Chen F, Lu Y, Kuhn D C, Maki M, Shi X, Sun S C, Demers L M
Department of Pathology, Pennsylvania State University Collage of Medicine, Milton S. Hershey Medical Center, Hershey 17033, USA.
Arch Biochem Biophys. 1997 Jun 15;342(2):383-8. doi: 10.1006/abbi.1997.0132.
Both silica and lipopolysaccharide (LPS) induce a rapid degradation of I kappa B alpha, an intracellular inhibitor of the nuclear factor (NF)-kappa B transcription factor. In this report, we demonstrate that MG132, a relatively specific proteasome inhibitor, is capable of suppressing LPS-induced I kappa B alpha degradation and NF-kappa B activation in mouse macrophage line RAW 264.7 cells, but is unable to influence the same induction produced by silica. In contrast, the lysosome inhibitor chloroquine has little effect on I kappa B alpha degradation induced by either silica or LPS. In fact, chloroquine enhances the signal-induced nuclear expression of NF-kappa B p50/p65 heterodimer by inhibiting the resynthesis of I kappa B alpha. With the use of transient transfection of a plasmid that expresses calpastatin, a natural inhibitor for calpain, the silica-induced degradation of I kappa B alpha and NF-kappa B activation was attenuated. In contrast, no inhibition of LPS-induced I kappa B alpha degradation and NF-kappa B activation was observed by the overexpression of calpastatin. This suggests that calpain contributes to silica-induced I kappa B alpha degradation and NF-kappa B activation but not to LPS-induced I kappa B alpha degradation and NF-kappa B activation.
二氧化硅和脂多糖(LPS)均可诱导核因子(NF)-κB转录因子的细胞内抑制剂IκBα迅速降解。在本报告中,我们证明,相对特异性的蛋白酶体抑制剂MG132能够抑制LPS诱导的小鼠巨噬细胞系RAW 264.7细胞中IκBα的降解和NF-κB的激活,但无法影响二氧化硅产生的相同诱导作用。相反,溶酶体抑制剂氯喹对二氧化硅或LPS诱导的IκBα降解几乎没有影响。实际上,氯喹通过抑制IκBα的再合成增强了信号诱导的NF-κB p50/p65异二聚体的核表达。通过瞬时转染表达钙蛋白酶抑制蛋白(一种钙蛋白酶的天然抑制剂)的质粒,二氧化硅诱导的IκBα降解和NF-κB激活减弱。相反,钙蛋白酶抑制蛋白的过表达未观察到对LPS诱导的IκBα降解和NF-κB激活的抑制作用。这表明钙蛋白酶促成了二氧化硅诱导的IκBα降解和NF-κB激活,但对LPS诱导的IκBα降解和NF-κB激活没有作用。