| Purpose:Verify the safety and effectiveness of acne lung heat syndrome,explore the influence on facial flora and the inhibition of propionibacterium acne,and network pharmacology to explore the target of acne treatment,for the clinical treatment of new treatment,for further experimental research to build solid research foundation,make acne can be better applied in clinical,benefit patients.Materials and methods:1.Clinical study:60 patients with acne vulgaris lung syndrome of wind heat were enrolled in the Affiliated Hospital of Liaoning University of traditional Chinese medicine and randomly assigned to the experimental group and the control group.After enrollment,all patients were orally publicized and educated by the researchers and signed the informed consent form.The clinical part of this study was approved by the ethics committee of the Affiliated Hospital of Liaoning University of traditional Chinese medicine.The patients in the experimental group were given 150 ml acne formula orally three times a day,and the control group were given 0.8 g Qingre acne capsule orally three times a day.Subjects in both groups were combined with chloramphenicol tincture for external use twice a day.The course of treatment was 4weeks,and the evaluation method of skin lesions was based on the Guiding Principles for Clinical Research of Traditional Chinese Medicine New Drugs.2.Clinical study:ten subjects were randomly selected from each of the two groups of the first part.Samples were taken before the drug intervention and after the last administration.16sr DNA high-throughput sequencing technology was uesd to abundance and structural changes of microbial flora in skin samples.3.The standard strain of Propionibacterium acnes was revived and subcultured by aseptic operation.The subcultured strains were observed morphologically and stained.After successful subculture,the strain concentration was adjusted to 5×10~5CFU/ml,dilute the traditional Chinese medicine solution by multiple dilution method.The liquid of traditional Chinese medicine and bacteria were added into 96 well plate for anaerobic culture,and the minimum inhibitory concentration was observed.4.Network pharmacology research:firstly,the main components of acne prescription:Angelica sinensis,licorice,Platycodon grandiflorum,forsythia forsythia,tangerine peel,coix seed,Hedyotis diffusa,honeysuckle,wild chrysanthemum,Prunella vulgaris,loquat leaf,saponin thorn,Poria cocos and Coptis chinensis.The targets of the effective components of acne formula were collected and converted into corresponding Gene symbols.Through the disease target database OMIM,GENECARD,disgenet,drugbank,etc.,search for relevant disease targets with"acne","acne cornea","cytic acne","oil acne",etc.Venn map will be drawn through Venn map website and intersection genes will be found.Through Cytoscape 3.7.2,construct the visual"drug active ingredient target"network,and use string database system for reasonable comprehensive calculation to construct the interaction network between proteins.Through the metascape website,the enrichment analysis of go and KEGG pathways of interactive targets was carried OTU.Results:1.A total of 60 subjects were included in this clinical trial,and no adverse reactions and cases were reported during the trial.The safety evaluation index in the two groups after treatment did not different from before treatment.Acne can effectively reduce the form of skin damage,the number of skin lesions,skin damage itching,skin damage color and traditional Chinese medicine symptoms score,effectively improve the state of acne skin damage.2.The bacteria with decreased species abundance after acne intervention were Firmicutes and Bacteroidetes;Actinobacteria and Proteobacteria.At the genus level,the abundance of Propionibacterium,Staphylococcus,Lactobacillus,Rhamnobacter,and Bacteroides was decreased;Corynebacterium,Lawsonella,Acinetobacter,and Streptococcus were increased.The bacteria groups with large differences before and after autotreatment of individual samples were Bacteroides,Streptococcus,Borreliaceae,Acinetobacter,Lawsinella,Rhammicella,Lactobacillus,Corynebacterium,Staphylococcus,and Propionibacterium.3.The colony of Propionibacterium acnes is round or semi-circular,milky white,with neat edges and raised on the agar surface.After Gram staining,it is purple,one is blunt and round,the other is slightly pointed,short rod-shaped or rod-shaped,slightly curved and arranged in single or pairs.Acne formula has obvious inhibitory effect on Propionibacterium acnes in vitro,and the minimum inhibitory concentration of Acne formula is 125mg/m L.4.Through the network pharmacological research,it is known that the main targets of acne prescription are quercetin,kaempferolβ-Sitosterol,luteolin,stigmasterol,epicatechin,farnesol,tetraglyceride,(R)-linalool,isorhamnetin.The main signals related to acne are IL-17 signal pathway,tumor necrosis factor signal pathway,HIF-1signal pathway,PI3K Akt signal pathway,toll like receptor signal pathway,T cell receptor signal pathway,JAK STAT signal pathway,MAPK signal pathway,nod like receptor signal pathway and NF-κB signal path.Conclusion:1.The treatment of lung meridian wind heat syndrome of acne vulgaris is safety and no obvious adverse by use acne formula and chloramphenicol tincture.2.Acne prescription has a certain impact on the diversity and abundance of facial skin flora in patients with acne vulgaris.The main intervention flora of acne prescription above family level is Corynebacteriaceae and Corynebacteriales;Cutibacterium,Propionibacteriales,Propionibacteriaceae;Actinobacteria,Actinobacteria.The main intervention flora of Qingre acne capsule is Firmicutes;Clostridiales,Clostridia;Proteobacteria,Gammaproteobacteria.3 Acne formula has obvious inhibitory effect on Propionibacterium acnes in vitro.4.The main drug active components of acne prescription may achieve the clinical effect of acne vulgaris by affecting the expression of Toll-like receptors and the expression of T o l l-like gene targets. |
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