HBV对健康人PBMC中内质网应激蛋白的影响研究

Study on the effect of HBV on endoplasmic reticulum stress proteins in healthy human PBMC

目的研究乙型肝炎病毒(HBV)对外周血单个核细胞(PBMC)中内质网应激(ERS)蛋白GRP78、GRP94及CHOP的影响,探讨HBV感染与PBMC ERS的关系。方法分离健康体检者PBMC,与不同量的慢性乙型肝炎(CHB)患者血清共培养,通过荧光定量聚合酶链式反应(PCR)法检测CHB患者和共培养后健康体检者PBMC中的HBV-DNA水平。采用荧光定量PCR法检测共培养后健康体检者PBMC中GRP78、GRP94、CHOP核酸水平的表达量。结果 22例CHB患者PBMC中,有16例检测到HBV-DNA,阳性检出率为72.7%。外周血HBV-DNA拷贝数越高,PBMC中HBV-DNA阳性率越高。健康体检者PBMC中加入CHB患者血清共培养,PBMC中HBV-DNA水平会随着加入的CHB患者血清量增加而升高。CHB患者血清与健康体检者PBMC共培养不同时间,PBMC中ERS蛋白GRP78、GRP94和CHOP在基因水平的表达均升高,差异均有统计学意义(P<0.05)。结论 CHB患者PBMC中可检测到HBV-DNA,HBV感染可以引起PBMC中ERS相关蛋白水平升高。

Objective To study the effects of hepatitis B virus(HBV)on endoplasmic reticulum stress(ERS)proteins GRP78,GRP94 and CHOP in peripheral blood mononuclear cells(PBMC),and explore the relationship between HBV infection and PBMC ERS.Methods PBMC from healthy subjects was isolated and co-cultured with sera from patients with chronic hepatitis B(CHB).Quantitative quantitative polymerase chain reaction(PCR)was used to detect HBV-DNA levels in PBMC of CHB patients and healthy subjects after co-culture.The expression of GRP78,GRP94,and CHOP nucleic acid levels in PBMCs of healthy subjects after co-culture were detected by fluorescent quantitative PCR.Results HBV-DNA was detected in 16 of 22 PBMC from CHB patients,and the positive detection rate was 72.7%.The higher the number of copies of HBV-DNA in peripheral blood,the higher the HBV-DNA positive rate in PBMC.When sera from CHB patients were added to PBMC of healthy subjects and co-cultured,HBVDNA levels in PBMC increased with the increase of serum levels of CHB patients.The serum of CHB patients was co-cultured with PBMC from healthy subjects for different time periods.The expression of ERS proteins GRP78,GRP94 and CHOP at the gene level in PBMC increased,and the differences were statistically significant(P<0.05).Conclusion HBV-DNA can be detected in PBMC of patients with CHB,and HBV infection can cause elevated levels of ERS-related proteins in PBMC.