2. Academic Validation
  3. AAD-2004 through clearing H2O2 reduces astrocyte proliferation and promotes neural regeneration after spinal cord injury

AAD-2004 through clearing H2O2 reduces astrocyte proliferation and promotes neural regeneration after spinal cord injury

  • Sci Rep. 2025 Dec 26. doi: 10.1038/s41598-025-33322-x.
Xiaodan Mi 1 2,  Keming Shan 1,  Xiangming Ye 1,  Ruidong Cheng 3
Affiliations

Affiliations

  • 1 Center for Rehabilitation Medicine, Department of Rehabilitation Medicine, Zhejiang Provincial People's Hospital (Affiliated People's Hospital, Hangzhou Medical College), Zhejiang Engineering Research Center for Digital-Intelligent Rehabilitation Equipment, Hangzhou, 310014, Zhejiang, China.
  • 2 Hangzhou First People's Hospital Xiasha Campus, Hangzhou Rehabilitation Hospital, Hangzhou, 310018, Zhejiang, China.
  • 3 Center for Rehabilitation Medicine, Department of Rehabilitation Medicine, Zhejiang Provincial People's Hospital (Affiliated People's Hospital, Hangzhou Medical College), Zhejiang Engineering Research Center for Digital-Intelligent Rehabilitation Equipment, Hangzhou, 310014, Zhejiang, China. [email protected].
PMID: 41454086 DOI: 10.1038/s41598-025-33322-x
Abstract

To assess the effect of AAD-2004 on spinal cord injury (SCI) and to explore its mechanism, we employed an in vitro model using OGD/R-challenged astrocytes to investigate the effects of AAD-2004 against cell death (terminal deoxynucleotidyl transferase dUTP nick-end labeling, tunel), oxidative stress (H2O2 level), and the expression of the key neuroprotective factor MAP2.AAD-2004[2-hydroxy-5-[2-(4-trifluoromethylphenyl)-ethylaminobenzoic acid] is a hydrogen peroxide(H2O2) scavenger primarily used for the treatment of amyotrophic lateral sclerosis and Alzheimer disease that has demonstrated certain neuroprotective properties. In parallel, modified allen's method was adopted, further exploring the potential molecular mechanism in vivo. Based on these conditions, histological and behavioral analysis were performed by Nissl staining, basso mouse scale and footprint analysis. The level of molecules associated with glial scar formation, nerve regeneration, axonal regeneration and H2O2 level were analyzed using western blot, immunofluorescence staining and H2O2 kit. AAD-2004 significantly improved the movement function after SCI and inhibited the proliferation of astrocytes, thus preventing the formation of glial scar by inhibiting of H2O2. At the same time, AAD-2004 promoted nerve regeneration, and the effect was due to neuronal regeneration and axonal regeneration pathways. The expression levels of GFAP and vimentin were significantly downregulated in AAD-2004-treated, and the expression level of Ki67 and PH3 were downregulated. The mean fluorescence intensity of neuronal regeneration (Neun+and MAP2+) and axonal regeneration-related (NF+ and GAP43+) were significantly upregulated after AAD-2004 treatment. Scavenging H2O2 level is a viable therapeutic strategy, and that AAD-2004 is prospective, and that scavenging H2O2 facilitated nerve regeneration and inhibited glial scar formation for SCI.

Keywords

Glial scar; Hydrogen peroxide; Neural regeneration; OGD/R; Spinal cord injury.

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