Dramatic increase in readthrough acetylcholinesterase in a cellular model of oxidative stress
Dramatic increase in readthrough acetylcholinesterase in a cellular model of oxidative stress
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DOI：
10.1111/j.1471-4159.2010.07164.x
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发表时间：
2011-03-01
影响因子：
4.7
通讯作者：
Zimmermann, Martina
中科院分区：
文献类型：
--
作者：
Haertl, Ricarda;Gleinich, Anne;Zimmermann, Martina
P>使用叔丁基过氧化氢作为氧化剂，在SH-SY 5 Y细胞中实现中度、短暂的氧化应激。在24小时的恢复期内，监测乙酰胆碱酯酶（AChE）剪接变体加尾AChE（AChE-T）和通读AChE（AChE-R）的酶活性和蛋白水平。评估了它们与促凋亡因子和抗凋亡因子（即分别为胱天蛋白酶3和Bcl-2）以及乳酸脱氢酶释放（作为细胞活力的量度）的时间依赖性相关性。与AChE-R相比，记录到AChE-T的明显不同的表达模式，因为AChE-T在6小时的时间段内仅显示出非常轻微的增加。相反，AChE-R在恢复期间持续上升，在应激后3-4小时达到细胞内峰值水平，比对照水平高出6倍，并从细胞中大量释放。此外，抗凋亡Bcl-2显著增加，在该AChE-R峰发生后2-3小时达到峰值。我们相信，这项研究提出了第一个工作，证明-不依赖于技术的过度表达-凋亡过程和相关的救援机制，涉及乙酰胆碱酯酶亚型在神经元细胞系之间的时间依赖性相关性。
P>Moderate, transient oxidative stress is achieved in SH-SY5Y cells using tertiary butylhydroperoxide as oxidant. Over a recovery period of 24 h, the enzymatic activity and protein levels of the acetylcholinesterase (AChE) splice variants tailed AChE (AChE-T) and readthrough AChE (AChE-R) are monitored. Their time-dependent correlation to pro- and anti-apoptotic factors, namely caspase 3 and Bcl-2, respectively, as well as lactate dehydrogenase release as a measure of cell viability is assessed. A distinctly different expression pattern of AChE-T as compared with AChE-R is recorded, in that AChE-T shows only a very slight increase over a 6 h time period. In contrast, AChE-R rises continuously during the recovery period, reaching peak intracellular levels that are up to six times higher than control levels 3-4 h post-stress, and is released from cells in substantial amounts. Moreover, anti-apoptotic Bcl-2 increases significantly, peaking 2-3 h after this AChE-R peak has occurred. We believe this study presents the first work that demonstrates - without relying on techniques of over-expression - the time-dependent correlation between apoptotic processes and related rescue mechanisms involving AChE isoforms in a neuronal cell line.