氟马西尼联合肾上腺素对钩吻中毒的解救作用

doi:10.11843/j.issn.0366-6964.2022.03.026

摘要/Abstract

摘要： 旨在研究氟马西尼联合肾上腺素对钩吻中毒大鼠的解救效果,并验证钩吻可能的毒性作用靶点,为钩吻中毒机制探讨及解救钩吻中毒病例提供参考。构建钩吻大鼠中毒模型,将60只大鼠随机分为4组,分别为对照组、氟马西尼解救组、肾上腺素解救组、联合用药(氟马西尼+肾上腺素)解救组,记录各组大鼠中毒及死亡情况,并监测各组大鼠血压、心率,评价各组药物的解救效果。采用分子对接、全细胞膜片钳技术,预测钩吻主要毒性物质钩吻素己与解救药物作用的靶点,阐释其可能的解毒机制。结果显示:钩吻灌胃大鼠的半数致死量(LD₅₀)为0.25 g·kg^-1,氟马西尼联合肾上腺素使用时对钩吻中毒的大鼠有显著的解救效果,存活率由25%提高到92%(P<0.01),且平均中毒症状出现时间由16.56 min延长至25.98 min (P<0.01),平均死亡时间由40.13 min延长至58.05 min (P<0.01),解救效果显著高于氟马西尼和肾上腺素单独解救组。联合用药组还可显著缓解中毒所致的大鼠血压、心率下降的临床表征。分子对接计算机模拟结果显示,氟马西尼与钩吻素己都与GABA_A受体同一活性位点有较好的结合效果,且氟马西尼的结合能-85.47 kJ·mol^-1,强于钩吻素己结合能-77.15 kJ·mol^-1。全细胞膜片钳结果显示,钩吻素己可显著延长GABA_A受体离子通道的开放时间,而氟马西尼可拮抗此作用。综上所述,氟马西尼和肾上腺素联合使用对钩吻中毒大鼠具有明显的解救效果,钩吻神经毒性靶点与氟马西尼作用的GABA_A受体密切相关,肾上腺素对调节中毒所致的血压下降起到关键作用,联合用药可起到标本兼治效果。

关键词: 钩吻, 氟马西尼, 肾上腺素, 毒性机制, GABA_A受体

Abstract: This study aimed to assessment the detoxification of flumazenil combined with epinephrine on Gelsemium elegans Benth (G. elegans) poisoning rats, providing reference for G. elegans toxic mechanism and poisoning cases. G. elegans poisoning rat model was constructed, and 60 rats were randomly divided into 4 groups:control group, flumazenil rescue group, epinephrine rescue group and combined drug (flumazenil + epinephrine) rescue group. The poisoning and death of the groups' rats were recorded; the blood pressure and heart rate were monitored. Then evaluate the rescue effect of drugs in each group. Molecular docking and whole cell patch clamp technique were used to predict the target sites of gelsenicine (main toxic substance of G. elegans) and rescue drugs, and elucidate their possible detoxification mechanisms. We found that the LD₅₀ of G. elegans was 0.25 g·kg^-1 by gavage in rats; flumazenil combined with epinephrine had a significant detoxification effect on G. elegans poisoning rats; the survival rate increased from 25% to 92% (P<0.01), and the mean time to poisoning symptoms was extended from 16.56 to 25.98 min (P<0.01); the mean time to death was prolonged from 40.13 to 58.05 min (P<0.01); the rescue effect was significantly higher than that of the flumazenil and epinephrine alone rescue group. The combination of flumazenil and epinephrine significantly alleviated the clinical manifestations of decreased blood pressure and heart rate in poisoning rats. The results of molecular docking computer simulation showed that both flumazenil and gelsenicine had better binding effect to the same active site of GABA_A receptor. The binding energy of flumazenil was stronger than gelsenicine (-85.47 vs -77.15 kJ·mol^-1), which could reverse the toxicity effect of G. elegans. The whole cell patch clamp results showed that gelsenicine significantly prolonged the opening time of GABA_A receptor ion channels, and this effect was antagonized by flumazenil. In conclusion, the combination of flumazenil and epinephrine had a significant detoxification effect in G. elegans poisoning rats. The target of G. elegans neurotoxicity may relate to the GABA_A receptor that flumazenil action in the CNS; epinephrine was essential to prevent the decrease of blood pressure and heart rate caused by poisoning. Combination of drugs could use a therapy for both the causes and symptoms.

Key words: Gelsemium elegans, flumazenil, epinephrine, toxicity mechanism, GABA_A receptor

中图分类号:

S859.87

李玉娟, 肖宁, 贾文丹, 李霞荣, 郑晓峰, 孙志良. 氟马西尼联合肾上腺素对钩吻中毒的解救作用[J]. 畜牧兽医学报, 2022, 53(3): 938-946.

LI Yujuan, XIAO Ning, JIA Wendan, LI Xiarong, ZHENG Xiaofeng, SUN Zhiliang. The Effectiveness of Flumazenil-Epinephrine Combination on Treatment for Acute Gelsemium elegans Poisoning[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(3): 938-946.

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