Pro-inflammatory mediators and apoptosis correlate to rt-PA response in a novel mouse model of thromboembolic stroke.
BACKGROUND
A recent study suggests that patients with persistent occlusion of the middle cerebral artery (MCA) following treatment with recombinant tissue plasminogen activator (rt-PA) have better outcomes than patients with MCA occlusion not receiving rt-PA. We performed a study to elucidate possible mechanisms of this finding in a new model of thromboembolic stroke closely mimicking human pathophysiology.


METHODS
Thromboembolic stroke was induced by local injection of thrombin directly into the right MCA of C57 black/6J mice. Rt-PA was administered 20 and 40 min after clot formation. The efficiency of rt-PA to induce thrombolysis was measured by laser Doppler. After 24 h, all animals were euthanized and interleukin (IL)-6, tumor necrosis factor-alpha (TNF-α), matrix metalloproteinase (MMP)-9, Caspase-3, hsp 32 and hsp 70 protein levels were investigated by immunofluorescence. Presence of hemorrhage was verified and infarct volume was measured using histology.


RESULTS
Thrombin injection resulted in clot formation giving rise to cortical brain infarction. Early rt-PA treatment starting at 20 min after the clot formation resulted in 100% recanalization. However, rt-PA-induced thrombolysis dissolved the clot in only 38% of the animals when administered 40 min after clot formation. Protein levels of IL-6, TNF-α, MMP-9, Caspase-3, hsp 32 and hsp 70 were increased after MCAO, whereas treatment with rt-PA attenuated the expressions of inflammatory markers in those animals where the thrombolysis was successful. In addition, the infarct size was significantly reduced with rt-PA treatment compared to non-treated MCAO, regardless of whether MCA thrombolysis was successful.


CONCLUSIONS
The present study demonstrates a clear correlation of the protein expression of inflammatory mediators, apoptosis and stress genes with the recanalization data after rt-PA treatment. In this model rt-PA treatment decreases the infarct size regardless of whether vessel recanalization is successful.
Department of Neurology, University Medicine Mannheim, Heidelberg University, Heidelberg, Germany.
Saema
S
Ansar
Department of Neurology, University Medicine Mannheim, Heidelberg University, Heidelberg, Germany.
Eva
E
Chatzikonstantinou
Department of Neurology, University Medicine Mannheim, Heidelberg University, Heidelberg, Germany.
Rushani
R
Thiagarajah
Department of Neurology, University Medicine Mannheim, Heidelberg University, Heidelberg, Germany.
Laurent
L
Tritschler
Department of Neurology, University Medicine Mannheim, Heidelberg University, Heidelberg, Germany.
Marc
M
Fatar
Department of Neurology, University Medicine Mannheim, Heidelberg University, Heidelberg, Germany.
Michael G
MG
Hennerici
Department of Neurology, University Medicine Mannheim, Heidelberg University, Heidelberg, Germany.
Stephen
S
Meairs
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D001921:Brain / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D053148:Caspase 3 / Q000378:metabolism; D004195:Disease Models, Animal; D005343:Fibrinolytic Agents / Q000494:pharmacology / Q000627:therapeutic use*; D006360:Heat-Shock Proteins / Q000378:metabolism; D015850:Interleukin-6 / Q000378:metabolism*; D008297:Male; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D020521:Stroke / Q000188:drug therapy / Q000378:metabolism* / Q000473:pathology; D013923:Thromboembolism / Q000188:drug therapy / Q000378:metabolism* / Q000473:pathology; D010959:Tissue Plasminogen Activator / Q000494:pharmacology / Q000627:therapeutic use*; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism*
D005343:Fibrinolytic Agents; D006360:Heat-Shock Proteins; D015850:Interleukin-6; D014409:Tumor Necrosis Factor-alpha; D010959:Tissue Plasminogen Activator; D053148:Caspase 3; D020780:Matrix Metalloproteinase 9