Involvement of caspase-3 and p38 mitogen-activated protein kinase in cobalt chloride-induced apoptosis in PC12 cells
Involvement of caspase-3 and p38 mitogen-activated protein kinase in cobalt chloride-induced apoptosis in PC12 cells
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DOI：
10.1002/jnr.10168
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发表时间：
2002-03-15
影响因子：
4.2
通讯作者：
Liu, XY
中科院分区：
文献类型：
--
作者：
Zou, WG;Zeng, JP;Liu, XY
我们的前期研究表明氯化钴（CoCl_2）可诱导PC 12细胞凋亡，CoCl_2处理的PC 12细胞可作为研究缺氧相关神经元疾病机制的一个简单的体外模型。本研究旨在探讨氯化钴诱导PC 12细胞凋亡的机制。半胱氨酸天冬氨酸蛋白酶参与多种细胞类型中由各种刺激诱导的凋亡。为了研究caspase在CoCl 2诱导的PC 12细胞凋亡中的作用，我们构建了稳定表达caspase抑制基因p35的PC 12细胞，并分析了p35对CoCl 2诱导的细胞凋亡过程的影响。我们还研究了半胱天冬酶的细胞渗透性肽抑制剂的作用。结果表明，杆状病毒p35基因和caspase抑制剂Z-VAD-FMK能显著阻断CoCl 2诱导的细胞凋亡，证实caspase参与了CoCl 2诱导的细胞凋亡。进一步的研究表明，在这个过程中，半胱天冬酶-3样活性增加，如细胞切割荧光肽底物Ac-Asp-Glu-Val-Asp-7-AMC和降解DNA修复酶聚（ADP-核糖）聚合酶（PARP）（内源性半胱天冬酶-3底物）的能力所示。同时，caspase-3特异性抑制剂，即肽Ac-DEVD-CHO，Ac-DEVD-FMK，部分抑制CoCl 2诱导的细胞凋亡。这些结果表明，caspase-3或caspase-3样蛋白酶参与CoCl 2诱导的PC 12细胞凋亡。此外，我们已经观察到，另一个凋亡标志物，p38丝裂原活化蛋白激酶（MAPK），是显着激活，在这个过程中，在一个时间依赖性的方式和选择性p38 MAPK抑制剂，SB 203580，部分抑制这种细胞死亡。SB 203580的加入也部分抑制半胱天冬酶-3样活性。所有这些结果证实，CoCl 2处理的PC 12细胞是一个有用的体外模型，研究缺氧相关的神经元疾病。此外，结果表明，杆状病毒p35基因和半胱天冬酶抑制剂具有显着的能力，以拯救PC 12细胞从氯化钴诱导的细胞死亡可能有影响未来的神经保护治疗方法的缺氧相关的疾病。(C)2002 Wiley-Liss，Inc.
Our previous study showed that cobalt chloride (CoCl2) could induce PC12 cell apoptosis and that the CoCl2- treated PC12 cells may serve as a simple in vitro model for the study of the mechanism of hypoxia-linked neuronal disorders. The aim of this study is to elucidate the mechanism of CoCl2-induced apoptosis in PC12 cells. Caspases are known to be involved in the apoptosis induced by various stimuli in many cell types. To investigate the involvement of caspases in CoCl2-induced apoptosis in PC12 cells, we generated PC12 cells that stably express the viral caspases inhibitor gene p35 and analyzed the effect of p35 on the process of apoptosis induced by CoCl2. We also examined the effect of cell-permeable peptide inhibitors of caspases. The results showed that the baculovirus p35 gene and the general caspases inhibitor Z-VAD-FMK significantly block apoptosis induced by CoCl2, confirming that caspase is involved in CoCl2-induced apoptosis. Further investigation showed that in this process the caspase-3-like activity is increased, as indicated by the cells' ability to cleave the fluorogenic peptide substrate Ac-Asp-Glu-Val-Asp-7-AMC and to degrade the DNA-repairing enzyme poly(ADP-ribose) polymerase (PARP), an endogenous caspase-3 substrate. At the same time, caspase-3-specific inhibitors, namely, the peptide Ac-DEVD-CHO, Ac-DEVD-FMK, partially inhibit CoCl2-induced apoptosis. These findings suggested that caspase-3 or caspase-3-like proteases are involved in the apoptosis induced by CoCl2 in PC12 cells. Additionally, we have observed that another apoptotic marker, p38 mitogen-activated protein kinase (MAPK), is significantly activated in this process in a time-dependent manner and that a selective p38 MAPK inhibitor, SB203580, partially inhibits this cell death. The addition of SB203580 also partially suppresses caspase-3-like activity. All these results confirm that the CoCl2-treated PC12 cell is a useful in vitro model with which to study hypoxia-linked neuronal disorders. Furthermore, the results showing that the baculovirus p35 gene and caspase inhibitors possess a remarkable ability to rescue PC12 cells from CoCl2- induced cell death may have implications for future neuroprotective therapeutic approaches for the hypoxia-associated disorders. (C) 2002 Wiley-Liss, Inc.