Sathasivam S, Grierson A J, Shaw P J
Laboratory of origin: Academic Neurology Unit, Section of Neuroscience, Division of Genomic Medicine, The Medical School, Beech Hill Road, University of Sheffield, Sheffield, UK.
Neuropathol Appl Neurobiol. 2005 Oct;31(5):467-85. doi: 10.1111/j.1365-2990.2005.00658.x.
There is increasing evidence that apoptosis or a similar programmed cell death pathway is the mechanism of cell death responsible for motor neurone degeneration in amyotrophic lateral sclerosis. Knowledge of the relative importance of different caspases in the cell death process is at present incomplete. In addition, there is little information on the critical point of the death pathway when the process of dying becomes irreversible. In this study, using the well-established NSC34 motor neurone-like cell line stably transfected with empty vector, normal or mutant human Cu-Zn superoxide dismutase (SOD1), we have characterized the activation of the caspase cascade in detail, revealing that the activation of caspases-9, -3 and -8 are important in motor neurone death and that the presence of mutant SOD1 causes increased activation of components of the apoptotic cascade under both basal culture conditions and following oxidative stress induced by serum withdrawal. Activation of the caspases identified in the cellular model has been confirmed in the G93A SOD1 transgenic mice. Furthermore, investigation of the effects of anti-apoptotic neuroprotective agents including specific caspase inhibitors, minocycline and nifedipine, have supported the importance of the mitochondrion-dependent apoptotic pathway in the death process and revealed that the upstream caspase cascade needs to be inhibited if useful neuro-protection is to be achieved.
越来越多的证据表明,凋亡或类似的程序性细胞死亡途径是肌萎缩侧索硬化症中运动神经元变性的细胞死亡机制。目前,关于不同半胱天冬酶在细胞死亡过程中的相对重要性的认识尚不完整。此外,关于死亡途径中细胞死亡过程变得不可逆转的关键点的信息很少。在本研究中,我们使用稳定转染空载体、正常或突变型人铜锌超氧化物歧化酶(SOD1)的成熟NSC34运动神经元样细胞系,详细表征了半胱天冬酶级联反应的激活,发现半胱天冬酶-9、-3和-8的激活在运动神经元死亡中很重要,并且突变型SOD1的存在会导致在基础培养条件下以及血清剥夺诱导的氧化应激后凋亡级联反应成分的激活增加。在G93A SOD1转基因小鼠中已证实了细胞模型中鉴定出的半胱天冬酶的激活。此外,对包括特异性半胱天冬酶抑制剂、米诺环素和硝苯地平在内的抗凋亡神经保护剂的作用的研究支持了线粒体依赖性凋亡途径在死亡过程中的重要性,并表明如果要实现有效的神经保护,需要抑制上游半胱天冬酶级联反应。