Correlation Between Differentiation Of T-lymphocyte And Regulation Mechanism Of HMGB1-PTEN Pathway In Chronic Hepatitis B With Liver Depression And Spleen Deficiency Syndrome And Splenogastric Hygropyrexia Syndrome

ObjectiveThe clinical manifestations of chronic hepatitis B with liver depression and spleen deficiency syndrome and splenogastric hygropyrexia syndrome are different due to their pathogenesis.The purpose of this study is to explore the nature of the immunological differences of liver depression and spleen deficiency syndrome and splenogastric hygropyrexia syndrome based on the differentiation difference of T-lymphocyte subpopulation between the two,and to further explore the correlation between T-lymphocyte difference and HMGB1-PTEN pathway axis regulation and mediation on the basis of previous literature research,so as to find the biological basis for different syndromes of chronic hepatitis B,to better promote the objectification of TCM clinical syndrome and the accuracy of syndrome differentiationMethodsAccording to the standard of traditional Chinese and Western medicine,36 cases of chronic hepatitis B were collected,18 cases of liver depression and spleen deficiency syndrome and 18 cases of splenogastric hygropyrexia syndrome respectively,and 14 healthy volunteers were included as the control group.Three peripheral venous blood samples(3-5ml/sample)were collected under the condition of fasting.ALT,AST,TB,ALB and other liver function indexes were detected by biochemical analyzer HBV-DNA load was detected by quantitative polymerase chain reaction(PCR)CD4+T,CD8+T,Th17,Treg and Tfh cells ratio were detected by flow cytometry HMGB1,PTEN,PI3K,PDK1 and Akt were detected by ELISA.Spss22.0 was used for data statistical analysis,STRING database was used to construct protein interaction network,GeneMANIA database was used to further analyze the functional expression correlation among HMGB1,PTEN,PI3K,PDK1 and Akt.A multiple regression model was constructed to analyze the correlation between HMGB1-PTEN pathway protein and T lymphocytesResults:1.There was no significant difference in ALT,AST,TB,ALB,HBV-DNA between chronic hepatitis B with liver depression and spleen deficiency syndrome and splenogastric hygropyrexia syndrome(P>0.05)2.Compared with the healthy control group,the CD8+T cells level of chronic hepatitis B with liver depression and spleen deficiency syndrome and splenogastric hygropyrexia syndrome was lower than that of the healthy control group,with statistical significance(P<0.05),and the Th17,Treg and Th17/Treg cells level were significantly higher than that of the healthy control group,with statistical significance(P<0.01);the CD4+T cells level of the splenogastric hygropyrexia syndrome was lower than that of the healthy control group,with statistical significance(P<0.05).There was no significant difference in the level of CD4+T cells between the liver depression and spleen deficiency syndrome and healthy control group(P>0.05);Compared with the splenogastric hygropyrexia syndrome,the level of CD4+T cells and Th17 cells of the liver depression and spleen deficiency syndrome were higher than that in the splenogastric hygropyrexia syndrome,with statistical significance(P<0.05).There was no statistical significance in the level of CD8+T,Treg and Th17/Treg cells between the two syndrome types(P>0.05).There was no statistical significance in the level of CD4+T/CD8+T and Tfh cells among the three groups(P>0.05)3.Compared with the healthy control group,the levels of HMGB1,PI3K,PDK1,and Akt in chronic hepatitis B with liver depression and spleen deficiency syndrome and splenogastric hygropyrexia syndrome were significantly higher than those of the healthy control group,with statistical significance(P<0.01).The PTEN level of liver depression and spleen deficiency syndrome was significantly higher than that of the healthy control group,with statistical significance(P<0.01).The PTEN level of splenogastric hygropyrexia syndrome was significantly lower than that of the healthy control group,with statistical significance(P<0.01).Compared with the splenogastric hygropyrexia syndrome,the level of HMGB1,PTEN and PI3K in the liver depression and spleen deficiency syndrome was higher than that in the splenogastric hygropyrexia syndrome,with statistical significance(P<0.05);There was no statistical significance between the level of PDK1 and Akt in the two syndrome types(P>0.05)4.A protein interaction network map was constructed through the STRING database.There are multiple intrinsic relationships between HMGB1,PTEN,PI3K,PDK1,and Akt.The GeneMANIA database was used to further analyze the functional expression correlations between HMGB1,PTEN,PI3K,PDK1,and Akt,and found that they are mainly related through physical interactions.HMGB1 is linked to PTEN,PI3K,and Akt through PDK1 to form a complex network interaction mechanism5.HMGB 1-PTEN pathway protein is linearly correlated with T lymphocyte levels,and the linearly related immune cell factors of different syndrome types are not consistentConclusion1.The cellular immune function of chronic hepatitis B with liver depression and spleen deficiency syndrome and splenogastric hygropyrexia syndrome is lower than normal,but the cellular immune level of splenogastric hygropyrexia syndrome is lower,there is a certain immune disorder in both syndrome types,and the immune disorder of liver depression and spleen deficiency syndrome is more obvious,which may be related to the different degree of deficiency of positive Qi between syndrome types2.At HMGB 1-PTEN pathway axis,the expression level of most proteins in the liver depression and spleen deficiency syndrome is higher than that in the splenogastric hygropyrexia syndrome,indicating that the inflammatory state of the liver depression and spleen deficiency syndrome is more active,and the inflammatory response of the splenogastric hygropyrexia syndrome is relatively not obvious due to its low autoimmune function,inactive immune response3.There is a certain correlation between T-lymphocyte differentiation and HMGB 1-PTEN pathway regulation in chronic hepatitis B with liver depression and spleen deficiency syndrome and splenogastric hygropyrexia syndrome,which reveals that the immune inflammatory response of liver depression and spleen deficiency syndrome is stronger than that of splenogastric hygropyrexia syndrome.At some extent,it reflects the biological essence of the same disease and different syndromes of chronic hepatitis B.