Intracerebral hemorrhage (ICH) is a devastating stroke. The damage that occurs in brain from ICH is due to the growing hematoma and clot formation that damages the adjacent brain through edema and apoptosis. Though these are distinct pathologic entities that occur following ICH, we have data that they share some common mechanisms. We found that apoptosis in peri-hematoma brain is mediated in part by glutamate excitotoxicity. This led us to the non-receptor tyrosine kinase, pp60-Src (Src) because it potentiates function of NMDA receptors through direct phosphorylation of the NR2A subunit. We found that Src kinase activity increases 4 fold following experimental ICH in rats, the Src family kinase (SFK) member Lyn increases over 21 fold following ICH, and Src inhibitors decrease apoptosis and improve behavioral outcome following ICH. We hypothesize that Src plays a central role in ICH mediated apoptosis and edema: ICH causes thrombin receptor activation of Src that activates NR2A subunits that mediate apoptosis; and ICH activates thrombin receptors which activates HIF and MMPs via Src to produce edema and poor behavioral outcome. These hypotheses are based upon previous studies showing that thrombin mediates the acute brain edema following ICH, and that thrombin activates Src via the thrombin receptor. Therefore, these studies will examine: (a) the effect of ICH on Src; (b) thrombin activation of Src; (c) ICH induced phosphorylation of NMDA receptors by Src; (d) Src activation of HIF-1, HIF-1 target genes and MMPs; and (e) the effect of Src blockade on cell survival, edema and behavioral outcome using rat models of intracerebral hemorrhage (ICH). The following aims will be addressed:
Aims #1 a-d: Demonstrate that Src mRNA, Src protein, Src phosphorylation and Src activity increase following ICH. Determine whether thrombin antagonists block these ICH induced changes of Src and whether thrombin and thrombin receptor agonists reproduce the ICH induced changes of Src.
Aim #2. Demonstrate changes of HIF-1, of the HIF-1 target gene VEGF, and the Matrix Metalloproteinases and the phosphorylation state of NMDA receptor subunits after ICH in rats with and with out pharmacological blockade of Src.
Aim #3. Demonstrate that pharmacological inhibition of Src improves cell survival, decreases brain edema and improves behavioral outcome following ICH in rats (a) using a blood infusion model of ICH and (b) using a suture induced vessel rupture model of ICH. ? ? ?
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