Evidence for the involvement of 5-lipoxygenase products in the regulation of the expression of the proenkephalin gene in cultured astroglial cells.

Abstract

Cultured astroglial cells secrete eicosanoids which are produced by the cyclooxygenase and lipoxygenases. These cells also transcribe the proenkephalin gene. In the present study, it was investigated whether agents which inhibit the metabolism of arachidonic acid affect the basal and stimulated expression of the gene. Tetradecanoyl phorbol acetate (TPA; 1-1000 nmol/l) increases the concentration of proenkephalin mRNA in these cells by activating protein kinase C. The enhancement in proenkephalin mRNA caused by TPA (10 nmol/l) was not affected by the cyclooxygenase inhibitor indomethacin (5 mumol/l). However, nordihydroguaiaretic acid, which blocks cyclooxygenase and lipoxygenases, potentiated the effect of TPA on proenkephalin mRNA, when used at concentrations of 0.5-50 mumol/l. Two selective inhibitors of 5-lipoxygenase, i.e. MK886 (5 mumol/l) and BAY X1005 (1 mumol/l), also enhanced the effect of TPA (10 nmol/l) without affecting the basal expression of the gene. When added to the incubation medium, leukotriene E4 (10-1000 nmol/l) diminished in a dose-dependent manner the basal and TPA-induced expression of the proenkephalin gene. It is concluded that in astroglial cells derived from cortex of new-born rats products of 5-lipoxygenase can diminish the action of protein kinase C on the proenkephalin gene.