Based On Metabolomics And PI3K/AKT Signaling Pathway To Explore The Mechanism Of Longchai Fang On CHB And Efficacy Evaluation

Chronic Hepatitis B(CHB)is a chronic infectious disease caused by hepatitis B virus(HBV)which causing liver damage.Longchai Fang(LCF)is one of the effective prescriptions for the treatment of CHB.This paper is on the therapeutic effect of Longchai Fang on CHB.Firstly,the theory and literature review were carried out,and a systematic evaluation was carried out.Then the animal experiments were carried out to verify the relevant pathways and analyze the metabolites,and comprehensively expounded the curative effect of the drug on CHB and the possible mechanism of the curative effect.Part I:Theoretical research.Firstly,four aspects of drug basis,theoretical mechanism,clinical and pharmacological research,the related research literature of Longchai Fang was retrospectively analyzed.Secondly,the research status of network pharmacology and metabolomics in CHB is summarized.Literature summary research shows that polysaccharide content is the highest,total saponins is second,and the decoction is the content of the index components is slightly higher,and the quality of Longchai Fang is stable.Longchai Fang has the functions of regulating the liver and spleen,clearing heat and dampness,activating blood and dissolve stasis,and has a certain anti-HBV effect.It can improve liver function and symptoms of CHB patients.Oxidative index activity,inhibits cell cycle,reduces cell viability,inhibits the secretion of HBV antigens,has good antiviral activity,and has certain advantages in inhibiting viral replication and rebound after drug withdrawal.The existing database of network pharmacology in the study of traditional Chinese medicine compound treatment of CHB is relatively limited in scale,and the relevant literature is mainly theoretical exposition,and there are few experimental studies.Metabolomics also shows great potential in the study of HBVrelated liver diseases,and will have great value in pathogenesis,early diagnosis,efficacy evaluation,and prognosis.Part II:Meta-analysis of Longchai Fang in the treatment of CHB.In order to further objectively evaluate the efficacy of Longchai Fang in the treatment of CHB,a Meta-analysis method was used to screen out a number of clinical research literatures on the treatment of CHB through the database and conduct a review.The results showed that Longchai Fang was better in improving ALT,AST,TBIL,HBV-DNA,HBeAg,HBsAg,symptoms and overall curative effect,which is better than the control drug,the effect is more remarkable by prolonging the treatment time,and its single use is better than the single use of matrine capsules,Yiganning tablets,compound Yiganling tablets and Yiganjiedu decoction,and the combined use is compared with other drugs.Has a therapeutic advantage.However,due to the relatively small number of clinical research reports,it is necessary to conduct standardized large-scale clinical trials,combined with research reports with negative conclusions,in order to improve the quality of the trials and obtain more rigorous,rich,comprehensive and accurate evaluation results to provide sufficient evidence for the treatment of CHB with Longchai Fang.Part III:Network pharmacology research.Based on network pharmacology,analyze and explore the molecular mechanism of Longchai Fang in the treatment of CHB.The potential active ingredients of Longchai Fang were screened through the TCMSP database,and the target of action was predicted by using the Swiss database,which was mapped with the CHB disease targets obtained from the Drugbank,Disgenet,and Genecards databases to obtain the potential targets of Longchai Fang for the treatment of CHB.Cytoscape software was applied in building the "Longchai Fang-ingredient-disease-target" and protein interaction network,and conducts GO biological function and KEGG pathway enrichment analysis on the targets of Longchai Fang,finally constructs a "disease-ingredient-target".The results show that the main active components of Longchai Fang in the treatment of CHB may be medipterin,isorhamnetin,licorice chalcone,licorice isoflavones,etc.Targets such as PIK3CA and Hepatitis B,PI3K-Akt,AGE-RAGE and other signaling pathways play a therapeutic role,providing an objective basis and new ideas for the molecular basis of Longchai Fang in the treatment of CHB.PART IV:Experimental Research1.Cell experiment.Based on network pharmacology research,in order to further verify whether the antiviral effect of Longchai Fang is related to PI3K-Akt-related signaling,we cultured Hepg2.2.15 cells in vitro and added different concentrations of Longchai Fang and Lamivudine(3-TC)containing serum,the cell viability was measured and the Western blot experiment was performed.The results confirmed that the Longchai Fang-containing serum could effectively inhibit the growth of Hepg2.2.15 cells and up-regulate the proteins of PI3K,AKT and PTEN in hepatocytes.This suggests that the treatment of CHB with Longchai Fang may be achieved by up-regulating the expression of PTEN through the PI3K/AKT signaling pathway.2.Metabolomics research.To explores the regulatory effect of Longchai Fang on the abnormal metabolism of DHBV Through animal experiments,based on metabolomics technology,the drug-containing serum of DHBV hemp duck was analyzed.Phospholipid metabolism pathway closely related to the therapeutic effect of Longchai Fang,may be its mechanism of action and potential therapeutic target.Part Ⅴ:Research on the rules and experience of Professor Zhu Fangshi in the treatment of CHBIn order to learn and inherit the tutor’s experience,data mining techniques such as "drug use frequency,cluster analysis,factor analysis,and association rule analysis" were used to summarize and analyze the clinical medication rules.To soothe the liver and strengthen the spleen,nourish qi and nourish yin.The core drugs are Solanum nigrum,Sophora flavescens,Hedyotis diffusa,Scutellaria baicalensis,Ligustrum lucidum,etc.The commonly used drug pairs are Sophora Radix+Solanum nigrum,Solanum nigrum+Scutellaria baicalensis,Hedyotis diffusa+Solanum nigrum,Solanum nigrum+Ligustrum lucidum,Ligustrum lucidum+Sophora flavescens,the basic side is Longchai side.The research results show that ① Longchai Fang has its theoretical basis for the curative effect of chronic liver disease.②The clinical efficacy of Longchai Fang in the treatment of CHB is positive.③Longchai Formula can inhibit the growth of Hepg2.2.15 cells by inhibiting the PI3K/AKT signaling pathway,which may be one of the molecular mechanisms of Longchai Fang in the treatment of CHB.④Phospholipid metabolism pathway may be another main aspect of Longchai Fang participating in the repair of liver cells and playing a therapeutic role.⑤The principle of the instructor’s treatment of CHB is mainly to clear heat and detoxify,supplemented by soothing liver and spleen,nourishing qi and nourishing yin,and the basic formula is also Longchai formula.