Role of PKC in BSA-AGEs-mediated inducible nitric
oxide synthase expression in RAW 264.7 macrophages
林建煌
Wu CH;Chang CH;Lin HC;Chen CM;Lin CH and Lee HM
Abstract
In the present study, the roles of protein kinase C (PKC) in BSA-derived advanced glycosylation end products (BSA-AGEs)-induced nitric oxide (NO) production and inducible nitric oxide synthase (iNOS) expression were investigated. Treatment of RAW264.7 cells with BSA-AGEs caused dose- and time-dependent increases in NO release and iNOS expression in RAW264.7 cells, whereas BSA alone had no effect on iNOS induction. The tyrosine kinase inhibitor (genistein), the phosphatidylinositol-specific phospholipase C inhibitor (U-73122), the phosphatidylcholine-specific phospholipase C inhibitor (D-609), and the PKC inhibitors (staurosporine, Ro 31-8220, and Go 6976) all inhibited BSA-AGE-induced NO release and iNOS expression in RAW264.7 cells. Stimulation of RAW264.7 cells with BSAAGEs resulted in the formation of inositol monophosphate; the response was attenuated by U-73122 and genistein. BSA-AGEs stimulated PKC-a, -bI, -d, and -Z but not -z translocation from the cytosol to the membrane. However, incubation of RAW 264.7 cells with BSAAGEs increased phosphorylation of PKC-z at threonine-410, which reflects activation of PKC-z, indicating the possible involvement of these PKC isoforms in AGE-mediated effects. Pretreatment of RAW 264.7 cells with U-73122, D-609, and genistein reduced the AGEstimulated
translocation of PKC-a, -bI, -d, and -Z and activation of PKC-z. Taken together, these data suggest that BSA-AGEs might activate PKC and subsequently induce iNOS expression and NO release.
