比格犬致命鹅膏急性肝毒性实验研究

Experimental study on acute hepatotoxicity in beagles induced by Amanita exitialis

目的建立比格犬致命鹅膏（Amanita exitialis）急性肝毒性模型，探讨含鹅膏肽类毒素蘑菇急性中毒性肝功能衰竭的特点，以期为含鹅膏肽类毒素的蘑菇中毒实验研究提供帮助。方法UPLC-MS／MS（超高效液相色谱串联质谱）法测定致命鹅膏干粉中鹅膏肽类毒素含量，胶囊喂饲比格犬60mg／kg体质量致命鹅膏干粉，观察其变化，通过检测凝血及肝肾功能、肝脏病理以及血浆和尿液中鹅膏肽类毒素含量，用于比格犬致命鹅膏急性肝毒性模型观测指标。结果本研究所用致命鹅膏干粉中鹅膏肽类毒素总含量为（3482．6±124．94）mg／kg。模型犬在12～48h出现呕吐、腹泻等症状，染毒后24h，模型犬ALT、AST、TBIL、ALP、PT和APTT水平出现明显升高，染毒后36h，ALT、AST、PT和Am水平达到峰值[（ALT（283．2±112．9），AST（223．9±93．8），PT（132．9±152．6）s，Am（131．4±153．9）s，染毒后48h，TBIL和ALP值达到峰值[（TBIL（23．3±14．6）μmol／L，ALP（274．5±115．5）U／L]，模型犬TBIL、TP和Am在染毒后1周恢复至正常水平，ALT、AST和ALP在染毒后3周恢复至正常水平。染毒后24～72h死亡3只，肝脏病理检查显示弥漫性肝细胞出血性坏死。染毒后24h内，血浆中可检测到鹅膏肽类毒素；染毒后96h内，尿液中可检测到鹅膏肽类毒素。结论鹅膏肽类毒素可引起肝细胞出血性坏死，导致急性肝功能衰竭，该模型符合含鹅膏肽类毒素蘑菇致中毒性肝功能衰竭临床病理生理特点，可应用于含鹅膏肽类毒素蘑菇中毒的诊断和治疗研究。

Objective To establish acute hepatotoxic model induced by Amanita exitialis and to study the characteristics of acute toxic liver failure induced by mushrooms containing peptide toxins, in hope for providing some help to experimental research on poisoning induced by mushrooms containing peptide toxins. Methods UPLC-MS/MS （Ultra performance liquid chromatographytandem mass spectrometry） method was used to detect peptide toxins in Amanita exitialis. To establish acute toxic liver hepatic failure model, the beagles were fed with 60 mg/kg of lyophilized powder of Amanita exitialis fungus which encapsulated in starch capsules. Toxic sighs were observed, coagulation function, hepatic and renal function, liver histopathological morphology, peptide toxin concentration in plasma and urine were detected during the experiment. Results Total peptide toxins in Amanita exitialis was （3 482. 6 ± 124. 94） mg/ kg. All the beagles had toxic signs including vomiting and diarrhea in 12-48 h after ingestion. On 24 h after ingestion, the beagles＇ ALT, AST, TBIL, ALP, PT and APTT levels increased obviously. On 36 h after ingestion, the beagles＇ ALT, AST, PT and AYTT values reached their peaks （ALT： 283.2 ± 112. 9 Kallmann unit; AST： 223.9 ±93.8 Kallmann units; PT： 132. 9 ± 152. 6 s; AFFF： 131.4 ± 153.9 s） . On 48 h after ingestion, the beagles＇ TBIL and ALP levels reached their peaks （TBIL： 23.3 ± 14. 6 tool/L; ALP： 274. 5 ±115.5 U/L） . The beagles＇ TBIL, TP and APTT returned to normal 1 week after ingestion, their ALT, AST and ALP levels returned to normal 3 weeks after ingestion. Three dogs died during 24-72 h after ingestion. Liver histopathological morphology study showed hemorrhagic necrosis of hepatoeytes. Peptide toxins can be detected in plasma within 24 h after ingestion. Peptide toxins can be detected in urine within 96 h after ingestion. Conclusion Amanita peptide toxins can cause hemorrhagic necrosis of liver cells and lead to acute liver failure. This model is consistent with clinical pathophysiologieal process of acute toxic liver failure induced by mushrooms containing peptide toxins, and it can be applied to the study of diagnosis and treatment of poisoning induced by mushrooms containing peptide toxins.