【摘要】 目的探讨模拟缺血再灌注引起神经元凋亡的途径和褪黑素(m elaton in,MT)抗凋亡的作用机理。方法建立原代培养大鼠小脑颗粒细胞的体外模拟缺血(Oxygen G lucose Deprivation,OGD)再灌注模型,测定培养液LDH活性和细胞DNA琼脂糖凝胶电泳;利用Rhodam ine123和激光共聚焦显微镜观察线粒体膜电位的变化;ELISA检测细胞浆中细胞色素C水平;用同样指标观察MT对其损伤的保护作用。结果在体外模拟缺血再灌注模型中培养液LDH活性增加(P<0.05),琼脂糖凝胶电泳DNA出现梯状条带,线粒体膜电位明显降低,细胞浆中细胞色素C含量增加(P<0.05),MT对上述现象有明显的抑制作用。结论(1)缺血再灌注引起的神经元凋亡机理之一是通过线粒体凋亡途径;(2)MT可通过阻止线粒体凋亡途径而保护模拟缺血再灌注诱导小脑颗粒细胞的凋亡。更多还原

【Abstract】 Objective To explore whether melatonin(MT) can protect neurons from apoptosis induced by ischemia-reperfusion injury and its mechanism.Methods Cerebellar granule cells in SD rats were islated and cultured,and mimic ischemia-reperfusion model was established by oxygen-glucose-deprivation(OGD) in vitro.The mensuration of LDH activity and DNA gel electrophresis were performed to determine the cell damage.The measurement of mitochondrial membrane potentiometer was used Rhodamine 123 and laser co-focus microscope.Cytochrome c was assayed by ELISA.The effects of MT on the neurodegeneration induced by OGD was observed.Results In the mimic ischemia-reperfusion model of cerebellar granule cells,LDH activity in the culture increased,DNA fragmentation was observed.The mitochondrial membrane potentiometer was depressed obviously and cytochrome c was released from mitochondria to cytosol.However,MT could restrain the effects that we describe above.Conclusions(1) The neuronal apoptosis induced by OGD is partly mediated by the mitochondrial apoptosis pathway.(2) MT can suppress the apoptosis of cerebellar granule cells by preventing the mitochondrial apoptosis pathway.更多还原