Purchase this article with an account.
L. J. Faia, Z. Li, P. Becerra, J. Amaral, B. Liu, R. Nussenblatt; The Effects of Pigment Epithelial Derived Factor on Murine Macrophages. Invest. Ophthalmol. Vis. Sci. 2009;50(13):4283.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
To assess the effects of pigment epithelial derived factor (PEDF) on murine macrophages, specifically on nitric oxide (nitrate) and cytokine production upon activation.
RAW 264.7 macrophages were maintained in phenol-free DMEM with 10% FBS. The cells were re-plated in 96-well plates at a concentration of 3.3 x 106 cells per well and stimulated with or without 1) LPS and/or 2) IFNγ in the absence or presence of PEDF (500ng/ml). All stimulation was done in duplicates. After 24 hours of stimulation, the supernatants were harvested, pooled and divided into 2 parts. Half the supernatant was assessed for nitrate production by a Nitric Oxide Assay Kit (Pierce, Rockford, IL) and the other half for cytokine analysis (IL-1b, IL-6, IL-10, IL-12, TNF and SDF-1b).
Both LPS and IFNγ increased nitrate production, while PEDF alone had minimum effect. Interestingly, PEDF suppressed IFNγ-induced NO production but had minimum effect on that by LPS (INFγ alone 35.8µM, INFγ +PEDF 19.2µM, LPS alone 25.3µM , LPS+PEDF 23.1µM). In addition, there was no synergistic effect of LPS+IFNγ in inducing NO production. PEDF had no effect when macrophages were stimulated with both LPS and IFNγ. For IL-10 production, IFNγ showed some effect on increasing its production (31%), but in the presence of PEDF, the induction of IL-10 by IFNγ increased dramatically, by almost 200%. Meanwhile, PEDF had a very minimal effect on induction of TNF by IFNγ. In addition, IFNγ completely abrogated SDF-1b production by macrophages but PEDF reversed this effect of IFNγ.
Our data suggested that PEDF may be an important factor in modulating inflammation by promoting anti-inflammatory macrophages through the increase of IL-10 and the decrease in NO production. The finding that PEDF completely reversed SDF-1b production abrogated by IFNγ is of particular interest. Whether a direct effect on IFNγ signaling or indirectly by PEDF, this finding is worthy of further investigation.
This PDF is available to Subscribers Only