The Study Of Traditional Chinese Medicine Compound And Its Drug Targets Based On The Zebrafish Drug Screening Platform

ObjectiveThe purpose of this project is to observe the effects of Tusi decoction on the circadian behavioral phenotype of zebrafish sleep by using a zebrafish digital drug screening platform,and through Zebrafish efficacy fingerprinting do behavioral phenotypic comparison.To investigate Tusi decoction different compound and compound disassembly of different drug combinations for common effects and differential effects on insomnia,in order to provide an objective basis for "different treatment of the same disease" in Chinese medicine。Through cluster analysis,correlation analysis,exploring the laws of the formula of Tusi decoction,at the same time,target fitting is performed through a database of compounds with known targets,to explore possible targets and mechanisms of action of and single drug.It provides theoretical basis for prescription optimization and discovery of new drugs,and explores new ways for Chinese herbal compound and single drug research.MethodsBy using the zebrafish drug screening platform,with 4 Dpf of wild-type zebrafish juveniles were added to the 96-well plates one by one with a pipetting gun,and then a set concentration of the intervention drug was added to each well by a pipette according to the drug grouping.In the blank group,an equal amount of Holt Buffer medium containing methyl blue was added and placed in the juvenile fish observation behavior apparatus to be observed.Open zebrafish behavior recorder and constant temperature water bath,set the bath temperature to a constant temperature of 28.5 degrees Celsius,set up the relevant parameters of the zebrafish behavior recorder and start the experiment,Zebrafish Automatic Imaging System automatically records relevant data.After 48 hours,the experiment is over and the recorded data is output.After processing through the R software,the data arranged by the six behavioral indicators is obtained.Through data processing methods such as one-way analysis of variance,cluster analysis,and correlation analysis,the zebrafish’s behavioral phenotypes,clustering patterns,and correlation strengths were analyzed to analyze its possible targets and possible mechanisms.ResultsThe Tusi decoction,s effects of different dosages and combinations of different drugs on the six behavioral indices of zebrafish showed diversity,but their normal sleep/wake rhythms existed.In the cluster analysis,the low-dose group of Fuling decoction was firstly grouped with the high-dose group of Radix Polygala Zaoren,together with melatonin group,decoction medium-dose and high-dose group,and low-dose and middle-dose group of Radix Polygala Zaoren.The ginseng angelica licorice middle-dose and high-dose group has successively formed a large class.The high-middle-low-dose group of dodder Deer cream and the low-dose group of ginseng angelica licorice licorice constitute another major category.The low-dose group of tusijian had high similarity with the middle and high-dose group of fuling yuanzhi zaoren.The middle and high-dose group of tusijian had a high similarity with the middle and high-dose group of fulingyuanzhizaoren and the high-dose group of renshendangguishanyaozhigancao.The low-dose group of lujiaoshuang and high dose of renshendanguishanyaozhigancao had higher similarity.Dose correlation analysis shows,except for the strong correlation between the decoction medium dose group and the high dose group of tusi decoction,there is no obvious correlation.There are seven main types of nerve receptors involved in the compounding of the target and the various groups,adrenergic receptors,serotonin receptors,dopamine receptors,gamma hydroxybutyrate receptors,melatonin receptors,antihistamine receptors,and glutamate receptors,respectively,there are 24 kinds of similar drugs involved,drugs with similar have sedative effects,small part has bidirectional regulation and dose dependence.ConclusionsThe efficacy of different doses of the compound,drug group and single drug is the result of comprehensive action through different targets,and there are differences in the target of each compound and drug group for different doses,which is effective after boiling with different drug combinations.It is related to the analysis of changes.The diversity of zebrafish juvenile behavior and the clustering analysis results are based on the characteristics of multi-component and multi-targets of Chinese herbal compound.The mechanism of Qishao decoction in treating insomnia may be related to its role in many biological targets with sedation.The low-dose decoction group was inhibited at night and had no significant effect during the day,which was in accordance with the biological rhythm characteristics of human eel nighting.Therefore,the clinical performance of its treatment of insomnia is great.