Effect of PPARγ overexpression on the inhibition of IL-1β-induced responses by 15d-PGJ

<p><b>Copyright information:</b></p><p>Taken from "Contrasting effects of peroxisome-proliferator-activated receptor (PPAR)γ agonists on membrane-associated prostaglandin Esynthase-1 in IL-1β-stimulated rat chondrocytes: evidence for PPARγ-independent inhibition by 15-deoxy-Δprostaglandin J"</p><p>Arthritis Research & Therapy 2005;7(6):R1325-R1337.</p><p>Published online 22 Sep 2005</p><p>PMCID:PMC1297580.</p><p>Copyright © 2005 Bianchi et al.; licensee BioMed Central Ltd.</p> Chondrocytes in six-well plates were transfected with pcDNA3.1 peroxisome-proliferator-activated receptor γ (PPARγ) construct (500 ng) for 36 hours. Thereafter, cells were pretreated for 4 hours with 10 μM 15-deoxy-Δprostaglandin J(15d-PGJ), then stimulated with 10 ng/ml IL-1β for 24 hours before extraction of total RNA and collection of culture supernatant. PGElevels assayed by ELISA in culture supernatant; relative abundance of microsomal prostaglandin E synthase-1 (mPGES-1) mRNAs analysed by real-time PCR and normalized to S29 mRNA; western blot control experiment of PPARγ and β-actin expression; modulation of adiponectin (a PPARγ target gene) mRNAs by PPARγ agonists and pcDNA3.1 PPARγ construct, analysed by real-time PCR and normalized to S29 mRNA. Results are expressed as means ± SD for at least three independent experiments. Statistically significant differences (< 0.05): *, comparison with non-stimulated controls; , comparison with IL-1β-stimulated cells; , comparison with PPARγ agonists alone or in combination with PPARγ plasmid

Effect of PPARγ agonists on IL-1β-induced prostaglandins levels, COX-2 and mPGES-1 mRNAs

<p><b>Copyright information:</b></p><p>Taken from "Contrasting effects of peroxisome-proliferator-activated receptor (PPAR)γ agonists on membrane-associated prostaglandin Esynthase-1 in IL-1β-stimulated rat chondrocytes: evidence for PPARγ-independent inhibition by 15-deoxy-Δprostaglandin J"</p><p>Arthritis Research & Therapy 2005;7(6):R1325-R1337.</p><p>Published online 22 Sep 2005</p><p>PMCID:PMC1297580.</p><p>Copyright © 2005 Bianchi et al.; licensee BioMed Central Ltd.</p> After 4 hours of pretreatment with 10 μM 15-deoxy-Δprostaglandin J(15d-PGJ) or rosiglitazone, chondrocytes were incubated with 10 ng/ml IL-1β for 12 or 24 hours. PGEand 6-keto-PGFlevels assayed by ELISA in culture supernatant; relative abundances of cyclo-oxygenase-2 (COX-2) and microsomal prostaglandin E synthase-1 (mPGES-1) mRNAs, analysed by real-time PCR and normalized to S29 mRNA COX-2 and mPGES-1 protein levels assessed by western blotting and normalized to β-actin level. Results are expressed as means ± SD for at least three independent experiments. Statistically significant differences (< 0.05): *, comparison with non-stimulated controls; , comparison with IL-1β-stimulated cells

Time course of prostaglandins production, COX-2 and mPGES-1 mRNA expression, in IL-1β-stimulated chondrocytes

<p><b>Copyright information:</b></p><p>Taken from "Contrasting effects of peroxisome-proliferator-activated receptor (PPAR)γ agonists on membrane-associated prostaglandin Esynthase-1 in IL-1β-stimulated rat chondrocytes: evidence for PPARγ-independent inhibition by 15-deoxy-Δprostaglandin J"</p><p>Arthritis Research & Therapy 2005;7(6):R1325-R1337.</p><p>Published online 22 Sep 2005</p><p>PMCID:PMC1297580.</p><p>Copyright © 2005 Bianchi et al.; licensee BioMed Central Ltd.</p> Rat cells were exposed to 10 ng/ml IL-1β for 6, 12, 24, 36 or 48 hours before total RNA extraction and collection of culture supernatant. Prostaglandin levels (PGE, 6-keto-PGF) assayed by ELISA in culture supernatant; relative abundances of cyclo-oxygenase-2 (COX-2) and microsomal prostaglandin E synthase-1 (mPGES-1) mRNAs, analysed by real-time PCR and normalized to S29 mRNA. Prostaglandin levels and PCR COX-2/S29 or mPGES-1/S29 mRNA ratios presented in histograms are expressed as means ± SD for at least three independent experiments. Statistically significant differences (< 0.05) from controls: * for PGEor COX-2; for 6-keto-PGFor mPGES-