目的 探讨哈巴苷对急性脑缺血小鼠的神经保护作用及线粒体保护机制。方法 采用脑中动脉阻断复制急性脑缺血模型,将ICR小鼠随机分为假手术组、模型组、尼莫地平组,哈巴苷(5、10、15 mg·kg-1)组;造模后立即腹腔注射给药,考察各组小鼠神经功能评分、脑含水量、脑指数、脑缺血体积及大脑组织结构病理变化;测定脑中动脉阻断小鼠大脑缺血区细胞线粒体内Ca2+-Mg2+- ATPase的活性及缺血区caspase-3蛋白表达水平;透射电镜(TEM)下观察脑中动脉阻断小鼠大脑缺血区细胞线粒体超微结构变化。结果 与模型组比较,哈巴苷各剂量组,均能不同程度降低脑中动脉阻断小鼠神经功能评分、脑含水量、脑指数及脑缺血体积(P<0.05,P<0.01);哈巴苷10 mg·kg-1能明显增强Ca2+-Mg2+-ATPase的活性(P<0.01),明显下调脑缺血区caspase-3蛋白的表达(P<0.01);哈巴苷各剂量能不同程度改善脑中动脉阻断小鼠大脑缺血区皮层及海马齿状回病理结构及线粒体超微结构变化,减少线粒体水肿。结论 哈巴苷对急性脑缺血小鼠具有明显的保护作用,其作用机制与保护线粒体损伤、下调细胞凋亡蛋白途径有关。
Abstract
OBJECTIVE To observe the neuro-protective effects of harpagide on acute cerebral ischemic injury in mice and its mechanism involving mitochondria. METHODS Acute cerebral ischemia were achieved by operation of MCAO in the left brain, random allocation was taken to divide ICR mice into sham group, model group, nimodipine group and harpagide(5, 10, 15 mg·kg-1) groups. Mice were intraperitoneal injected harpagide immediately after surgery. Nerve function score, content of brain water, brain index and the common changes of brain pathological structure in HE staining were measured; Ability of mitochondria′ Ca2+-Mg2+-ATPase and protein expression level of caspase-3 in the MCAO mice′ brains was determined; Ultrastructure change of mitochondria under the TEM was observed. RESULTS Compared with model group, the harpagide groups could decreased the nerve function score, the content of brain water, brain index and the volume of ischemia in mice with different degrees in MCAO mice(P<0.05,P<0.01); 10 mg·kg-1 of harpagide could increased the activity of Ca2+-Mg2+-ATPase obviously(P<0.01); And significantly decreased the protein expression level of caspase-3(P<0.01); harpagide groups could protect the pathogeny structure and the ultrastructure of mitochondria with different degrees in MCAO mice, decrease edema of mitochondria obviously. CONCLUSION Harpagide could obviously protect acute cerebral ischemia in mice, its therapeutical effects are approached to protecting the activity of brain mitochondria and decreasing protein expression level of caspase-3.
关键词
哈巴苷 /
急性脑缺血 /
线粒体超微结构 /
caspase-3 /
保护作用
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Key words
harpagide /
acute cerebral ischemia /
mitochondrial ultrastructure /
caspase-3 /
protective effect
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中图分类号：
R965
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参考文献
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脚注
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基金
浙江省自然基金资助项目(LY14H280005;LY15H280004);浙江省公益研究计划项目(2013C31106);
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