摘要
目的探讨线粒体分裂抑制剂（Mdivi-1）在大鼠脑缺血再灌注损伤线粒体凋亡过程中的保护作用及其机制。方法按随机数字表法将成年健康雄性Wistar大鼠60只分为假手术组、模型组和Mdivi-l预处理组．每组各20只，其中模型组和Mdivi一1预处理组大鼠采用线栓法经左侧颈外一颈内动脉插线建立脑缺血再灌注损伤模型，并于脑缺血前15min预先尾静脉给予等容量二甲基亚砜和Mdivi-1（1．2mg／kg）。再灌注24h后应用TUNEL法观察各组大鼠脑组织神经细胞凋亡情况，SABC免疫组化染色检测细胞色素C（CytC）阳性细胞表达，Westernblotting检测CytC蛋白表达，RT．PCR检测CytCmRNA表达。结果与假手术组比较，模型组神经细胞凋亡率、CvtC阳性细胞率、CvtC蛋白及mRNA表达水平（4．16％±0-25％0s45．49％±O．77％；4．22％±0．20％口555．12％±1．47％；0．395＋0．002／）S0．932＋0．00l；0．467±0．003口s0．789±0．009）显著升高，差异有统计学意义（P〈0．05）。应用Mdivi-1预处理后神经细胞凋亡率、CvtC阳性细胞率、CvtC蛋白及mRNA表达水平（30．76％±0．51％；35．07％±1．42％：0．59．4＋-0．002；0．523±0．004）较模型组明显降低，差异有统计学意义（P〈0．05）。结论Mdivi．1通过阻断线粒体一CytC途径从而抑制细胞凋亡，减轻脑缺血再灌注损伤。
Objective To observe the effect ofmitochondrial division inhibitor （Mdivi-1） on the neuronal apoptosis via mitochondria signaling pathways in rats after cerebral ischemia/reperfusion （I/R） and its possible mechanism. Methods Sixty male adult healthy Wistar rats were chosen and randomly divided into sham-operated group, vehicle group and Mdivi-1 pre-conditioning group （n=20）; animal models of middle cerebral artery occlusion/reperfusion in the later 2 groups were established by a filament method in the left external-internal carotid artery. Rats were injected with 1.2 mg/kg Mdivi-1 in the Mdivi-1 pre-conditioning group and dimethyl sulfoxide in the vehicle group 15 minutes before the cerebral ischemia. After 24 h of reperfusion, apoptotic cell death was assessed by TUNEL staining, the cytochrome C expression （Cyt C） was detected by SABC immunohistochemical staining, the protein level ofCyt C in the cerebral issues was detected by Western blotting and the mRNA expression was measured by semiquantitative reverse transeriptase polymerase chain reaction （RT-PCR）. Results The apoptosis rate （45.49%~0.77%）, Cyt C positive cell rate （55.12%~1.47%） and protein and mRNA expressions of Cyt C （0.932 ~0.001, 0.789 ~0.009） in the vehicle group were significantly higher that in the sham-operated group （4.16%_＋0.25%, 4.22%_＋0.20%, 0.395~0.002, 0.467~0.003, P〈0.05）. The apoptosis rate （45.49%~0.77%）, Cyt C positive cell rate （55.12%~1.47%） and protein and mRNA expressions of Cyt C （0.594＋0.002, 0.523~0.004） in the Mdivi-1 pre-conditioning group were significantly decreased as compared with those in the vehicle group （P〈O.05）. Conclusion Mitochondrial division inhibitor Mdivi-1 might reduce neuronal apoptosis by inhibiting mitochondria cytochrome C apoptotic pathway to protect cerebral against ischemia/reperfusion injury.
出处
《中华神经医学杂志》
CAS
CSCD
北大核心
2013年第4期354-358,共5页
Chinese Journal of Neuromedicine
基金
国家自然科学基金（30972855）
关键词
脑缺血再灌注
线粒体分裂抑制剂
细胞色素C
细胞凋亡
Cerebral ischemia/reperfusion
Mitochondrial division inhibitor
Cytochrome C
Apoptosis
