Qiang Li1, Wei Li1, Suju Ding2, Jianping Tang1, Jing Fang1, Benqiang Deng2, Tao Wu2

1Department of Neurology, Ninth People’s Hospital of Shanghai Jiao Tong University, Shanghai  200011, China
2Department of Neurology, Changhai Hospital, Second Military Medical University of Chinese PLA, Shanghai  200043, China

Abstract
BACKGROUND: Activated clotting factor VII has been demonstrated to exhibit obvious anti-apoptosis effects.
OBJECTIVE: To observe the effect of activated clotting factor VII on 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 factor VIIa (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, 80.0 μL 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 sepa-rately: 6, 24, 72 hours and 7 days following ICH. The rats in the rFVIIa group were injected with 160 ?g/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 in-creased 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 signifi-cantly 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.

Key Words: intracerebral hemorrhage; apoptosis; activated clotting factor VII; caspase-3; in situ nick-end labeling

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