ObjectiveBased on the previous application of protein chip technology and iTRAQ combined with LC-MS/MS technology to study the differentially expressed proteins of dififerent TCM syndromes of chronic hepatitis B,this study will use ELISA method to detect the expression levels of I-309,MCP-1,SDF-1,FAP,M-CSF,IGF-1,VCAM-1 and ApoM proteins related to the liver and gallbladder damp-heat of chronic hepatitis B in this syndrome and chronic hepatitis B without TCM syndrome,so as to obtain the proteins material basis related to the characteristics of liver and gallbladder damp-heat of chronic hepatitis B.MethodsThe study subjects include 30 patients of the liver and gallbladder damp-heat of chronic hepatitis B,15 patients of chronic hepatitis B without TCM syndrome and 15 healthy volunteers.Peripheral venous blood samples were collected from each group,and the expression level of target proteins in each group was detected by ELISA,GO enrichment analysis and KEGG signaling pathway enrichment analysis of tifferentially expressed proteins using STRING database.All the data in this study were analyzed by SPSS 22 software.MedCalc 19.0.3 software was used to drawing ROC curve.Results1.Compared with healthy group,the expression of I-309,MCP-1,SDF-1,FAP,M-CSF,IGF-1 and VCAM-1 proteins in the liver and gallbladder damp-heat syndrome and without TCM syndrome of chronic hepatitis B were all up-regulated with statistically significant(P<0.05),and the expression of ApoM protein were down-regulated with statistically significant(P<0.05),2.Compared with the chronic bepatitis B without TCM syndrome group,the TBIL level in the liver and gallbladder damp-heat syndrome of chronic hepatitis B increased without statistically significant(P>0.05);The levels of ALT,AST and HBV DNA decreased without statistically significant(P>0.05);the expression of I-309,MCP-1,SDF-1,FAP,M-CSF,IGF-1,VCAM-1 proteins decreased,and the difference was statistically significant(P<0.05);ApoM protein expression was up-regulated,but there was no statistically significant.(P>0.05).3.GO enrichment analysis and KEGG signaling pathway enrichment analysis of I-309,MCP-1,SDF-1,FAP,M-CSF,IGF-1,VCAM-1 proteins showed that they were mainly involved in the positive regulation of immune system,the positive regulation of leukocyte chemotaxis and T cell activation,the positive regulation of cell adhesion,migration and proliferation,response to stimuli,inflammatory response and signal receptor activity in biological processes.In molecular function,they were mainly involved in signal receptor binding,integrhn binding,receptor ligand activity,cytokine activity,chemokine activity,etc.In cell location,they were mainly located in extracellular space,extracellular area,etc.They were mainly concentrated in cytokine-cytokine receptor interaction signaling pathway,rheumatoid arthritis signaling pathway,TNF signaling pathway,chemokine signaling pathway,etc.4.ROC curve analysis showed that the sensitivity,specificity and the area under the curve of the combination of I-309,MCP-1,SDF-1,FAP,M-CSF,IGF-1 and VCAM-1 proteins for differentiating the liver and gallbladder damp-heat syndrome of chronic hepatitis B and without TCM syndrome of Chronic Hepatitis B were 96.67%,93.33%and 0.960 respectively.conclusions1.The changes of I-309,MCP-1,SDF-1,FAP,M-CSF,IGF-1,VCAM-1 and ApoM expression level are related to chronic hepatitis B.2.The changes of I-309,MCP-1,SDF-1,FAP,M-CSF,IGF-1 and VCAM-1 expression level are related to the characteristics of the liver and gallbladder damp-heat syndrome of chronic hepatitis B.The function of differentially expressed proteins mainly involves the regulation of T lymphocyte and monocyte,which provides a direction for the study of biological regulation mechanism of syndrome occurrence. |