gw-7845 and ciglitazone

Peroxisome proliferator-activated receptor gamma (PPARgamma) possesses general beneficial effects on the cardiovascular system, such as inhibition of vascular lesion formation and atherosclerosis. However, molecular mechanisms for these effects are yet to be fully defined. The aim of this study is to elucidate whether interferon regulatory factor-1 (IRF-1), a transcriptional factor with anti-proliferative and pro-apoptotic properties, mediates PPARgamma-induced apoptosis in vascular smooth muscle cells (VSMCs).. Using Northern and Western blot analyses, we documented that PPARgamma ligands, including ciglitazone, troglitazone, and GW7845, significantly increased IRF-1 expression in VSMCs; however, the PPARalpha ligand (Wy14643) and PPARdelta ligand (GW0742) did not affect its expression. PPARgamma-induced IRF-1 expression was abrogated by pretreatment with the PPARgamma antagonist GW9662. In contrast, adenoviral expression of PPARgamma in VSMCs dramatically increased IRF-1 level. Furthermore, PPARgamma activation increased IRF-1 promoter activity but did not affect IRF-1 mRNA stability. Finally, reducing IRF-1 expression by antisense technology attenuated PPARgamma-induced VSMC apoptosis through decreasing cyclin-dependent kinase inhibitor p21(cip1) and caspase-3 activity.. Our data demonstrate that IRF-1 is a novel PPARgamma target gene and mediates PPARgamma-induced VSMC apoptosis.

Topics: Adenoviridae; Animals; Aorta; Apoptosis; Cells, Cultured; Chlorocebus aethiops; Chromans; DNA-Binding Proteins; Dose-Response Relationship, Drug; Gene Expression Regulation; Humans; Interferon Regulatory Factor-1; Ligands; Muscle, Smooth, Vascular; Oxazoles; Phosphoproteins; Promoter Regions, Genetic; Receptors, Cytoplasmic and Nuclear; RNA Stability; RNA, Messenger; Thiazolidinediones; Time Factors; Transcription Factors; Troglitazone; Tyrosine; Up-Regulation

Inhibitor of DNA binding (Id2) is a member of the helix-loop-helix family of transcription regulators that is known to play important roles in the proliferation and differentiation of many cell types. Overexpression of Id2 has been reported to result in significant enhancement of vascular smooth muscle cell growth via increased S phase entry. We hypothesized that downregulation of Id2 gene expression by thiazolidinediones (TZDs), a class of anti-diabetic drugs and peroxisome proliferator-activated receptor gamma (PPARgamma) activators, might contribute to the anti-atherosclerotic and anti-hypertensive effects of the PPARgamma. Here we document that TZDs, including troglitazone and ciglitazone, repress Id2 gene expression in a doses- and time-dependent manner. However, GW7845, a high-affinity and non-TZD PPARgamma activator, had no inhibitory effect on Id2 gene expression. In addition, PPARgamma antagonist GW9662 did not rescue TZD-induced Id2 repression. Taken together, our data suggest that TZDs repress Id2 expression through a PPARgamma-independent pathway.

Topics: Anilides; Aorta; Chromans; DNA-Binding Proteins; Dose-Response Relationship, Drug; Down-Regulation; Gene Expression Regulation; Humans; Hypoglycemic Agents; Inhibitor of Differentiation Protein 2; Muscle, Smooth, Vascular; Oxazoles; Receptors, Cytoplasmic and Nuclear; Repressor Proteins; RNA, Messenger; Thiazoles; Thiazolidinediones; Time Factors; Transcription Factors; Troglitazone; Tyrosine

The role of peroxisome proliferator-activated receptor gamma (PPARgamma) in adipocyte physiology has been exploited for the treatment of diabetes. The expression of PPARgamma in lymphoid organs and its modulation of macrophage inflammatory responses, T cell proliferation and cytokine production, and B cell proliferation also implicate it in immune regulation. Despite significant human exposure to PPARgamma agonists, little is known about the consequences of PPARgamma activation in the developing immune system. Here, well-characterized models of B lymphopoiesis were used to investigate the effects of PPARgamma ligands on nontransformed pro/pre-B (BU-11) and transformed immature B (WEHI-231) cell development. Treatment of BU-11, WEHI-231, or primary bone marrow B cells with PPARgamma agonists (ciglitazone and GW347845X) resulted in rapid apoptosis. A role for PPARgamma and its dimerization partner, retinoid X receptor (RXR)alpha, in death signaling was supported by 1) the expression of RXRalpha mRNA and cytosolic PPARgamma protein, 2) agonist-induced binding of PPARgamma to a PPRE, and 3) synergistic increases in apoptosis following cotreatment with PPARgamma agonists and 9-cis-retinoic acid, an RXRalpha agonist. PPARgamma agonists activated NF-kappaB (p50, Rel A, c-Rel) binding to the upstream kappaB regulatory element site of c-myc. Only doses of agonists that induced apoptosis stimulated NF-kappaB-DNA binding. Cotreatment with 9-cis-retinoic acid and PPARgamma agonists decreased the dose required to activate NF-kappaB. These data suggest that activation of PPARgamma-RXR initiates a potent apoptotic signaling cascade in B cells, potentially through NF-kappaB activation. These results have implications for the nominal role of the PPARgamma in B cell development and for the use of PPARgamma agonists as immunomodulatory therapeutics.

Topics: Animals; Apoptosis; B-Lymphocytes; Bone Marrow Cells; Cell Line; Cell Line, Transformed; Male; Mice; Mice, Inbred C57BL; NF-kappa B; Oxazoles; Peroxisome Proliferators; Peroxisomes; Receptors, Cytoplasmic and Nuclear; Receptors, Retinoic Acid; Retinoid X Receptors; Signal Transduction; Stem Cells; Thiazoles; Thiazolidinediones; Transcription Factors; Tyrosine