and GPx4. Hyperoside also reduced cleaved Caspase-3 and corrected the Bax/Bcl-2 ratio, improving cell viability under basal and oxidative stress conditions. In vivo, hyperoside modestly prolonged survival, increased cerebral hemisphere length, and significantly preserved cortical neuronal numbers in AAV9-EGFP-GR50 mice.

Conclusion: Hyperoside mitigates poly-GR-induced neurotoxicity by alleviating excessive mitochondrial fission, strengthening Nrf2-dependent antioxidant defenses, and suppressing Apoptosis. These findings support hyperoside as a promising multi-target therapeutic candidate for C9orf72-associated ALS.

Keywords
C9orf72-associated ALS; Hyperoside; Mitochondrial dysfunction; Nrf2; Oxidative stress.
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