In situ expression of platelet-activating factor (PAF)-receptor gene in rat skin and effects of PAF on proliferation and differentiation of cultured human keratinocytes.

ABSTRACT

Platelet-activating factor (PAF) is a potent lipid mediator that exhibits versatile biologic activities in many diverse systems by binding to a specific cell-surface receptor (PAFR). Although the production of PAF in cultured keratinocytes and fibroblasts has been reported, physiologic roles of this mediator in skin remain unclear. In this study, we examined in situ expression of PAFR gene in rat skin and the effects of PAF on the proliferation and differentiation of cultured human keratinocytes. In rat epidermis, PAFR mRNA expression was found from the basal cells to the granular cells, and strong signals were seen in the stratum spinosum. In cultured human keratinocytes, a 3.8 kb PAFR mRNA expression was demonstrated by northern blotting, and two distinct type transcripts driven by different promoters were detected by reverse transcriptase polymerase chain reaction analysis. Addition of PAF (30-100 nM) to cultured keratinocytes during a growth phase inhibited the proliferation. This effect was receptor dependent, because the inhibition was completely blocked by a PAFR antagonist, WEB 2086 (100 nM). On the other hand, whereas PAF (30-100 nM) alone did not affect the cornified envelope formation during the process of keratinocyte differentiation, WEB 2086 (30-300 nM) accelerated it in a concentration-dependent manner. Addition of PAF (100 nM) reversed the effect of WEB 2086, suggesting that WEB 2086 induced cornification by inhibiting PAF endogeneously produced by keratinocytes in an autocrine manner. Thus, we propose that PAF is an intrinsic regulator of keratinocyte during proliferation and differentiation.