# Combining Network Pharmacology and Experimental Verification to Investigate the Protective Effect of Melatonin on Fluoride-Induced Brain Injury

## TL;DR

This study shows that melatonin protects against brain damage caused by fluoride exposure by improving mitochondrial function and regulating key signaling pathways.

## Contribution

The study combines network pharmacology and experimental validation to reveal melatonin's protective mechanism against fluoride-induced neurotoxicity.

## Key findings

- Melatonin improved cognitive impairments and structural hippocampal damage in rats and cells exposed to fluoride.
- Melatonin upregulated SIRT3 and downregulated HIF-1α, restoring mitochondrial function and ATP levels.
- The protective effect of melatonin is linked to the SIRT3/HIF-1α signaling pathway.

## Abstract

Excessive fluoride exposure induces developmental neurotoxicity, but effective preventive strategies are currently scarce. Melatonin (Mel), a lipophilic hormone secreted by the pineal gland, exerts antioxidant, anti-inflammatory, and neuroprotective properties. This study aimed to explore Mel’s protective effect and mechanism against fluoride-induced developmental brain injury. We employed a network pharmacology approach to screen the common targets of Mel and fluoride-induced brain injury and performed enrichment analysis. A total of 189 common targets were identified, and these targets were mainly enriched in the HIF-1 signaling pathway and oxidative stress-related pathways. In vivo, Sprague Dawley rats were subjected to perinatal sodium fluoride (NaF) exposure with/without Mel; in vitro, HT22 cells were subjected to NaF and/or Mel. The results showed that Mel improved cognitive impairments and alleviated structural damage to hippocampal neurons and mitochondria. Furthermore, Mel upregulated SIRT3 and downregulated HIF-1α, thereby restoring mitochondrial oxidative phosphorylation and ATP content. This study demonstrates that Mel alleviates fluoride-induced developmental neurotoxicity by improving mitochondrial function through regulating the SIRT3/HIF-1α signaling pathway. This not only offers a novel perspective for elucidating the underlying molecular mechanisms of fluoride-induced developmental neurotoxicity but also provides a theoretical foundation for Mel as a potential protective candidate against fluoride exposure.