Suppression by cyclosporin A of interleukin 1 beta-induced expression of group II phospholipase A2 in rat renal mesangial cells.
1. We investigated whether cyclosporin A, a potent immunosuppressive drug, affects group II phospholipase A2. (PLA2; EC 3.1.1.4) induction in rat renal mesangial cells. 2. Previously we showed that the expression of group II PLA2 in rat renal mesangial cells is triggered by exposure of the cells to inflammatory cytokines such as interleukin 1 beta (IL-1 beta) or tumour necrosis factor alpha and agents that elevate cellular levels of cyclic AMP. Treatment of mesangial cells with IL-1 beta for 24 h induced PLA2 activity secreted into cell culture supernatants by about 16 fold. Incubation of mesangial cells with cyclosporin A inhibited IL-1 beta-induced PLA2 section in a dose-dependent fashion, with an IC50 value of 4.3 microM. Cyclosporin A did not directly inhibit enzymatic activity of PLA2. 3. Immunoprecipitation of radioactively labelled PLA2 protein from mesangial cell supernatants revealed that the inhibition of PLA2 activity is due to a suppression of PLA2 protein levels. This effect was preceded by a reduction of PLA2 mRNA steady state levels, as demonstrated by Northern blot analyses of total cellular RNA isolated from stimulated mesangial cells. 4. In order to evaluate whether cyclosporin A would affect the transcriptional activity of the PLA2 gene, we performed nuclear run on transcription experiments and provided evidence that the transcription rate of the PLA2 gene is reduced by cyclosporin A. 5. Previously we found that the nuclear transcription factor kappa B (NF kappa B) is an essential component of the IL-1 beta-dependent upregulation of PLA2 gene transcription. By electrophoretic mobility shift analysis, we demonstrated that cyclosporin A diminishes the formation of NF kappa B DNA-binding complexes, thus suggesting that this transcription factor is a target for cyclosporin A-mediated repression of PLA2 gene transcription. 6. The data presented in this study strongly suggest that the cellular mechanism involved in the IL1 beta-dependent transcriptional upregulation of the PLA2 gene in mesangial cells is a target for the action of cyclosporin A.
Walker
G
G
Department of Pharmacology, Biozentrum, University of Basel, Switzerland.
Kunz
D
D
Pignat
W
W
van den Bosch
H
H
Pfeilschifter
J
J
D000818:Animals; D016572:Cyclosporine / Q000494:pharmacology*; D004790:Enzyme Induction / Q000187:drug effects; D005920:Glomerular Mesangium / Q000187:drug effects*; D007165:Immunosuppression Therapy; D007166:Immunosuppressive Agents / Q000494:pharmacology*; D007375:Interleukin-1 / Q000235:genetics / Q000378:metabolism; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism*; D010741:Phospholipases A / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D054467:Phospholipases A2; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats
D007166:Immunosuppressive Agents; D007375:Interleukin-1; D016328:NF-kappa B; D012333:RNA, Messenger; D016572:Cyclosporine; D010741:Phospholipases A; D054467:Phospholipases A2