| Zaoren Anshen Prescription(ZAP),the classic formulae,is simplified from Tianwang Buxin Dan.It consists of Semen Ziziphi Spinosae(Suanzaoren,Fried),Salviae Miltiorrhizae Radix et Rhizoma(Danshen,vinegar)and Fructus Schisandrae Chinensis(Wuweizi,vinegar).It has the effect of buxin and tranquilizing,and is mainly used to treat insomnia caused by heart,liver and blood deficiency in Traditional Chinese medicine.Abnormal changes in monoamine neurotransmitters can be used as endogenous indicators for insomnia patients.Therefore,this paper took 7 active components and 4 monoamine neurotransmitters in zaoren anshen prescription as indicators and adopted uplc-tsq-ms /MS method for synchronous analysis.The obtained experimental results are as follows:1.An Ultra performance liquid chromatography-tandem triple quadrupole mass spectrometry(UPLC-TSQ-MS/MS)method was established for the simultaneous determination of 7 effective components and 4 monoamine neurotransmitters in plasma and tissues of normal and insomnia rats.The results of the methodological investigation showed that the linear relationship of the 11 components was good,the precision values were less than 14.92%,the accuracy values were within ﹣18.12%-19.88%,the extraction recovery and matrix effect were in the range of 73.94%-101.06 and 80.06%-105.70%,respectively.The results were consistent with the requirements of biological samples.The established method can be applied to determine the content of 11 components in plasma and various tissues,and applied to the comparative study of pharmacokinetics of Zaoren Anshen Formula in normal and insomnia rats.2.The rats were divided into normal group and model group,the tail with chronic stimulation combined with intraperitoneal injection of chlorobenzene alanine(PCPA)method was adopted to make insomnia model,the same dose of ZAP extract(15 g/kg)was given to normal rats and insomnia rats by intragastric administration of seven consecutive days at the morning of each day,and the changes of plasma pharmacokinetics of seven active components and four monoamine neurotransmitters were investigated.The results showed that the peak concentration(C max)and area under the blood-drug concentration-time curve(AUC)of seven active components in insomnia model rats were both higher than that in normal rats(P < 0.05).The rats after administrated,the plasma levels of noradrenaline(NA)and dopamine(DA)in normal and insomnia rats showed a downward trend,while the levels of 5-hydroxy tryptamine(5-HT)and 5-hydroxyindoleacetic acid(5-HIAA)showed an upward trend,which was the same as the content of seven active components.And compared with normal rats,the level of NA and DA decreased more strongly and the level of 5-HT and 5-HIAA increased more strongly in plasma of insomnia model rats.The results provided a supporting role for further research on the mechanism of ZAP.3.The extract of ZAP was given to normal and insomnia rat,the results of tissue distribution showed that the levels of NA and DA decreased from 5 min to 7 h in brain tissue of normal and insomnia rats after administrated,and the concentration of salvianolic acid B was the same,while the 5-HT level showed an upward trend,which was the same as that of spinosin,danshensu,protocatechualdehyde,schisandrin and deoxyschisandrin.Compared with the normal state,the level of DA was decreased to a higher degree in the brain tissue of rats with insomnia,and the level of 5-HT was also increased to a higher degree.Meanwhile,the brain content of the protocatechualdehyde,spinosin,salvianolic acid B,schisandrin and deoxyschisandrin was higher in the insomnia state.In addition,the comparison between the model administration compound group and the model administration positive drug(esazolam)group showed that there was no significant difference between the positive drug and zhongren anshen prescription in terms of the improvement effect of brain 5-HT level in the whole collection time.The results provide a reference for elaborating the material basis of ZAP. |
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