兴安杜鹃3种主要成分的HPLC测定及其对大鼠气管平滑肌的舒张作用  

HPLC Determination of Three Main Components of Rhododendron dauricum L.and Their Relaxing Effects on Rat Tracheal Smooth Muscle

作　　者：张颖慧 王晨蔚 周苑婷 陈思淼 张怡然 孙宇婷 张琳[1,2,4,5] ZHANG Ying-hui;WANG Chen-wei;ZHOU Yuan-ting(Key Laboratory of Forest Plant Ecology,Ministry of Education,College of Chemistry,Chemical Engineering and Resource Utilization,Northeast Forestry University,Harbin,Heilongjiang 150040;Heilongjiang Provincial Key Laboratory of Ecological Utilization of Forestry-based Active Substances,Harbin,Heilongjiang 150040;College of Chemistry,Chemical Engineering and Resource Utilization,Northeast Forestry University,Harbin,Heilongjiang 150040)

机构地区：[1]东北林业大学化学化工与资源利用学院森林植物生态学教育部重点实验室,黑龙江哈尔滨150040 [2]黑龙江省林源活性物质生态利用重点实验室,黑龙江哈尔滨150040 [3]东北林业大学化学化工与资源利用学院,黑龙江哈尔滨150040 [4]东北林业大学化学化工与资源利用学院林业生物制剂教育部工程研究中心,黑龙江哈尔滨150040 [5]东北林业大学化学化工与资源利用学院生物资源生态利用国家地方工程中心,黑龙江哈尔滨150040

出　　处：《安徽农业科学》2024年第3期163-170,共8页Journal of Anhui Agricultural Sciences

基　　金：东北林业大学大学生省级创新训练计划项目(S2022102252-55);国家林草局野生植物保护管理项目(2019073060)。

摘　　要：[目的]建立同时测定兴安杜鹃中金丝桃苷、槲皮素和杜鹃素含量的高效液相色谱法(HPLC),并探讨兴安杜鹃提取物及3种主要成分对大鼠气管平滑肌舒张作用的影响及其作用机制。[方法]采用HPLC法测定兴安杜鹃中金丝桃苷、槲皮素和杜鹃素的含量并进行方法学考察。采用离体气管灌流法取大鼠气管环,将气管环固定于麦氏浴皿中,使用扑尔敏、吲哚美辛、格列苯脲、左旋硝基精氨酸甲酯(L-NAME)、硝苯地平和普萘洛尔进行预孵育,待气管环张力稳定后,加入60 mmol/L KCl溶液收缩气管环,进行兴安杜鹃对气管环的舒张试验,观察大鼠气管平滑肌张力变化。[结果]兴安杜鹃中3种主要成分HPLC测定方法考察结果显示,3种成分均在2~80μg/mL线性关系良好,平均回收率分别为金丝桃苷100.3%(RSD=2.63%)、槲皮素102.7%(RSD=2.89%)、杜鹃素102.1%(RSD=2.36%)。在吲哚美辛、格列本脲和普萘洛尔孵育组织中,兴安杜鹃提取物及3种单体成分的舒张作用与未孵育组相比没有显著差异;在扑尔敏、L-NAME和硝苯地平孵育的组织中,兴安杜鹃及金丝桃苷与未孵育的组织相比有显著差异,舒张作用有所降低。这种现象可能是通过抑制组胺H1受体、一氧化氮合酶途径和钙离子通道介导的钙离子内流实现的。[结论]该试验建立的方法简单、准确,能快速测定兴安杜鹃中金丝桃苷、槲皮素和杜鹃素的含量,为其质量评价和控制提供了有效的分析手段;并且这3种成分与兴安杜鹃舒张气管的作用有关,可作为支气管扩张剂开发的理论依据。[Objective]To establish a method for the simultaneous determination of hyperin,quercetin and farrerol in Rhododendron dauricum by HPLC,and to discuss the effects of Rhododendron dauricum extract and its three main components on the relaxation of rat tracheal smooth muscle and the mechanism.[Method]The contents of hypericin,quercetin and farrerol in Rhododendron dauricum were determined by HPLC and the methodology was investigated.Rat tracheal rings were obtained by in vivo air tube perfusion method,and fixed in a Mack’s bath.The tracheal rings were pre-incubated with chlorphenamine,indometacin,glibenclamide,L-nitroarginine methyl ester(L-NAME),nifedipine and propranolol.After the tension of the tracheal ring stabilized,60 mmol/L KCl solution was added to shrink the tracheal ring.The relaxation experiment of tracheal ring by Rhododendron dauricum was conducted to observe the changes of tracheal smooth muscle tension in rats.[Result]The investigation results of the HPLC determination method for three main components in Rhododendron dauricum showed that the linear relationship between the three components was good in the range of 2-80μg/mL.The average recovery rates were 100.3%(RSD=2.63%)of hypericin,102.7%(RSD=2.89%)of quercetin and 102.1%(RSD=2.36%)of farrerol.The diastolic effects of the extract and the three main components of Rhododendron dauricum in the incubated tissue of indometacin,glibenclamide and propranolol were not significantly different from those in the unincubated group.The diastolic effects of Rhododendron dauricum and hypericin in the tissues incubated with chlorphenamine,L-NAME and nifedipine were significantly different from those in the unincubated tissues.This phenomenon may be achieved through inhibition of histamine H1 receptors,nitric oxide synthase pathways,and calcium channel mediated calcium influx.[Conclusion]The established method is simple,accurate,and can quickly determine the content of hypericin,quercetin and farrerol in Rhododendron dauricum,providing an effective analytical m

关 键 词：兴安杜鹃 HPLC 金丝桃苷 槲皮素 杜鹃素 含量测定 气管平滑肌 舒张作用 

分 类 号：R285[医药卫生—中药学]