Viswanath V, Wu Y, Boonplueang R, Chen S, Stevenson F F, Yantiri F, Yang L, Beal M F, Andersen J K
Buck Institute for Age Research, Novato, California 94945, USA.
J Neurosci. 2001 Dec 15;21(24):9519-28. doi: 10.1523/JNEUROSCI.21-24-09519.2001.
Parkinson's disease (PD) and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) toxicity are both associated with dopaminergic neuron death in the substantia nigra (SN). Apoptosis has been implicated in this cell loss; however, whether or not it is a major component of disease pathology remains controversial. Caspases are a major class of proteases involved in the apoptotic process. To evaluate the role of caspases in PD, we analyzed caspase activation in MPTP-treated mice, in cultured dopaminergic cells, and in postmortem PD brain tissue. MPTP was found to elicit not only the activation of the effector caspase-3 but also the initiators caspase-8 and caspase-9, mitochondrial cytochrome c release, and Bid cleavage in the SN of wild-type mice. These changes were attenuated in transgenic mice neuronally expressing the general caspase inhibitor protein baculoviral p35. These mice also displayed increased resistance to the cytotoxic effects of the drug. MPTP-associated toxicity in culture was found temporally to involve cytochrome c release, activation of caspase-9, caspase-3, and caspase-8, and Bid cleavage. Caspase-9 inhibition prevented the activation of both caspase-3 and caspase-8 and also inhibited Bid cleavage, but not cytochrome c release. Activated caspase-8 and caspase-9 were immunologically detectable within MPP(+)-treated mesencephalic dopaminergic neurons, dopaminergic nigral neurons from MPTP-treated mice, and autopsied Parkinsonian tissue from late-onset sporadic cases of the disease. These data demonstrate that MPTP-mediated activation of caspase-9 via cytochrome c release results in the activation of caspase-8 and Bid cleavage, which we speculate may be involved in the amplification of caspase-mediated dopaminergic cell death. These data suggest that caspase inhibitors constitute a plausible therapeutic for PD.
帕金森病(PD)和1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)毒性均与黑质(SN)中的多巴胺能神经元死亡有关。凋亡被认为与这种细胞损失有关;然而,它是否是疾病病理学的主要组成部分仍存在争议。半胱天冬酶是参与凋亡过程的一类主要蛋白酶。为了评估半胱天冬酶在PD中的作用,我们分析了MPTP处理的小鼠、培养的多巴胺能细胞和死后PD脑组织中的半胱天冬酶激活情况。发现MPTP不仅能引发效应半胱天冬酶-3的激活,还能引发起始半胱天冬酶-8和半胱天冬酶-9的激活、线粒体细胞色素c释放以及野生型小鼠SN中的Bid裂解。在神经元中表达通用半胱天冬酶抑制剂蛋白杆状病毒p35的转基因小鼠中,这些变化有所减弱。这些小鼠对药物的细胞毒性作用也表现出更强的抵抗力。发现在培养物中与MPTP相关的毒性在时间上涉及细胞色素c释放、半胱天冬酶-9、半胱天冬酶-3和半胱天冬酶-8的激活以及Bid裂解。半胱天冬酶-9抑制可阻止半胱天冬酶-3和半胱天冬酶-8的激活,还可抑制Bid裂解,但不影响细胞色素c释放。在经MPP(+)处理的中脑多巴胺能神经元、经MPTP处理的小鼠的多巴胺能黑质神经元以及该疾病晚发性散发病例的尸检帕金森病组织中,可通过免疫检测到激活的半胱天冬酶-8和半胱天冬酶-9。这些数据表明,MPTP通过细胞色素c释放介导的半胱天冬酶-9激活导致半胱天冬酶-8的激活和Bid裂解,我们推测这可能参与了半胱天冬酶介导的多巴胺能细胞死亡的放大过程。这些数据表明,半胱天冬酶抑制剂可能是一种合理的PD治疗方法。