ABSTRACT
Matrix metalloproteinase-9 (MMP-9) secreted from macrophages plays an important role in tissue destruction and inflammation through degradation of matrix proteins and proteolytic activation of cytokines/chemokines. Whereas the MEK-ERK and PI3K-Akt pathways up-regulate MMP-9 expression, regulation of MMP-9 by JNK remains controversial. Presently, we aimed to determine the role of JNK in MMP-9 regulation in Raw 264.7 cells. Inhibition of JNK by the JNK inhibitor SP600125 induced MMP-9 in the absence of serum and suppressed the expression of TNF-alpha, IL-6 and cyclooxygenase-2 in LPS-treated Raw 264.7 cells. In a knockdown experiment with small interfering RNA, suppression of JNK1 induced MMP-9 expression. Interestingly, mouse serum suppressed SP600125-mediated MMP-9 induction, similar to IFN-gamma. However, the inhibitory activity of mouse serum was not affected by pyridone 6, which inhibits Janus kinase downstream to IFN-gamma. In addition to mouse serum, conditioned media of Raw 264.7 cells contained the inhibitory factor(s) larger than 10 kDa, which suppressed SP600125- or LPS-induced MMP-9 expression. Taken together, these data suggest that JNK1 suppresses MMP-9 expression in the absence of serum. In addition, the inhibitory factor(s) present in serum or secreted from macrophages may negatively control MMP-9 expression.
Subject(s)
Animals , Mice , Anthracenes/metabolism , Cell Line , Culture Media, Conditioned/chemistry , Enzyme Activation , Enzyme Induction , Enzyme Inhibitors/metabolism , Extracellular Signal-Regulated MAP Kinases/genetics , Gene Expression Regulation, Enzymologic , MAP Kinase Signaling System/physiology , Macrophages/cytology , Matrix Metalloproteinase 9/genetics , Mitogen-Activated Protein Kinase 8/genetics , NF-kappa B/genetics , Proto-Oncogene Proteins c-akt/genetics , Tumor Necrosis Factor-alpha/metabolism , p38 Mitogen-Activated Protein Kinases/geneticsABSTRACT
Using spectrophotometric and spectrofluorometric techniques, the interaction of iodine and 2-anthracene sulfonate (ANS) with the phospholipids (PL) isolated from four genetically correlated Salmonella minnesota isolates viz., a smooth form (S), a deeply rough mutant (Rc) and two intermediate forms (Ra and Rb) were studied. Appearance of an isosbestic point and a new band in absorption spectra indicated charge-transfer (C-T) interaction of iodine with the PL through the formation of 1:1 complex. Stern-Volmer type fluorescence quenching of PL was observed with the addition of iodine to PL, while PL enhanced the fluorescence of anionic dye ANS. The values of the binding constants between iodine/PL and ANS/PL, measured by using suitable equations, showed a systematic gradation in the molecular properties of the PL in the membrane structure in smooth (S) and rough (Ra, Rb and Rc) mutants of Salmonella minnesota.