Effect of prostaglandin E2 on PMA-induced macrophage differentiation

https://doi.org/10.1016/j.prostaglandins.2004.08.001Get rights and content

Abstract

Major trauma such as severe burns and extensive surgery could result in accelerated macrophage differentiation and hyperactivation causing an excessive release of proinflammatory cytokines and prostaglandin E2 (PGE2) with consequent severe impairment of immunologic reactivity. HL-60 cells stimulated with phorbol 12-myristate 13-acetate (PMA) have been used as a model to asses the PGE2 role in the macrophage differentiation observed after major trauma. Cell adhesion, matrix metalloproteinase-9 (MMP-9) and tumor necrosis factor-α (TNF-α) production were measured after 24 h of PMA treatment in the presence of PGE2 (lnM-l μM). PGE2 increased both the PMA-induced cell adhesion and MMP-9 production via EP2/EP4 receptors while it had no effect on the induced TNF-α release. The cAMP/PKA pathway, usually linked to EP2/EP4 activation, was not involved in the phenomenon, suggesting that an alternative signalling pathway could be linked to a PKC-activated enzyme. In fact PGE2 activity was partially inhibited by Wortmannin, a phosphoinositide-3 kinase (PI-3K) inhibitor indicating that PGE2 act as a co-factor able to increase macrophage differentiation in vitro via a PI-3K dependent pathway that could be also involved in the immunosuppression observed in the aftermath of trauma.

Introduction

Trauma that induce a large volume of tissue injury and hypovolemic hemorrhagic shock such as burns, severe fractures and extensive surgical procedures, could result in severe impairment of immunologic reactivity that in turn leads to the development of serious infections [1]. A sequence of states of cellular activation has been described in the aftermath of trauma and, to a certain degree, they can also be associated with consecutive clinical states such as the acute phase response, anergy, infection, and organ failure [2]. The monocyte/macrophage lineage plays a key role in this immunological derangement as macrophage participates to a number of regulatory loops with T cells [2]. In fact, under stressful conditions macrophage show an immediate hyperactivation characterized by an excessive release of proinflammatory cytokines [3] and prostaglandin E2 (PGE2), the most powerful endogenous immune suppressant [4], [5], followed by a paralysis of cell functions, which is then overcome in few days with the recruitment of immature macrophage [6].

While the role of PGE2 in the after trauma impairment of the immune response has been extensively studied [7], [8], [9], [10], its possible role in the accelerated macrophage differentiation remains elusive [11], [12].

Human HL-60 myeloid leukemia cells retains the ability to differentiate along the monocyte, macrophage, or granulocyte pathway [13] in response to a wide variety of differentiation inducers and this cell line is a suitable model to study the modulation of macrophage differentiation.

In fact, the phorbol 12-myristate 13-acetate (PMA) induces HL-60 terminal differentiation toward the macrophage lineage [14], [15] characterized by the appearance of cell adhesion and spreading [16], [17], matrix metalloproteinase-9 (MMP-9) [17], [18], [19] and tumor necrosis factor-α (TNF-α) production [18], [19]. The present study was initiated to investigate the effect of PGE2 on the cell adhesion, MMP-9 and TNF-α production in untreated and PMA-treated HL-60 cells in order to clarify if PGE2 could participate in the onset of accelerated macrophage differentiation observed in the patients victims of major trauma.

Section snippets

Materials

The PKC activator phorbol 12-myristate 13-acetate (PMA), prostaglandin E2 (PGE2), forskolin and all the material for cell culture and substrate zymography were obtained from Sigma (St. Louis, MO, USA). Sulprostone, Misoprostol, Butaprost and 16,16-dimethyl PGE2 were purchased from Cayman Chemical (Ann Arbor, MI, USA). The MAP kinase kinase inhibitor PD 98,059 (PD), the PI-3K inhibitor Wortmannin (WRT), the inhibitor of adenilyl cyclase SQ 22,536 (SQ) and the Adenosine 3′,5′-cyclic

PGE2 increases the PMA-induced macrophage differentiation in HL-60 via a TNF-α independent pathway

The macrophage differentiation of HL-60 cells was monitored measuring the number of adherent cells and the activity of the MMP-9 (gelatinase B) in the cell medium for 24 h. The addition of PGE2 (1 μM) to the HL-60 cell in the presence of 10% FCS (Fig. 1B) did not altered significantly the spontaneous cell adhesion that occurs also in the undifferentiated HL-60 cells (Fig. 1A). In fact the mean adherent cell number ± standard error (S.E.) scored in 10 random fields was 81 ± 3 for in the control

Acknowledgements

The authors thank Dr. M. De Andrea from the Microbiology Laboratory, for the kind gift of HL-60 cells, Dr. C. Campa and Dr. D. Speretta from the Human Anatomy Laboratory, for the precious and skillful technical assistance.

References (38)

Cited by (8)

  • The role of PGE2 and EP receptors on lung's immune and structural cells; possibilities for future asthma therapy

    2023, Pharmacology and Therapeutics
    Citation Excerpt :

    Mediated by stimulation of the EP2 and EP4 receptors, cell adhesion and matrix metalloprotease (MMP)-9 production in the PMA-stimulated HL-60 cell model were enhanced, whereas TNFα release remained unaffected in vitro (Renò & Cannas, 2005). Interestingly, these effects involved PI3K dependent pathways, but not adenylyl cyclase/PKA dependent pathways (Renò & Cannas, 2005). However, PGE2 was found to reduce expression of TNFα in LPS-stimulated human lung-derived macrophages (Gill et al., 2016) and murine bone marrow-derived macrophages (Saleh, Vanderheyden, Frederickson, & Bryant, 2020) mediated by EP2 and primarily by EP4 signaling (Gill et al., 2016; Saleh et al., 2020).

  • Prostaglandin E2 induces matrix metalloproteinase 9 expression in dendritic cells through two independent signaling pathways leading to Activator Protein 1 (AP-1) activation

    2011, Journal of Biological Chemistry
    Citation Excerpt :

    In addition, a direct correlation between induction of PGE2 and MMP-9 was reported in a number of cell types, including monocytes (32, 33). Exogenous and endogenous PGE2 was also shown to play a role in MMP-9 expression in macrophages stimulated with PMA, LPS, or exposed to ECM (10, 13). We showed previously that exogenous PGE2 induced MMP-9 expression in immature DCs, up-regulated MMP-9 in DCs treated with TNF-α and IFN-α, and that PGE2-induced MMP-9 was essential for in vitro migration of DCs through Matrigel and for the in vivo migration of DCs in wild-type and MMP-9-deficient hosts (3).

  • PGE2-induced metalloproteinase-9 is essential for dendritic cell migration

    2008, Blood
    Citation Excerpt :

    PGE2 inhibits MMP-9 expression in the breast cancer cell line MCF-7, in peritoneal macrophages isolated from women with endometriosis, and in interleukin-1 (IL-1)–stimulated rabbit articular chondrocytes.32TTT–34 In contrast, exogenous PGE2 promotes MMP-9 expression in macrophages stimulated with phorbol myristate acetate (PMA) or plated on ECM and in LPS-stimulated trophoblasts.35TTTTTT–38 In addition, endogenous PGE2 was shown to play an important role in MMP-9 expression in macrophages exposed to ECM or stimulated with LPS.35,39

View all citing articles on Scopus
View full text