# Anti-Inflammatory and Synaptic Protective Effects of TNF-α Inactivation in the MDX Mouse Model

## TL;DR

Blocking TNF-α with Etanercept reduces inflammation and protects synapses in a mouse model of Duchenne muscular dystrophy.

## Contribution

This study demonstrates that Etanercept reduces neuroinflammation and synaptic damage in dystrophic mice by blocking TNF-α signaling.

## Key findings

- Etanercept at 12 mg/Kg reduced astrogliosis and microglial activation in MDX mice.
- Treatment restored synaptic markers like synaptophysin and GAD-65 in the spinal cord.
- Etanercept improved locomotor function and downregulated Th1 lymphocyte polarization.

## Abstract

Background: Duchenne muscular dystrophy (DMD) is a severe neuromuscular disorder caused by the absence of functional dystrophin, leading to progressive muscle degeneration, inflammation, and alterations in the central nervous system. The sustained inflammatory response in DMD increases glial activation and the release of tumor necrosis factor alpha (TNF-α), which contributes to muscle fiber damage. Here, we investigated the anti-inflammatory and neuroprotective effects of Etanercept, a TNF-α receptor-blocking therapeutic, on the spinal cord of MDX mice, a widely used model of DMD. Methods: Adult male MDX and control C57BL/10 mice received vehicle or Etanercept (3, 6, or 12 mg/Kg, intraperitoneally (i.p.)) every 72 h for two weeks, along with daily gait assessment. At the end of treatment, flow cytometry and immunolabeling analyses were performed in the lumbar spinal cord. Results: Etanercept at 12 mg/Kg reduced astrogliosis and microglial activation; restored synaptic markers, including synaptophysin, glutamic acid decarboxylase 65 (GAD-65), and vesicular glutamate transporter 1 (VGLUT-1); and decreased pro-inflammatory cytokines. The treatment reduced GFAP+/TNF-α+ astrocytes and significantly downregulated Th1 lymphocyte polarization in treated MDX mice. These cellular effects were accompanied by improvements in locomotor function. Conclusions: Together, our findings indicate that TNF-α blockade by Etanercept exerts neuroprotective and anti-inflammatory actions in the spinal cord of dystrophic mice, providing new insights into the impact of TNF-α signaling on neuroinflammatory processes in DMD.