Lipopolysaccharide-induced protein kinase D activation mediated by interleukin-1beta and protein kinase C.
Protein kinase D (PKD), a newly described serine/threonine kinase, has been implicated in many signal transduction pathways. The present study was designed to determine whether and how PKD is activated in inflammation. The results demonstrated that lipopolysaccharide (LPS, 30 microg/ml) stimulated PKD and protein kinase C (PKC) phosphorylation in spinal neurons within 0.5 h, and the activation reached a maximum at 3 or 8 h and declined at 12 h. The phosphorylation could be inhibited by the selective inhibitors for PKC (100 nM), mainly for PKCalpha and PKCbeta, suggesting the involvement of the PKC pathway. Particularly, PKCalpha might be critical for LPS-induced PKD activation since the PKCbeta inhibitor (100 nM) observed no effect on the phosphorylation of PKD. Furthermore, the expression of interleukin-1beta (IL-1beta) was significantly induced by LPS within 0.5 h, and reached a maximum at 8 h. IL-1 receptor antagonist inhibited PKD and PKCs activation induced by LPS at a concentration of 50 nM and achieved maximum at 1000 nM. These results demonstrated for the first time that PKD could be activated by LPS in spinal neurons, might via the IL-1beta/PKCalpha pathway. Additionally, immunostaining showed an increase in number of phosphorylated PKD-immunoreactive cells of adult spinal dorsal horn induced by intraplantar injected carrageenan (2 microg/100 microl), and antisense oligodeoxynucleotide to IL-1 receptor type I (50 microg/10 microl, intrathecal injected) inhibited the PKD activation, suggesting an involvement of IL-1beta/PKD pathway in inflammation in adult spinal cord.
Song
Ming-Juan
MJ
Department of Integrative Medicine and Neurobiology, Institute of Acupuncture Research, Shanghai Medical College, Fudan University, Shanghai 200032, China.
Wang
Yan-Qing
YQ
Wu
Gen-Cheng
GC
D000818:Animals; D002351:Carrageenan / Q000494:pharmacology; D002478:Cells, Cultured; D004789:Enzyme Activation / Q000187:drug effects / Q000276:immunology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D018836:Inflammation Mediators / Q000494:pharmacology; D053590:Interleukin 1 Receptor Antagonist Protein / Q000378:metabolism / Q000494:pharmacology; D053583:Interleukin-1beta / Q000187:drug effects / Q000276:immunology / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000494:pharmacology; D008297:Male; D009187:Myelitis / Q000139:chemically induced / Q000201:enzymology* / Q000276:immunology; D009474:Neurons / Q000187:drug effects / Q000201:enzymology / Q000276:immunology; D016376:Oligonucleotides, Antisense / Q000494:pharmacology; D010766:Phosphorylation / Q000187:drug effects; D011493:Protein Kinase C / Q000187:drug effects / Q000276:immunology / Q000378:metabolism*; D051571:Protein Kinase C-alpha / Q000187:drug effects / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D017472:Receptors, Interleukin-1 / Q000187:drug effects / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D013116:Spinal Cord / Q000187:drug effects / Q000201:enzymology* / Q000276:immunology; D015854:Up-Regulation / Q000187:drug effects / Q000276:immunology
D004791:Enzyme Inhibitors; D018836:Inflammation Mediators; D053590:Interleukin 1 Receptor Antagonist Protein; D053583:Interleukin-1beta; D008070:Lipopolysaccharides; D016376:Oligonucleotides, Antisense; D017472:Receptors, Interleukin-1; D002351:Carrageenan; C089212:protein kinase D; D011493:Protein Kinase C; D051571:Protein Kinase C-alpha