# Inhalation of Ultrafine Carbon Black-Induced Mitochondrial Dysfunction in Mouse Heart Through Changes in Acetylation

## TL;DR

Repeated inhalation of ultrafine carbon black harms heart mitochondria by disrupting energy production and increasing oxidative stress through protein acetylation.

## Contribution

This study reveals a novel mechanism of mitochondrial dysfunction in the heart caused by ultrafine carbon black exposure through acetylation changes.

## Key findings

- Repeated CB inhalation impairs fatty acid oxidation and disrupts ETC supercomplex assembly in the heart.
- CB exposure increases mitochondrial protein hyperacetylation and reduces SOD2 activity, contributing to oxidative stress.
- Elevated GCN5L1 levels and lipid peroxidation are linked to mitochondrial dysfunction and antioxidant defense weakening.

## Abstract

Air pollution, particularly from fine and ultrafine particulate matter (PM), has been increasingly associated with cardiovascular diseases. Ultrafine carbon, a component of ultrafine PM widely used in industrial settings, is both an environmental and occupational hazard. But the cardiac toxicity of repeated inhalation exposure to ultrafine carbon black (CB) remains unclear. In this study, we investigated how repeated inhalation of CB affects cardiac mitochondrial function, focusing on metabolic pathways and regulatory mechanisms involved in energy production. Male C57BL/6J mice were exposed to either filtered air or CB aerosols (10 mg/m3) for four consecutive days. Cardiac tissues were collected and analyzed to assess changes in metabolic enzyme activity, protein expression, and mitochondrial function using Western blotting, enzymatic assays, and immunoprecipitation. Despite there being few changes in overall protein expression levels, we observed significant impairments in fatty acid oxidation, increased glucose oxidation, and disrupted electron transport chain (ETC) supercomplex assembly, particularly in Complexes III and IV. These changes were accompanied by increased hyperacetylation of mitochondrial proteins and elevated levels of GCN5L1, a mitochondrial acetyltransferase. We also found increased lipid peroxidation and hyperacetylation of antioxidant enzyme SOD2 at the K-122 site, which reflects reduced enzymatic activity contributing to oxidative stress. Our findings suggest that repeated CB inhalation leads to mitochondrial dysfunction in the heart by dysregulating substrate utilization, impairing ETC activities, and weakening antioxidant defenses primarily through lysine acetylation. These findings reveal a potential role of key post-translational mechanisms in environmental particulate exposure to mitochondrial impairment and provide a potential therapeutic target for CB-induced cardiotoxicity.