15-deoxyprostaglandin-j2 and Arteriosclerosis

The Nrf2-Keap1 system coordinately regulates cytoprotective gene expression via the antioxidant responsive element (ARE). The expression of several ARE-regulated genes was found to be up-regulated in endothelial cells by laminar shear stress, suggesting that Nrf2 contributes to the anti-atherosclerosis response via the ARE. To gain further insight into the roles that Nrf2 plays in the development of atherosclerosis, we examined how Nrf2 regulates gene expression in response to anti-atherogenic laminar flow (L-flow) or pro-atherogenic oscillatory flow (O-flow). Exposure of human aortic endothelial cells (HAECs) to L-flow, but not to O-flow, induced the expression of cytoprotective genes, such as NAD(P)H quinone oxidoreductase 1 (NQO1) by 5-fold and heme oxygenase-1 by 8-fold. The critical contribution of Nrf2 to the expression induced by L-flow was ascertained in siRNA-mediated knock-down experiments. Two cyclooxygenase-2 (COX-2) specific inhibitors attenuated Nrf2 nuclear accumulation in the acute phase of L-flow exposure. A downstream product of COX-2, 15-deoxy-Delta(12,14)-prostaglandin J2 (15d-PGJ2), activated the Nrf2 regulatory pathway in HAECs through binding to the cysteines of Keap1. These results demonstrate that 15d-PGJ2 is essential for L-flow to activate Nrf2 and induce anti-atherosclerotic gene expression. Whereas both L-flow and O-flow induced the nuclear accumulation of Nrf2 to comparable levels, chromatin immunoprecipitation analysis revealed that Nrf2 binding to the NQO1 ARE was significantly diminished in the case of O-flow compared with that of L-flow. These results suggest that O-flow inhibits Nrf2 activity at the DNA binding step, thereby suppressing athero-protective gene expression and hence predisposing the blood vessels to the formation of atherosclerosis.

Topics: Aorta; Arteriosclerosis; Cell Nucleus; Cells, Cultured; DNA-Binding Proteins; Endothelium, Vascular; Gene Expression Regulation; Humans; Intracellular Signaling Peptides and Proteins; Kelch-Like ECH-Associated Protein 1; NAD(P)H Dehydrogenase (Quinone); NF-E2-Related Factor 2; Prostaglandin D2; Proteins; Response Elements; RNA, Small Interfering; Stress, Mechanical; Trans-Activators

Atherogenesis represents a type of chronic inflammation and involves elements of the immune response, eg, the expression of proinflammatory cytokines. In advanced atherosclerotic lesions, lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) is expressed in endothelial cells, macrophages, and smooth muscle cells (SMCs). In vitro, the expression of LOX-1 is induced by inflammatory cytokines like TNF-alpha and transforming growth factor (TGF)-beta. Therefore, LOX-1 is thought to be upregulated locally in response to cytokines in vivo.. We determined by reverse-transcription polymerase chain reaction (PCR) and Western blot analysis whether the mediators of the acute phase response in inflammation, IL-1alpha, IL-1beta, and TNF-alpha, regulate LOX-1 expression in cultured SMC, and whether this regulation is influenced by peroxisome proliferator-activated receptor gamma (PPARgamma). We studied by immunohistochemistry whether these cytokines are spatially correlated with LOX-1 expression in advanced atherosclerotic lesions. We found upregulation of LOX-1 expression in SMC in a dose- and time-dependent manner after incubation with IL-1alpha, IL-1beta, and TNF-alpha. Simultaneous incubation with these cytokines at saturated concentrations had an additive effect on LOX-1 expression. The PPARgamma activator, 15d-PGJ(2), however, inhibited IL-1beta-induced upregulation of LOX-1. In the intima of atherosclerotic lesions regions of IL-1alpha, IL-1beta, and TNF-alpha expression corresponded to regions of LOX-1 expression.. We suppose that upregulated LOX-1 expression in SMC of advanced atherosclerotic lesions is a response to these proinflammatory cytokines. Moreover, the proinflammatory effects of these cytokines can be decreased by the antiinflammatory effect of PPARgamma.

Topics: Aorta; Arteriosclerosis; Cells, Cultured; Cytokines; Humans; Interleukin-1; Lasers; Microdissection; Muscle, Smooth, Vascular; PPAR gamma; Prostaglandin D2; Receptors, LDL; Receptors, Oxidized LDL; RNA, Messenger; Scavenger Receptors, Class E; Tumor Necrosis Factor-alpha; Tunica Intima; Tunica Media

Atherosclerotic lesions occur as a result of excess lipid deposition within the vascular tissues. The peroxisome proliferator-activated receptors (PPARs) present in adipose and hepatic tissues have been shown to promote fatty acid oxidation and lipid storage. An immunohistochemical assessment of PPARalpha and PPARgamma revealed both proteins were also present in the medial and intimal layers of human arteries, predominately in regions containing smooth muscle cells. In agreement with this observation, smooth muscle cells isolated from these vessels were found by RT-PCR to express both PPARalpha and PPARgamma1. The functionality of these receptors was tested with selective PPAR agonists. Mitogenic stimulation of smooth muscle cell proliferation was blocked by 15d-PGJ2, a PPARgamma agonist, as well as by WY14643, a PPARalpha agonist. These data indicate PPAR activation by selective agonists could influence lesion progression directly, as well as indirectly through reductions in serum lipoprotein and triglyceride levels.

Topics: Arteriosclerosis; Base Sequence; Cell Division; Cells, Cultured; Humans; Immunohistochemistry; Muscle, Smooth, Vascular; Prostaglandin D2; Pyrimidines; Receptors, Cytoplasmic and Nuclear; RNA, Messenger; Transcription Factors