慢性乙肝感染者外周血单个核细胞乙肝病毒X基因表达与氧化应激的相关性

Expression of HBV X gene in peripheral blood mononuclear cell of HBsAg chronic infection patients and its relationship with oxidative stress

目的:探讨慢性乙肝感染者外周血单个核细胞(peripheral blood mononuclear cell,PBMC)乙肝病毒X基因(hepatitis B virus X gene,HBVX)表达与机体氧化应激的相关性。方法:采用RT-PCR检测正常对照组、轻度患者组、中度患者组、重度患者组外周血PBMC中HBVX m RNA的表达;采用流式细胞仪检测以上各组细胞内ROS含量;采用紫外分光光度仪检测各组血清MDA含量。结果:各乙肝患者组PBMC中HBVX m RNA与正常对照组比较,差异有统计学意义(正常对照组HBVX m RNA相对表达量为0,轻度患者组为0.48±0.24,中度患者组为0.71±0.15,重度患者组为0.89±0.27;P<0.001),组间比较,轻度患者组与中度患者组和重度患者组比较,差异有统计学意义(P<0.001);中度患者组与重度患者组比较,差异有统计学意义(P<0.001)。正常对照组ROS相对表达量为24.63±8.93,轻度患者组为25.35±5.66,中度患者组为38.24±11.64,重度患者组为43.45±14.33。PBMC中ROS含量,随着乙肝程度加重,ROS含量增加,轻度患者组与正常对照组比较,差异无统计学意义(P>0.05);中度患者组和重度患者组与正常对照组及轻度患者组比较,差异有统计学意义(P<0.001)。正常对照组血清MDA含量为(5.31±1.24)nmol/L,轻度患者组为(6.03±1.45)nmol/L,中度患者组为(12.27±3.53)nmol/L,重度患者组为(19.72±4.66)nmol/L,MDA含量随着患病程度加重而增加,轻度患者组与正常对照组比较,差异无统计学意义(P>0.05);中度患者组和重度患者组与正常对照组及轻度患者组比较,差异有统计学意义(P<0.001)。HBVX m RNA表达与ROS,MDA含量呈正相关(Spearman’s r=0.382,r=0.467,P<0.05)。结论:慢性乙肝感染者外周血PBMC中HBVX m RNA表达随疾病程度加重而增加,其与患者体内氧化应激反应呈正相关。

Objective： To explore the expression of HBVX gene in PBMC in the patient of HBV chronic infection, and the correlation between it and oxidative stress. Methods： The expression of HBVX m RNA in PBMC was detected by RT-PCR. ROS in PBMC was detected by flow cytometer. MDA in blood serum was detected by ultraviolet spectrophotometer. Results： There were four groups through the degree of chronic hepatitis, including a normal group, a mild degree group, a midrange degree group and a severe degree group. There was significant difference of gene expression of HBVX m RNA in PBMC between the normal group and the chronic hepatitis group（HBVX m RNA expression of the normal group, the mild degree group, the midrange degree group and the severe degree group was 0, 0.48±0.24, 0.71±0.15 and 0.89±0.27, respectively, P〈0.001）. There was significant difference of gene expression of HBVX m RNA of PBMC between the mild degree group and the midrange degree group or the severe degree group（P〈0.001）. The content of ROS in the the normal group, the mild degree group, the midrange degree group and the severe degree group was 24.63±8.93, 25.35±5.66, 38.24±11.64 and 43.45±14.33, respectively. There was no significant difference of the content of ROS in PBMC between the normal group and the mild degree group（P〈0.05）. There was significant difference of the content of ROS in PBMC between the midrange degree group or the severe degree group and the normal group or the mild degree group（P〈0.001）. The content of MDA in the the normal group, the mild degree group, the midrange degree group and the severe degree group was（5.31±1.24）,（6.03±1.45）,（12.27±3.53） and（19.72±4.66） nmol/L. There was no significant difference of the content of MDA in blood serum between the normal group and the mild degree group（P〈0.05）. There was significant difference of the content of MDA in blood serum between the midrange degree group or the severe degree group and the normal group or the mild degree group（P〈0.001）. There was positive correlation between the expression of HBVX m RNA and the content of ROS and MDA（Spearman＇s r=0.382, r=0.467, P〈0.05）. Conclusion： The expression of HBVX m RNA in PBMC in the patient of Hbs Ag chronic infection is increased with degree of chronic hepatitis. There is positive correlation between the expression of HBVX m RNA and oxidative stress in the patient chronic hepatitis.