Protein kinase C-δ is involved in the inflammatory effect of IL-6 in mouse adipose cells

• DOI: 10.1007/s00125-010-1668-1
• Publication Date: 2010-02-11T13:25:03Z
• Authors: E. Wallerstedt; U. Smith; C. X. Andersson
• Keywords: Inflammation; STAT3 Transcription Factor; Transcriptional Activation; 0301 basic medicine; Serum Amyloid A Protein; Interleukin-6; Reverse Transcriptase Polymerase Chain Reaction; Blotting, Western; Acetophenones; Gene Expression; Statistics, Nonparametric; Cell Line; Mice; Protein Kinase C-delta; 03 medical and health sciences; Adipocytes; Animals; Benzopyrans; Phosphorylation; Cells, Cultured; Signal Transduction

The aim of the study was to address the role of protein kinase C-delta (PKCdelta) on phosphorylation of signal transducer and activator of transcription 3 (STAT3) and activation of inflammatory genes in response to IL-6 in adipose cells.Differentiated mouse 3T3-L1 adipocytes preincubated with the PKCdelta inhibitor rottlerin and mouse embryonic fibroblasts (MEFs) lacking PKCdelta were incubated with IL-6 and/or insulin. RNA was extracted and the gene expression was analysed by real-time PCR, while the proteins from total, nuclear and cytoplasmic lysates were analysed by immunoblotting.Inhibition of PKCdelta by rottlerin significantly reduced both Ser-727 and Tyr-705 phosphorylation of STAT3. Consequently, nuclear translocation of STAT3 and the IL-6-induced gene transcription and protein release of the inflammatory molecule serum amyloid A 3 (SAA3) were reduced. Similarly, the IL-6-regulated gene transcription of Il-6 (also known as Il6) to Hp and the feedback inhibitor of IL-6, Socs3, were also attenuated by rottlerin. Furthermore, PKCdelta was found to translocate to the nucleus following IL-6 treatment and this was also reduced by rottlerin. In agreement with the effect of rottlerin, Pkcdelta (also known as Prkcd) ( -/- ) MEFs also displayed a markedly reduced ability of IL-6 to activate the transcription of Saa3, Hp, Socs3 and Il6 genes compared with wild-type MEFs. These results correlated with a reduced nuclear translocation and phosphorylation of STAT3.These results show that PKCdelta plays a key role in the inflammatory effect of IL-6 in adipose cells and may be a suitable target for novel anti-inflammatory agents.