The Study On The Treatment Of Influenza A With Xiaochaihu Decoction Based On Autophagy Pathway

BackgroundInfluenza A virus is a highly infectious and pathogenic virus,and it is highly mutagenic.The influenza vaccine is prone to failure and antiviral drug treatment has certain limitations.Therefore,treatments that target host factors have become emerging therapies and future trends for anti-influenza viruses.This change is the manifestation of the relationship between pathogy and positive in Chinese medicine,and it is also the advantage of Chinese medicine treatment.Autophagy is one of the important mechanisms of the interaction between influenza virus and host.Influenza A virus can induce a large number of autophagosomes to accumulate and prevent the fusion of autophagosomes and lysosomes,resulting in an unsmooth flow of autophagy.The unsmooth flow of autophagy happens to be very similar to that of typhoid and Shaoyang cardinal disadvantage.Therefore,based on this level,in view of the disadvantages of Shaoyang cardinals,choosing Shaoyang prescriptions(such as Xiao Chaihu Tang)may promote the completion of effective autophagy,thereby cutting the depth of the disease and preventing the occurrence of pneumonia.Xiao Chai Hu Decoction is the main prescription for Shaoyang disease in Treatise on Febrile Diseases,and it is also a representative prescription for strengthening the body and eliminating evil.It has been reported that Xiao Chaihu Decoction has obvious anti-influenza virus effect,and can improve the body’s immunity and reduce inflammation..However,the effect of Xiao Chai Hu Tang in regulating autophagy is rarely reported.Therefore,this study conducted a preliminary discussion on the efficacy,mechanism and compatibility of Xiao Chaihu Decoction in the treatment of influenza A from four aspects: scientometrics,theoretical research,network pharmacology,and experimental research,with a view to preventing and treating viral infectious diseases.Drug targets provide new ideas.Objective1.To analyze the current situation and frontier hotspots of influenza research,explore the joint pathways of influenza and autophagy,and predict future research trends with citespace.2.By tracing the original meaning of Shaoyang,combined with the original text of "Treatise on Febrile Diseases",expound the theory and clinical practice of Xiao Chai Hu Decoction in the treatment of influenza A from aspects such as the pathogenesis of Shaoyang disease,the prescription of Xiao Chai Hu Decoction,and the pharmacology spectrum of Xiao Chai Hu Decoction.in accordance with.3.Based on the network pharmacology study of Xiao Chaihu Decoction in the treatment of influenza,the main mechanism of action is predicted from the level of "Traditional Chinese Medicine-Ingredients-Target-Pathway" and the main pathways that reflect the significance of drug compatibility.4.Combining the principles of formulating prescriptions and the results of online medicines,to study the in vitro inhibitory effects of Xiao Chaihu Decoction and its separate prescriptions on H1N1 influenza virus,whether its mechanism is related to autophagy,and to explain its compatibility significance.Method1.Taking "influenza" as the research theme,using citespace software to search the 2005-2020 documents of the Web of Science(Wo S)database,and the results from keyword co-occurrence,keyword clustering,keyword emergence,double-image overlay,etc.Visual analysis in four aspects.2.Screen the effective ingredients of Xiaochaihu Decoction and correspond to target genes,and search for influenza disease targets.The Cytoscape 3.2.6 software was used to construct the Xiao Chai Hu Tang-influenza-target-pathway network,screen the core target genes,perform GO function and KEGG enrichment analysis,predict the main mechanism of action and the main pathways of the compound compatibility significance.3.Prepare three preparations of Xiao Chaihu Decoction and the water-extracted and alcohol-precipitated extracts,granules and medicated serum of the disassembled recipes.The cytotoxicity and neuraminidase activity of human A549 and dog kidney cell(MDCK)cell lines are measured respectively.Determine and screen the optimal preparation plan and the most suitable cell line.4.Based on the optimal cell line,Xiao Chaihu Decoction and the optimal preparation plan of the decomposed formulas,Western blotting was used to detect the expression and expression of LC3-Ⅰ,LC3-Ⅱ,p62,m TOR,phosphorylation and beclin protein,and explore the effect of Xiao Chaihu Decoction and the decomposed formulas The regulation and mechanism of influenza virus-induced autophagy,and the significance of interpretation and compatibility.Results1.A total of 7223 papers on influenza mechanism were included.Keyword clustering is mainly divided into influenza pathogenicity and pathogenesis research;host immune response and inflammatory response research;anti-influenza drug and vaccine research.The common keywords in the research of influenza mechanism and autophagy are autophagy,replication,infection,apoptosis,NF-κB,natural immune response and so on.2.There are 51 targets of Xiaochaihu Decoction components and influenza disease targets,the main components of which are quercetin,kaempferol,stigmasterol,isorhamnetin,β-sitosterol,dimethoxyflavone,and ginseng Saponins Rh2,cyanocyanine,and leaves contain nine types of kaurein A.The core genes are VEGFA,AKT1,IL6,TNF,MMP9,JUN,IL10,and IL18.The enrichment analysis is mostly related to inflammation.Various metabolisms are related.3.In MDCK cells,the antiviral selection index of Xiao Chaihu Decoction and core disassembled water extract is about 1;mouse serum shows strong anti-influenza virus activity in in vitro(cell)antiviral experiments.The index is above 128.2;the antiviral selection index of Xiao Chaihu Decoction and Chai-Qin-Shen granules is about 1,and the anti-viral selection index of Chai-Qin granules is 2,which has certain anti-viral efficacy.In addition,the overall survival rate of Xiao Chaihu Decoction and disassembled prescription granules on human A549 cells is not high,especially the low concentration of Chai-Qin granules shows certain toxicity to A549 cells,and the survival rate is less than 85%.In the detection of neuraminidase inhibitory activity,it was shown that Xiao Chaihu Decoction and Split Recipe Granules had no inhibitory effect on H1N1 influenza virus and no SI.4.Compared with the blank control group,the expression level of LC3-II protein in the virus infection group was significantly increased(P<0.01).Compared with the virus-infected control group,Xiao Chaihu Decoction and disassembled granules had the effect of down-regulating the expression of LC3-Ⅱ protein(P<0.01),especially in the Xiao Chaihu Decoction group.Compared with the virus-infected group,Xiao Chaihu Decoction and Disassembled Granules both had the effect of up-regulating the expression of p62 protein.Among them,the Chai-Qin group was the most obvious(P<0.01),followed by the Chai-Qin-Shen group(P<0.05).),Xiao Chai Hu Tang is the last time.Compared with the virus-infected group,Xiao Chaihu Decoction and the three groups of disassembled prescriptions had a tendency to up-regulate the expression of p-m TOR protein(P>0.05),and the Chai-Qin group had the most significant up-regulation effect.Compared with the virus-infected group,the Xiao Chaihu Decoction group had the most significant down-regulation of beclin-1 protein expression(P<0.05),and the Chai-Qin-Shen group had the least down-regulation effect,with no statistical difference(P>0.05).Conclusion1.Citespace and theoretical studies have shown that the occurrence,development and severity of influenza are inseparable from the relationship between the virus and the host.The treatment of the host as the target is an important change in the field of research,and this change reflects the characteristics of TCM treatment of strengthening the body and eliminating evil.Autophagy is one of the important mechanisms of the interaction between influenza virus and host.In the view of traditional Chinese medicine,the unsmooth flow of autophagy caused by influenza A virus is very similar to the disadvantage of typhoid and Shaoyang cardinal,which is also consistent with the prescription of Xiao Chaihu Decoction.2.The results of network pharmacology show that Xiao Chaihu Decoction can regulate nerve-immune-inflammatory response and treat influenza A.Its main mechanism is related to the three medicines of Bupleurum,Scutellaria and Ginseng.3.The results of the experiment show that only Bupleurum-Scutellaria baicalensis showed a certain inhibitory effect in terms of inhibiting the activity of H1N1 neuraminidase,reflecting its ability to eliminate evil.Xiao Chai Hu Decoction has no inhibitory effect on NA activity,but it has a significant effect on autophagy regulation,and is more prominent in down-regulating the expression of LC3-Ⅱ/LC3-I and inhibiting the level of Beclin-1 protein.Its mechanism may be related to Beclin-1.However,there are differences in the Bupleurum-Scutellaria-Ginseng group,which also has the meaning of strengthening the body and eliminating evils,suggesting that the four herbs of summer,ginger,jujube,and grass may play a role in promoting autophagy degradation in the compatibility.In addition,in view of the fact that Xiao Chaihu Decoction itself cannot inhibit NA activity,it means that the target of Xiao Chaihu Decoction may not directly act on the virus,and it may be mainly due to host factors,which are related to the regulation of autophagy and the promotion of autophagy metabolism.Therefore,the regulation effect of Xiaochaihu Decoction on autophagy,in a sense,expands the new idea of the clinical application of Xiaochaihu Decoction,and also reveals to a certain extent the mechanism of Xiaochaihu Decoction for strengthening the body and eliminating pathogenic factors and the compatibility of the drug monarch and adjuvant.The significance of this will lay the foundation for further in-depth research in the future.