Kiss K, Kiss J, Rudolf E, Cervinka M, Szeberényi J
Department of Medical Biology, Faculty of Medicine, University of Pécs, Szigeti u. 12., H-7624 Pécs, Hungary.
J Biochem Biophys Methods. 2004 Oct 29;61(1-2):229-40. doi: 10.1016/j.jbbm.2004.06.003.
Sodium salicylate (NaSal) is an effective analgetic and antiinflammatory drug. Beside its well-known inhibitory effect on the cyclooxigenase enzymes, it influences the activity of other signal transduction proteins including nuclear factor kappa B (NF-kappaB) transcription factor. NF-kappaB is found in the cytoplasm bound to an inhibitory protein, inhibitory kappa B (IkappaB). After its phosphorylation, IkappaB is degraded and the released NF-kappaB translocates into the nucleus. Sodium salicylate inhibits the degradation of IkappaB, thus, NF-kappaB activation cannot occur. According to previous observations, the inhibition of this activation can lead to apoptosis. The main goals of this study were to demonstrate that inhibition of NF-kappaB by sodium salicylate decreases the viability of rat phaeochromocytoma PC12 cells and to investigate the nature of cell damage and death. PC12 cells were treated with different concentrations of sodium salicylate (1-20 mM). Higher concentrations (10-20 mM) killed PC12 cells in a dose-dependent manner. The assessments were done by direct cell counting in a Burker chamber and by the WST-1 cytotoxicity assay. We also found a decreased NF-kappaB activity after sodium salicylate treatment by electrophoretic mobility shift assay (EMSA). The cells treated with sodium salicylate were undergoing apoptosis as seen on our records obtained by time-lapse videomicroscopy as well as shown by DNA fragmentation experiments. The decreased DNA binding activity of NF-kappaB indicates that the inhibition of NF-kappaB can play a role in these processes.
水杨酸钠（NaSal）是一种有效的镇痛和抗炎药物。除了其对环氧化酶众所周知的抑制作用外，它还影响包括核因子κB（NF-κB）转录因子在内的其他信号转导蛋白的活性。NF-κB存在于细胞质中，与一种抑制蛋白，即抑制性κB（IkappaB）结合。磷酸化后，IkappaB降解，释放的NF-κB转移到细胞核中。水杨酸钠抑制IkappaB的降解，因此，NF-κB的激活无法发生。根据先前的观察，这种激活的抑制可导致细胞凋亡。本研究的主要目的是证明水杨酸钠对NF-κB的抑制会降低大鼠嗜铬细胞瘤PC12细胞的活力，并研究细胞损伤和死亡的性质。用不同浓度的水杨酸钠（1-20 mM）处理PC12细胞。较高浓度（10-20 mM）以剂量依赖性方式杀死PC12细胞。通过在Burker计数板中直接细胞计数和WST-1细胞毒性测定进行评估。我们还通过电泳迁移率变动分析（EMSA）发现水杨酸钠处理后NF-κB活性降低。如我们通过延时视频显微镜获得的记录以及DNA片段化实验所示，用水杨酸钠处理的细胞正在发生凋亡。NF-κB的DNA结合活性降低表明NF-κB的抑制可能在这些过程中起作用。
