Li Zhi-qing, Huang Yue-sheng, Yang Zong-cheng, Wang Jia-han
Department of Burns, Nanfang Hospital, The First Military Medical University, Guangzhou 510515, P.R. China.
Zhonghua Shao Shang Za Zhi. 2004 Oct;20(5):265-7.
To investigate the effects of burn serum on nuclear translocation of monocytic NF-kappaB heterodimers p50/p65 and the degradation of inhibiting kappaB (IkappaBalpha), so as to further explore the role of burn serum on the activation of monocytes.
Peripheral blood monocytes (PBMCs) isolated from healthy volunteers were employed as the target cells. The cells were stimulated by the serum from healthy volunteers and burn patients, and by burn serum together with pyrrolidine dithiocarbamate (PDTC). Sera from normal healthy volunteers were taken as control. The nuclear translocation of monocytic p50 and p65 at 30th, 60th, 120th and 480th post stimulation minutes (PSM) was observed with laser confocal microscopy. The degradation of monocytic IkappaBalpha protein at 30th, 60th, 90th and 120th PSM was determined by Western blot.
Compared to that in control group, the nuclear translocation of monocytic p50 and p65 took place 30 min after the PBMCs were stimulated by burn serum, peaking at 30 to 60 min, but it gradually recovered to pre-stimulation state at 2 hrs with decreased intra-nuclear collection. Meanwhile, the IkappaBalpha degradation occurred within 30 min after PBMCs being stimulated by burn serum, and it peaked at 60 mins. However, IkappaBalpha gradually reappeared in the cytoplasm after 2 hrs of stimulation. PDTC (an antioxidants) could effectively inhibit monocytic IkappaBalpha degradation and nuclear translocation of NF-kappaB induced by burn serum.
Burn serum could induce nuclear translocation of p50 and p65 components of NF-kappaB in monocytes into the nucleus and degradation of IkappaBalpha, leading ultimately to the secretion of cytokines from the PBMCs.
研究烧伤血清对单核细胞NF-κB异二聚体p50/p65核转位及抑制性κB（IkappaBα）降解的影响，以进一步探讨烧伤血清在单核细胞激活中的作用。
以从健康志愿者分离的外周血单核细胞（PBMCs）为靶细胞。用健康志愿者和烧伤患者的血清以及烧伤血清联合吡咯烷二硫代氨基甲酸盐（PDTC）刺激细胞。取正常健康志愿者的血清作为对照。用激光共聚焦显微镜观察刺激后第30、60、120和480分钟（PSM）时单核细胞p50和p65的核转位情况。用蛋白质免疫印迹法测定刺激后第30、60、90和120分钟时单核细胞IkappaBα蛋白的降解情况。
与对照组相比，PBMCs受烧伤血清刺激30分钟后单核细胞p50和p65发生核转位，30至60分钟时达到峰值，但2小时时逐渐恢复到刺激前状态，核内聚集减少。同时，PBMCs受烧伤血清刺激30分钟内发生IkappaBα降解，60分钟时达到峰值。然而，刺激2小时后IkappaBα逐渐重新出现在细胞质中。PDTC（一种抗氧化剂）可有效抑制烧伤血清诱导的单核细胞IkappaBα降解和NF-κB核转位。
烧伤血清可诱导单核细胞中NF-κB的p50和p65成分核转位至细胞核并导致IkappaBα降解，最终导致PBMCs分泌细胞因子。
