Essential Role of NF-KB Activation in Silica-Induced Inflammatory Mediator Production in Macrophages

"In this report, we demonstrate that NF-KB, a ubiquitous transcription factor, plays an essential role in silica-induced inflammatory mediator production in the mouse macrophage cell line RAW 264.7. Compared to the effect of lipopolysaccharide (LPS), silica mediated a stronger activation of NF-KB p50/p50 homodimer at early phase of poststimulation. Furthermore, activation of NF-xB by silica and LPS appears to be mediated by different signal transduction pathways. Both silica and LPS increased mRNA expression in these cells for cyclooxygenase II, inducible nitric oxide synthase, tumor necrosis factor-a and interleukin- I a. This expression was attenuated along with the inhibition of NF-KB activation. It is well documented that exposure of alveolar macrophages to silica can initiate excessive production of inflammatory mediators and cytokines from several early response genes (1-3). At the DNA level, a common structural characteristic of these genes is the presence of one to five NF-lCB binding sequences (KB) in the 5'-flanking region (4-6). In resting cells, NF-KB is retained in the cytoplasm in an inactive form by binding to inhibitory protein known as hcBcx, hcB~. p 105 (precursor of p50) and p 100 (precursor of p52) (7-9). Upon cellular stimulation, these inhibitory proteins are proteolytically degraded or processed by certain proteases, which allows the liberated NF-KB to translocate into the nucleus as an activated form to initiate or regulate early response gene transcription by binding to its consensus element (GGGRNNYYCC)(7).In this report, we demonstrate that silica can efficiently induce NF-lCB activation in RAW 264.7 cells, and that silica displays different characteristic from LPS in the activation of NF-KB, both with respect to NF-KB subtype composition and the signal transduction pathway that leads to the activation of NF-KB. In addition, abrogation of NF-KB activation by inhibiting the protease can dramatically reduce the mRNA expression of cyclooxygenase II, inducible nitric oxide synthase, tumor necrosis factor-ex (TNF-o.) and interleukin-lex (IL-lex). These findings strongly suggest that NF-KB plays an essential role in silica-induced inflammatory mediator production in the monocyte/ macrophage cells."