Integrating network pharmacology and molecular docking to reveal the mechanism of Baihu Decoction in alleviating myocardial injury of heat stressed chicks

Baihu Decoction (BHD) is a classic traditional Chinese medicine formula used clinically for febrile diseases. Previous studies have demonstrated that BHD can prevent and alleviate heat stress in mice, chickens, and beef cattle, although its mechanisms remain unclear. In this study, chicks were used as experimental animals, and a heat stress injury model was established by exposing them to high temperature and humidity. UHPLC-Q-Exactive Orbitrap MS was employed to analyze and identify the chemical components of BHD, while the SwissADME platform was utilized to screen for active components. Through network pharmacology, the core targets and core components associated with heat stress were identified. Subsequently, potential targets were further analyzed through GO functional and KEGG pathway enrichment analyses, followed by molecular docking and animal experiments to validate the finding. The result showed that 169 active substances were identified and screened in BHD. KEGG pathway analysis revealed a strong correlation between the MAPK pathway and the intersecting targets. Molecular docking further confirmed the high binding affinity between the core components of BHD and the key targets in the MAPK pathway. These findings indicate that the MAPK pathway is critically involved in heat stress-induced myocardial injury in chicks as well as the cardioprotective effects of BHD. Animal experiments demonstrated that administering BHD via drinking water significantly alleviated myocardial pathology injury in heat stressed chicks exposed to high temperature and humidity. BHD treatment improved cardiac function and enhanced antioxidant capacity. Furthermore, it regulated the phosphorylation levels of ERK1/2, p38, and JNK1/2 in myocardial tissues, thereby inhibiting MAPK pathway activation. Additionally, BHD decreased the levels of Bax and cleaved caspase-3 while increasing Bcl2 expression. In conclusion, BHD effectively alleviates heat stress-induced myocardial injury and cardiac dysfunction in chicks, and its mechanism is closely associated with the inhibition of MAPK signaling pathway-mediated apoptosis. This study identifies BHD as a potential therapeutic drug for heat stress in animals and offers experimental support for its application in heat stress prevention and treatment.