目的 观察半胱氨酸蛋白酶(Caspase)抑制剂z-VAD-fmk对于缺血缺氧性脑损伤(hypoxic-ischemic brain damage, HIBD)大鼠脑组织的保护作用, 并探讨其机制。方法 7 d龄Wistar大鼠60只, 随机分为对照组、HIBD组和阻断组, 各20只。HIBD组及阻断组制作HIBD模型, 对照组大鼠分离但不结扎左侧颈总动脉, 术后亦不进行缺氧处理。阻断组在建模后给予z-VAD-fmk, 连续7 d。第8天处死全部动物。分别采用TUNEL法和RT-PCR法检测三组动物脑组织中神经细胞凋亡指数(apoptosis index, AI)和Caspase-3 mRNA。结果 对照组、HIBD组、阻断组AI值分别为0.15±0.02、21.36±2.45、10.42±1.62, 三组相比差异有统计学意义(P均＜0.05)。三组脑组织中Caspase-3 mRNA的相对表达量分别为0.016±0.003、0.561±0.067、0.216 ±0.036, 三组相比差异有统计学意义(P均＜0.05)。AI值随Caspase-3 mRNA的升高而升高, 随其降低而降低, 二者之间具有正相关关系(r=0.628, P＜0.05)。结论 z-VAD-fmk对HIBD大鼠脑组织有一定的保护作用, 其机制可能是降低脑组织中Caspase-3 mRNA的表达, 减轻神经细胞凋亡。
Abstract
Objective To study the effect of hypoxia ischemia brain damage (HIBD) of neonatal rats with cerebral cortex apoptosis by gene inhibiter so as to explore the possible mechanism. Methods Model of HIBD was built in neonatal rats, 60 Wistar rats were randomly divided in:control group;HIBD group;obstruct group(HIBD+z-VAD-fmk).Adopting HE dyeing, three animal apoptosis index (AI) in brain and Caspase-3mRNA were detected by TUNEL and RT-PCR. Results Express of Caspase-3mRNA nanomolar in control group was significant heighten in HIBD group(P＜0.01).There was significantly lower express between obstruct group and HIBD group(P＜0.05).AI value changed with Caspase-3mRNA.They had significance positive correlation (r=0.628, P<0.05). Conclusion Express heigten of Caspase-3mRNA leads to histiocytic apoptosis in HIBD group, and lower express of Caspase-3mRNA by z-VAD-fmk, could protect brain of neonatal rats.
关键词
缺血缺氧性脑损伤 /
大鼠 /
细胞凋亡 /
半胱氨酸蛋白酶-3
Key words
hypoxia ischemia damage /
rat /
apoptosis /
Caspase-3
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