Clinical Study And Mechanism Discussion On Chronic Hepatitis B With Liver Fibrosis Treated By Shugan-Jianpi-Huoxue Method

At present,chronic hepatitis B(CHB)is still a global public health problem,seriously endangering people's health.Liver fibrosis is a necessary intermediate link in the development of CHB to cirrhosis.If it is not blocked in time,it can further progress to cirrhosis,portal hypertension and liver cancer,and even to end-stage liver disease,which is life-threatening.Therefore,blocking and reversing liver fibrosis and preventing disease progression are another important prevention and treatment targets of CHB besides antiviral treatment.However,the treatment of liver fibrosis is still a difficult problem in the prevention and treatment of CHB.At present,the vast majority of all kinds of specific anti fibrosis drug preparations in modern medicine are still in the experimental research stage.Modern medical treatment of CHB-fibrosis is mainly aimed at the cause of treatment,namely antiviral treatment.Entecavir is a first-line antiviral therapy recommended by domestic and foreign guidelines.It has strong antiviral effect and low drug resistance.However,HBe Ag conversion rate is low,and even after antiviral treatment,a considerable number of patients still have liver fibrosis progression.Therefore,how to prevent the progress of liver fibrosis and improve the HBe Ag conversion rate is a difficult problem in the treatment of CHB-fibrosis.A large number of studies have confirmed that traditional Chinese medicine has good curative effect and no side effects in the treatment of liver fibrosis.Its efficacy and advantages have been widely recognized by scholars at home and abroad.In the past few decades,many researchers have carried out a large number of clinical and experimental studies on chronic hepatitis B liver fibrosis.Although the syndrome differentiation and classification of CHB-fibrosis is not unified in clinical practice,the method of soothing the liver,strengthening the spleen and activating blood circulation has been more and more recognized by doctors and scholars throughout the recent ten years.My department in the past clinical practice found that the combination of Shugan Jianpi Huoxue method can significantly improve liver fibrosis,improve HBe Ag conversion rate,but lack of objective clinical evaluation and mechanism of action.ObjectiveThe firstd part uses the method of literature research,based on frequency analysis,complex network analysis and association rules and other data mining methods to analyze and summarize the drug compatibility law of CHB-fibrosis treated by Shugan Jianpi Huoxue Method,in order to provide medication reference for clinical application of Shugan Jianpi Huoxue Method in the treatment of CHB-fibrosis,and also for the next step of Shugan Jianpi Huoxue Method in the treatment of CHB-fibrosis.In the second part of this project,the effect of Shugan Jianpi Huoxue Method on pathological liver fibrosis stage,liver hardness value and serum liver fibrosis indexes of patients with CHB-fibrosis was observed by retrospective study method,so as to objectively evaluate its clinical efficacy and provide an objective basis for clinical application.In the third part,the network pharmacology method was used to study the top five herbs with the highest frequency found in the second part of data mining,and to explore the target and signal pathway of Shugan Jianpi Huoxue Method in the treatment of CHB-fibrosis,in order to provide an objective reference for the follow-up study of Shugan Jianpi Huoxue Method in the treatment of CHB-fibrosis.MethodsPart one: Based on data mining method,the article summarizes the law of the treatment of CHB-fibrosis by dredging liver,invigorating spleen and activating blood circulationTaking "China zhinet","China Biomedical Journal Citation Database","Chongqing VIP Chinese Journal Database","Wanfang medical network" as the retrieval database,the modern literature of liver dredging,spleen and blood circulation method for treating CHB-fibrosis from the establishment of the database to September 2020 was searched.The modern literature research database of liver and spleen invigoration and blood circulation method for CHB-fibrosis was established,and frequency was further used The data mining methods such as data mining such as data mining,such as data mining,such as data mining,such as data mining,summarized the law of the treatment of CHB-fibrosis by the method of clearing liver,invigorating spleen and activating blood.Part two: clinical study on the treatment of CHB-fibrosis by dredging liver,invigorating spleen and activating blood circulationThe patients with CHB-fibrosis were included in the outpatient and inpatient department of liver disease department of Guangdong Hospital of traditional Chinese medicine from January 2010 to September 2020 by retrospective cohort study.The patients were divided into treatment group and control group for retrospective observation.The control group was treated with entecavir,and the treatment group was treated by dredging liver,invigorating spleen and activating blood,and collecting treatment course to The clinical data of patients with less than 144 weeks were compared.The difference of pathological stage,hardness value,serum hepatic fibrosis index and HBe Ag conversion rate of patients treated with liver fibrosis by the combination of "Shugan Jianpi Huoxue" and "entecavir" were compared.The anti fibrosis effect and disease resistance of the treatment of hepatitis B liver fibrosis were evaluated objectively Toxic effect.Part three: Based on the network pharmacology,the mechanism of the treatment of CHB-fibrosis by the method of clearing liver,invigorating spleen and activating blood circulationThe main effective components and targets of core drugs of the method of clearing liver,invigorating spleen and activating blood circulation were collected and screened by TCM System Pharmacology database and analysis platform(TCMSP).The gene of target genes related to CHB-fibrosis was collected and sorted by searching genecards,OMIM and Pharm GKB databases,and then Venn map of drug disease target was drawn by venn2.1 online tool to obtain the interaction between traditional Chinese medicine and disease Target.Then,PPI map of TCM disease interaction target was drawn by string database and cytoscape3.8.1 software,and core genes were screened.Finally,the enrichment analysis of go and KEGG channels is carried out by using David database ResultsPart 1: data miningA total of 257 articles related to liver fibrosis of hepatitis B treated by Shugan Jianpi Huoxue Method were preliminarily retrieved.97 qualified literatures were screened according to the title,and then the contents of each literature were manually screened according to the inclusion and exclusion criteria.After eliminating the unqualified literatures,48 qualified papers were finally included,involving 48 prescriptions,96 traditional Chinese medicines,and the total frequency of drug use It was 537 times,involving 17 kinds of traditional Chinese medicine.A total of 48 modern literatures on CHB-fibrosis treated by Shugan Jianpi Huoxue Method were retrieved.The results showed that 96 herbs were involved in the treatment of CHB-fibrosis.the top five Chinese medicines were Radix Bupleuri,radix salviaemiltiorrhizae,Radix Paeoniae Alba,Atractylodis Macrocephalae and Astragalus membranaceus.There are 17 kinds of drugs involved,and the top four kinds of drugs are: tonifying deficiency Medicine > promoting blood circulation and removing blood stasis Medicine > regulating qi Medicine >diuresis and dampness drugs,with the cumulative frequency of 88.3%.In the method of soothing liver,invigorating spleen and activating blood circulation,tonifying deficiency medicine is the most frequently used,followed by promoting blood circulation and removing blood stasis.Among them,Astragalus membranaceus,Atractylodesmacrocephala,Radix Paeoniae Alba,turtle shell,Angelica sinensis,dangshen,etc.Through the complex network analysis,the core drugs of soothing the liver,strengthening the spleen and activating blood circulation in the treatment of liver fibrosis of hepatitis B include bupleurum,Salvia miltiorrhiza,Radix Paeoniae Alba,Atractylodes Macrocephalae,Astragalus membranaceus,turtle shell,Poriacocos,red peony,Angelica sinensis,Fructus aurantii Immaturus,etc.Among them,Qi regulating drugs,tonifying deficiency drugs and promoting blood circulation to remove blood stasis drugs are the most commonly used drugs in the treatment of CHB-fibrosis.Part 2: clinical researchA total of 385 patients with hepatitis B liver fibrosis who received ETV treatment for 144 weeks were enrolled.Among them,81 patients completed the second liver pathological evaluation before and after treatment,83 patients completed Fibro Scan detection before and after treatment,and 114 patients completed serum liver fibrosis index detection before and after treatment.The liver fibrosis reversion rate of the treatment group was significantly higher than that of the control group(95.5% vs 18.2%,P < 0.001).In the progress of liver fibrosis,there was no case of liver fibrosis progress in the treatment group,while 50% of the patients in the control group still had liver fibrosis progress.The level of hyaluronic acid(HA)in the control group had no significant change(40.9 vs 46.6,P = 0.744),while the level of HAin the treatment group was significantly lower than that before treatment(55.2 vs41.4,P = 0.003),and the difference between before and after treatment in the treatment group was significantly higher than that in the control group(P = 0.02).At the same time,the serum levels of procollagen ?(PC?),laminin(LN)and type ? collagen(C?)in the two groups were decreased compared with those before treatment(P < 0.05).Compared with the control group,the serum levels of PC?,LN and C? in the treatment group decreased more significantly than those before treatment,but there was no significant difference between the two groups before and after treatment(P > 0.05).In terms of curative effect evaluation of Fibro Scan liver hardness value,the liver hardness value of the two groups decreased significantly compared with that before treatment(P < 0.05),but compared with the control group,the difference of LSM value in the treatment group was larger than that before treatment,but there was no significant difference between the two groups before and after treatment(P = 0.142).In order to further observe the effect of Shugan Jianpi Huoxue Method on the antiviral efficacy for CHB-fibrosis,70 cases of HBe Ag positive patients with ALT < 2ULN were selected as the research objects.The results showed that the HBe Ag seroconversion rate of the two groups increased year by year,but the increasing trend of HBe Ag conversion rate in the treatment group was higher than that in the control group,and the HBe Ag conversion rate of the treatment group at 48 weeks,96 weeks and 144 weeks was respectively higher than that of the control group The HBe Ag conversion rate was 17.1%,26.5% and 28.6% in the control group,which was higher than 2.9%,12.9% and 10.3% in the control group;the HBe Ag conversion rate of the two groups at 48 weeks was statistically different(P = 0.046).Part 3: network pharmacology researchObjective to collect the effective chemical components and targets of Radix Bupleuri,radixsalviaemiltiorrhizae,Radix Paeoniae Alba,Atractylodis Macrocephalae,Astragalus membranaceus and other high-frequency drugs,which are the top five high-frequency drugs of soothing the liver,strengthening the spleen and promoting blood circulation.Results a total of205 interactive target genes were found between the core drug of Shugan Jianpi Huoxue Method and hepatitis B liver fibrosis disease.PPI protein interaction network analysis showed that the key targets of the core drugs of soothing the liver,strengthening the spleen and promoting blood circulation in the treatment of hepatitis B liver fibrosis were AKT1,TP53,CDK1,ccna2,ccnb1,TOP2 A,BIRC5,VEGFA,EGFR,CASP3 and other core genes.Go analysis revealed six most relevant biological processes,including DNA transcription,promoting RNA polymerase II promoter transcription,promoting DNA transcription,inflammatory reaction,inhibiting apoptosis process and promoting cell proliferation;and 7 most relevant cell components,ConclusionLiterature studies have found that there are certain compatibility rules in the treatment of CHB-fibrosis with Shugan Jianpi Huoxue Method.The most frequently used drugs are tonifying deficiency drugs,followed by promoting blood circulation and removing blood stasis drugs and regulating qi drugs.There are Astragalus membranaceus,Atractylodesmacrocephala,Radix Paeoniae Alba,turtle shell,Angelica sinensis,Codonopsispilosula,etc.;the drugs for promoting blood circulation and removing blood stasis are Salvia miltiorrhiza,radix paeoniae rubra,Radix et Rhizoma Chuanxiong,semen persicae,etc.;the medicines for regulating qi include bupleurum,Fructus aurantii,Pericarpium Citri Reticulatae,Rhizoma Cyperi.The top five core Chinese medicines of soothing liver,strengthening spleen and activating blood circulation are bupleurum,Salvia miltiorrhiza,Paeonia alba,Atractylodesmacrocephala and Astragalus membranaceus.Clinical studies have found that the traditional Chinese medicine therapy of Shugan Jianpi Huoxue can synergistically improve the anti fibrosis effect and HBe Ag seroconversion rate of entecavir in the treatment of chb-fibrosis.Network pharmacology research found that the core drugs of Shugan Jianpi Huoxue method can participate in the anti fibrosis process through a variety of cell molecules,biological processes and multiple signal pathways.It mainly acts on the cytoplasmic,extracellular space,nuclear and other components,and regulates HIF-1,PI3 K Akt,toll like receptor and NF kappa B.Apoptosis,p53,nod like receptor,MAPK,VEGF,FOXO and other signaling pathways are involved in homeodimer protein activation,sequence specific DNA binding,chromatin binding,ATP binding and other molecular functions,regulating DNA transcription,promoting RNA polymerase II promoter transcription,participating in inflammatory reaction,inhibiting apoptosis process and promoting cell proliferation and other biological processes,so as to play an anti fibrosis therapy efficacy.However,its specific mechanism still needs to be verified by further experimental research.