不同来源牛黄与雄黄配伍的牛黄解毒片对小鼠毒性的影响

目的:比较不同来源牛黄与雄黄配伍的牛黄解毒片对小鼠的毒性反应,探讨临床上牛黄解毒片引起慢性砷中毒的机制。方法:按2015年版《中国药典》的配方用天然牛黄、人工牛黄、体外培育牛黄分别制成牛黄解毒片,并用同法制成无牛黄成分、无雄黄成分的缺味组,另设同剂量单味雄黄作为对照,每种牛黄解毒片组再分为0.5,1.0,2.0 g·kg~(-1)剂量组,单味雄黄组分为0.06,0.12,0.24 g·kg~(-1)剂量组,对ICR小鼠进行灌胃给药28 d,观察毒性反应;给药结束后解剖取血,分离血清检测肝肾功能,取心、肝、脾、肺、肾做组织病理学检查和电镜下超微结构观察。结果:给药期间各给药组未观察到毒性反应,给药结束亦未观察到大体内脏及体表皮肤病变,血液生化指标检查未能明确雄黄对肝肾的毒性作用。组织病理学和电镜下超微结构检查各牛黄解毒片组未发现异常毒性病理改变,但苏木素-伊红(HE)染色显示单味雄黄高剂量组肝脏出现小叶中心性肝细胞肥大和糖原聚集,过碘酸希夫(PAS)染色表明肝细胞内糖原聚集,电镜下超微结构检查发现肝细胞内局灶性胞浆溶解,内有数量不等糖原颗粒,提示雄黄可致小鼠肝脏毒性。结论:用不同来源牛黄与雄黄配伍的牛黄解毒片对小鼠进行28 d灌胃给药,均未发现对小鼠有明显的毒性反应。而用单味雄黄却引起肝脏毒性,提示牛黄解毒片中的其他成分与雄黄配伍能显著降低可溶性砷和价态砷的含量,从而降低雄黄的毒性。

Toxicity of Niuhuang Jiedu Tablets Containing Calculus Bovis from Different Sources in Mice

Objective: To conduct a comparative study on the toxic reaction of Niuhuang Jiedu tablets containing Calculus Bovis (Niuhuang) from different sources in mice, in order to explore the mechanism of chronic arsenic poisoning clinically caused by Niuhuang Jiedu tablets. Method: Test sample Niuhuang Jiedu tablets were respectively made by using natural Calculus Bovis, Bovis Calculus Artificial, and Bovis Calculus Sativus, with no Calculus Bovis element and no Realgar element, according to Chinese Pharmacopoeia (Version 2015). The same dose of test sample single Realgar was used for control. Each Niuhuang Jiedu tablets group was divided into low, middle and high-dose groups, ICR mice were intragastrically administered with drugs for 28 days, then the toxic reaction was observed during the period of drug administration. After drug administration, these mice were put to death. Their blood samples were collected to test liver and kidney function, and tissues of heart, liver, spleen, lung and kidney were collected for histopathological examination and ultrastructure observation under electronic microscopy. Result: In each test group, no toxic reaction was observed during drug administration. No gross lesion was observed in viscera and skins after the mice were put to death. And no clear toxic effect of Realgar on liver and kidney functions was found in blood biochemistry examination. In each Niuhuang Jiedu tablets group, no change in histopathology and ultrastructure under electronic microscopy was found. But in high-dose single Realgar group, the histopathological examination showed that in hepatic lobule, hepatocytes became enlarged, and glycogen accumulated in cytoplasma according to hematoxylin-eosin (HE) staining, which were further confirmed by periodic acid Schiff reaction (PAS) staining. Ultrastructure under electronic microscopy showed in hepatocytes focal cytoplasm dissolutions, and glycogen granules were found inside. Conclusion: No obvious toxic reaction was found in ICR mice after administration with Niuhuang Jiedu tablets containing bovis calculus from different sources for 28 days, but the same dose of single Realgar could cause liver toxicity. It is suggested that other ingredients in Niuhuang Jiedu tablets combined with Realgar can significantly reduce the concentrations of soluble arsenic and arsenic valence state, thus reducing the toxicity of Realgar.