Li Zhiqing, Huang Yuesheng, Yang Zongceng
Institute of Burn Research, Southwestern Hospital, The Third Military Medical University. Chongqing 400038, P.R. China.
Zhonghua Shao Shang Za Zhi. 2002 Oct;18(5):265-7.
To investigate the effects of burn sera on the nuclear translocation of endothelial NF-kappaB heterodimers p50/p65 and on the degradation of inhibiting kappaB (IkappaBalpha), in order to explore the role of burn sera on activation of the endothelium.
Cultured human umbilical vein endothelial cells (HUVECs) (ECV-304 strain) were employed as the target cells. The cells were stimulated by sera from healthy volunteers and from burn patients and burn sera together with PDTC (pyrrolidine dithiocarbarnate). The normal cultured cells were taken as the control. The nuclear translocation of endothelial p50/p65 at 30, 60, 120 and 480 mins after the stimulation was observed with laser confocal microscopy, and the endothelial IkappaBalpha protein degradation at 30, 60, 90 and 120 mins after the stimulation was determined by Western blotting.
When compared to that in control group, the nuclear translocation of p50/p65 took place 30 mins after the endothelial cells were stimulated by burn sera, and it reached the summit at 30 - 60 mins, but recovered to pre-stimulation state at 2hrs. In addition, IkBalpha degradation occurred 30 mins after the cells were stimulated by burn sera (P < 0.01) and peaking at 45 - 60 mins after the stimulation and recovered at 2hrs after the stimulation. The nuclear translocation of endothelial p50/p65 and IkBalpha degradation at 30 and 60 mins after the stimulation by burn sera could be effectively inhibited by PDTC.
Burn sera might induce the nuclear translocation of endothelial NF-kappaB p50/p65 and IkappaBalpha degradation and activate NF-kappaB, which ultimately lead to the secretion of cytokines from the endothelium.
研究烧伤血清对内皮细胞NF-κB异二聚体p50/p65核转位及抑制性κB（IkappaBα）降解的影响，以探讨烧伤血清在内皮细胞激活中的作用。
以培养的人脐静脉内皮细胞（HUVECs，ECV-304株）为靶细胞。用健康志愿者血清、烧伤患者血清及烧伤血清联合吡咯烷二硫代氨基甲酸盐（PDTC）刺激细胞，以正常培养的细胞作为对照。用激光共聚焦显微镜观察刺激后30、60、120和480分钟时内皮细胞p50/p65的核转位情况，并用蛋白质免疫印迹法检测刺激后30、60、90和120分钟时内皮细胞IkappaBα蛋白的降解情况。
与对照组相比，烧伤血清刺激内皮细胞30分钟后p50/p65发生核转位，30 - 60分钟时达到高峰，但2小时后恢复到刺激前状态。此外，烧伤血清刺激细胞30分钟后IkBα发生降解（P < 0.01），45 - 60分钟时达到峰值，刺激后2小时恢复。PDTC可有效抑制烧伤血清刺激后30和60分钟时内皮细胞p50/p65的核转位及IkBα的降解。
烧伤血清可能诱导内皮细胞NF-κB p50/p65核转位及IkappaBα降解，激活NF-κB，最终导致内皮细胞分泌细胞因子。
