Explore On Professor Yang Suqing’s Core Prescription And Mechanism Of Acne Based On Data Mining And Network Pharmacology

Objective:1.Through the Traditional Chinese Medicine Inheritance Support System(TCMISS)and data mining technology,this paper makes a retrospective study on Professor Yang Suqing clinical medical records in the treatment of acne,analyzes the medication law,prescription characteristics and syndrome differentiation ideas in the treatment of acne,and refines her clinical experience.2.To preliminarily predict the core drug combination of the most representative TCM syndrome type of acne and the potential molecular mechanisms by which they exert their therapeutic effects.3.by network pharmacology and molecular docking methods,Combined with the results of traditional Chinese medicine inheritance support system and network pharmacology,the core prescriptions of acne key syndromes were used in the leading theoretical pathway of network pharmacology for experimental verification.To establish an animal model of acne with combination of disease and syndrome,and to analyze the micro targets of core prescriptions.Methods:1.According to the diagnosis,inclusion and exclusion criteria,751 valid acne medical records of Professor Yang Suqing outpatient clinic in the the First Affiliated Hospital of Heilongjiang University of traditional Chinese medicine from September 2013 to December 2021 were collected.After preliminary processing,the collected acne medical records were uploaded to the traditional Chinese medicine inheritance support system(V2.5)by double input.The common TCM syndrome types,high-frequency Chinese medicine,nature and taste tropism,core drug combination,medication rules and clinical experience of the tutor’s diagnosis and treatment of acne were summarized.2.Network pharmacology and molecular docking analysis were performed to select the core drug combination of acne with phlegm blocking collaterals.TCMSP database was used to retrieve and obtain the chemical composition and potential targets of the drugs in the prescription,and Cytoscape 3.7.1 was used to network display their interaction.The acne targets were obtained from Genecards,OMIM,TTD and Drugbank databases.Protein-protein interaction network analysis was performed using the STRING database to construct the PPI network.Cytoscape 3.7.1 software was used to analyze the network topology to screen the core targets of the formula in the treatment of acne.The DAVID database was used to perform GO and KEGG pathway enrichment analysis of intersection targets.Autodock software was used to perform molecular docking between the main active ingredients and the core targets to predict the potential mechanism of action of the core recipe in the treatment of acne.3.A rat model of phlegm and blood stasis syndrome acne was established by oleic acid smearing,Bacillus acnes injection,swimming in ice water and highfat diet.The rats were randomly divided into 6 groups,with 5 rats in each group.The rats were divided into blank control group,model control group,western medicine control group(isotretinoin soft capsule),Chinese medicine low-dose group(core prescription low dose),Chinese medicine medium-dose group(core prescription medium dose),Chinese medicine high-dose group(core prescription high dose).After drug gavage,the apparent morphology of skin lesions in each group was observed,and the changes of skin lesions during gavage were recorded.HE staining was used to compare the micropathological changes of skin lesions in each group;ELISA was used to detect the levels of mTOR,HIF-1α,and VEGF in serum of each group;RT-PCR was used to detect the expression of mTORmRNA,HIF-1αmRNA,VEGFmRNA,and Western Blot was used to detect the expression of mTOR,HIF-1α,and VEGF protein in the lesions.The expressions of mTOR,HIF-1α and VEGF proteins in the skin lesions were detected by immunohistochemical staining,and the expression of Th17 cells in the serum of rats in each group was detected by flow cytometry.The target effects of the core prescription of traditional Chinese medicine on mTOR/HIF-1α/VEGF pathway and Th17 cells were analyzed.Results:1.Related results of traditional Chinese medicine inheritance support systemAmong the 751 cases hace 617 female patients and 134 male patients.Age distribution:20-29 years old has 352 cases;30-39 years old has 203 cases;1019 years old has 132 cases;other age groups have 64 cases in total.Patients with family history accounted for about 56%of the total.602 patients felt that there were predisposing factors,among which diet,mood,staying up late,menstruation,cosmetics and other factors accounted for a high proportion.IIn terms of the relationship between the degree classification and TCM syndrome types,the phlegm turbidity Blocking Collaterals type has the most moderate and severe grade Ⅳ;the liver depression transforming fire type has the most moderate grade Ⅲ;the Chong Ren disharmony type has the most moderate grade Ⅲ;the blood hot wind type has the most moderate grade Ⅱ;the cold coagulation and qi stagnation type has the most severe grade Ⅳ;and the heat toxin accumulation type has the most moderate grade Ⅲ;spleen deficiency dampness excess type and rheumatism heat accumulation type are moderate grade Ⅱ;The 10 most frequent symptoms from high to low were nodules,pustules,greasy face,poor appetite,slippery pulse,late menstruation,irritability,abdominal distension,cysts,white and greasy fur.Statistical results of TCM syndrome types:Among 751 cases of acne,354 cases(47.14%)were phlegm blocking collaterals;144 cases(19.17%)were liver stagnation and fire;108 cases(14.38%)were Chong Ren disharmony;48 cases(6.39%)were blood hot wind;42 cases(5.59%)were cold coagulation and qi stagnation;23 cases(3.06%)were heat toxin accumulation;17 cases(2.26%)were rheumatic heat accumulation;15 cases(2.00%)were spleen deficiency and dampness accumulation.The four qi frequencies of drugs from high to low were warm,cold,flat,hot and cool.The frequency of the five flavors of the drugs used was from high to low,followed by spicy,bitter,sweet,salty and sour.The frequency of the drugs used was liver meridian,spleen meridian,lung meridian,stomach meridian,kidney meridian,heart meridian,gallbladder meridian,bladder meridian,large intestine meridian,small intestine meridian,pericardium meridian and Sanjiao meridian from high to low.The top 10 drugs with medication frequency are licorice,Pinellia ternata,tangerine peel,Fritillaria thunbergii,bupleurum,Scutellaria baicalensis,black medicine,green bark,fried Atractylodes macrocephala with bran and Ligusticum chuanxiong.Based on the study of frequency,clustering,association rules,and complex network analysis,the core drug combinations for the diagnosis and treatment of acne were determined as Scutellaria baicalensis,Radix aconiti,Pinellia pinellia,Radix bupleurum,Radix phellodendri,Radix codonopsis,Radix glycyrrhizae,Rhizoma atractylodes rhizoma,Fritillaria fritillaris,Phellodendri officinalis,Ligusticum chuanxiong.2.Research Results on Network Pharmacology and Molecular Docking:Through network pharmacology analysis,228 medicinal chemical components in the core recipe of phlegm-turbidium-blocking collateralblocking pattern were screened,280 potential targets were obtained,1466 disease targets of acne and 113 drug-disease intersection targets were retrieved.The core targets included STAT3,RELA,JUN,TNF,IL-6,AKT1,MAPK1,TP53,IL10,ESR1,MYC,IL4,IL1B,MAPK8,IL2,IL1A,CXCL8,etc.GO gene annotation entries involved 492 BP,93 MF and 44 CC(P<0.01).114 KEGG signaling pathways,Involved in TNF signaling pathway,HIF-1 signaling pathway,FoxO signaling pathway,Toll receptor signaling pathway,T cell receptor signaling pathway,PI3K-AKT signaling pathway,NF-kB signaling pathway,MAPK signaling pathway,ErbB signaling pathway,VEGF signaling pathway,mTOR signaling pathway,steroid hormone biosynthesis,Through nutrient signaling pathway,estrogen signaling pathway,Jak-STAT signaling pathway,p53 signaling pathway,AMPK signaling pathway,etc.The two active ingredients with the highest visibility value of molecular docking results had good binding with the core targets AKT1,JUN,MAPK1,TNF,and IL-6.3.Related research results of core prescription intervention on mTOR/HIFlα/VEGF pathway and Th17 cell mechanism:Under macroscopic observation,after the core prescription intervention,the acne lesions of rats showed a significant repair process,and western medicine control group≈Chinese medicine high dose group≈Chinese medicine medium dose group>Chinese medicine low dose group.After gavage,the pathological changes of microscopic skin lesions in each group showed that there were still obvious abnormal keratinization of epidermis,spinous layer thickening,inflammatory cell infiltration,telangiectasia and other pathological changes in the model control group,while the skin lesions of rats in the middle dose and high dose of Chinese medicine groups and the western medicine control group basically returned to normal,with complete structure and regular and neat arrangement of epidermal cells.Compared with the blank control group,mTOR/HIF-1α/VEGF pathway and Th17 cells were highly expressed in the above different serum and skin lesions.The levels of mTOR,HIF-1α,VEGF and CD4+IL-17+in serum were significantly decreased,and the expressions of mTORmRNA,HIF-1αmRNA and VEGFmRNA in skin lesions were inhibited in the TCM middle-dose group,the TCM high-dose group and the western medicine control group.At the same time,the high dose of traditional Chinese medicine group and the western medicine control group could significantly inhibit the expression levels of mTOR protein,HIF-1α protein and VEGF protein in skin lesions.For the expression of the above positive proteins located in the nucleus and cytoplasm,the TCM middle-dose group,the TCM high-dose group and the western medicine control group could significantly inhibit the positive expression rate of their protein content.The low dose of traditional Chinese medicine also inhibited the related indicators of mTOR/HIF-1α/VEGF pathway and CD4+IL17+expression.Therefore,the core prescription of traditional Chinese medicine can significantly affect the dynamic and continuous process of mTOR/HIF-1α/VEGF pathway and CD4+IL-17+from serum distribution,to skin lesion genes,and then to skin lesion protein synthesis.Conclusion:1.The tutor’s treatment of acne is mainly based on Yang deficiency and phlegm obstruction,emphasizing warming Yang and eliminating phlegm.The mentor’s clinical syndrome usually divides acne into 8 TCM syndrome types."Phlegm-stasis mutual knot type" is the most common syndrome type for the teacher to distinguish and treat,and is often treated with "warming Yang strengthening spleen,Qi removing phlegm,promoting blood circulation and removing blood stasis".2.Through the analysis of the network pharmacological mechanism of"phlegm-stasis internode type" acne core prescription,it is preliminatively speculated that it can achieve the effect of acne treatment by participating in the regulation of inflammatory response,immune mechanism,hormone level,sebum metabolism,oxidative stress and other biological processes.3.In this study,the acne rat model of "phlegm-stasis interjunction type"was established,and the model was systematically evaluated from the perspective of skin lesions and TCM syndromes,which provided a scientific and reasonable basis for the construction of the acne rat model of "phlegmstasis interjunction type".4.Based on network pharmacology,this study found the intervention mechanism target of "Phlegm-stasis Hu-jietype" acne core prescription on the acne rat model of "phlegm-stasis Hu-jietype".It may inhibit mTOR/HIFlα/VEGF pathway conduction and Th17 cell expression in serum and skin tissue,thus improving the environment of sebaceous glands,inhibiting inflammatory response,affecting angiogenesis and so on,and finally achieving the purpose of acne treatment.