The expression pattern of ADP-ribosyltransferase 3 in rat traumatic brain injury
The expression pattern of ADP-ribosyltransferase 3 in rat traumatic brain injury
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ADP-核糖基转移酶3在大鼠脑外伤中的表达模式
DOI：
10.1007/s10735-011-9366-y
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发表时间：
2012-02-01
影响因子：
3.2
通讯作者：
Chen, Jian
中科院分区：
文献类型：
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作者：
Shi, Wei;Gong, Peipei;Chen, Jian
哺乳动物胞外ADP-核糖基转移酶（ARTs）通过催化NAD（+）上的ADP-核糖部分转移到靶蛋白的特定氨基酸上，调节多种细胞的生物学功能。ART 3是已知的ART家族的成员，其参与细胞分裂、DNA修复和炎症反应的调节。为了阐明ART 3在中枢神经系统损伤和修复中的表达、细胞定位和可能的功能，我们在成年大鼠中建立了急性创伤性脑损伤模型。Western blot结果显示，创伤性脑损伤（TBI）后，同侧大脑皮质中ART 3的表达增加，第3天达到高峰，随后逐渐下降。而对侧大脑皮层无明显改变。免疫组织化学显示，与对侧大脑皮质相比，ART 3在同侧大脑中高度显著的积累。免疫荧光双标记显示，ART 3主要定位于神经元的质膜，而不是在星形胶质细胞或小胶质细胞在3毫米内的损伤部位在伤后3天。此外，我们还检测了该TBI模型中caspase-3和生长相关蛋白43（GAP-43）的表达谱，其变化与ART 3的表达谱相关。此外，在NeuN阳性细胞中分别检测到ART 3/活性caspase-3和ART 3/GAP 43的共定位。此外，用H2 O2处理嗜铬细胞瘤（PC 12）细胞，以建立凋亡模型。结果显示，ART 3的表达呈浓度和时间依赖性增加。为了进一步检查ART 3在PC 12细胞凋亡中的参与，将3-甲氧基苯甲酰胺用于用膜联蛋白V和PI染色的凋亡细胞的流式细胞术分析。与未处理组相比，使用3-甲氧基苯甲酰胺的实验组具有相对低水平的凋亡指数。结合以往的报道，我们推测ART 3可能在TBI后的CNS病理生理学中发挥重要作用，需要进一步研究以更好地了解其功能和机制。
Mammalian ecto ADP-ribosyltransferases (ARTs) can regulate the biological functions of various types of cells by catalyzing the transfer of single ADP-ribose moiety from NAD(+) to a specific amino acid in a target protein. ART3 is a member of the known ART family which is involved in cell division, DNA-repair and the regulation of the inflammatory response. To elucidate the expression, cellular localization and possible functions of ART3 in central nervous system (CNS) lesion and repair, we performed an acute traumatic brain injury model in adult rats. Western blot analysis showed that the expression of ART3 in ipsilateral brain cortex increased, then reached a peak at day 3 after traumatic brain injury (TBI), and gradually declined during the following days. But in the contralateral brain cortex, no obvious alterations were observed. Immunohistochemistry revealed the highly significant accumulation of ART3 at the ipsilateral brain in comparison to contralateral cerebral cortex. Double immunofluorescence labeling suggested that ART3 was localized mainly in the plasmalemma of neurons, but not in astrocytes or microglias within 3 mm from the lesion site at day 3 post-injury. In addition, we detected the expression profiles of caspase-3 and growth associated protein 43 (GAP-43) whose changes were correlated with the expression profiles of ART3 in this TBI model. Besides, co-localization of ART3/active caspase-3 and ART3/GAP43 were detected in NeuN-positive cells, respectively. Moreover, Pheochromocytoma (PC12) cells were treated with H2O2 to establish an apoptosis model. The results showed that the expression of ART3 was increased in the concentration and time dependence way. To further examine the involvement of ART3 in apoptosis of PC12, 3-Methoxybenzamide was used in flow cytometry analysis of apoptotic cells stained with Annexin V and PI. The experimental group in which 3-Methoxybenzamide used had a relative low level of apoptotic index compared with the untreated group. Together with previous reports, we hypothesize that ART3 may play important roles in CNS pathophysiology after TBI and further research is needed to have a good understanding of its function and mechanism.