Original Contribution
PGD2 and PGE2 regulate gene expression of Prx 6 in primary macrophages via Nrf2

https://doi.org/10.1016/j.freeradbiomed.2011.05.022Get rights and content

Abstract

Peroxiredoxin 6 (Prx 6) is a bifunctional enzyme with both glutathione peroxidase and acidic Ca2+-independent phospholipase A2 activities. We have recently shown that exposure of murine bone marrow-derived macrophages to LPS and IFN-γ leads to induction of COX-2 expression and secretion of PGE2, up-regulating Prx 6 mRNA levels. This study was designed to investigate various prostaglandins (PGs) for their ability to induce gene expression of Prxs, in particular Prx 6, and to determine the underlying regulatory mechanisms. We provide evidence that both conventional and cyclopentenone PGs enhance Prx 6 mRNA expression. Treatment with either activators or inhibitors of adenylate cyclase as well as cAMP analogs indicated that Prx 6 gene expression is regulated by adenylate cyclase in response to PGD2 or PGE2. Furthermore, our study revealed that JAK2, PI3K, PKC, and p38 MAPK contribute to the PGD2- or PGE2-dependent Prx 6 induction. Using stimulated macrophages from Nrf2-deficient mice or activators of Nrf2 and PPARγ, we found that Nrf2, but not PPARγ, is involved in the PG-dependent increase in Prx 6 mRNA expression. In summary, our data suggest multiple signaling pathways of Prx 6 regulation by PGs and identified Nrf2 as a critical player mediating transcriptional induction.

Section snippets

Materials

PGA1, PGA2, PGE2, PGF, 15d-PGJ2, dibuturyl-cAMP, forskolin, IBMX, H-89, KT5720, MDL 12,330A, LY294002, Ro31-8220, Gö6796, SB202190, SP600125, PD98059, AG490, MAFP, AACOCF3, sulforaphane, caffeic acid phenethyl ester (CAPE), tert-butylhydroquinone (tBHQ), triglitazone, ciglitazone, GW-9662, and L-NIL were obtained from Enzo Life Sciences (Lörrach, Germany). PGD2 and PGE1 were from Calbiochem (Darmstadt, Germany). Griess reagent (modified) and LPS (Escherichia coli serotype 055:B5) were from

Gene expression of Prx 6 is increased by prostaglandins in bone marrow-derived macrophages

To elucidate the role of PGs in Prx gene expression, BMMs of C57BL/6 mice were stimulated with conventional PGs, i.e., PGD2 (50 μM), PGE1 (50 μM), PGE2 (50 μM), PGF (50 μM); cyclopentenone PGs, i.e., PGA1 (50 μM), PGA2 (50 μM), 15d-PGJ2 (10 μM); or corresponding vehicle (DMSO, methyl acetate). After 18 h of stimulation, nitrite was quantified and mRNA expression of Prxs 1–6 was measured by qRT-PCR. In agreement with Itoh et al. [13], gene expression of Prx 1 was significantly increased by 15d-PGJ2,

Discussion

Most recently, we demonstrated that stimulation of primary murine macrophages with LPS and IFN-γ leads to COX-2-mediated PGE2 secretion resulting in increased Prx 6 gene expression. Various COX-1 and -2 inhibitors diminished Prx 6 mRNA induction, suggesting that LPS- and IFN-γ-induced gene expression of Prx 6 is regulated by COX enzymes [10]. Further inhibition experiments revealed that, in addition to COX-2, cPLA2 plays an important role in the up-regulation of Prx 6 in response to LPS and

Acknowledgments

We are grateful to Gabi Würtz, Claudia Weber, and Eylin Topfstedt for excellent technical assistance and to Thomas Herdegen and Yuet W. Kan for providing the Nrf2-deficient mice. S.E. was supported by a grant from the Graduate College 840 (Deutsche Forschungsgemeinschaft) to I.S. and R.W.

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    These authors contributed equally to this work.

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