摘要
山楂具有化浊降脂的功效,在临床上用于治疗高脂血症,但其功效成分尚不清楚.该文采用"谱效"关系结合体内药效评价的方式筛选、验证山楂化浊降脂功效成分,并采用液相色谱质谱联用(liquid chromatography-mass spectrometry,LC-MS)技术建立山楂质量控制方法.首先,复制高脂血症大鼠模型,灌胃给予山楂提取物不同极性部位及其组合,比较其对模型大鼠血清中总胆固醇(total cholesterol,TC)、甘油三酯(triglycerides,TG)和低密度脂蛋白胆固醇(low-density lipoprotein-cholester-ol,LDL-C)的影响;采用正交偏最小二乘(orthogonal partial least squares,OPLS)算法建立山楂提取物不同极性部位及组合中24 个化学成分与药效指标间的"谱效"关系模型,筛选山楂化浊降脂功效成分,并通过体内实验对功效成分进行验证;最后,以枸橼酸、奎尼酸等 10 个化学成分为指标,采用LC-MS建立山楂的质量控制方法.结果表明,山楂提取物不同极性部位及其组合对高脂血症模型大鼠血清中TG、TC和LDL-C均有不同程度的调节作用,其中乙酸乙酯部位的效果最优;通过OPLS模型筛选山楂化浊降脂功效成分为牡荆素-4″-O-葡萄糖苷、牡荆素-2″-O-鼠李糖苷、芦丁、枸橼酸、苹果酸和奎尼酸;灌胃给予高脂血症模型大鼠功效成分混悬液能显著降低模型大鼠血清中TC和TG水平;50 批山楂中枸橼酸、奎尼酸、芦丁、没食子酸、牡荆素-4″-O-葡萄糖苷、牡荆素-2-O″-鼠李糖苷、苹果酸、香草酸、新绿原酸和延胡索酸的平均质量分数分别为 38、11、0.018、0.009 5、0.037、0.017、8.1、0.009 5、0.073、0.98 mg·g-1.该文初步阐明了山楂化浊降脂的功效成分,并建立了山楂的含量测定方法,为科学评价山楂的质量提供了数据和方法支撑.
Abstract
Hawthorn has the efficacy of eliminating turbidity and lowering the blood lipid level,and it is used for treating hyperlipi-demia in clinic.However,the bioactive components of hawthorn are still unclear.In this study,the spectrum-effect relationship was employed to screen the bioactive components of hawthorn in the treatment of hyperlipidemia,and then the bioactive components screened out were verified in vivo.Furthermore,the quality control method for hawthorn was developed based on liquid chromatogra-phy-mass spectrometry(LC-MS).The hyperlipidemia model of rats was built,and different polar fractions of hawthorn extracts and their combinations were administrated by gavage.The effects of different hawthorn extract fractions on the total cholesterol(TC),tri-glycerides(TG),and low-density lipoprotein-cholesterol(LDL-C)in the serum of model rats were studied.The orthogonal projections to latent structures(OPLS)algorithm was used to establish the spectrum-effect relationship model between the 24 chemical components of hawthorn and the pharmacodynamic indexes,and the bioactive components were screened out and verified in vivo.Finally,10 chemical components of hawthorn,including citric acid and quinic acid,were selected to establish the method for evaluating hawthorn quality based on LC-MS.The results showed that different polar fractions of hawthorn extracts and their combinations regulated the TG,TC,and LDL-C levels in the serum of the model rats.The bioactive components of hawthorn screened by the OPLS model were vitexin-4″-O-glucoside,vitexin-2″-O-rhamnoside,rutin,citric acid,malic acid,and quinic acid.The 10 chemical components of hawthorn,i.e.,citric acid,quinic acid,rutin,gallic acid,vitexin-4″-O-glucoside,vitexin-2″-O-rhamnoside,malic acid,vanillic acid,neochlo-rogenic acid,and fumaric acid were determined,with the average content of 38,11,0.018,0.009 5,0.037,0.017,8.1,0.009 5,0.073,and 0.98 mg·g-1,respectively.This study provided a scientific basis for elucidating the material basis of hawthorn in treating hyperlipidemia and developed a content determination method for evaluating the quality of hawthorn.
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维普
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