HOECHST33342荧光.doc

【摘要】目的　探讨乙型肝炎患者外周血淋巴细胞(peripheral blood lymphocytes, PBL)凋亡在乙型肝炎病毒(HBV)持续感染中的作用。方法　分离15例慢性乙型肝炎患者PBL，体外单独或与金黄色葡萄球菌肠毒素B(SEB)或重组HBcAg(rHBcAg)共培养，用荧光素YOPRO-1和Hoechst33342对PBL染色，观察PBL中凋亡细胞比例。结果　慢性乙型肝炎患者PBL体外培养48小时后，无论自发性还是由SEB或rHBcAg诱导的PBL凋亡率均明显高于正常对照组，P0.01。其中rHBcAg诱导的PBL凋亡率最高，达24.6%。血清HBeAg阳性组患者PBL凋亡率明显高于HBeAg阴性组患者。慢性轻度和中度肝炎PBL凋亡率又高于慢性重度或慢性重型肝炎患者。结论　慢性乙型肝炎PBL凋亡可能是HBV持续感染的重要因素之一。　　【关键词】乙型肝炎　淋巴细胞　凋亡 Apoptosis of peripheral blood lymphocytes of patients with chronic hepatitis B and clinical significance JI Wei, WANG Huifen, FENG Chuanqian, et al.302 Hospital of PLA, Beijing 100039 　　【Abstract】 Objective To explore the effects of apoptosis of peripheral blood lymphocytes (PBLs) of patients with chronic hepatitis B (CHB) on the persistent infection of hepatitis B virus (HBV). Methods The PBLs of 15 patients with CHB were isolated and cultured with or without Staphylococcus aureus enterotoxin B (SEB; 0.2mg/L), or in the presence or absence of recombinant HBcAg (rHBcAg; 1.0mg/L) for 48 hours in vitro. After incubations, the cells were harvested by centrifugation and then apoptosis of the PBLs was studied by staining with fluorescent dyes YOPRO-1 and Hoechst33342. Results The percentage of apoptotic cells of PBLs of patients with CHB was significanlty higher than in normal controls (P0.01) in the presence or absence of SEB or rHBcAg, but the ratio of apoptotic cells in rHBcAg-stimulated PBLs of the patients was the highest, reaching (24.6±6.1)%. The patients with seropositive for HBeAg had higher percentage of apoptotic cells in their cultured PBLs than those with seronegative for HBeAg had, but the ratio of apoptotic cells in cultured PBLs of the patients with chronic heavy and severe hepatitis B was lower than that of the patients with chronic mild and moderate hepatitis B (P0.01). Conclusion Activation-induced cell death of PBLs of the patients with CHB may be related to persistent infection of HBV, and assay of apoptosis of PBLs of patients with CHB may provide valuable