Study On The Effect Of Danhong Injection On Neurotransmitters In The Brain And Its Metabolism In Rats

Danhong injection (DI), derived from the classicherb pair of Radix Salviae Miltiorrhiae and Flos Carthamitinctorii, is a Chinese medicine compound preparation with function of promoting blood circulation and removing blood stasis. It has significant effect on the treatment of cardiovascular and cerebrovascular diseases in clinic. Previous study showed that the content of monoamineneurotransmitter change dremarkably and it made serious damage to different regions of brain, especially the hippocampus when ischemic cerebrovascular diseases occurred. To date, there is few literature about the effect of DI on the brain neurotransmitter of cerebrovascular diseases and its metabolic pattern in vivo. This paper has focused on the research from the following three aspects by using microdialysis(MD) combined with HPLC-ECD/Q-TOF-MS with high sensitivity and resolution.(1) By optimizing the detection conditions of microdialysis rate, working voltage, types and proportion of mobile phase, a MD-HPLC-ECD method for the determination of primary neurotransmitters such as epinephrine, levodopa, dopamine,5-hydroxyindole acetic acid and tyrosine was established. The results showed that good chromatographic separation and specificity for the five components was achieved with other interfering substances in the brain. The lower limits of detection were 3.3,4.0,3.1,4.0 and 6.0 ng/mL respectively. All of the five ingredients have a good linear relationship during 10?1000 ng/mL. The percentage of relative standard deviation of precision and stability in vivo study was within 5.0%.The above results manifested that the validated method is accurate, sensitive and stable, and it can be applied to the quantitative detection of trace neurotransmitters in the brain.(2) The effect of DI on the content of the neurotransmitter on the hippocampus in ratwas studied by using the MD-HPLC-ECD method mentioned above. Compared with control group, the content of epinephrine, levodopa,5-hydroxyindole acetic acid and tyrosine in drug-treated group has changed significantly. The content of epinephrine and dopamine firstly increased, and then decreased. The content of 5-hydroxyindole acetic acid firstly declined, then increased and finally reduced. The content of the three components above returned to normal level after 9 h, while the content of tyrosine presented a declined trend. Meanwhile, the prototype of DSS was found within 0-1.5 h after administration in the rat brain, and this suggested DSS may penetrate the blood brain barrier.(3) The study on the metabolism of DI in rat brain, heart and liver was carried out preliminarily in rats by HPLC-Q-TOF-MS.11,18 and 19 new compounds were found respectively in brain, heart, liver in rats of drug-treated group, and the structures of 5,14 and 8 compounds were identified by the utilization of MS/MS spectrum. The results exhibited that cryptotanshinone, alkannic acid, DSS, salvianolic acid A and D could pass the blood brain barrier, and the metabolic patterns of the effective components of DI mainly involved in methylation and glucuronidatio in vivo. Synthesize the results of 2nd part, it suggested DSS could penetrate the blood brain barrier.