摘要目的 探讨慢性乙型重型肝炎(CSHB)患者外周血单个核细胞衍生的树突状细胞核因子(NF)-κB、肿瘤坏死因子(TNF)α、白细胞介素(IL)-6与疾病严重程度及预后的相关性.方法 采集37例CSHB患者、20例慢性乙型肝炎(CHB)患者和20例健康对照者的外周血,分为CSHB组、CHB组与对照组并分别用密度梯度离心法分离外周血单个核细胞,用免疫磁珠细胞分选法获得纯化的单核细胞,体外培养6 d为未成熟的树突状细胞,用聚肌胞刺激48 h为成熟的树突状细胞(mDC).用实时定量PCR检测mDC NF-κB p50表达;用酶联免疫吸附试验测定培养mDC上清液中TNF α、IL-6的分泌水平.两组间比较采用t检验,3组间比较采用One-Way ANOVA分析法,相关分析采用直线相关分析法.结果 3组研究对象mDC表达NF-κB p50,表达量CSHB组为2.63±0.94、CHB组为2.03±0.39、对照组为1.41±0.40,3组比较,F=19.01,P<0.01,差异有统计学意义.mDC分泌TNF α,CSHB组、CHB组、对照组分别为(15 317.69±4124.90)pg/ml、(9670.29±3654.68)pg/ml、(6547.43±1027.20)pg/ml,3组比较,F=45.77,P<0.01,差异有统计学意义; IL-6分泌量分别为(1423.78±375.14)pg/ml、(862.68±93.68)pg/ml、(567.26±167.04)pg/ml,3组比较,F=67.60,P<0.01,差异有统计学意义.3组研究对象mDCNF-κB p50表达与TNF α、IL-6水平呈正相关(r=0.52,P<0.01;r=0.65,P<0.01).CSHB组mDC表达TNF α、IL-6与凝血酶原活动度呈负相关(r=-0.41,P=0.01;r=-0.40,P=0.01),与ALT、总胆红素、HBV DNA无相关性(r=-0.03,P=0.85,r=0.01,P=0.93,r=0.01,P=0.95;r=-0.09,P=0.58,r=0.16,P=0.34,r=0.09,P=0.59).CSHB 存活组与死亡组mDC表达TNF α、IL-6差异无统计学意义(t=0.42,P=0.67;t=0.76,P=0.45).结论 CSHB患者树突状细胞衍生的NF-κB及其激活产物TNF α、IL-6表达增加,可能与CSHB疾病严重程度相关.

abstractsObjective To investigate the correlation of nuclear factor- κ B (NF- κ B) level and its regulated cytokines in monocyte-derived dendritic cells (MoDC) with illness severity and prognosis in patients with chronic severe hepatitis B (CSHB). Methods Peripheral blood mononuclear cells (PBMC) were isolated by Ficoll density gradient separation from 37 patients with CSHB, 20 chronically HBV-infected patients, and 20 normal controls. Monocytes were acquired using immunomagnetic anti-CD14-beads. Next,monocytes were induced into immature DC (iDC) in vitro. On day six, polyI:C was added to induce DC maturation. Then mature DCs (mDCs) were collected at 48h after polyI:C treatment to test the expression of NF- κB p50 by real time PCR. The supernatants were also collected respectively. The expression of TNF α and IL-6 in the supernatants were measured by ELISA. Results The expression of NF- κB p50 in mDCs of patients with CSHB was the highest in three groups (F = 19.01, P < 0.01). The secretion of TNF α and IL-6 in mDCs from group CSHB was extremely high when compared with the other two groups, the secretions of TNF α in groups CSHB, CHB and NC were(15317.69 ± 4124.90) pg/ml, (9670.29 ± 3654.68) pg/ml and (6547.43 ± 1027.20) pg/ml (F = 45.77, P < 0.01) respectively, and the productions of IL-6 in groups CSHB,CHB and NC were (1423.78 ± 375.14) pg/ml, (862.68 ± 93.68) pg/ml and (567.26 ± 167.04) pg/ml (F=67.60, P < 0.01), respectively. NF- κ B p50 showed significant correlations with TNF α (r= 0.52, P < 0.01)and IL-6 (r = 0.65, P < 0.01) in mDCs. Furthermore, the secretions ofTNF α and IL-6 in mDCs from group CSHB were negatively associated with PTA (r = -0.41, P = 0.01; r = -0.40, P = 0.01), but not with ALT, TBil and virus loads (r= -0.03, P = 0.85, r = 0.01, P = 0.93, r = 0.01, P = 0.95; r = -0.09, P = 0.58, r = 0.16, P =0.34, r = 0.09, P = 0.59). No significant difference found in the expression of TNF α and IL-6 between the survival subgroup and the death subgroup in group CSHB (t = 0.42, P = 0.67; t = 0.76, P =0.45). Conclusions The expression levels of NF- κ B and its regulated cytokines in monocytederived DCs in patients with chronic severe hepatitis B were up-regulated and perhaps associated positively with the illness severity.