Analyze Of Professor Yu Renhuan's Law Of Formula For Treating Idiopathic Membranous Nephropathy And The Network Pharmacology Of The Core Prescription

Objective:The purpose of this study was to explore Professor Yu Renhuan's law of formula with Jianpi Qushi Heluo method for treating Idiopathic Membranous Nephropathy(IMN),and to explore the potential mechanism of the core prescription for treating IMN.Methods:Clinical data of 86 patients with IMN of spleen-kidney qi-deficiency type were collected from June 2016 to January 2019 after treatment for 12 months by professor Yu Renhuan in Xiyuan hospital China Academy of Chinese Medical Sciences clinic with complete remission or partial remission.The patient information was input into the software of "TCM inheritance assistant platform(V2.5)" to explore the core prescription and law of formula of treating IMN with Jianpi Qushi Heluo method by Yu Renhuan professor.The active ingredient information and pharmacologic information of drugs in the core prescription can be obtained by inquiring in the TCMSP database.The effective components of the drug were screened according to the standard of oral bioavailability 30%or higher and drug-likeness or higher,and the corresponding targets were found in the TCMSP database(http://lsp.nwu.edu.cn/tcmsp.php)and the website of SwissTargetPrediction.IMN-related targets were retrieved from the Human Phenotype Ontology(HPO),Comparative Toxicogenomics Database(CTD)and Therapeutic Target Database(TTD).The data in the three databases were integrated to build the IMN-related target database.The corresponding targets of the screened core prescription active ingredients were matched with the disease-related targets to find the repeated targets in the two sets.The matching targets were used to build a protein interaction network in the STRING database(https://string-db.org/).Then topological analysis of drug targets was carried out with Cytoscape 3.6.0(http://www.cytoscape.org/)software and core targets were sceened.Finally,David v6.8 database was used to perform gene enrichment analysis of Gene Ontology Biological Process(GOBP)and pathway enrichment analysis of Kyoto Encyclopedia of Genes and Genomes(KEGG)on the proteins in the protein interaction network to speculate the mechanism of the core prescription for the treatment of IMN.Results:1.Distribution of sex and age:of the 86 patients,49(57%)were males and 37(43%)were females.24%were aged 30-39,21%were aged 40-49,23%were aged 50-59,and 20%were aged 60-69.2.Among the 86 prescriptions screened,there were 116 traditional Chinese medicines.According to the frequency and proportion of use,the top 10 drugs were:Raw radix astragali,Atractylodes,Fangji,Tuckahoe,Perilla leaf,Dioscorea nipponica,Coix seed,Radix angelica,Codonopsis,and Licorice.From high to low,the proportion of the four gases used was warm,cold,flat,cool and hot.From high to low,the proportion of the five flavors of the medicine was sweet,bitter,spicy,sour and salty.From high to low,the proportions of the meridians were spleen,lung,liver,kidney,stomach,heart,bladder,large intestine,bile,small intestine,pericardium,and three focal meridians3.Using association rules to mine formulas and core prescriptions,the results showed that there were 20 groups of high-frequency drug pattern combinations(drug combination occurs more than 60 times).There were 30 drug combinations with a confidence of 0.9 or above.Among them,there were 6 groups of drug combinations with the correlation coefficient of more than 98%,including Raw radix astragali,Atractylodes,Radix Stephaniae Tetrandrae,Tuckahoe,Perilla leaf and Dioscorea nipponica.Set the support level to 40 and the confidence level to 0.9,and the medicines included Raw radix astragali,Atractylodes,Fangji,Tuckahoe,Perilla leaf,Dioscorea nipponica,Coix seed,Radix angelica,Codonopsis,and Licorice.Combined with the results of statistical analysis and data mining,the core prescription consisted of Raw radix astragali,Atractylodes,Radix Stephaniae Tetrandrae,Tuckahoe,Perilla leaf,Dioscorea nipponica,Coix seed,Radix angelica,Codonopsis,and Licorice.4.Using entropy clustering method to mine potential new drug combinations in addition to core prescriptions and core medication laws,the results showed that the new drug combinations with the correlation coefficient greater than 3%were Lotus seed and Cicada slough,Codonopsis and Hedyotis diffusa,Lotus seed and Rhizoma chuanxiong,Licorice and Alisma,Licorice and Hedyotis diffusa,Lotus seed and Raw rehmannia,Cornus officinalis and Licorice,Lotus seed and Codonopsis,Lotus seed and Radix cyathulae,and then the potential drug combinations of 3?4 flavors were analyzed by entropy clustering,such as Lotus seed,Radix scutellariae and Pyrrosia Leaf;Euryale seed,Rosa laevigata Michx,and Agrimonia pilosa;Antain seed,the root bark of white mulberry,Tangerine peel,etc.Finally,two groups of potential drug combinations were clustered to obtain new 10 groups of new drug combinations,including Lotus seed,Radix scutellariae,white hyacinth bean and Pyrrosia Leaf;Lotus seed,Radix angelica,Dioscorea nipponica,white hyacinth bean and Red peony root;Euryale seed,Saposhnicovia divaricate,Rosa laevigata Michx,and Agrimonia pilosa;Cicada slough,Cassia twig,Licorice and Rhizoma chuanxiong;Radix Stephaniae Tetrandrae,Polygonatum Kingianum,Semen cassia,Ligustrum lucidum Ait and Perilla stem;Tangerine peel,Forsythia,the root bark of white mulberry,Antain seed and Pericarpium arecae;Licorice,Indirubin,Dictamni Cortex and Hives;Red peony root,Pyrrosia Leaf,Codonopsis and Rhizoma chuanxiong;Paeonia suffruticosa,Cornus officinalis,Tangerine peel and Raw rehmannia;Cornus officinalis,Raw rehmannia,Ripe rehmannia,Wolfberry and Coptis.5.The results of network pharmacology showed that the main effective ingredients of core prescription for treating IMN were quercetin contained in Licorice and Raw radix astragali,luteolin contained in Codonopsis and Perilla leaf,tuckahoe acid contained in Tuckahoe.There were 7 core targets in the core prescription,including:protein kinase B(PKB/Akt),Tumor protein p53(TP53),Albumin(ALB),Caspase 3(CASP3),Interleukin 6(IL-6),Mitogen-activated protein kinase 3(MAPK3),Tumor necrosis factor(TNF).Through GOBP enrichment analysis,it was found that the core prescription had biological functions such as regulating cell proliferation,aging,apoptosis and response to other substances,and mainly focused on the biological function of regulating cell apoptosis.The results of KEGG pathway enrichment analysis indicated that the PI3K-Akt and p38MAPK signaling pathways may be the main pathways of the core prescription.Conclusions:Through data mining,Yu renhuan professor with Jianpi Qushi Heluo method of the treatment of IMN core prescription consisted of Raw radix astragali,Atractylodes,Fangji,Tuckahoe,Perilla leaf,Dioscorea nipponica,Coix seed,Radix angelica,Codonopsis,Licorice 10 drug.Based on network pharmacology,the main effective ingredients of core prescription for treating IMN were quercetin contained in Licorice and Raw radix astragali,luteolin contained in Codonopsis and Perilla leaf,tuckahoe acid contained in Tuckahoe.There were 7 core targets in the core prescription,including:Akt?TP53?ALB?CASP3?IL-6?MAPK3?TNF.Through GOBP enrichment analysis,it was found that the core prescription had biological functions such as regulating cell proliferation,aging,apoptosis and response to other substances,and mainly focused on the biological function of regulating cell apoptosis.The results of KEGG pathway enrichment analysis indicated that the PI3K-Akt and p38MAPK signaling pathways may be the main pathways of the core prescription.