HBV慢性感染患者PBMC凋亡与血清病毒载量

Relationship between PBMC apoptosis and serum HBV level in HBV positive patients

目的:研究HBV慢性感染患者血清HBV载量与外周血单个核细胞(PBMC)凋亡的关系. 方法:无菌条件下分离42例HBV阳性患者及健康献血员10 名外周血单个核细胞(PBMC),在PHA的刺激下培养72 h 后收集细胞.采用脱氧核糖核酸末端标记方法(terminal deoxyribonucleotidyl transferase-mediated biotinylated dUTP nick-end labeling,TUNEL)、流式细胞术(flow cytometry,FCM)检测HBV慢性感染患者PBMC凋亡,并用HBV-PCR定量试剂盒检测血清HBV-DNA的含量.对不同组的PBMC凋亡及血清HBV-DNA含量进行单因素方差分析,并对凋亡情况及血清HBV-DNA含量进行相关性分析. 结果:TUNEL法与FCM检测HBV阳性患者PBMC,证实PBMC存在凋亡,慢性乙型肝炎轻度组、中度组与肝硬化组凋亡率明显升高分别为(29.3±9.3%,24.8±10.8%, 28.8±5.0%),与健康对照(18.0±6.4%)比较凋亡率显著升高(P<0.05).血清HBV-DNA含量在慢性乙型肝炎轻度组最高(14.3±24.8×107cp/L),与其他组比较病毒含量明显增高(P<0.05).HBV慢性感染患者血清病毒载量与其PBMC 凋亡率有一定的相关性,r=0.338,P=0.014(P<0.05). 结论:HBV慢性感染患者PBMC存在凋亡,其凋亡率较健康组明显增加.慢性乙型肝炎轻度患者血清病毒含量最高, 其凋亡率亦最高.凋亡状况与乙型肝炎的慢性化、机体的免疫状态相关.HBV慢性感染患者PBMC凋亡与其血清病毒载量有相关性.

AIM: To investigate the relationship between the serum HBV DNA quantification and the PBMCs apoptosis of the chronic hepatitis B. METHODS: Peripheral blood mononuclear cells (PBMCs) were isolated from 10 healthy blood donors and 42 patients with HBV positive. PBMCs were cultured with PHA for 72 h in vitro. Then we used TUNEL and flow cytometry to assay the apoptosis. The HBV-DNA extracted from the serum in patients by PCR was quantified by ELISA. We used the one way ANOVA to analysis the apoptosis and the HBV-DNA quantity, and made correlation analysis between the apoptosis and the HBV-DNA quantity. RESULTS: We identified the apoptosis by TUNEL and flow cytometry in cultured PBMCs. The apoptosis rate in mild type of chronic hepatitis B, moderate type of chronic hepatitis B and cirrhosis were (29.3±9.3%, 24.8±10.8%, 28.8±5.0%), compared to the healthy group (18.0±6.4%). Serum HBV-DNA level was the highest in mild type of chronic hepatitis B (14.3±24.8×107cp/L), which was higher than other groups (P<0.05). The serum HBV-DNA level in mild type of chronic hepatitis B is the highest. The serum HBV-DNA level had the correlation with the apoptosis in PBMC (r =0.338, P =0.014). CONCLUSION: There is apoptosis in PBMC of the HBV chronic infections. The serum virus level is the highest in mild type of chronic hepatitis B, while its apoptosis is also the highest. The immune and HBV virus persistent condition are related to the apoptosis. The percentage of the apoptosis in PBMC has the correlation with the virus level in serum of the HBV chronic infections.