HPLC conditions.

Naja atra bite is one of the most common severe snakebites in emergency departments. Unfortunately, the pathophysiological changes caused by Naja atra bite are unclear due to the lack of good animal models. In this study, an animal model of Naja atra bite in Guangxi Bama miniature pigs was established by intramuscular injection at 2 mg/kg of Naja atra venom, and serum metabolites were systematically analyzed using untargeted metabolomic and targeted metabolomic approaches. Untargeted metabolomic analysis revealed that 5045 chromatographic peaks were obtained in ESI+ and 3871 chromatographic peaks were obtained in ESI-. Screening in ESI+ modes and ESI- modes identified 22 and 36 differential metabolites compared to controls. The presence of 8 core metabolites of glutamine, arginine, proline, leucine, phenylalanine, inosine, thymidine and hippuric acid in the process of Naja atra bite was verified by targeted metabolomics significant difference (P<0.05). At the same time, during the verification process of the serum clinical samples with Naja atra bite, we found that the contents of three metabolites of proline, phenylalanine and inosine in the serum of the patients were significantly different from those of the normal human serum (P<0.05). By conducting functional analysis of core and metabolic pathway analysis, we revealed a potential correlation between changes in key metabolites after the Naja atra bite and the resulting pathophysiological alterations, and our research aims to establish a theoretical foundation for the prompt diagnosis and treatment of Naja atra bite.

舟山眼镜蛇（Naja atra）咬伤是急诊科最常见的重症蛇咬伤类型之一。遗憾的是，由于缺乏优质动物模型，舟山眼镜蛇咬伤引发的病理生理变化尚未明确。本研究通过肌内注射2mg/kg剂量的舟山眼镜蛇毒液，构建了广西巴马小型猪的舟山眼镜蛇咬伤动物模型，并采用非靶向代谢组学（untargeted metabolomics）与靶向代谢组学（targeted metabolomics）技术对血清代谢物进行系统性分析。非靶向代谢组学分析显示，在ESI+模式下获得5045个色谱峰，在ESI-模式下获得3871个色谱峰。与对照组相比，在ESI+与ESI-模式下分别筛选得到22种和36种差异代谢物。靶向代谢组学验证了谷氨酰胺、精氨酸、脯氨酸、亮氨酸、苯丙氨酸、肌苷、胸苷与马尿酸这8种核心代谢物在舟山眼镜蛇咬伤过程中存在显著差异（P<0.05）。同时，在对舟山眼镜蛇咬伤患者的血清临床样本进行验证时，我们发现脯氨酸、苯丙氨酸与肌苷这三种代谢物在患者血清中的含量与正常人体血清存在显著差异（P<0.05）。通过对核心代谢物进行功能分析及代谢通路富集分析，本研究揭示了舟山眼镜蛇咬伤后关键代谢物的变化与后续病理生理改变之间的潜在关联，本研究旨在为舟山眼镜蛇咬伤的快速诊断与治疗奠定理论基础。