牛黄解毒片全方及方中其他中药对雄黄配伍减毒机制的研究

Niuhuang Jiedu Tablet (NJT) is an effective formula of traditional Chinese medicine (TCM) used in clearing heat and detoxicating. Officially listed in the Chinese Pharmacopeia (Volume I, 2015), NJT is composed of Realgar (As2S2), Bovis Calculus Artifactus, Borneolum Synthecticum, Gypsum Fibrosum, Rhei Radix et Rhizoma, Scutellariae Radix, Platycodonis Radix and Glycyrrhizae Radix et Rhizoma. As a component of NJT, the dosage of realgar recommended by the Chinese Pharmacopoeia ranges from 0.2 to 0.3 g per day, two or three times higher than the highest dosage regulated in the Chinese Pharmacopoeia when it is used alone. Modern medical research indicated realgar had toxicological effects on liver and kidney and the toxicity of realgar depends on the total quantities and the speciation of the soluble arsenic. Based on the relevant literatures and our previous researches, it was more secure and less toxic for combined realgar in NJT, but the relationship between toxicity alleviation effects of NJT and the other TCMs in the prescription to realgar and the total quantities and the speciation of the soluble arsenic influenced by NJT and the other TCMs in the prescription need to be solved urgently. This project is to systematically study on the ways and mechanisms of toxicity alleviation effects of NJT and the other TCMs in the prescription to realgar by evaluation of arsenic concentrations and species in gastrointestinal fluids, arsenic distribution and pharmacokinetics in rats, and the accumulation of arsenic concentrations and species in rats using ICP-MS and HPLC-ICP-MS. The study could provide references for the safe and rational use of realgar and realgar containing formulae.

牛黄解毒片是著名的清热解毒中药复方，由雄黄、大黄、黄芩、桔梗、甘草、人工牛黄、冰片和石膏等八味中药组成，被2015年版《中国药典》一部收载。牛黄解毒片每日服用量中雄黄的含量高达0.2～0.3g，相当于《中国药典》规定的雄黄最高服用量的2～3倍。现代药学研究表明雄黄对动物肝、肾等器官的毒性很强，其毒性主要取决于可溶性砷的含量和形态。相关文献及课题前期研究表明，牛黄解毒片全方配伍可降低雄黄的砷毒性，而配伍减毒作用与影响雄黄可溶性砷含量和形态的相关性问题亟待研究。本课题拟采用电感耦合等离子体质谱（ICP-MS）及其与高效液相色谱联用技术HPLC-ICP-MS，从胃肠道溶出、吸收、分布、代谢和排泄等环节，系统研究牛黄解毒片全方及方中其他中药对雄黄可溶性砷含量和形态体内外过程的影响，揭示全方及方中其他中药对雄黄配伍减毒作用可能的方式和机制，为雄黄及含雄黄中药复方砷毒性的控制和临床合理使用提供参考。

牛黄解毒片是著名的清热解毒药物，由雄黄（二硫化二砷）、人工牛黄、冰片、石膏、大黄、黄芩、桔梗和甘草等八味中药组成，被2020年版《中国药典》一部收载。牛黄解毒片每日服用量中有毒矿物中药雄黄的含量高达0.2～0.3g，相当于《中国药典》规定的雄黄最高服用量的2～3倍。相关其他研究及本课题前期研究结果均表明，牛黄解毒片配伍可降低雄黄的砷毒性。然而，其可能的配伍减毒机制尚不明确。可溶性砷是雄黄毒性的主要来源，本课题从全方及方中其他中药对雄黄可溶性砷的体内外过程的影响的角度，探讨其中可能的配伍减毒机制。.经研究得到以下结果：（1）牛黄解毒片全方及方中的大黄、黄芩、桔梗和甘草在体外可显著降低雄黄中可溶性砷的生物可给性。（2）牛黄解毒片全方配伍可显著降低雄黄中砷在小鼠体内的生物利用度。（3）牛黄解毒片全方配伍可显著降低雄黄在小鼠体内的砷蓄积，从而降低其砷毒性。大黄、黄芩、桔梗和甘草各单味中药可不同程度地降低雄黄在小鼠体内的砷蓄积，上述四味中药配伍后可达到全方对雄黄的配伍减毒效果。（4）肠道菌群在体外可降低雄黄中砷的生物可给性。（5）肠道菌群可降低雄黄在小鼠体内的砷蓄积量。.本课题进一步明确了牛黄解毒片对雄黄的配伍减毒作用与对其可溶性砷体内外过程影响的相关性，同时初步发现肠道菌群作为靶点在防止雄黄砷毒性方面具有潜在作用和研究价值，为雄黄及含雄黄中药复方的砷毒性的控制提供参考和依据。