Protective Effect of Butylated Hydroxytoluene on the Hydrogen Peroxide-Induced Skeletal Muscle Atrophy in Cultured Myotubes

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Vikas Dutt^1,2,a#, Nirmal Jeet Kaur^1,b#, Onkar Sharma^1,c, and Ashwani Mittal^1,d*

¹Skeletal Muscle Lab, Institute of Integrated & Honors Studies, Kurukshetra University, Kurukshetra, Haryana, 136119, India

²Department of Life Sciences, School of Bio-Science and Technology, Sharda University, Greater Noida, Uttar Pradesh, 201310, India

^* To whom correspondence should be addressed.

^# These authors contributed equally to the work.

Received: March 3, 2025; Revised: September 27, 2025; Accepted: October 13, 2025

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Butylated hydroxytoluene (BHT) is a well-known synthetic antioxidant and a commonly used synthetic food additive. It is prominently employed in pharmaceutical, rubber, oil, and petroleum industries. However, the evidence supporting its role in preventing skeletal muscle atrophy is lacking. In this study, the effect of BHT on the oxidative stress (100 µM H₂O₂)-induced atrophy was investigated in C2C12 myotubes. The antioxidative potential of BHT was compared to that of β-carotene. BHT demonstrated a superior free radical-scavenging ability compared to β-carotene in both DPPH and ABTS assays. Furthermore, pretreatment with 25 μg/ml BHT for 4 h preserved myotube morphology and membrane integrity and promoted creatine kinase activity in the oxidative stress-induced atrophy model. BHT also prevented the degradation of myosin heavy chain (a key structural protein) by downregulating the activity of calpain and suppressing expression of MuRF-1 mRNA (ubiquitin proteasome system), as well as reduced lipid peroxidation and ROS levels and increased lactate dehydrogenase activity, indicating improved cellular resilience. This study provides the first direct evidence of the protective effects of BHT against H₂O₂-induced atrophy in cultured myotubes and highlights a therapeutic potential of BHT in the mitigation of oxidative stress-related muscle atrophy.

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KEY WORDS: butylated hydroxytoluene (BHT), muscle atrophy, oxidative stress

DOI: 10.1134/S0006297925603351

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