Thrombomodulin as a new marker of lesion-induced astrogliosis: Involvementof thrombin through the G-protein-coupled protease activated receptor-1

Because injury of the CNS causes an astrogliosis, characterized by cell swe lling and proliferation, similar to the effects of the serine protease thro mbin on astrocytes, we hypothesized that a high level of thrombin at the si te of injury might initially induce an astrocyte reaction and later increas e the expression of its specific inhibitor, thrombomodulin. Thrombomodulin could then stabilize the astroglial scar through its adhesive properties. Here, we studied the in vivo injury response of astrocytes in the anterior medullary velum of adult rat by immunostaining and in situ hybridization of thrombomodulin. Thrombomodulin was poorly expressed on astrocytes in norma l tissue, increased up to 2 d after injury, and was still highly expressed at 6 d. To check that thrombin had a direct effect on thrombomodulin expression by astrocytes, we used brain cortical astrocyte primary cultures treated with either thrombin or the agonist peptide thrombin receptor-activating peptide -6, known to activate directly the thrombin G-protein-coupled receptor (GPC R) protease-activated receptor-1 (PAR-1). Modification of thrombomodulin ex pression was studied by Western blotting and quantitative reverse transcrip tion-PCR. There was a dose-dependent increase in thrombomodulin after 48 hr of treatment, with gene expression peaking at 24 hr but falling to control levels by 48 hr. Together, these results show the following: (1) injury increases astrocyte thrombomodulin expression; (2) thrombin might mediate thrombomodulin expres sion via the specific receptor PAR-1; and (3) serine proteases, their inhib itors, and the new family of GPCR, PARs, are active on astrogliosis.