葡萄籽原花青素可能通过AKT-Caspase-3途径对深低温缺血再灌注脑起保护作用

Influence of Grape seed proanthocyanidin extract on deep hypothermia cerebral ischemia-reperfusion in mice

目的探讨葡萄籽原花青素（GSPE）预处理对小鼠脑缺血再灌注（I／R）损伤的保护作用机制。方法将60只4周龄C57／BL6小鼠随机平均分为假手术组、模型组和GSPE组建立深低温I／R模型，再灌注24h后处死小鼠取脑，Westernblot检测丝氨酸．苏氨酸蛋白激酶（AKT）的表达，逆转录-聚合酶链反应（1it—PCR）技术检测小鼠脑组织半胱天冬氨酸蛋白酶（Caspase）-3mRNA，TUNEL法检测小鼠大脑皮层细胞凋亡。结果与假手术组比较模型组和GSPE组于再灌注24h后p-AKT显著激活（P〈0．05），Caspase-mRNA表达明显增加（P〈0．05）并且出现明显的病理凋亡（P〈0．05）；GSPE组p-AKT表达较模型组显著增高[A值（1．90±0．02）〉（1．6±0．01），P〈0．05]，且Caspase-3mRNA、病理凋亡均显著低于模型组[（0．43±0．02）〈（0．86±0．03）；（40．92±3．18）％〈（79．86±4．68）％，P〈0．05]。结论葡萄籽原花青素可能通过AKT-Caspase-3途径对深低温缺血再灌注脑起保护作用。

Objective To determine the mechanism of Grape seed proanthocyanidin extract on a model of brain injury induced by deep hypothermic cerebral ischemia/reperfusion （I/R）. Methods Sixty four-week-old C57BL6 mice were randomly and equally divided into sham operation group, model group and grape seed proanthocyanidin extract （GSPE） group. The mice were sacrificed 24 h after reperfusion for sampling of brain tissues. The expression of serine/threonine kinase （AKT） protein was detected by using Western blotting, Caspase-3 mRNA was detected by reverse transeription-polymerase chain reaction （ RT- PCR） technique, and apoptosis of cerebral cells was examined by using TUNEL stalling. Results The expression of p-AKT protein and Caspase-3 mRNA, and apoptotic level in model group and GSPE group were significantly increased as compared with sham operation group （ P 〈 0.05 ）. The expression of p-AKT protein [ A （ 1.9 ± 0. 02） 〉 （ 1.6 ± 0.01 ） ,P 〈 0. 05 ] was increased, and expression of Caspase-3 mRNA and apoptosis were significantly decreased in GSPE group as compared with model group [ （0.43 ± 0. 02） 〈 （0.86±0.03） ; （40.92 ±3.18）% 〈 （79.86 ±4.68）% ,P〈0.05]. Conclusion The resuhs suggest that GSPE has a protective effect against deep hypothermic cerebral I/R injury through AKT-Caspase-3 pathway.