刺五加叶药物成分脑内代谢动力学及治疗脑缺血作用的在线微透析-质谱分析

摘要

考察了刺五加叶中金丝桃苷和1,5-二咖啡酰奎宁酸入脑及脑内代谢动力学过程,同时评价刺五加叶对脑缺血大鼠脑内神经递质的影响。结扎大鼠双侧颈总动脉构建急性不完全性脑缺血模型,利用在线微透析-液相色谱-串联质谱法,ACE 5 C18-AR液相色谱柱等度洗脱,MRM负离子模式下监测金丝桃苷和1,5-二咖啡酰奎宁酸在健康及缺血大鼠脑内代谢动力学差异,正离子模式检测不同组别大鼠脑内谷氨酸( Glu)、天冬氨酸(Asp)、γ-氨基丁酸(GABA)、乙酰胆碱(Ach)、5-羟色胺(5-HT)、多巴胺(DA)6种神经递质含量。实验结果表明,在线方法线性良好(R2>0.99),准确度、精密度满足分析要求；金丝桃苷和1,5-二咖啡酰奎宁酸均能透过血脑屏障到达脑部,代谢迅速,在缺血大鼠脑内吸收降低,消除加快；相比模型大鼠,预防性刺五加叶给药7天后,缺血大鼠脑内氨基酸类神经递质Glu、Asp、GABA和单胺类神经递质DA的水平显著降低(p<0.01), Ach水平显著升高(p<0.05)。%The brain pharmacokinetics of hyperoside and 1 , 5-dicaffeoylquinic acid and the effects of acanthopanax senticosus leaves on neurotransmitters in cerebral ischemic rat brain were investigated. In the study, acute incompleteness cerebral ischemia model was developed by ligating the bilateral common carotid arteries of rats. The differences of brain pharmacokinetics of hyperoside and 1,5-dicaffeoylquinic acid between control and cerebral ischemic rats were determined by online microdialysis coupled with MS/MS method. At the same time, the contents of glutamic acid (Glu), aspartic acid (Asp), γ-aminobutyric acid (GABA), serotonin (5-HT) , dopamine ( DA) , and acetylcholine( Ach) in rat hippocampus among different groups were determined simultaneously in MRM mode. ACE 5 C18-AR column was used for separation. Results showed that the online analytical method had excellent linearity (R2>0. 99). The accuracy and precision could meet the analysis requirement. Besides, the hyperoside and 1,5-dicaffeoylquinic acid penetrated the blood-brain barrier successfully and metabolized quickly, but the absorption reduced and elimination became faster in cerebral ischemic rats; the contents of Glu, Asp, GABA and DA in brain issue of cerebral ischemic rats decreased significantly ( p<0 . 01 ) and the level of Ach increased remarkably ( p<0 . 05 ) after a preventive medication with the extract of acanthopanax senticosus leaves for one week.