Cuzzocrea Salvatore, Pisano Barbara, Dugo Laura, Ianaro Angela, Ndengele Michael, Salvemini Daniela
Department of Clinical and Experimental Medicine and Pharmacology, Torre Biologica, Policlinico Universitario, 98123 Messina, Italy.
Antioxid Redox Signal. 2004 Aug;6(4):699-704. doi: 10.1089/1523086041361659.
The nuclear factor-kappaB (NF-kappaB) is a transcription factor that plays a pivotal role in the induction of genes involved in physiological processes, as well as in the response to inflammation. In this study, we used a selective nonpeptidyl superoxide dismutase mimetic, M40403, to investigate the role of superoxide anion in NF-kappaB activation during acute inflammation in mice. Injection of carrageenan into the pleural cavity of mice induced an acute inflammatory response characterized by fluid accumulation in the pleural cavity that contained a large number of neutrophils, as well as an increased production of tumor necrosis factor-alpha and interleukin-1beta. All parameters of inflammation were attenuated by M40403 (10 mg/kg i. p., 30 min prior to carrageenan administration). These inflammatory events were associated with the activation of NF-kappaB in the lung. In particular, the appearance of inhibitory protein kappaB-alpha (IkappaB-alpha) in homogenates of lung tissues was investigated by immunoblot analysis at 4 h after carrageenan administration. IkappaB-alpha levels were substantially reduced in the lung tissue from carrageenan-treated mice in comparison with sham-treated mice. Furthermore, to detect NF-kappaB/DNA binding activity, whole extracts from lung tissue of each mouse were analyzed by electrophoretic mobility-shift assay. The DNA binding activity significantly increased in whole extracts obtained from lung tissues of vehicle-treated mice 4 h after carrageenan administration. Treatment of mice with M40403 caused a significant inhibition of carrageenan-induced IkappaB-alpha degradation and NF-kappaB/DNA binding activity. These data confirm that M40403 exerts a potent antiinflammatory activity and clearly demonstrate that the reduction of the inflammatory process is associated with modification of the activation of signal transduction pathways.
核因子-κB（NF-κB）是一种转录因子，在参与生理过程的基因诱导以及炎症反应中起关键作用。在本研究中，我们使用了一种选择性非肽类超氧化物歧化酶模拟物M40403，来研究超氧阴离子在小鼠急性炎症期间NF-κB激活中的作用。向小鼠胸腔内注射角叉菜胶可诱导急性炎症反应，其特征为胸腔内液体蓄积，其中含有大量中性粒细胞，同时肿瘤坏死因子-α和白细胞介素-1β的产生增加。M40403（10 mg/kg腹腔注射，在给予角叉菜胶前30分钟）可减轻所有炎症参数。这些炎症事件与肺中NF-κB的激活有关。特别是，在给予角叉菜胶后4小时，通过免疫印迹分析研究了肺组织匀浆中抑制蛋白κB-α（IkappaB-α）的出现情况。与假处理小鼠相比，角叉菜胶处理小鼠的肺组织中IkappaB-α水平显著降低。此外，为了检测NF-κB/DNA结合活性，通过电泳迁移率变动分析对每只小鼠肺组织的全提取物进行了分析。在给予角叉菜胶后4小时，从载体处理小鼠的肺组织获得的全提取物中，DNA结合活性显著增加。用M40403处理小鼠可显著抑制角叉菜胶诱导的IkappaB-α降解和NF-κB/DNA结合活性。这些数据证实M40403具有强大的抗炎活性，并清楚地表明炎症过程的减轻与信号转导途径激活的改变有关。
