CAJ | 학술논문

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.
목적 탐토만성을간(CHB)환자혈청HBVDNA재량여외주혈단개핵세포(PBMC)조망급Caspase-8활성적관계.방법 선취30례CHB환자위실험조,병안단위혈청중HBVDNA재량고저분위A(고병독재량)、B(중병독재량)、C(저병독재량)삼소조,선취10례정상성년인위대조조,용림파세포분리액분리량조적PBMC,체외여식물혈응소(PHA)공동배양72 h,용PI대PBMC진행염색병이류식세포의검측PBMC적조망백분솔,동시이비색법검측PBMC세포내Caspase-8적활성.결과 실험조PBMC적조망솔(26.88±7.37)%고우건강대조조(14.95±2.53)%(P<0.01);실험조중A、B、C삼개소조PBMC적조망솔의차체감,분별위(34.75±4.59)%,(25.63±3.55)%,(18.91±3.81)%;실험조Caspase-8적활성2.99±0.82고우건강대조조1.43±0.91(P<0.01),차A、B、C삼개소조적Caspase-8활성역의차체감,분별위3.87±0.35,2.95±0.36,1.95±0.29.실험조PBMC조망솔여Caspase-8활성정정상관(r=0.825,P<0.01).결론 CHB환자PBMC존재활화유도세포사망(AICD)현상,조망매단백Caspase-8재차과정중활성승고,가능삼여료조망신호적전도과정;HBVDNA재량여PBMC조망솔급Caspase-8적활성상관,가능시유발PBMC조망적원인지일.
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.