Study On Therapeutic Mechanism Of Qingfei Chengqi Decoction On ARDS Caused By Abdominal Infection

Objective:This study aimed to clarify the potential molecular mechanism of Qingfei Chengqi Decoction in the treatment of patients with ARDS caused by abdominal infection.Firstly,through the analysis of multi-center clinical data,it was initially shown that Qingfei Chengqi Decoction had a definite therapeutic effect in clinical treatment.Further,by constructing a network pharmacology model of Qingfei Chengqi Decoction to treat ARDS,the correlation between potential gene targets and signaling pathways for clear treatment effects was evaluated;finally,the lungs were screened by constructing a model of ARDS rats caused by abdominal infection Chengqi Decoction treats differential microRNAs of ARDS caused by intra-abdominal infection,evaluated downstream gene targets and signal pathways of differential microRNAs,and cross-analyzes the underlying molecular mechanism.Methods Part One: A Multicenter Study on the Protective Mechanism of Qingfei Chengqi Decoction for ARDS Patients Caused by Acute Abdominal Infection:27 of patients with ARDS due to celiac infection were selected from 3 top three hospitals,who were Randomly divided into 14 cases of basic treatment with Qingfei Chengqi Decoction and 13 cases of basic treatment.Alveolar lavage fluid was taken twice 3 days before and 3 days after treatment,and secretion of IL-4,IL-6,IL-10 was detected in two groups by microsphere detection,alveolar Macrophage Apoptosis was analyzed by Flow Cytometry in Two Groups.Part Two: Network Pharmacology Analysis of Qingfei Chengqi Decoction in Treating Acute Respiratory Distress Syndrome:Retrieved the chemical components and targets in Qingfei Chengqi Decoction through the TCMSP database,compared with the ARDS targets in the Gene Cards database and OMIM database to obtain drug-disease targets,and then by Cytoscape3.7.1 software established a "compound-predicted target" network,predicted the protein interaction(PPI)network between targets,KEGG enrichment analysis of predicted targets,and combined the clinical results of the first part to evaluate the treatment of ARDS Possible gene targets and signaling pathways.Part Three: Genomics Study of Qingfei Chengqi Decoction on ARDS Rats Caused by Abdominal Infection:12 of ARDS rat models were constructed by cecal ligation(CLP)to create abdominal infections.The rat models were divided into a control group(n = 6)and a traditional Chinese medicine treatment group(n = 6)by random number method.Three rats were selected to take lung tissue for HE staining,and the differential microRNA was screened by high-throughput gene chip method.GO and KEGG analysis of downstream genes of differential microRNA was performed by GO database and KEGG database,and compared with the second part of the results analysis.Results Part One: A Multicenter Study on the Protective Mechanism of Qingfei Chengqi Decoction for Patients with ARDS Caused by Acute Abdominal Infection:1.General information of patients: There was no statistically significant difference in the general conditions of the two groups of patients.2.Comparison of alveolar macrophage apoptosis between the two groups: there was no significant difference between the two groups before treatment(P> 0.05).After treatment,there was a significant difference in the proportion of alveolar macrophage apoptosis between the two groups(both P <0.05)According to the difference statistics before and after treatment,the area of apoptosis in the treatment group decreased significantly(P <0.05).3.Comparison of alveolar signal factors between the two groups of patients: There was no significant difference in cytokine levels between the three groups before treatment(P> 0.05),and there were differences between the treatment group and the control group after treatment(P <0.05)0.05),IL-6 secretion in the treatment group increased significantly after treatment with traditional Chinese medicine(P <0.05),while IL-10 secretion showed a downward trend(P <0.05),IL-4 had a certain downward trend,but P> 0.05 It means that the downward trend is not significant.Part Two: Network Pharmacology Analysis of Qingfei Chengqi Decoction in Treating Acute Respiratory Distress Syndrome:1.Screening of pharmacologically active compounds: A total of 78 compounds were screened out,including 16 species of rhubarb,22 species of Fructus Aurantii,2species of Magnolia officinalis,14 species of Coptis chinensis,13 species of Pinellia,and 11 species of melon.2.Screening of potential targets for Qingfei Chengqi Decoction in the treatment of ARDS: 476 ARDS-related gene targets were obtained,225 corresponding targets of active compounds in Qingfei Chengqi Decoction were screened,and cross-screening of disease and drug targets was performed to obtain common potential targets 63 points.3."Compound-Predicted Target" composite network diagram of Qingfei Chengqi Decoction for the treatment of ARDS: A network diagram was constructed based on Qingfei Chengqi Decoction containing active compounds and their corresponding targets and ARDS potential targets.It consists of 110 nodes(44 compound nodes and63 gene targets)and 324 edges.Quercetin is the compound with the highest correlation value.BCL2,ESR1,and AR are combined with 7,16,and 19 compounds,respectively.They play a key role in cell proliferation,inflammation,and apoptosis..4.PPI network diagram of Qingfei Chengqi Decoction for the treatment of ARDS potential targets: Use the String database to construct a PPI network diagram for 63disease-drug potential targets.The PPI diagram has 68 nodes and 1078 edges.The PPI diagram correlation value The highest is IL-6.5.Analysis of KEGG pathway of potential targets: KEGG pathway statistics of 63 potential targets of Qingfei Chengqi Decoction in the treatment of ARDS,63 potential targets mapped to 78 KEGG pathway species,TNF pathway,NF-κB pathway and The T cell receptor pathway belongs to the top signaling pathway.Part Three: Genomics Study of Qingfei Chengqi Decoction on ARDS Rats Caused by Abdominal Infection:1.Screening of microRNA: After screening,16 groups of microRNAs with obvious differences,including rno-mi R-338-5p,rno-mi R-20a-5p,rno-mi R-350, rno-mi R-199a-5p,were obtained,Rno-mi R-218a-1-3p,rno-mi R-224-5p,rno-mi R-362-3p,rno-mi R-678,rno-mi R-3584-5p,rno-mi R-1839-5p,rno-mi R-6216,rno-mir-15 b,rno-mir-27 b,rno-mir-30c-1,rno-mir-320,rno-mir-665.Qingfei Chengqi Decoction regulates the body’s reactivity by separately enhancing or inhibiting the expression of these 16 microRNAs.2.The pathological effect of Qingfei Chengqi Decoction on ARDS rats caused by abdominal infection: HE staining of lung tissue in the traditional Chinese medicine treatment group compared with the control group: the alveolar size of the stem cell treatment group was more uniform,inflammatory cell infiltration,pulmonary hemorrhage,interstitial edema,Alveolar interval thickening was significantly reduced.3.GO and KEGG analysis of microRNA: KEGG analysis obtained Wnt signaling pathway,T cell receptor signaling pathway,Protein processing in endoplasmic reticulum,Gn RH signaling pathway,Erb B signaling pathway,which are pathways that directly or indirectly interfere with apoptosis,while T cell receptor The signaling pathway and p53 signaling pathway are key pathways in immune regulation.According to GO analysis,the biological activities of Qingfei Chengqi Decoction mainly focus on IL-12 secretion and cell electrical activity;the cell components mainly focus on gene assembly,cell membrane composition,vesicle membrane formation,and myelin sheath axon area Formation;in terms of molecular function,it focuses on virus binding,receptor inhibitor activity,and acetylcholine receptor binding.Conclusions:In summary,through multi-center clinical data,network pharmacological model evaluation and animal experiments,this experiment initially explored the potential molecular mechanism of Qingfei Chengqi Decoction in the treatment of ARDS caused by abdominal infection,in terms of apoptosis of lung macrophages,Qingfei Chengqi Decoction is regulated by microRNA-20a-BCL2 axis system and microRNA-350-JNK axis system;in terms of inflammation regulation,Qingfei Chengqi Decoction regulates IL-6,IL by microRNA-350 and microRNA-20a-5p-10,AR,CXCL8,PPARγ and the interleukin family pathway,NF-κ B pathway and TNF pathway mapped by them affect the inflammatory response.