| ObjectivesAccording to the property theory of cold/heat herbs in traditional Chinese medicine,the system biology techniques such as network pharmacology and metabonomics were used on the projects.As a bottom-up researching strategies,the dry experiment was used firstly to predict and then the wet experiments were used to validate.Regarding these,we would like to explore the possible molecular mechanisms corresponding to the specific biological effects of the typical cold/heat drug group,and to create conditions for further revealing the scientific connotation of cold and hot herbs of Chinese herbal medicine.Material and MethodsDry experiment:To explore the molecular mechanism of the specific biological effects of herbs with typical cold/heat properties based on network pharmacological technique.Ten commonly-used herbs were selected,including five typical cold property(Rheum L,Scutellaria baicalensis Georgi,Coptis chinensis Franch.CortexPhellodendriChinensis,Gentians scabra Bunge)and five typical heat properties(Aconitum carmichaeli Debx,Cinnamomum cassia Presl,Lonicera japonica Thunb,Evodia rutaecarpa Juss,Benth,Curculigo glabrescens),then retrieve all the chemical compositions of each herbal medicine in the ’Chinese Dictionary(Second Edition)’,and screen the active ingredients according to the ADME parameter based on TCMSP database.The human target proteins which can be docking with the selected active ingredients were searched in the PubChem and TCMSP database and visualized by Cytoscape 3.5.1 software.The specific target proteins related in only cold or heat propertied medicines were filtered based on frequencies count then imported into network analysis platform to build a molecular pharmaceutical network of herbal medicines with cold/heat properties.By using the function of the pathway analysis module,the classic pathways involved in specific target proteins of the cold drug group and the heat drug group were obtained.A comparative analysis was made of the effect of the cold drug group and the hot drug group,and the molecular mechanism of the cold drug effect of Chinese medicine was predicted through the comparison of the action sites.Wet test:The healthy Sprague-Dawley Sprague-Dawley(SD)SD rats were used for the study.Each group consisted of 8 rats and 4 males and 4 females each:the blank group,the cold drug C1-C5 group,and the hot drug H1-H5 group.Record baseline data to ensure comparable between the groups at baseline.The rats were fed adaptive for 1 week.Choose 5 flavors of typical cold(rhubarb,astragalus,berberine,gardenia,gentian),heat(Fuzi,dried ginger,ebony,cinnamon,curculigo),and use traditional Chinese medicine granules,according to the maximum dose of adult pharmacopoeia Per rat drug dose,the powder was dissolved in distilled water,the rats were given intragastric administration once a day,each time 1ml/100g animal body weight(each group of rats fed a traditional Chinese medicine);blank group orally distilled water 1ml/100g animal body weight.21 days after continuous gavage,the material was taken.Observed indicators include:general conditions(rat hair,stool,tongue color,mental status,etc.),food intake,water intake,rectal temperature,behavioral tests(open field test,lane tending),organ coefficient(liver weight,liver weight/weight,kidney weight,kidney weight/body weight,spleen weight,spleen weight/body weight,thymus weight,thymus weight/body weight,heart weight,heart weight/body weight,adrenal weight/body weight),biochemical indicators of whole blood,each The serum metabolism of the rats in the whole spectrum.It is used to compare the biological effects produced by normal and cold drugs on macroscopic and microscopic aspects.Results(1)Although the active ingredients in the cold and hot drug groups are different,they share a large part of the target protein,which is consistent with the results of previous studies on cold drugs and hot drugs;(2)In the selected herbal medicines,the cold specific molecular included glycogen synthase kinase(GSK3),mitogen activated protein kinase(Mapk)3,G-protein coupled receptor(Gpcr),cyclin-dependent kinase(CDK),the related biological processes were glycogen synthesis,autonomic,inflammation,apoptosis.Heat specific molecular consisted of matrix metalloproteinases(MMP-2,MMP-9),y-aminobutyric acid(of GABA),transcriptional regulatory protein(BACHE1),cytochrome P450 3A4 enzyme(of CYP3A4),the related regulated functional pathways were central nervous system,cardiovascular system function has played its pharmacological function;(3)In the pharmacological network of cold and hot drugs,the direction of action and strength information of the drugs are added,and the qualitative and quantitative vector pharmacology network can better use the network pharmacology method to discriminate the cold and heat properties of traditional Chinese medicines.Now it is widely used.Metabolomics techniques can provide technical support for the achievement of this goal.Conclusion(1)Cold/hot drugs have great differences in active ingredients and biological behaviors.Even if the cold drug group and the hot drug group share the same target,there are also differences in the direction and intensity of the specific control of the drug.(2)Prediction of GSK3,Mapk,G protein-coupled receptors(Gpcr),CDK,MMP-2,MMP-9,gamma-aminobutyric acid(GABA),transcriptional regulatory proteins,BACHE1,and CYP3A4 as a means of distinguishing between traditional Chinese medicine and cold by network pharmacology Potential drug targets.It can serve as an important entry point for follow-up verification of molecular biology.(3)In the pharmacological network of cold and hot drugs,the direction of action and strength information of the drugs are added,and a qualitative and quantitative vector pharmacology network can be constructed to better use the network pharmacology method to discriminate the cold and heat properties of traditional Chinese medicines.Currently widely used metabolism Metaomics technology can provide technical support for the realization of this goal. |
PDF Full Text Request