Recombinant-activated factorⅦand neuronal apoptosis in a rat model of intracerebral hemorrhage

摘要

BACKGROUND:Activated clotting factorⅦhas been demonstrated to exhibit obvious anti-apoptosis effects. OBJECTIVE:To observe the effect of activated clotting factorⅦon neuronal apoptosis at different time points following rat intracerebral hemorrhage(ICH). DESIGN,TIME AND SETTING:A randomized,controlled,animal experiment was performed at the Neurobiological Laboratory of Second Military Medical University from October 2005 to April 2006. MATERIALS:Recombinant-activated clotting factorⅦa(rFVIIa) was purchased from Danish Novo Nordisk,Denmark.In situ cell death detection kit-POD kit was purchased from Roche,Switzerland. Caspase-3 activity determination kit from Biovision,USA. METHODS:A total of 72 healthy,male,Sprague Dawley rats,aged 5-8 months,were randomly assigned to three groups(n = 24):sham-operated,ICH model,and rFVIIa.In the ICH model and rFVIIa groups,80.0μL autologous non-clotting blood from rat tails was injected into the right caudate putamen to establish the ICH.The empty microinjector was inserted into the caudate putamen in the sham-operated group.The ICH model and rFVIIa groups were subdivided into four subsets separately:6,24,72 hours and 7 days following ICH.The rats in the rFVIIa group were injected with 160μg/kg rFVIIa via the dorsal vein of the penis. MAIN OUTCOME MEASURES:Apoptotic cells were detected in the right caudate putamen by TUNEL;caspase-3 activity by spectrophotometry;and rat neurological function was evaluated by neurological functional impairment scales. RESULTS:Rat neurological function was deteriorated at 24,72 hours,and 7 days following ICH. The TUNEL-positive cells and caspase-3 activity in the right caudate putamen was significantly increased in the ICH rats(P<0.05);rFVIIa treatment reduced the number of TUNEL-positive cells and caspase-3 activity in the right caudate putamen(P<0.05),and neurological function was significantly improved(P<0.05). CONCLUSION:rFVIIa was applied within 72 hours after ICH,which reduced the amount of neuronal apoptosis and promoted neurological function restoration by possibly inhibiting caspase-3 activity.