附子水煎液在正常和虚寒小鼠体内血药浓度差异研究

【作者】 杨媛；

【导师】 黄建梅；

【作者基本信息】 北京中医药大学 ， 中医学， 2008， 硕士

【摘要】 中药毒性反应发生的机制与中药本身的性质有关,也与人体机体状态有关。人体在生理状态和病理状态下对中药的反应有很大的差异,未辨证使用毒性药物是引起中毒的重要原因之一,因此理性认识中药的毒性,积极探求中药毒性及其与机体状态的相关性,对于以中药现代化为契机加强对中药毒性的研究,使中药作为天然药物、绿色疗法更好地服务于人类有着重要的意义。附子为毛茛科植物乌头Aconitum carmichaeli Debx.的子根的加工品,具有回阳救逆、补火助阳、逐风寒湿邪的功效。附子生用有大毒,多炮制加工降毒后入药。本实验选取附子作为有毒中药的代表,研究有毒中药毒性与机体状态的关系。本课题是国家“973”计划课题《基于“药性构成三要素”的中药药性实质研究》(课题编号2007CB512605)的一部分,主要通过对虚寒状态小鼠和正常状态小鼠血清中次乌头碱的检测,结合附子水煎剂对虚寒状态小鼠和正常小鼠体重、毛色等生理状态、生化指标以及存活率的影响,以明确不同生理病理状态下机体对有毒中药的不同反应。从而探讨中药的毒性及其与机体状态的相关性。本课题主要进行了以下2个方面的研究:一建立了附子水煎液中乌头碱,次乌头碱,新乌头碱的高效液相色谱分析检测方法,对实验室中按照临床方法制备的附子水煎液进行检测,明确其中每毫升乌头碱、次乌头碱、新乌头碱分别为0.149mg、1.382mg、1.089mg,为后期的动物实验提供质量可控的药物;二建立了小鼠血清中次乌头碱的高效液相色谱分析检测方法,对比研究虚寒状态和正常状态小鼠血清中次乌头碱浓度差异。复制小鼠虚寒状态模型,连续给予附子水煎液14天,检测第1、7和14天虚寒状态及正常状态小鼠血清中次乌头碱的浓度,结合在给药过程中小鼠体重毛色等生理指标及生化指标的变化,分析讨论机体在不同状态对毒性中药的不同反应,结果分析表明,附子对虚寒状态小鼠机体代谢起调节作用,使其恢复或接近正常水平;附子水煎液干扰正常状态小鼠代谢环节(机体内代谢酶受到抑制、靶器官(代谢器官)损害),对机体表现出毒性作用。从小鼠附子水煎液灌胃后第7天及第14天的存活情况看,虚寒状态小鼠的存活率明显高于正常小鼠(2组:正常小鼠存活率87.5%,虚寒状态小鼠存活率100%;3组:正常小鼠存活率70.0%,虚寒状态小鼠存活率92.5%),说明虚寒状态的小鼠对附子水煎液毒性的耐受力较强。这与传统中医药理论中的“辨证论治,对症下药”、“毒药之于常人为有毒,对病患则治病”相符合,证明临床强调合理用药的正确性。本实验的创新性在于,首次采用虚寒动物模型与正常动物相比较检测血药浓度,研究药物与机体状态的相互关系,为研究毒性中药提供新的思路与方法。更多还原

【Abstract】 The mechanism of the Chinese Traditional medicine toxicity responds to have something to do with the property of Chinese traditional medicine, also with human body function. The human body has obvious difference to the reaction of medicine under the pHysiology appearance and the pathologic appearance, whether the toxicity medicine is fit of the dialectics usage is also an important reason to cause poison. So reasonableness understanding Chinese Traditional medicine of toxicity, investigating a Chinese Traditional medicine toxicity actively and the body relativity of the appearance, which for with Chinese Traditional medicine modern change into a chance to strengthen toxic research of the Chinese Traditional medicine and bitterly for make Chinese Traditional medicine as the natural medicine, green therapy to serve in mankind has important meaning.Aconitum carmichaeli is a kind of processed product of Aconitum carmichaeli Debx roots in the secondary root of buttercup, its functions are to cause restoration from collapse, supplement body fire and reinforce yang, dispel wind, cold and damp. The un-processed product of Aconitum carmichaili has strong toxicity; it can use when it was preprocessed to reduce toxicity. This study selects Aconitum carmichaili as the representative of poisonous Chinese Traditional medicine to study the relation between poisonous Chinese herbal medicine toxicity and body appearance.This study is a part o《fBased on the "three elements of properties constitute" real Chinese medicine research》—“973”The national research development of the point foundation plans (NO: 2007CB512605) which about the Plasma Concentration of Aconitum carmichaili, mainly by researching the difference between falsely cold appearance mice and the normal in the Plasma Concentration and pHysiology appearances, such as weight and hair color...etc., descend the influence of the different pHysiology pathologic appearance by definitely poisonous Chinese Traditional medicine and inquiry the toxicity of Chinese Traditional medicine and the body about the relativity of the appearance. In this study, the following fields of research were conducted:Firstly, While building up a HPLC examination method to analyze Aconitum carmichaili water fried liquid in aconitine, mesaconitine, hypaconitine in our laboratory, according to replication water of the clinical method making fried the liquid carry on an examination, definite among them the Content of aconitine, hypaconitine, mesaconitine is 0.112 mgs, 1.034 mgs, 0.541 mgs, which can be used to the next animal experiment;Secondly,we have built up a HPLC examination method to analyze hypaconitine in the mice serum efficiently. Making duplicate mice falsely cold appearance model, carrying on an examination to analyze hypaconitine in the mice serum in the falsely cold appearance and normal appearance mice with weight and hair color etc, is in the medicine process the variety of the pHysiology index sign to discuss the relation that body in the different appearances having different responds to the toxicity Chinese Traditional medicine .Test result enunciates that falsely cold appearance mice bear to be subjected to the toxicity which attaches Aconitum carmichaili to fry better.The mice attached Aconitum carmichaili water fried liquid after infusing the stomach to pass a survival a circumstance to see for 14 day on the 7th day , the livability of falsely cold appearance mice were obviously higher than the normally mice(2 set:The normally mice livability is 87.5%, falsely cold ppearance mice livability 100%;3 set:The normally mice livability is 70.0%, falsely cold ppearance mice livability 92.5%), also explains the mice of falsely cold appearance is stronger towards infusing Aconitum carmichaili water a fried liquid toxically of bearing and being subjected to dint.This agrees with traditional"the dialectical theory cure, prescribe medicine for specific ailment", "poison in ordinary people is poisonous, but cures illness of the patient" within theory of the Chinese medicine pHarmacology to match, proving clinical empHasize reasonable to use medicine accurately.The innovation of this experiment lies in adopting a method to compare Plasma Concentrationan between falsely cold animal model and normal animal for the very first time density, studys the correlation between medicine and body appearance, for providing a new way to study toxicity Chinese Traditional medicine.更多还原