• Postgraduate Institute of Southern Medical University. Guangzhou, Guangdong, 510515, ChinaCorresponding Author: LI-li, E-mail: lili_China@ 163. com;
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Objective  To establish a cell culture model in vitro of acute lung injury and investigate the effects of NF-κB p65 on the inflammation and oxidative stress in TNF-α-activated type Ⅱ alveolar epithelial cells.
Methods  A549 cells were treated with TNF-α ( 10 ng/mL, 24 h) in the absence or presence of NF-κB p65 siRNA ( 50 nmol /L) . RT-PCR and Western blot were performed to analyze the silence efficiency of RNAi targeting NF-κB p65. The contents of IL-1β, IL-4, and IL-6 in the culture supernatant were measured by ELISA. The concentration of MDA and SOD were detected by colorimetric method. The survival rate of cell was assessed by the methyl thiazolyl tetrazolium ( MTT) assay.
Results  P65 RNAi significantly decreased the transcription and translation of NF-κB p65 induced by TNF-α( P  lt; 0. 05) . The levels of IL-1β, IL-4, and IL-6 were significantly lower in the supernatants of A549 cells pretransfected with NF-κB p65 siRNA ( P  lt;0. 05) , while the concentration of MDA markedly decreased ( P  lt; 0. 05) , and the activation of SOD increased dramatically ( P  lt; 0. 05) . Consequently, the survival rate of A549 in the p65 siRNA group improved( P  lt; 0. 05) .
Conclusions  NF-κB p65 plays a key role in the oxidative stress induced by TNF-α. NF-κB p65 silencing can down-regulate the inflammation and oxidative stress induced by TNF-αand enhance the proliferation of alveolar epithelial cells.

Citation: WU Weijing ,LI Li,YUAN Weifeng,LI Weifeng,HUANG Wenjie. The Role of NF-κB p65 in Oxidative Stress Induced by TNF-αin Type Ⅱ Alveolar Epithelial Cells. Chinese Journal of Respiratory and Critical Care Medicine, 2012, 11(1): 46-51. doi: Copy

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