Basic Research On The Biological Material Of The Transcriptome Of Chronic Hepatitis B And Non-alcoholic Fatty Liver

Objective:(1)Based on transcriptome data and molecular biology verification,screen the potential characteristic m RNA of chronic hepatitis B(CHB)and non-alcoholic fatty liver disease(NAFLD).(2)Calculate the dynamic relationship between TCM syndrome characteristics and core genes of CHB and NAFLD liver and gallbladder damp-heat syndrome,and explore the mechanism of CHB and NAFLD "different disease syndrome",and provide objective basis for syndrome differentiation of CHB and NAFLD.Methods:(1)According to the characteristics of transcriptome expression data,the random variance model RVM algorithm was designed to screen differentially expressed genes.The existing expression spectrum data in GEO database was used to train RVM algorithm,and the optimal parameters of differential expression gene screening were obtained.(2)Collected TCM syndrome information of CHB and NAFLD patients with dampness-heat in liver and gallbladder,collected CHB,NAFLD and healthy volunteer blood lymphocyte samples,detect sample transcriptome(m RNA)expression profile data,and using RVM algorithm to screen differentially expressed genes,the expressed genes were analyzed by functional enrichment of gene ontology(GO)and KEGG to obtain the characteristic biomaterial basis of CHB and NAFLD dampness-heat in liver and gallbladder.(3)Cytoscape program was used to construct a gene regulatory network for CHB and NAFLD hepatobiliary damp-heat syndrome.Use Cluster ONE to filter and reconstruct the core Cluster network,calculate the network topology parameters and screen the core nodes of the Cluster network as important characteristic biological substances(m RNA);(4)Core features of CHB and NAFLD dampness-heat in liver and gallbladder respectively.The biological substances(m RNA)were verified by q RT-PCR,and the multi-step regression and ROC analysis methods were used to determine the core characteristics of CHB and NAFLD.The DFAA association algorithm was designed to analyze the dynamic association between the characteristics of biological substances and TCM syndromes of CHB and NAFLD hepatobiliary damp-heat syndrome.The connotation and relationship mechanism of CHB and NAFLD hepatobiliary damp-heat syndrome "different disease syndrome" were revealed from the material basis and syndrome characteristics.Results:(1)Implementation of a random variance model of high-throughput transcriptome data by RVM algorithm.The differential expression model RVM was designed for the expression of spectral data.The RVM algorithm was trained by GSE121248 data in GEO database.The results showed that RVM algorithm could effectively improve the screening efficiency of differentially expressed genes.(2)CHB and NAFLD dampness-heat in liver and gallbladder characteristics of biological basis.The differentially expressed genes of CHB and NAFLD hepatobiliary damp-heat syndrome were screened by RVM algorithm.Under the condition of p<0.001,there were 1618 differentially expressed genes in CHB hepatobiliary damp-heat syndrome,and 622 genes with up-regulated expression were down-regulated.There are 961 genes;343 differentially expressed genes were obtained in the NAFLD hepatobiliary damp-heat syndrome,of which 227 were up-regulated and 116 were down-regulated.Co-expression gene analysis showed that there were 305 co-expressing genes in CHB and NAFLD,among which 134 genes showed the same trend in the two groups,and 171 showed inconsistent expression trends in the two groups.Gene ontology(GO)and KEGG pathway analysis showed that these co-expressed genes play an important role in the mechanism of CHB and NAFLD "isomorphism".(3)Use Cytoscape software to construct CHB and NAFLD hepatobiliary damp-heat syndrome co-expressing gene regulatory network,using Cluster ONE method to screen the core cluster in the network and reconstruct the Cluster network,calculate the degree distribution(Degree)and topological coefficient of the Cluster network(Topological Coefficient),Clustering Coefficient,Betweenness Centrality,Closeness Centrality and other parameters.Eight core nodes were screened according to the mean value of the parameters as CHB and NAFLD hepatobiliary damp-heat syndrome.The core characteristics of biological substances are AK1,BLVRB,GLUL,NFIX,PTPRC,RIOK3,SHARPIN and SPTB.(4)The q RT-PCR method was used to verify the core characteristics of CHB and NAFLD dampness-heat in liver and gallbladder.The results of multiple stepwise regression and ROC analysis showed that these 8 core nodes are important in the mechanism of CHB and NAFLD.(5)Characteristic biological material.Using the DFAA algorithm to correlate the information of CHB(n=385)and NAFLD(n=355)hepatobiliary dampness syndrome with 109 co-expressed genes and extract 25 common syndromes with CHB and NAFLD.TCM syndrome characteristics closely related to the relationship mechanism.Conclusion: In this study,the differentially expressed genes of CHB and NAFLD hepatobiliary damp-heat syndrome were screened by random variance model RVM algorithm,and 305 co-expressed genes were obtained.Cytoscape software was used to construct CHB and NAFLD hepatobiliary damp-heat syndrome to express gene regulatory network and obtain 8 core genes,namely AK1,BLVRB,GLUL,NFIX,PTPRC,RIOK3,SHARPIN and SPTB.QRT-PCR validation and data analysis showed that these core genes may be important biological substances in the mechanism of CHB and NAFLD "different disease syndrome".Furthermore,the dynamic correlation analysis of 109 TCM four diagnosis information and 8 co-expressed genes of CHB(n=385)and NAFLD(n=355)liver and gallbladder dampness syndrome were carried out,and 25 TCM characteristics information was extracted as CHB and NAFLD.The TCM syndrome characteristics of the disease-associated relationship mechanism,together with the eight core co-expressed genes,participate in the regulation of the relationship between CHB and NAFLD dampness-heat in liver and gallbladder.