Activation of P2X(7) purinoceptor-stimulated TGF-beta 1 mRNA expression involves PKC/MAPK signalling pathway in a rat brain-derived type-2 astrocyte cell line, RBA-2

The present study investigates the receptor and mechanisms involved in ATP-stimulated transforming growth factor-beta 1 (TGF-beta1) mRNA expression of a type-2 astrocyte cell line, RBA-2. RT-PCR analysis revealed that RBA-2 type-2 astrocytes possess abundant P2X(4) and P2X(7) receptors. ATP and P2X(7) receptor-sensitive agonist, BzATP, both stimulated TGF-beta1 mRNA expression in a time and dose-dependent manner. The stimulation required a minimum of 500 muM ATP; BzATP was much more potent that ATP, and P2X(7)-selective antagonist, oATP, inhibited the effects. In addition, ATP metabolites ADP, AMP and adenosine were ineffective in stimulation of TGF-beta1 mRNA expression. Thus, the effect of ATP was mediated through the P2X(7) receptors. To investigate further the mechanisms by which the P2X(7) receptor mediated the TGF-beta1 mRNA expression, the cells were treated with inhibitors for mitogen-activated kinase (MAPK) or protein kinase C (PKC), PD98059 or GF109203X, respectively. Both PD98059 and GF109203X inhibited the ATP-stimulated TGF-beta1 mRNA expression. Furthermore, ATP and BzATP stimulated ERK1/2 activation and the activation was inhibited by PKC inhibitors, GF109203X and Go6976. In conclusion, activation of P2X(7) receptors enhanced TGFbeta1 mRNA expression and the effect involved PKC/MAPK signalling pathway in RBA-2 type-2 astrocytes. (C) 2003 Elsevier Inc. All rights reserved.