l-663536 and 20-hydroxy-5-8-11-14-eicosatetraenoic-acid

Diabetic nephropathy (DN) is one of the most severe complications of diabetes mellitus. The pathomolecular events behind DN remain uncertain. Peroxisome proliferator-activated receptors (PPARs) play essential functions in the development of DN. Meanwhile, 20-hydroxyeicosatetraenoic acid (20-HETE) also plays central roles in the regulation of renal function. However, the relationship between PPARs and 20-HETE is rarely studied in DN. It was revealed in our study that both PPARs expression and CYP4A-20-HETE level were decreased under DN conditions in vivo and in vitro. Supplementation with bezafibrate, a PPAR pan-agonist, improved the damage of kidney in DN mice and in high glucose-induced NRK-52E cells, following the up-regulation of PPARs and the increase of CYP4A-20-HETE. PPARα antagonist (MK886), PPARβ antagonist (GSK0660), and PPARγ antagonist (GW9662) reversed the protection of bezafibrate in NRK-52E, and abrogated the up-regulation of CYP4A-20-HETE produced by bezafibrate. Noteworthily, 20-HETE synthetase inhibitor, HET0016, also blocked the bezafibrate-mediated improvement of NRK-52E, and abolished the up-regulation of PPARs expression. Collectively, our data suggest that the concurrent down-regulation and interaction of PPARs and 20-HETE play crucial roles in the pathogenesis process of DN, and we provide a novel evidence that PPARs/20-HETE signaling may be served as a therapeutic target for DN patients.

Topics: Amidines; Anilides; Animals; Cell Line; Cytochrome P-450 CYP4A; Diabetic Nephropathies; Down-Regulation; Epithelial Cells; Gene Expression Regulation; Glucose; Hydroxyeicosatetraenoic Acids; Indoles; Kidney Tubules; Male; Mice; PPAR alpha; PPAR gamma; PPAR-beta; Rats; Sulfones; Thiophenes

20-Hydroxyeicosatetraenoic acid (20-HETE), formed from arachidonate by cytochrome P450, regulates vascular smooth muscle cell (VSMC) function. Because 20-HETE may activate peroxisome proliferator activator receptors (PPARs) and may participate in inflammatory responses, we asked whether 20-HETE may inhibit cyclooxygenase 2 (COX-2) expression by activating PPARs in VSMC.. Quiescent neonatal VSMC (R22D cell line), were incubated with 20-HETE, synthetic ligands of PPARs, or inhibitors of the extracellular signal regulated kinase (ERK1/2), c-jun N-terminal kinase and the transcription factor activated protein-1 before adding ATPγS. mRNA and protein expression of COX-2 and the promoter luciferase activity of COX-2 and PPAR response element were determined.. Pretreatment with 20-HETE (5-10 µM) significantly inhibited ATPγS-induced COX-2 mRNA and protein expression in VSMC. The inhibitory effect of 20-HETE on COX-2 expression was mimicked by WY14643, a PPARα ligand and inhibited by MK886, a PPARα inhibitor or by transfection of shRNA for PPARα. Both 20-HETE and WY14643 significantly increased the PPAR-response element luciferase activity. Furthermore, ATPγS-induced activation of the COX-2 promoter containing the activated protein-1 site was also inhibited by pretreatment with 20-HETE, which was reversed by MK886 or by transfection with shRNA for PPARα.. The PPARα may mediate the inhibitory effects of 20-HETE on COX-2 expression through a negative cross-talk between PPARα and the COX-2 promoter.

Topics: Adenosine Triphosphate; Animals; Cell Line; Cyclooxygenase 2; Hydroxyeicosatetraenoic Acids; Indoles; Luciferases; Mitogen-Activated Protein Kinase Kinases; Muscle, Smooth, Vascular; PPAR alpha; Promoter Regions, Genetic; Pyrimidines; Rats; RNA, Small Interfering; Transcription Factor AP-1