不同附子炮制品四逆汤化学成分及毒性差异分析_刘洪.pdf

中草药 中草药 2023 年年 3 月月 第第 54 卷卷 第第 5 期期 Chinese Traditional and Herbal Drugs 2023 March Vol.54 No.5 1397 不同附子炮制品四逆汤化学成分及毒性差异分析 刘 洪，钟凌云*，邓延文，童恒力，陈 浩，王 硕，卢兴美 江西中医药大学，江西 南昌 330004 摘 要：目的 比较生附片、淡附片、阴附片、阳附片 4 种附子炮制品入方四逆汤化学成分以及毒性差异，为临床安全用药提供科学的依据。方法 分别以阴附片、阳附片、生附片、淡附片制备单味药以及四逆汤汤剂，经 UPLC 法检测汤剂中乌头碱、新乌头碱、次乌头碱、苯甲酰乌头原碱、苯甲酰新乌头原碱、苯甲酰次乌头原碱、甘草苷、甘草酸、6-姜酚含量，通过系统聚类分析（hierarchical cluster analysis，HCA）、主成分分析（pricipal component analysis，PCA）和正交偏最小二乘-判别分析（orthogonal partial least squares discriminant analysis，OPLS-DA）对 4 种四逆汤进行分类分析，再经急性毒性试验和长期毒性试验比较 4 种附子炮制品四逆汤毒性差异。结果 4 种四逆汤中成分含量以及毒性大小存在显著性差异。双酯型生物碱：乌头碱、新乌头碱、次乌头碱含量及其总量高低排序为生附片四逆汤阴附片四逆汤阳附片四逆汤淡附片四逆汤；单酯型生物碱：苯甲酰乌头原碱、苯甲酰新乌头原碱、苯甲酰次乌头原碱含量及其总量高低排序为阴附片四逆汤阳附片四逆汤淡附片四逆汤生附片四逆汤；在急性毒性试验中仅有生附片四逆汤可使小鼠产生轻微明显毒性反应，4 种四逆汤均无致死性。经计算阴附片四逆汤、阳附片四逆汤小鼠最大给药量倍数均相当于人临床每日用药量的 200 倍左右，和淡附片四逆汤以及生附片四逆汤比较无差异；在长期毒性试验中，4 种四逆汤对慢性心衰大鼠均不会致使肝肾损害，无肝肾毒性。结论 综合分析阴附片四逆汤、阳附片四逆汤、生附片四逆汤、淡附片四逆汤中双酯型生物碱以及单酯型生物碱含量差异，并结合毒性试验结果，得出 4 种附子炮制品入方经典名方四逆汤均具有合理性，其中以阴附片、阳附片入方四逆汤在一定程度上更具优势，阴附片更佳。关键词：四逆汤；生附片；淡附片；阴附片；阳附片；毒性差异；合理性；双酯型生物碱；单酯型生物碱；乌头碱；新乌头碱；次乌头碱；苯甲酰乌头原碱；苯甲酰新乌头原碱；苯甲酰次乌头原碱；系统聚类分析；主成分分析；正交偏最小二乘-判别分析；急性毒性；长期毒性 中图分类号：R283.6 文献标志码：A 文章编号：0253-2670(2023)05-1397-14 DOI:10.7501/j.issn.0253-2670.2023.05.006 Analysis on chemical constituents and toxicity of Sini Decoction of different Aconiti Lateralis Radix Praeparata processed products LIU Hong,ZHONG Ling-yun,DENG Yan-wen,TONG Heng-li,CHEN Hao,WANG Shuo,LU Xing-mei Jiangxi University of Chinese Medicine,Nanchang 330004,China Abstract:Objective To compare the differences in chemical composition and toxicity of four kinds of Fuzi(Aconiti Lateralis Radix Praeparata,Aconite)processed products of unprocessed Aconite piece(UAP),Glycyrrhiza cooked Aconite piece(GCAP),dried ginger steamed Aconite piece(DGSAP)and sand fried Aconite piece(SFAP)in Sini Decoction,so as to provide scientific basis for subsequent clinical safe medication.Methods Single-flavor Aconiti Decoction and Sini Decoction were prepared by UAP,GCAP,DGSAP and SFAP.The contents of aconitine,mesaconitine,hypaconitine,benzoylaconine,benzoylmesaconine,benzoylhypaconine,liquiritin,glycyrrhizic acid,6-gingerol in the decoction were detected by UPLC.Hierarchical cluster analysis(HCA),pricipal component analysis(PCA)and orthogonal partial least squares-discriminant analysis(OPLS-DA)were used to classify and analyze four kinds of Sini Decoction,which forming unprocessed Aconite piece Sini Decoction(UAPSND),Glycyrrhiza cooked Aconite piece Sini Decoction(GCAPSND),dried ginger steamed Aconite piece Sini Decoction(DGSAPSND)and sand fried aconite piece(SFAPSND),and then the toxicity of four kinds of Aconite processed products Sini Decoction were compared by acute toxicity test and long term toxicity 收稿日期：2022-09-13 基金项目：国家自然科学基金资助项目（81860703）；中药炮制技术传承创新团队（CXLD22003）作者简介：刘 洪，硕士研究生，从事中药炮制机制及其药效研究。Tel:15270310313 E-mail:*通信作者：钟凌云，博士生导师，教授，从事中药炮制机制及其药效研究。Tel:(0791)87118939 E-mail: 1398 中草药 中草药 2023 年年 3 月月 第第 54 卷卷 第第 5 期期 Chinese Traditional and Herbal Drugs 2023 March Vol.54 No.5 test.Results There were significant differences in the contents and toxicity of the four kinds of Aconite processed products Sini Decoction.Diester alkaloids:The content and total amount of aconitine,mesaconitine,hypaconitine were in the order of UAPSND DGSAPSND SFAPSND GCAPSND.Monoester alkaloids:The content and total amount of benzoylaconine,benzoylmesaconine,bnzoylhypaconine were in the order of DGSAPSND SFAPSND GCAPSND UAPSND.In the acute toxicity test,only the UAPSND could cause mild and obvious toxicity in rats,and the four kinds of Aconite processed products Sini Decoction were not fatal.According to the calculation,the maximum dosage of DGSAPSND and SFAPSND for rats are equivalent to about 200 times of the clinical daily dosage of human,which is not different from that of GCAPSND and UAPSND.In the long term toxicity test,four kinds of Sini decoction did not cause liver and kidney damage and had no liver and kidney toxicity in chronic heart failure rats.Conclusion By comprehensive analysis of the differences in the content of diester alkaloids and monoester alkaloids in DGSAPSND,SFAPSND,GCAPSND and UAPSND,combined with the results of toxicity test,it was concluded that the four kinds of Aconite processed products were reasonable in Sini Decoction of classic famous recipe.DGSAP and SFAP apply to the Sini decoction has more advantages to a certain extent,and DGSAP is better.Key words:Sini Decoction;unprocessed Aconite piece;Glycyrrhiza cooked Aconite piece;dried ginger steamed Aconite piece;sand fried aconite piece;differences of toxicity;rationality;diester alkaloids;monoester alkaloids;aconitine;mesaconitine;hypaconitine;benzoylaconine;benzoylmesaconine;benzoylhypaconine;hierarchical cluster analysis;principal component analysis;orthogonal partial least squares discriminant analysis;acute toxicity;long term toxicity 四逆汤为汉代张仲景所著伤寒论中用于治疗心肾阳衰寒阙证的主方，由附子、干姜、炙甘草组成。四逆汤原始方以生附片入方1，借生附子的峻烈强心作用发挥回阳救逆的作用，但因其主成分双酯型生物碱过高极易使机体中毒，难以把握其用药剂量，故后续各种校订本中多以炮制品入药2。附子经不同方法炮制后，不仅可以降低其毒性，还能改变