Objective <p>This study investigates the role of transient receptor potential vanilloid subtype 1 (TRPV1) in acute hypoxic exercise.</p> Methods <p>After acute hypoxia intervention, the mRNA expression levels of TRPV1 and 5-hydroxytryptamine 1&#xa0;A (5-HT1A) in the prefrontal cortex of rats were detected by real-time quantitative polymerase chain reaction (RT-PCR). Meanwhile, the content of 5-hydroxytryptamine (5-HT) in this area was determined by enzyme-linked immunosorbent assay (ELISA).</p> Results <p>Hypoxic conditions significantly reduced the duration of high-load exercise performance in rats, resulting in an earlier onset of fatigue and a pronounced decline in exercise capacity (<i>p</i> &lt; 0.05). Acute hypoxic exercise upregulated the expression of TRPV1, 5-HT, and 5-HT1A in the prefrontal cortex (<i>p</i> &lt; 0.05), which may lead to a decrease in exercise capacity. Pharmacological blockade of TRPV1 and 5-HT1A receptors extended the duration of high-load exercise under hypoxic conditions and improved exercise capacity (<i>p</i> &lt; 0.05).</p> Conclusion <p>Our findings indicate that the upregulation of TRPV1, 5-HT, and 5-HT1A is a key mechanism underlying the decline in exercise performance during acute hypoxia. Pharmacological blockade of these pathways effectively alleviates hypoxia-induced exercise fatigue, suggesting they represent promising therapeutic targets for enhancing performance under hypoxic conditions.</p> Significance <p>These findings provide a biological basis for developing nutritional strategies to counteract the initial decline in physical performance experienced by military personnel and adventurers during their ascent to high-altitude environments.</p>