IN VIVO ACUTE TOXICITY OF DETOXIFIED FUZI (LATERAL ROOT OF ACONITUM CARMICHAELI) AFTER A TRADITIONAL DETOXIFICATION PROCESS
IN VIVO ACUTE TOXICITY OF DETOXIFIED FUZI (LATERAL ROOT OF ACONITUM CARMICHAELI) AFTER A TRADITIONAL DETOXIFICATION PROCESS
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传统解毒工艺后解毒附子（乌头侧根）的体内急性毒性
DOI：
10.17179/excli2018-1607
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发表时间：
2018-01-01
期刊：
影响因子：
4.6
通讯作者：
Efferth, Thomas
中科院分区：
文献类型：
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作者：
Sun, Wan;Yan, Bo;Efferth, Thomas
许多中药不仅具有治疗功效，而且对正常组织也有毒性。中药毒性有时会因错误使用中药而导致严重不良事件甚至致命中毒。附子（Aconitum carmichaeli）的侧根就是这样一种草药，其有毒成分是附子。乌头碱、新乌头碱、次乌头碱是附子的主要有毒成分，经水煎剂水解成无毒衍生物。因此，长期煎煮是附子使用前解毒的传统方法。然而，最近的临床试验对长期煎煮附子引起的不良事件进行了预选，对传统解毒程序后附子的安全性提出了一些质疑。为了彻底确定长期煎煮附子是否安全，我们使用啮齿动物和斑马鱼模型进行了体内急性毒性测定，并使用 HPLC 和 UPLC-MS 进行了化学特征分析。 HPLC分析表明，随着煎煮时间的延长，有毒乌头碱成分水解为苯甲酰衍生物。煎煮 2 小时后的附子 (FZ-120) 中，HPLC 未检测到这些乌头碱，表明 FZ-120 似乎无毒性。与具有致死毒性的非煎附子（FZ-0）和60分钟煎附子（FZ-60）不同，130 g/kg的FZ-120在体重和生化参数方面不会导致小鼠任何死亡或副作用。这似乎证实了附子经过长时间煎煮后的安全性。然而，组织病理学观察显示，FZ-120 治疗后肝脏表型异常，肝脏指数显着下降，表明 FZ-120 具有潜在的肝毒性。通过使用斑马鱼模型，我们观察到FZ-120在288至896 μg/ml的剂量范围内引起了相当多的不良事件，包括心律失常、肝变性、卵黄囊吸收延迟、长度减少和鱼鳔丢失，这清楚地说明了对心血管、消化、发育和呼吸系统的急性毒性。 FZ-120的剂量范围低于人类临床应用的剂量范围。此外，UPLC-MS 显示 FZ-120 仍然含有 HPLC 检测不到的有毒乌头碱，这可能解释了它对斑马鱼的急性毒性。我们的结论是，即使经过长时间煎煮，附子也不够安全。斑马鱼模型与 UPLC-MS 检测相结合可能代表了揭示乌头碱相关急性毒性的适当测试系统。
Many herbs of traditional Chinese medicine (TCM) possess not only therapeutic efficacy, but also toxicity towards normal tissues. The herbal toxicities occasionally cause serious adverse events or even fatal poisoning due to the erroneous use of TCM herbs. Fuzi (lateral root of Aconitum carmichaeli) is such an herb with its toxic ingredient, aconites. Aconitine, mesaconitine, and hypaconitine are the main toxic components of Fuzi, which are hydrolyzed into non-toxic derivatives by water decoction. Therefore, long-time decoction was commonly applied as a traditional way to detoxify Fuzi before use. Nevertheless, recent clinical trials presorted on adverse events induced by long-time decocted Fuzi, putting some doubt on the safety of Fuzi after the traditional detoxification procedure. To thoroughly determine whether or not long-time decocted Fuzi was safe, we conducted in vivo acute toxicity assays using both rodent and zebrafish models and performed chemoprofile analyses using HPLC and UPLC-MS. The HPLC analysis showed that toxic aconitine components were hydrolyzed into benzoyl derivatives with increasing time of decoction. These aconitines were undetected by HPLC in Fuzi after 2 h-decoction (FZ-120), indicating seemingly non-toxicity of FZ-120. Unlike the non-decocted Fuzi (FZ-0) and 60 min-decocted Fuzi (FZ-60) with lethal toxicity, FZ-120 at 130 g/kg did not cause any deaths or side effects in mice regarding body weight and biochemical parameters. This seems to confirm safety of Fuzi after long-time decoction. However, histopathological observations revealed an abnormal liver phenotype and a significant decrease of the liver index following FZ-120 treatment, indicating a potential hepatoxicity of FZ-120. By using a zebrafish model, we observed that FZ-120 at a dose range from 288 to 896 μg/ml caused considerable adverse events including arrhythmia, liver degeneration, yolk sac absorption delay, length decrease, and swim bladder loss, which clearly speak for acute toxicity on cardiovascular, digestive, development, and respiratory systems. The dose range of FZ-120 was lower than that used for clinical application in human beings. Moreover, UPLC-MS revealed that FZ-120 still contained toxic aconitines that were not detectable by HPLC, which might explain its acute toxicity in zebrafish. We concluded that Fuzi is not sufficiently safe even after long-time decoction. The zebrafish model combined with UPLC-MS assay may represent an appropriate test system to unravel aconitine-related acute toxicity.