# Uncovering the Potential Mechanisms of Ergothioneine in Neuroinflammation Through Network Pharmacology, Molecular Docking, Molecular Dynamics Simulation, and In Vitro Validation

## TL;DR

This study explores how ergothioneine may reduce neuroinflammation by targeting key proteins and pathways, using computational and experimental methods.

## Contribution

The study provides a multi-omics and experimental validation of ergothioneine's anti-inflammatory mechanisms in neuroinflammation.

## Key findings

- EGT targets TNF, AKT1, CASP3, and IL-6 to regulate inflammation pathways.
- In vitro experiments showed EGT reduces pro-inflammatory cytokines in microglia cells.
- Molecular simulations confirmed stable binding between EGT and its targets.

## Abstract

Neuroinflammation is a critical pathological process implicated in several neurological disorders. It arises from complex interactions among immune cells and the excessive release of pro-inflammatory mediators, ultimately leading to neuronal damage. Ergothioneine (EGT), a naturally occurring antioxidant, has attracted attention for its potential anti-inflammatory role in neuroinflammation, although it remains poorly understood. We employed a comprehensive strategy combining network pharmacology, molecular docking, molecular dynamics simulations, and in vitro experiments to explore how EGT influences neuroinflammatory pathways. Computational analyses indicated that EGT might regulate several inflammation-related signaling cascades by targeting key molecules such as Tumor Necrosis Factor (TNF), AKT Serine/Threonine Kinase 1 (AKT1), Caspase 3 (CASP3), and Interleukin 6 (IL-6). Docking and dynamics simulations confirmed strong and stable binding between EGT and these targets. Experiments using lipopolysaccharide-stimulated BV2 microglia cells demonstrated that EGT significantly reduced pro-inflammatory cytokine production, primarily through modulation of the phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT) and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) signaling pathways. By integrating multi-omics approaches with cellular validation, this study sheds light on the molecular mechanisms underlying EGT’s anti-inflammatory effect and supports its potential application as a functional food ingredient for managing neuroinflammation.