目的 探讨慢性脑缺血大鼠脑组织线粒体蛋白质组与能量代谢相关性,以期揭示慢性脑缺血疾病的线粒体机制.方法 通过差速离心分离大鼠脑线粒体,检测脑线粒体能量代谢相关的氧化磷酸化指标,应用凝胶电泳技术构建大鼠脑线粒体蛋白质组表达图谱;通过结扎大鼠双侧颈总动脉建立大鼠慢性脑缺血模型,分析比较慢性脑缺血时脑线粒体蛋白质表达谱的改变,从脑线粒体蛋白质组角度阐述慢性脑缺血与能量代谢的关系.结果 实验发现,与正常对照组相比,慢性脑缺血大鼠脑线粒体ADP/O及氧化磷酸化效率明显降低,蛋白质组结果显示慢性脑缺血大鼠NADH 脱氢酶复合体亚基、细胞色素C 氧化酶亚基、丙酮酸脱氢酶(硫辛酰胺)β、乙酰辅酶A 乙酰转移酶、烯醇化酶、醛缩酶C等表达量降低,而3-含氧酸辅酶A转移酶、4-氨基丁酸转氨酶等表达量增加.结论 实验表明,慢性脑缺血大鼠能量代谢功能及能量代谢相关酶发生了很大变化,慢性脑缺血疾病机制与线粒体具有密切关系.%Aim To study the relationship between brain mitochondria proteome and energy metabolism in rats after chronic cerebral hypoperfusion. Methods Rats models of cognitive deficits were established with permanent bilateral common carotid arteries occlusion ( 2VO ). Cerebral cortex mitochondria was isolated to study the oxidative phosphorylation function. 2D elec-trophoresis was used to explore the difference of brain mitochondria proteome. MALDI-TOF MS was used tornprovide sensitive mass spectral data for 16 unique proteins that changed in abundance between sham and 2VO rats. Results Parameters of oxidative phosphorylation in 2VO rats changed, the ADP/O rate and OPR were significantly decreased in rat brains after 2VO. In these proteins, the expression of NADH dehy-drogenase ( ubiquinone ) 1 alpha subcomplex 5, cyto-chrome c oxidase, subunit Via, pyruvate dehydrogen-ase ( lipoamide ) beta, Acetyl-CoA acetyltransferase, enolase 2, aldolase C decreased, while 3-oxoacid CoA transferase 1 and 4-aminobutyrate aminotransferase increased in 2VO rats. Conclusion These findings indicate that the change of energy metabolism itself and the enzymes related to energy metabolism in 2VO rats isrninvolved in the chronic cerebral hypoperfusion in the clinic.
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