摘要
AIM: To determine the NF-kB activity in peripheral blood mononuclear cells (PBMC) in patients with acute cholangitis of severe type (ACST) and correlate the degree of NF-kB activation with severity of biliary tract infection and clinical outcome. METHODS: Twenty patients with ACST were divided into survivor group (13 cases) and nonsurvivor group (7 cases). Other ten patients undergoing elective gastrectomy or inguinal hernia repair were selected as control group. Peripheral blood samples were taken 24 hours postoperatively. PBMC were separated by density gradient centrifugation, then nuclear proteins were isolated from PBMC, and Electrophoretic Mobility Shift Assay (EMSA) used determined. The results were quantified by scanning densitometer of a Bio-Image Analysis System and expressed as relative optical density (ROD). The levels of TNF-alpha, IL-6, and IL-10 in the plasma of patients with ACST and healthy control subjects were determined by using an enzyme-linked immunoassay (ELISA). RESULTS: The NF-kB activity was 5.02 +/- 1.03 in nonsurvivor group, 2.98 +/- 0.51 in survivor group and 1.06 +/- 0.34 in control group. There were statistical differences in three groups (P【0.05). The levels of TNF-alpha and IL-6 in plasma were (498 +/- 53)ng.L(-1)and (587 +/- 64)ng.L(-1)in nonsurvivor group, (284 +/- 32)ng.L(-1) and (318 +/- 49)ng.L(-1)in survivor group and (89 +/- 11)ng.L(-1) and (102 +/-13)ng.L(-1)in control group. All patients with ACST had increased levels of TNF-alpha and IL-6, which were many-fold greater than those of control group, and there was an evidence of significantly higher levels in those of nonsurvivor group than that in survivor group (P【0.05). The levels of IL-10 in plasma were (378+/-32)ng.L(-1), (384+/-37)ng.L(-1) and (68+/-11)ng.L(-1) in three groups, respectively. All patients had also increased levels of IL-10 when compared with control group (P【0.05), but the IL-10 levels were not significantly higher in nonsurvivors than in survivors (P】0.05). CONCLUSION: NF-kB activity in PBMC in patients with ACST increases markedly and the degree of NF-kB activation is correlated with severity of biliary tract infection and clinical outcome.
瞄准:在外部血决定 NF-kB 活动在有严重类型(ACST ) 和相互关联的尖锐胆管炎的病人的单音的原子房间(PBMC ) 有胆汁的道感染和临床的结果的严厉的 NF-kB 激活的度。方法:有 ACST 的二十个病人被划分成幸存者组(13 个盒子) 并且非幸存者组(7 个盒子) 。经历选任的 gastrectomy 或腹股沟的脱肠修理的另外的十个病人作为控制组被选择。外部血样品手术后地被花 24 个小时。PBMC 被密度坡度 centrifugation 分开,然后原子的蛋白质从 PBMC 被孤立,并且 Electrophoretic 活动性移动试金(EMSA ) 使用了坚定。结果被扫描一个简历图象分析系统的 densitometer 确定并且表示了同样相对的光密度(杆) 。在有 ACST 和健康控制题目的病人的血浆的 TNF-alpha, IL-6,和 IL-10 的层次被使用连接酶的免疫分析(ELISA ) 决定。结果:NF-kB 活动是 5.02 +/- 1.03 在里面非幸存者组, 0.51 在幸存者组织的 2.98 +/- 和 1.06 +/- 0.34 在控制组。在三个组(P<0.05 ) 有统计差别。在血浆的 TNF-alpha 和 IL-6 的层次是( 498 +/- 53 ) ng.L ( -1)and ( 587 +/- 64 ) ng.L ( -1)in 非幸存者组,( 284 +/- 32 ) ng.L (-1)并且( 318 +/- 49 ) ng.L ( -1)in 幸存者组并且( 89 +/- 11 ) ng.L (-1)并且( 102 +/-13 ) ng.L ( -1)in 控制组。有 ACST 的所有病人增加了 TNF-alpha 和 IL-6 的层次,它是控制组的比那些大的许多褶层,并且有一条证据在那些显著地高级非幸存者组比那在幸存者组(P<0.05 ) 。在血浆的 IL-10 的层次是(378+/-32 ) ng.L (-1),(384+/-37 ) ng.L (-1) 并且(68+/-11 ) ng.L (在三个组的 -1) 分别地。当与控制相比组(P<0.05 ) ,而是 IL-10 层次不是显著地更高级的在时,所有病人也增加了 IL-10 的层次非幸存者比在幸存者(P>0.05 ) 。结论:在在有 ACST 的病人的 PBMC 的 NF-kB 活动显著地增加并且 NF-kB 激活的度与胆汁的道感染和临床的结果的严厉被相关。
基金
the National Natural Science Foundation of China,No.39970719,30170919