Reassessment of caspase inhibition to augment grafted dopamine neuron survival
Reassessment of caspase inhibition to augment grafted dopamine neuron survival
复制标题
DOI：
10.3727/000000004783983972
复制
发表时间：
2004-01-01
影响因子：
3.3
通讯作者：
Sortwell, CE
中科院分区：
文献类型：
--
作者：
Marchionini, DM;Collier, TJ;Sortwell, CE
帕金森病(PD)的一种实验性治疗方法是胚胎腹侧中脑组织移植。不幸的是。高达95%的移植神经元死亡，其中许多是通过细胞凋亡。激活的半胱氨酸天冬氨酸酶在细胞凋亡途径的执行中起着关键作用：因此，暴露于半胱氨酸天冬氨酸酶抑制剂可能提供一种有效的干预策略，以防止细胞凋亡。在本研究中，我们检测了两种不同的caspase抑制剂，caspase-1抑制剂Ac-YVAD-CMK和Caspase-3抑制剂Ac-DEVD-CMK的疗效。目的：增强大鼠中脑酪氨酸羟化酶免疫反应(TH-IR)神经元在培养和植入失神经纹状体后的存活。我们报道，在低密度培养的中脑培养中，Ac-YVAD-CMK对TH-ir神经元有部分但不显著的保护作用，而在高密度培养中，两种caspase抑制剂都没有促进TH-ir神经元的存活。模拟嫁接密度。我们证明，细胞培养程序(全井和微岛)和细胞密度直接影响血清撤除后TH-IR神经元所经历的损伤程度。这种不同程度的伤害直接影响到caspase抑制是否会增加TH-IR神经元的存活。我们的移植实验表明，AC-YVAD-CMK在移植前加入中脑细胞悬液或在移植前加入体外培养的中脑聚集体3天，并不能增加移植的TH-IR神经元的存活率。这些实验提供了进一步的证据，证明这些caspase抑制剂未能提高TH-ir神经元的存活率。更重要的是。我们建议，用于模拟移植范例的细胞培养范例必须更接近于中脑移植的细胞密度，以有效地筛选潜在的强化治疗。
One experimental therapy for Parkinson's disease (PD) is the transplantation of embryonic ventral mesencephalic tissue. Unfortunately. up to 95% of grafted neurons die, many via apoptosis. Activated caspases play a key role in execution of the apoptotic pathway: therefore, exposure to caspase inhibitors may provide an effective intervention strategy for protection against apoptotic cell death. In the present study we examined the efficacy of two different caspase inhibitors, caspase-1 inhibitor Ac-YVAD-CMK and caspase-3 inhibitor Ac-DEVD-CMK. to augment mesencephalic tyrosine hydroxylase-immunoreactive (TH-ir) neuron survival in culture and following implantation into the denervated striatum of rats. We report that treatment with Ac-YVAD-CMK provided partial but nonsignificant protection for TH-ir neurons against serum withdrawal in mesencephalic cultures plated at low density, while neither caspase inhibitor promoted TH-ir neuron survival in higher density cultures. simulating graft density. We demonstrate that plating procedures (full well vs. microislands) and cell density directly affect the degree of insult experienced by TH-ir neurons following serum withdrawal. This varying degree of insult directly impacts whether caspase inhibition will augment TH-ir neuron survival. Our grafting experiments demonstrate that Ac-YVAD-CMK does not augment grafted TH-ir neuron survival when added to mesencephalic cell suspensions prior to grafting or to mesencephalic reaggregates for 3 days in vitro prior to transplantation. These experiments provide further evidence of the failure of these caspase inhibitors to augment TH-ir neuron survival. Furthermore. we suggest that cell culture paradigms used to model grafting paradigms must more closely approximate the cell densities of mesencephalic grafts to effectively screen potential augmentative treatments.