Prof. Wei Junping's Research On Prescription Rules And Network Pharmacology In The Treatment Of Hashimoto's Disease With Liver Stagnation And Spleen Deficiency Syndrome

Objective:1.Using the software of "Traditional Chinese medicine inheritance support system(TCMISS)v2.5",this article analyzed the medication rules of Professor Wei Junping in the treatment of Hashimoto’s thyroiditis(HT)of liver depression and spleen deficiency syndrome,in order to excavate its core drugs and compatibility methods,to summarize Professor Wei’s medication experience,and to obtain the core prescription.2.To explore the potential key targets and pathways of the core prescription in the treatment of HT by using the method of network pharmacology,so as to lay a theoretical foundation for the study of the mechanism of the intervention of liver-soothing and spleen-strengthening method in HT.Methods:1.The medical records and prescriptions data of patients with HT of liver depression and spleen deficiency syndrome treated in the outpatient clinic of Director Wei Junping of Guang’anmen Hospital from September 1,2018 to December 31,2020 were collected.According to the inclusion and exclusion criteria,the medical record data were screened and standardized,and entered into the software of TCMISS v2.5,which was analyzed from the aspects of four qi,five flavors,meridian tropism,drug group association and so on.After that,I explored the prescription rules through association rules and mined potential new drug combinations based on entropy clustering and other methods by TCMISS v2.5.Then the core prescription was obtained by having an interview and discussion with Professor Wei Junping.2.Through TCMSP database,the main active components and action targets of each drug in the core prescription were screened according to the standards of oral bioavailability(OB)≥30%and drug-like(DL)≥0.18,and the information was standardized by Uniprot protein database.Through Genecards,OMIM,DisGeNET and other databases,with"Hashimoto’s thyroiditis","Hashimoto’s Disease" and "HT" as key words,the related targets of HT were obtained,after merged and removed,the core prescription-HT common targets were obtained.Then the PPI network was constructed by using STRING database.The KEGG signal pathway and GO bioaccumulation were analyzed by Metascape platform,and visualized by Cytoscape3.6.1 software,the "prescription-ingredients-disease-targets-pathways" network was constructed.Results:1.A total of 206 medical records were included in this study,of which 91.75%were female and 8.25%were male,and the youngest was 7 years old and the maximum was 76 years old.The distribution is as follows:2.43%of patients under 10 years old,1.46%between 10 and 19 years old,11.65%between 20 and 29 years old,11.65%between 30 and 39 years old,39,32%between 40 and 49 years old,16.50%between 40 and 49 years old,20.39%between 60 and 69 years old,2.43%,>70 years old,2.43%,The distribution of thyroid function in patients with liver depression and spleen deficiency syndrome was as follows:normal thyroid function accounted for 40.29%,subclinical hypothyroidism accounted for 19.42%,and hypothyroidism accounted for 40.29%.The frequency of symptoms from high to low is:fatigue,loose stool,poor sleep,palpitation,chills,irritability,abdominal distension,neck swelling,sweating,chest tightness,pharynx foreign body sensation,phlegm in pharynx,dry stool,less menstruation,stomach distension and so on.2.Among the 206 prescriptions,the top 15 frequently used herbs were Bupleurum,Poria cocos,tangerine peel,Magnolia,licorice,white peony,perilla leaf,Angelica,Atractylodes macrocephala,Prunella vulgaris,Pinellia ternata,perilla stem,Radix Astragali,turmeric and Fructus Aurantii.The distribution of the properties from high to low were warm,cold,plain,cool and hot medicine.The distribution of medicine tastes from high to low is bitter taste,pungent taste,sweet taste,sour taste,salty taste and astringent taste.And the top five meridians from high to low are spleen meridian,liver meridian,lung meridian,stomach meridian and heart meridian.3.After the analysis of association rules,three pairs of high frequency drug combination patterns were obtained:tangerine peel-Bupleurum,tangerine peel-Poria cocos and Bupleurum bupleurum-Poria cocos.Four groups of drug combinations with correlation degree>0.98 were obtained:Bupleurum,perilla stem->perilla leaf,Poria cocos,perilla stem->perilla leaf,Magnolia officinalis,perilla stem->perilla leaf.The core drugs were perilla leaf,white peony,Poria cocos,magnolia,Bupleurum,tangerine peel,perilla,grilled licorice and angelica.17 potential new drug combinations were obtained based on entropy clustering.After having an interview and discussion with Professor Wei,the core prescriptions were set as Perilla leaves.Radix Paeoniae Alba,Poria cocos,Magnolia,Bupleurum,Citrus,Radix Astragali and Pinellia ternata.4.Based on the network pharmacological analysis of the core prescription,a total of 86 active components were obtained,mainly include quercitrin,kaempferol.β-sitosterol,isorhamnetin,stigmasterol,etc.82 intersecting targets of prescription and disease were obtained,and TNF,JUN,TP53,MAPK1,AKT1,IL6,MAPK14,VEGFA,EGFR as the main targets.Enrichment analysis showed that there were 301 related pathways,including AGE-RAGE signal pathway,TNF signal pathway,MAPK signal pathway,IL-17 signal pathway,EGFR tyrosine kinase inhibitor resistance pathway,PI3K-Akt signal pathway,Toll-like receptor signal pathway,Th17 cell differentiation pathway and VEGF signal pathway,etcConclusions:1.Through data mining,it is found that the patients with HT of liver depression and spleen deficiency syndrome are mainly female,and their ages are mostly between 30 and 39 years old.The distribution of thyroid function is mainly normal thyroid function and clinical hypothyroidism,and some patients belong to subclinical hypothyroidism.2.When Professor Wei Junping treats HT patients of liver depression and spleen deficiency syndrome,he not only deal with liver meridian and spleen meridian,but also combines lung,stomach and heart to treat together.He usually chooses warm,cold and flat herbs to make yin and yang harmonious,and he mostly with the taste of pungent,bitter and sweet to regulate qi.Using data mining and interviews,the core prescriptions are:perilla leaf,white peony,Poria cocos,Magnolia,Bupleurum,tangerine peel,raw astragalus,Pinellia ternata3.Through the study of network pharmacology,it is concluded that the core prescription of the method of soothing the liver and invigorating the spleen is to treat HT in a comprehensive way of multi-components,multi-targets and multi-pathways.The main mechanism is to regulate 82 targets related to HT through the core active components with the functions of inflammation,oxidative stress and immune regulation,and to regulate many pathways such as AGE-RAGE signal pathway,TNF signal pathway,MAPK signal pathway,IL-17 signal pathway,PI3K-Akt signal pathway and so on,so as to intervene and treat HT.