慢性乙肝患者乙肝病毒载量与PBMC凋亡及Caspase-8的相关性研究

目的 探讨慢性乙肝（CHB）患者血清HBVDNA载量与外周血单个核细胞（PBMC）凋亡及Caspase-8活性的关系.方法 选取30例CHB患者为实验组,并按单位血清中HBVDNA载量高低分为A（高病毒载量）、B（中病毒载量）、C（低病毒载量）三小组,选取10例正常成年人为对照组,用淋巴细胞分离液分离两组的PBMC,体外与植物血凝素（PHA）共同培养72 h,用PI对PBMC进行染色并以流式细胞仪检测PBMC的凋亡百分率,同时以比色法检测PBMC细胞内Caspase-8的活性.结果 实验组PBMC的凋亡率（26.88±7.37）%高于健康对照组（14.95±2.53）%（P〈0.01）;实验组中A、B、C三个小组PBMC的凋亡率依次递减,分别为（34.75±4.59）%,（25.63±3.55）%,（18.91±3.81）%;实验组Caspase-8的活性2.99±0.82高于健康对照组1.43±0.91（P〈0.01）,且A、B、C三个小组的Caspase-8活性亦依次递减,分别为3.87±0.35,2.95±0.36,1.95±0.29.实验组PBMC凋亡率与Caspase-8活性呈正相关（r=0.825,P〈0.01）.结论 CHB患者PBMC存在活化诱导细胞死亡（AICD）现象,凋亡酶蛋白Caspase-8在此过程中活性升高,可能参与了凋亡信号的转导过程;HBVDNA载量与PBMC凋亡率及Caspase-8的活性相关,可能是诱发PBMC凋亡的原因之一.

Studies on the relationship between serum HBV DNA level and apoptosis of PBMC and Caspase-8 in patients with CHB

Objective To study the relationship between serum HBV DNA level and apoptosis of peripheral blood mononuclear cells (PBMC) , and the relationship between serum HBV DNA level and the activity of caspase-8 in the patients with chronic hepatitis B (CHB). Method 30 CHB patients were selected as experimental group, and it was divided into three subgroups according to the serum HBV DNA level, subgroup A (high serum HBVDNA), subgroup B (medium serum HBVDNA), and subgroup C (low serum HBVDNA). 10 healthy adults were random selected as control group. PBMC were isolated from two groups by separating medium of lymphocytic cell and culturing it with phytohemagglutinin (PHA) in vitro for 72 hours. The PBMC was stained with PI and the apoptosis was assayed with flow cytometry. At the same time, the aetivity of caspase-8 of PBMC was assayed by color matching. Results The apoptosis rate of PBMC of experimental group ( 26. 88 ± 7.37 ) % were higher than that of the control group ( 14. 95 ±2. 53)% ( P <0. 01 ). In the experimental group, the apoptosis rate of PBMC of subgroups A, B and C showed decreasing order (34. 75 ± 4. 59)%, (25.63 ± 3.55 )%, ( 18. 91 ± 3. 81 )%. The activity of caspase-8 of experimental group 2. 99 ±0. 82 were higher than that of the control group 1.43 ±0. 91 ( P <0. 01 ). The activity of caspase-8 of subgroup A, B and C showed the same decreasing order: 3. 87 ±0. 35,2. 95 ± 0. 36, 1.95 ± 0. 29. There was a positive correlation between the apoptosis level of PBMC and the activity of caspase-8 in experimen tal group ( r = 0. 610, P < 0. 01 ). Conclusion AICD of PBMC was found in patients with CHB. The activity of caspase-8 increased in that process, and it may participate in the transduction of apoptosis signal. Serum HBV DNA level was related with the apoptosis rate of PBMC and the activity of caspase-8, and it may be one of the reasons of apoptosis in PBMC.