Exploration Of Professor Li Zhuying’s Medication Pattern In The Stable Stage Of COPD Based On Date Mining And Network Pharmacology And Study On The Mechanism Of Shenqi Bufei Decotion In The Treatment Of COPD

Objective:We studied Professor Li Zhuying’s medical case of treating chronic obstr-uctive pulmonary disease during the stable period,summarized her medica-tion patterns and clinical experience,and provided ideas and guidance for clinical treatment;Based on network pharmacology,we summarized the chemical components of Chinese herbal medicines in Shenqi Bufei Decoction,Professor Li Zhuying’s empirical prescription for treating COPD in stable stages,predicted the potential targets,biological processes,and action pathways of Shenqi Bufei Decoction for treating COPD,and provided theoretical basis for the clinical application of empirical based formulas;Then we selected the AMPK/NF-κB pathway in network pharmacology and replicated a COPD rat model to detect the expression of AMPK/NF-κB pathway related factors in the lung tissue of the COPD rat model,in order to elucidate the anti-inflammatory mechanism of Shenqi Bufei Tang in treating COPD.Methods:1.Part One: We collected prescription medical records of patients with chronic obstructive pulmonary disease in stable phase who were treated by Professor Li Zhuying in the Respiratory Department of the First Affiliated Hospital of Heilongjiang University of Chinese Medicine from June 2020 to June 2022.We used the Traditional Chinese Medicine Inheritance Support System Platform(V3.0)to statistically analyze the syndrome types,medicati-on frequency,drug properties,taste,and meridians of Professor Li’s applied prescriptions,and conducted association rules and high-frequency drug cluster analysis,To summarize her medication patterns and characteristics.2.Part Two: We have summarized the chemical components of the drugs in Shenqi Bufei Decoction in the TCMSP database,using the Swiss Drug Design platform to predict target proteins and query the corresponding gene names.Afterwards,we searched for gene targets of COPD on platforms such as Gene Cards and OMIM,and used software and platforms such as Excel and String to intersect drug component action targets and disease-related targets to obtain common action targets.Afterwards,we used Cytoscape 3.9.0software to construct a drug compound target network diagram,and constr-ucted a protein interaction diagram on the String platform.Finally,David and Metascape platforms were used for GO and KEGG pathway enrichment analysis.3.Part Three: Based on the results of network pharmacology,we selected the possible action pathway AMPK/NF-κB signaling pathway of Shenqi Bufei Decoction in COPD rats to explore the anti-inflammatory mechanism of Shenqi Bufei Decoction.We randomly divided 60 rats into 4 groups,namely blank group,model group,Chinese medicine group,and western medicine group(15 rats/group).Except for the blank group,the other three groups were treated with LPS+smoking method to create COPD rat models.The blank group and model group were given physiological saline by gavage,while the Chinese medicine group and western medicine group were treated with Shenqi Bufei Tang(1.48g/100g)and Doxophylline(0.002g/100g),respectively.We observed the general state of the rats before and after treatment,and sacrificed them on the 29 th day,Then we collected rat lung tissue samples and observed the pathological changes under light microscopy.Immunohistoch-emical and Western Blot methods were used to detect the expression level of SIRT1,PGC-1α and FoxO3a in lung tissue;The Real Time PCR method was used to detect the expression level of AMPK mRNA,SYT1 and TLR4 mRNA in the lung tissue of the rat model,and the ELISA method was used to detect the expression level of p65 mRNA,IL-1β and IL-6 in the lung tissue homogenate.Results:1.We included 206 patient prescriptions in this study,including 134 male patients(65.05%)and 72 female patients(34.95%).We found that the patient population is mostly distributed between the ages of 71 and 80.The study involves four syndrome types,with lung kidney qi deficiency syndrome(33.01%)appearing the most frequently,followed by lung qi deficiency syndrome(26.70%)and lung spleen qi deficiency syndrome(25.24%).The results indicate that the main symptoms include shortness of breath,fatigue,wheezing,susceptibility to colds,coughing and soreness and weakness of waist and knees,etc.The treatment principles and methods mainly focus on tonifying the lungs,relieving cough,relieving asthma,absorbing qi,tonifying kidney qi,calming asthma and invigorate the spleen and replenish qi,etc.High frequency medication includes Codonopsis pilosula,perilla seed,astragalus,Dilong,tangerine peel,licorice,epimedium,bitter almond,magnolia,Atractylodes macrocephala,etc.The main properties of the medication are warm,sweet,pungent and bitter.The meridians mainly enter the lung and kidney meridians,followed by the spleen meridian.The main effects of the medication are tonifying deficiency,resolving phlegm,relieving cough,and relieving asthma.The core drug pairs are "Dangshen Huangqi","Baizhu Chenpi","Dangshen Huangqi Epimedium",etc.The core prescriptions are 1: Baibu,Zisu Zi,Dangshen,Dilong,Huangqi,and roasted licorice;2: Atractylodes macrocephala,Codonopsis pilosula,Poria cocos,bitter almonds,roasted licorice,Astragalus membranaceus;3: Codonopsis pilosula,Cornus officinalis,Fructus perilla,Fructus Psoraleae,Astragalus membranaceus,Epimedium.2.We selected 194 eligible chemical components from Shenqi Bufei Decoction,and obtained 2163 COPD related targets in Gene Cards,OMIM,and Drug Bank databases,of which 345 were regulated by Shenqi Bufei Decoction.CYP19A1,CDK2,ACHE,ESR2,CDK1,PTPN1,ESR1,AR,GABRG2,BACE,etc.are the main targets of action.Research has found that the GO and KEGG pathways are highly concentrated,with a total of 3020 biological processes and 191 signaling pathways involved in the mechanism of Shenqi Bufei Tang in treating COPD.Biological processes mainly involved inflammatory reactions,positive regulation of external stimulus responses,and responses to oxygen levels.Signal pathways mainly involve PI3K-Akt signaling pathway,Fox O signaling pathway,NF-κB signaling pathway,and AMPK signaling pathway.3.General state of rats: Research has found that the blank group rats mainly breathe normally during the observation period,have shiny hair,move freely,can eat and drink normally,and quickly avoid catching water;In the model group,the respiratory rate of rats was accelerated,and their breathing was short.Abdominal muscle twitching was seen when they were short of breath.The rats were irritable,and their hair was sparse and less shiny.The upper limbs often scratched the nose,and the extremities were slightly cyanotic;It was observed that the rats in the other two treatment groups behaved similarly to the model group at the initial stage.With the extension of drug treatment time,the avoidance degree,hair gloss,and respiratory rate of rats in the model group were slower and faster than those in the model group.The rats in both groups were better than those in the model group.4.Observation under light microscope: We found that after modeling,inflammatory cell infiltration,edema,congestion,and structural damage were observed in the lung tissue of each group.Multiple pulmonary bullae were observed,and varying degrees of bleeding were observed in the pulmonary interstitium.However,after treatment with Shenqi Bufei Tang and Doxophylline,compared with the model group,the formation of model type pulmonary bullae in the two groups was reduced,the infiltration of inflammatory cells was reduced,and the situation of emphysema in rats was improved.5.The study found that compared with the blank group,the model group showed a decrease in AMPK and PRKKA2 mRNA expression(p<0.05),while the expression levels of TLR4 and SYT1 mRNA increased(p<0.05);Compared with the model group,Shenqi Bufei Tang and Doxophylline can increase the expression of AMPK protein PRKKA2(p<0.05)and reduce the expression levels of TLR4 and SYT1(p<0.05);There was no difference between the two groups(p>0.05).6.Research has shown that in immunohistochemistry and Western blot detection,the expression level of SIRT1,PGC-1α and Fox O3 a protein in the model group rats decreased compared to the blank group(p<0.05);Compared with the model group,both Shenqi Bufei Tang and Doxophylline can increase the expression level of SIRT1,PGC-1α and FoxO3a(p<0.05);The comparison between Shenqi Bufei Tang and Doxophylline showed no statistical significance(p>0.05).7.The ELISA test results showed that compared with the blank group,the expression of pro-inflammatory cytokine IL-1β,IL-6 and NF-κB signaling pathway P65 protein in the model group rats increased(p<0.05);Compared with the model group,Shenqi Bufei Tang and Doxophylline can reduce the expression of P65,IL-1β and IL-6(p<0.05).There was no difference between the two treatment groups(p>0.05).Conclusion:1.Research has found that lung and kidney qi deficiency syndrome,lung qi deficiency syndrome,and lung spleen qi deficiency syndrome are common syndromes in the stable stage of COPD.During the stable phase of chronic obstructive pulmonary disease,the medication is mainly warm in nature,with a sweet,spicy,and bitter in taste.It mainly enters the lung and kidney meridians,followed by the spleen meridian.This medicine has the main effects of tonifying deficiency,resolving phlegm,relieving cough,and relieving asthma.The core high-frequency drugs mainly include Codonopsis pilosula,purple perilla seed,Astragalus membranaceus,earthworm,tangerine peel,roasted licorice,epimedium,bitter almond,magnolia officinalis,atractylodes macrocephala,etc.2.Shen Qi Bu Fei Tang exerts its positive therapeutic effect on COPD through multiple components and targeted effects.The signal pathway mainly involves multiple signal pathways such as PI3K-Akt,Fox O,NF-κB,and AMPK.3.Shenqi Bufei Tang can effectively control and treat inflammatory reactions,activate SIRT1,PGC-1α and FoxO3 a factor expression,promote AMPK pathway activation,and exert anti-inflammatory effects.At the same time,it reduces the expression and release of TLR4,P65,SYT1,and pro-inflammatory factor IL-1β,IL-6,inhibits NF-κB pathway and protein phosphorylation and ubiquitination,thereby inhibiting COPD inflammation.It may be through regulating the AMPK/NF-κB axis,achieving the therapeutic effect of COPD.