A Preliminary Study On Expression Level Of Endoplasmic Reticulum Stress Protein In Peripheral Blood Mononuclear Cells Of Patients With Chronic Hepatitis B

Objective:To assess the correlation between endoplasmic reticulum stress protein in PBMCs and the states of chronic hepatitis B(CHB)patients,the expression level of endoplasmic reticulum stress protein in patients with CHB are analyzed.This work will provide some evidences to predict the development of CHB patients.Method:(1)PBMCs were harvested from 41 healthy peripheral blood samples and 102peripheral blood samples from CHB patients.PBMCs were isolated by density gradient centrifugation.(2)Levels of HBV-DNA were detected in PBMCs of 22 patients with CHB by fluorescence probe PCR.(3)PBMCs from 6 healthy volunteers were inoculated in cell culture plate.After 24 hours,5μL,10μL,15μL,and 20μL serum from CHB patients and same volume serum from healthy people as the control group were added(The HBV-DNA level was 10~7copies/mL).HBV-DNA in PBMCs were detected by fluorescence probe quantitative PCR after another 24 hours.And Glucose regulator protein 78(GRP78),Glucose regulatory protein 94(GRP94)and C/EBP homologous protein(CHOP)expression levels were detected by real-time fluorescence qPCR.(4)35 PBMC samples from healthy people and 80 PBMC samples from chronic hepatitis B patients(30 mild chronic hepatitis B cases,25 moderate chronic hepatitis B cases,and 25 severe chronic hepatitis B cases)were detected by real-time quantitative PCR and western-blot(WB)techniques to measure expression of GRP78,GRP94 and CHOP levels.The correlation between AST,ALT and GRP78,GRP94,CHOP expression levels could be analyzed.Results:(1)HBV-DNA was found in 22 PBMCs of CHB patients.(2)HBV-DNA in PBMCs increased after adding serum of CHB patients.The CHOP gene expression levels increased in the experimental groups,5μL serum group excepted.(3)The expression levels of GRP78,GRP94,and CHOP in the mild chronic hepatitis B and severe chronic hepatitis B group increased significantly(P<0.05).The expression of GRP78,GRP94 group and CHOP were positively correlated with the level of ALT and AST in peripheral blood of CHB patients.Conclusion:The expression levels of ER stress protein GRP78,GRP94 and CHOP in PBMCs of CHB patients increased with the condition of the CHB,and are related with ALT and AST level in the peripheral blood.The increase of endoplasmic stress proteins may be related with the extent of liver inflammation.