大黄对大鼠解热作用及其药效物质基础研究

采用LC-MS技术与活性评价相结合的方法,阐明大黄解热药效物质基础。利用干酵母致大鼠发热模型,灌胃大黄乙醇提取物及不同极性萃取物,测定并记录给药后0.5,1,2,3,4,5 h的大鼠肛温,比较各部位对大鼠体温升高的抑制率,得到大黄解热活性部位。再采用UPLC-ESI-Orbitrap-MS/MS技术对活性部位化学成分进行分析,鉴定其活性成分。与模型组比较,大黄乙醇提取物组体温增高在各测定时间点均有减少且3 h后效果显著,给药后3,4,5 h,其对大鼠体温增高的抑制率分别为38.7%(P<0.05),78.2%(P<0.01),72.4%(P<0.01);正丁醇和水部位在早期也都表现出一定的解热活性,给药后1 h,二者的抑制率分别为28.1%(P<0.01),24.9%(P<0.05);而其他部位的解热作用均不明显。经LC-MS/MS分析,从正丁醇部位鉴定了33个化合物,水部位鉴定了12个,共包括10个鞣质、15个蒽醌苷、4个蒽酮苷、1个酚苷、1个萘衍生物、1个蒽醌和1个蔗糖。结果表明大黄具有明确的解热作用,而且鞣质和蒽醌苷类成分为其主要药效物质基础。

Study on antipyretic effect of rhubarb on rats and its antipyretic ingredients

A combination of LC-MS technology and activity evaluation was used to identify the antipyretic ingredients in rhubarb.The rat model of fever was established with dried yeast and then was administered ethanol extract and different polar fractions of rhubarb.Next,the anal temperature of these rats was measured and recorded at 0.5,1,2,3,4 and 5 h after administration,and the inhibition rate of each part on the rise of body temperature was calculated.The inhibition rate is higher and the antipyretic effect is better.The chemical composition of the effective fraction was analyzed with UPLC-ESI-Orbitrap-MS/MS technology.Compared with the model group,the increase of body temperature of ethanol extract group all reduced at each measurement time especially after 3 h,and the inhibition rate were 38.7%(P<0.05),78.2%(P<0.01)and 72.4%(P<0.01)at 3 h,4 h,and 5 h after administration,respectively.Both n-butanol and water fraction showed some antipyretic activity in the early stage,with the inhibition rate of 28.1%(P<0.01)and 24.9%(P<0.05)at 1 h after administration,respectively,while other fractions were not active.Thirty-three and twelve compounds were identified from n-butanol and water fraction by LC-MS/MS analysis,respectively,including ten tannins,fifteen anthraquinone glycosides,four anthrone glycosides,one phenolic glycoside,one naphthaline derivative,one anthraquinone and one sucrose.These results revealed that rhubarb had antipyretic activity on rats,and tannin and anthraquinone glycosides were the main active ingredients inside.