摘要
目的：探究麻黄-苦杏仁药对减轻大鼠气道损伤的物质基础,初步探讨其作用机制。方法：雄性SD大鼠适应性饲养3 d,筛选合格后随机分为9组[正常组、模型组、地塞米松组、麻黄-苦杏仁药对不同配比组（N1~N6组）]。上午9：00开始灌胃相应药物,灌胃1 h后立即用2%氯化乙酰胆碱＋0.4%磷酸组胺引喘并记录引喘潜伏期,连续给药7 d。末次引喘后即刻麻醉,取出主气管,一部分气管用10%甲醛固定,苏木精-伊红（HE）染色观察各组气道病理学特征,另一部分气管匀浆处理,运用酶联免疫吸附法测定表皮生长因子（EGF）,诱导型一氧化氮合酶（i NOS）,内皮素-1（ET-1）的质量浓度。同批6只大鼠适应性饲养后分离出空肠、回肠肠段,制备含药肠吸收液,运用超高效液相色谱-四极杆-飞行时间串联质谱法（UPLC-QTOF-MS/MS）分析麻黄-苦杏仁药对含药肠吸收液中主要差异性成分及其含量。采用多元线性回归分析效应指标与主要差异性成分的相关性。结果：N1～N6组肠吸收液的差异性成分为麻黄碱、伪麻黄碱、甲基麻黄碱和苦杏仁苷;效应指标分别为引喘潜伏期,ET-1,i NOS,EGF质量浓度以及气道病理学总评分。结论：麻黄-苦杏仁药对延长引喘潜伏期和修复气道损伤的主要物质基础为麻黄碱、伪麻黄碱、甲基麻黄碱和苦杏仁苷,其减少i NOS和ET-1含量、抑制EGF生成及维持较低水平NO/ET的主要有效成分为甲基麻黄碱,说明麻黄为主要药味,苦杏仁辅助其修复气道损伤,其中N3组治疗效果最佳。
Objective： To explore the material basis of couplet medicines of Ephedrae Herba and Armeniacae Semen Amarum in reducing airway damage in rats,and to explore its mechanism of action. Method：Male Sprague-Dawley（ SD） rats were adaptively reared for 3 days. After screening,they were randomly divided into 9 groups [normal group, model group, dexamethasone group, and Ephedrae Herba-Armeniacae Semen Amarum with different ratio groups（ N1-N6 groups） ]. At 9： 00 am, the corresponding drugs were intragastrically administered. Immediately after 1 hour of intragastric administration,2% acetylcholine chloride ＋0. 4% histamine phosphate was used to induce asthma and the incubation period was recorded for 7 days.Immediately after the last asthmatic anesthesia,the main trachea was taken out,some of the trachea were fixed with 10% formaldehyde,and hematoxylin-eosin（ HE） staining was used to observe the pathological features of each group. Another part of the tracheal homogenate was treated by enzyme-linked immunosorbent assay for the concentration of epidermal growth factor（ EGF）, inducible nitric oxide synthase（ i NOS） and endothelin-1（ ET-1）. After the adaptive feeding of 6 rats in the same batch,the jejunum and ileum intestine were separated to prepare the drug-containing intestinal absorption liquid. Ultra-high performance liquid chromatography-quadrupoletime-of-flight tandem mass spectrometry（ UPLC-Q-TOF-MS/MS） was used to analyze the major differential components and their content in the drug-containing intestinal absorption liquid of Ephedrae Herba-Armeniacae Semen Amarum. Multiple linear regression analysis was used to analyze the correlation between the effect indicators and the main differential components. Result： The differential components in the intestinal absorption liquid of N1-N6 groups were ephedrine, pseudoephedrine, methylephedrine and amygdalin; the effect indicators were asthmatic latency,ET-1,i NOS,EGF mass concentrations and total score of airway pathology. Conclusion： The main material basis of Ephedrae Herba-Armeniacae Semen Amarum for prolonging the incubation period and repairing airway damage are ephedrine, pseudoephedrine, methylephedrine and amygdalin. The main active ingredient that reduces i NOS and ET-1 levels,inhibits EGF production,and maintains lower levels of NO/ET is methylephedrine. It indicates that Ephedrae Herba was the main medicinal taste,and Armeniacae Semen Amarum assists repairing airway damage. The N3 group has the best therapeutic effect.
作者
周开放
黄刚
肖雄
张颖
敖波
李文宏
ZHOU Kai-fang;HUANG Gang;XIAO Xiong;ZHANG Ying;AO Bo;LI Wen-hong(School of Pharmacy,Jiangxi University of Traditional Chinese Medicine,Nanchang 330004,China;Quanzhou Medical College,Quanzhou 362100,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2018年第19期172-178,共7页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(81460610)
江西自然科学基金计划项目(20151BAB205066)
关键词
麻黄-苦杏仁
药对
引喘潜伏期
甲基麻黄碱
诱导型一氧化氮合酶
内皮素-1
表皮生长因子
Ephedrae Herba-Armeniacae Semen Amarum
couplet medicines
incubation period for asthma
methylephedrine
inducible nitric oxide synthase
endothelin-1
epidermal growth factor
作者简介
周开放，在读硕士，从事中药药理学研究，E-mail：812629849@qq．com;[通信作者]李文宏，博士，副教授，从事中药药理学研究，E-mail：liwhong0311@163．com
