中文摘要：

目的了解核因子（NF）kB活化对烧伤血清诱导单核细胞活化分泌细胞因子的作用，探讨烧伤血清激活单核细胞的机制方法收集体外培养的人外周血单核细胞（PBMC），分别用正常人血清、烧伤患者血清、烧伤患苦血清＋吡咯烷二硫代氨基甲酸盐（PDTC）刺激后依次分为对照组、烧伤血清组、PDTC组、采用电泳迁移率分析法检测刺激前及刺激0．5、1、0、2．0、4、0h时PBMC的NF-kB活性；酶联免疫吸附测定法和原位杂交法检测刺激前及刺激1．0、2．0、4．0、6．0h时PBMC培养上清液中肿瘤坏死因子（TNF）α、白细胞介素（IL）8水平及TNF-α mRNA、IL-8 mRNA的表达情况。结果血清刺激后，烧伤血清组PBMC NF-kB活性迅速升高，刺激1．0h时达峰值（30．2±3．5）×10^4积分灰度值，与对照组（4．4±0．8）×10^4积分灰度值比较差异有统计学意义（P〈0．01）。刺激2、0h后逐渐下降；而PDTC组NF-kB活性无明显升高，刺激1．0h时为（6．8±0．9）×10^4积分灰度值。烧伤血清组刺激PBMC 1．0h时，TNF-α mRNA表达量和培养上清液中TNF-α水平即升高达峰值，并明显高于对照组（P〈0．01）；IL-8 mRNA表达量和IL-8水平在刺激4．0h时达峰值，也明显高于对照组（P〈0．01）；而PDTC组PBMC培养上清液中TNF-α刺激1．0h时达峰值（0．52±0．06）μg／L；刺激4．0h时IL-8达峰值（239±20）ng／L，与对照组[（0．13±0．07）μg／L、〈156ng／L]比较，差异有统计学意义（P〈0．01）。结论烧伤血清可通过活化NF-kB，启动PBMC对细胞因子的合成和释放，提示NF-kB活化在烧伤血清诱导PBMC分泌细胞因子的过程中起重要作用。

英文摘要：

Objective To investigate the effects of NF-kB activation on the expression of cytokines in monocytes stimulated by burn serum, so as to explore the mechanism in monocyte activation by burn serum. Methods Peripheral blood monocytes（PBMCs） isolated from healthy volunteers were employed as the target cells. The cells were stimulated by serum from healthy volunteers （control） , by serum from burn patients （burn serum） , and by burn serum with addition of PDTC （ pyrrolidine dithioncarbamate）. Activation of monocytic NF-kB before stimulation and at 0. 5, 1. 0, 2.0 and 4.0 poststimulation hours（PSI-I） was assessed with electrophoretic mobility shift assay （EMSA） . Expression of TNF-α and IL-8 mRNA at 1. 0, 2. 0, 4.0, 6.0 PSHs was assayed with in situ hybridization（ISH）. Meanwhile, the contents of TNF-αand IL-8 in the supernatants were assayed by enzyme linked immunosorbent assay （ELISA）. Results The monocytic NF-kB activity in burn serum group increased significantly and reached the peak level at 1 PSH [ （30.2±3.5 ）×10^4 integration gray scale value] after the PBMCs were stimulated by burn serum, and it was obviously higher than that in control group [ （4.4±0. 8）×10^4 integration gray scale value], （ P 〈0.01）. It gradually decreased and returned to the pre-stimulation state at 2 PSH. The monocytic NF-kB activity in PDTC group dereased to [ （6.8±0. 9）×10^4 integration gray scale value at 1 PSH ] after the stimulation. The expression of TNF-α mRNA of the monocytesand the TNF-α level in the supernatant of the cultured PBMCs reached peak level at 1 PSHs after being stimulated by burn serum, and they were obviously higher than those in control group （ P 〈0.O1 ）. While the expression of IL-8 mRNA and the IL-8 level in the supernatant of the cultured PBMCs reached peak level at the 4 PSHs after being stimulated by burn serum, which were obviously higher than those in control group （ P 〈0. 01 ） too. In addition, the synthesis and release of TNF-ct （peaked