Study On Dialectic Typing Of Chronic Hepatitis B By Metabolic Analysis

Objectives:Chronic hepatitis B refers to the liver damage caused by hepatitis B virus,blood HBsAg positive 6 months or more clinical manifestations of fatigue,fear of food,nausea,bloating,liver pain and other symptoms.Due to the high pathogenicity of chronic hepatitis B,early detection of chronic hepatitis B infection is the key to disease treatment and disease prevention.Clinical diagnosis includes laboratory tests:(Alanine aminotransferase(Cerealthirdtransaminase,ALT),Aspartate transaminase(Aspartate transaminase,AST),Total bilirubin(Total bilirubin,TBIL),Prothrombin time(Prothrombin time,PT),Cholinesterase,hepatitis B virus markers,HBV DNA quantification,and imaging studies).At present,the effect of western medicine treatment is not satisfactory.The dialectical theory of traditional Chinese medicine has achieved good results.According to the consensus of experts on diagnosis of chronic hepatitis B in 2012,The patients with chronic hepatitis B were divided into three main syndromes: damp-heat accumulation type,liver stagnation and spleen deficiency type and collaterals type of blood stasis group.However,these syndromes are diagnosed by Chinese medicine practitioners based on their own experience.Molecular biomarkers are not yet clear,and definite diagnosis requires more biomarkers for joint application.Metabolomics is an emerging technology that is powerful and has been widely used in the diagnosis,treatment and prevention of diseases.Liver involved in a variety of metabolic processes,when the disease occurs in the liver,the body metabolites will change significantly.The purpose of this study was to evaluate the clinical efficacy of20 metabolites of dampness and heat accumulation,24 cases of stagnation of liver qi and spleen,20 cases of chronic obstruction of blood stasis syndrome Hepatitis B three syndromes of TCM syndromes of serum metabolites were compared and analyzed from the molecular biology aspects of the three groups of patients serum metabolites,screening can be used for TCM dialectical type of chronic hepatitis B,assist Low-grade Chinese medicine practitioners for the diagnosis of atypical disease symptoms,patients can reduce the financial burden.Methods: 1.Subjects and experimental groups: This experiment collected blood samples of 64 patients confirmed to be chronic hepatitis B in the laboratory of Dalian Sixth People's Hospital.For each case,three high-grade TCM syndromes were required.At the same time,Confirm to confirm the group.TCM dialectical type of three groups of patients were damp heat Accumulation type 20 cases,24 cases of liver depression and spleen deficiency,blood stasis type 20 cases of chronic hepatitis B patients.Collect the patient's plasma samples.All patients in this experiment within the past six months have not been Chinese medicine and interferon or nucleoside analog anti-virus treatment,and no other systemic complications.2.Sample Processing and Data Analysis: The collected samples were subjected to pretreatment to extract metabolites for liquid chromatography-mass spectrometry analysis.Briefly,mass spectrometry was performed after the metabolites were separated by liquid chromatography.3.Biomarker screening and identification: the experimental data obtained after the extraction,pretreatment and other operations,through the library search for metabolites qualitative and quantitative analysis.Then,Principal Component Analysis(PCA)and Partial least squares discriminant analysis(PLS-DA)were used for statistical analysis to screen differential metabolites.Results:(1)In this study,the method of liquid chromatography-mass spectrometry(LC-MS / MS)was used to analyze the plasma metabolic profile in patients with dampness and heat accumulation,liver depression and spleen deficiency syndrome and blood stasis syndrome type chronic hepatitis B patients.Multivariate statistical analysis was used to screen out the relevant differential metabolites among the three TCM syndrome types of chronic hepatitis B patients.(2)A total of 184 metabolites and 88 clinical indexes were detected in this preliminary screening test.After statistical analysis,it was found that ALT,total bilirubin and SM(33: 1)Chronic hepatitis B there are significant differences.Among them,the content of ALT in damp-heat accumulation type chronic hepatitis B was significantly higher than that in the other two groups.Total bilirubin and SM(33: 1)were lower in the group of chronic hepatitis B with liver depression and spleen deficiency syndrome.Conclusion: In summary,this study using LC-MS-based metabonomics research strategy screening potential clinical serum markers of serum markers for chronic hepatitis B can provide ideas for the identification of traditional Chinese medicine.