Study On The Mechanism Of Action Of Modified Chaihu Guizhi Decoction In The Treatment Of Chronic Obstructive Pulmonary Diseas

Purpose:Based on the data mining method,to explore the rule of drug use in the treatment of chronic obstructive pulmonary disease by Professor Lv Xiaodong;The mechanism of action of Chaihu Guizhi Decoction in treating chronic obstructive pulmonary disease was analyzed by means of network pharmacology.To evaluate the therapeutic effect of Chaihu Guizhitang Huazai Decoction on chronic obstructive pulmonary disease(COPD)in rats,and explore its mechanism based on PI3K/Akt/NF-κB p65 signaling pathway.Material and method:1 Clinical observation:Collect 260 outpatient medical records of Professor Lv Xiaodong’s application of traditional Chinese medicine compound in the treatment of chronic obstructive pulmonary disease patients from July 2019 to November 2022.Use statistical methods such as frequency analysis,cluster analysis,association rule analysis,and factor analysis to summarize and explore the nature,taste,and meridian of drugs,summarize prescription characteristics,and explore the compatibility relationship between drugs,Thus analyzing Professor Lv Xiaodong’s prescription and medication patterns for treating chronic obstructive pulmonary disease.2 Network pharmacological research:The active ingredients of Chaihu Guizhi Decoction were retrieved through TCMSP database and BATMAN database,and the target was corrected by UniProt database to complete standardized processing.Target genes related to chronic obstructive pulmonary disease were collected through GeneCards,OMIM and Disgenet databases,and the common targets of buplehuguizhi Decoction decoction and chronic obstructive pulmonary disease were obtained by drawing the Venn diagram.According to the common targets,the relationship diagram of "buplehuguizhi Decoction Decoction decoction-active ingredient-target" and PPI protein interaction network were constructed.The core target of treating chronic obstructive pulmonary disease with Chaihu Guizhi Decoction was obtained.Through the enrichment analysis of GO function and KEGG pathway,the relevant signaling pathways of Chaihu Guizhi Decoction Huazai Decoction in the treatment of chronic obstructive pulmonary disease were screened.3 Animal experiments:40 SPF SD rats were randomly divided into normal group,model group,Chaihu Guizhitang Huazai Fang group and aminophylline group,with 10 rats in each group.After 1 week of adaptive feeding,the other three groups except the normal group were prepared by lipopolysaccharide and smoking.After 4 weeks of continuous administration,alveolar lavage fluid and lung tissue were taken.The general state and body mass of rats were observed,pathological sections of j oints were prepared,and the levels of IL-1β,IL-6 and TNF-α in alveolar lavage fluid were detected.The expression of IL-6,p-P13K,P13K,p-AKT,AKT,p-NF-κB p65,NF-κB p6 and TNF-α,IL-1β,IL-6,PI3K and Akt in rat lung tissues were detected by immunohistochemistry,Western blot and QPCR,respectively The expression of mRNA.Results:1 Clinical study results:Professor Lv Xiaodong clinical treatment of COPD is based on Chaihu Guizhi Decoction Huazai prescription,with the addition and reduction of the syndrome,and pay attention to the treatment of phlegm and qi,attack and tonalization,specimen consideration.(1)Frequency analysis results:The single flavor drugs were statistically analyzed,and the drugs with higher frequency were glycyrrhiza,almond,bupleurum,laurel twig,scutellaria,ephedra,ophiopogon,Tuckaia,pinellia and platycodon.The statistical analysis of the four kinds of drugs showed that the most frequent drug was cold drug,followed by warm drug,and then calm drug and hot drug.The five tastes of drugs were analyzed statistically.Bitter,sweet and sweet were the most frequently used drugs.Statistical analysis was made on the traditional Chinese medicine’s meridian normalization,among which the most frequent drugs were lung,spleen and stomach channels.According to the statistics of the efficacy of traditional Chinese medicine,the proportion of phlegm-reducing cough and antiasthmatic drugs,vacuum-tonifying drugs and antipyretic drugs was>50%.(2)Association rule analysis results:Professor Lv Xiaodong’s clinical experience of high-frequency drug combinations:pinellia-Liquorice,cassia twig-Bupleurum,pinelliabupleurum,licorice-bupleurum,etc.;Horn medicine combination:pinellia,cassia twig,bupleurum;Liquorice,cassia twig,Bupleurum;Pinellia,cassia twig,licorice;Pinellia,bupleurum,licorice;Pinellia,scutellaria,licorice,etc.(3)Cluster analysis results:high-frequency drugs were divided into 14 categories:Cl:raw keel,raw oyster,almond;C2:Wood fragrance,jade gold,chinaberry seed;C3:Coptis coptis,Magnolia officinalis,Semen raphani;C4:Red peony,peach kernel,Angelica;C5:Salvia miltiorrhiza,Chuanxiong,ophiopogon;C6:Peony,Astragalus,white art,Codonopsis;C7:Poria,alisma,yam,turtle beetle;C8:Scutellaria,pinellia,licorice;C9:Loquat leaf,burdock seed,Perilla seed,cicada decidua,rigidopsis mori,Earth dragon,Schisandra;C10:Schizonepeta,Prehu,aster;C11:Bupleuri,cassia branch,dandelion,forsythia:C12:orange peel,poncho;C13:Asarum,prepared aconite,ephedra,dried ginger,platycodon.(4)Factor analysis results:A total of 17 groups of drug combination factors were obtained.2.Results of network pharmacology study:(1)The results of "Drug-active ingredient-disease-target" in the treatment of COPD showed that quercetin,kaferol,baicalein,isorhamnetin and β-sitosterol were the main active ingredients in the treatment of COPD in the treatment of Chaihuguizhi Decoction.(2)AKT1,MAPK3,IL-1,TP53,JUN,IL-6 and TNF were the key targets of the target interaction network regulated by Chaihuguizhu Decoction in the treatment of COPD,which may play a major role in the treatment of COPD.(3)The concentration of GO function of the key target of Chaihu Guizhi Decoction in treating COPD showed that Chaihuguizhi Decoction may treat COPD through positive regulation of RNA polymerase Ⅱ promoter transcription,positive regulation of DNA template transcription,positive regulation of gene expression,positive/negative regulation of apoptosis process,response to exogenous stimulation,response to hypoxia and other biological processes.Most of its targets are in cell fluid,plasma membrane,extracellular space,mitochondria and molecules Complex and membrane raft have molecular functions such as protein binding,enzyme binding,protein kinase binding,transcription factor activity and protein kinase activity.KEGG results showed that Chaihu Guizhu Decoction could treat COPD by regulating cancer pathway,lipid and atherosclerosis,PI3K-Akt signaling pathway,MAPK signaling pathway,IL-17 signaling pathway,TNF signaling pathway and other signaling pathways.(4)PI3K/AKT/NF-κB p65 signaling pathway is one of the key pathways in the treatment of COPD with Chaihuguizhitang Huazai prescription.3.Results of animal experiments(1)Chaihu Guizhi Decoction can significantly improve the mental state,cough,wheezing,weight loss and other general conditions of model rats,and alleviate the degree of infiltration of alveolar-inflammatory cells and other pathological manifestations.(2)The results of ELISA showed that the expression of IL-1β,IL-6 and TNF-α in alveolar lavage fluid of COPD rats could be reduced,suggesting that the decoction could play a therapeutic role by inhibiting inflammation.(3)According to the experimental results of immunohistochemistry,Western blot and Q-PCR,The expression levels of IL-6,p-P13K,P13K,p-AKT,AKT,p-NF-κB p65,NF-κB p6 protein and mRNA expression levels of TNF-α,IL-1β,IL-6,PI3K and Akt in lung tissue of COPD rats were decreased.It is suggested that Chaihu Guizhu Decoction can play a role in the treatment of COPD through the PI3K/Akt/NF-κB p65 signaling pathway.It is concluded that the inhibition of PI3K/Akt/NF-κB p65 signaling pathway by Chaihu Guizhu Decoction may be one of the anti-inflammatory mechanisms of COPD.Conclusion:1.Professor Lv Xiaodong clinical treatment of COPD is based on Chaihu Guizhi Decoction Huatai prescription,according to the different syndroms and clinical manifestations of patients,with the increase and decrease of the syndrome,in the treatment process to pay attention to the treatment of phlegm and qi,attack and supplement,specimen consideration.Through the data mining results,the commonly used drugs,drug pairs or drug groups of Professor Lv Xiaodong in the treatment of COPD can be summarized and quantified,providing data basis and research ideas for clinical drugs in the treatment of COPD.2.By using the method and technique of network pharmacology,it was predicted that quercetin,kaempferol,baicalin,isorhamnetin,β-sitosterol and so on might play an active role in Chaihuguizhi Decoction,and its target might be AKT1,MAPK3,IL-1,TP53,JUN,IL-6,TNF,etc.Through GO enrichment analysis and KEGG pathway analysis,it was found that the pathway of its action may be through the regulation of cancer,lipid and atherosclerosis,PI3K-Akt signaling pathway,MAPK signaling pathway,IL-17 signaling pathway,TNF signaling pathway and other pathways.Therefore,the treatment of COPD by Chaihu Guizhi Decoction is realized through multi-component,multi-target and multi-pathway.The PI3K/Akt/NF-κB p65 signaling pathway is one of the key pathways in the treatment of COPD with Chaihuguizhitang Huazai Prescription.3.Combined with the results of network pharmacology,the COPD rat model was prepared,and the mechanism of the treatment of COPD was finally verified by animal experiments.It is suggested that Chaihu Guizhi Decoction decoction can significantly improve mental state,cough,wheezing and weight loss in model rats.Meanwhile,the analysis of tissue,protein and gene expression suggests that Chaihu Guizhi Decoction decoction can act on PI3K/Akt/NF-κB p65 signaling pathway.Down-regulating the activity of PI3K/Akt/NF-κB p65 signaling pathway,thereby inhibiting the expression of inflammatory factors,alleviating inflammatory response,and achieving the purpose of treating COPD.To provide laboratory basis for clinical application of Chaihu Guizhi Decoction Huazai prescription in the treatment of COPD.