Potentially neuroprotective gene modulation in an in vitro model of mild traumatic brain injury
Potentially neuroprotective gene modulation in an in vitro model of mild traumatic brain injury
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DOI：
10.1007/s11010-012-1541-2
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发表时间：
2013-03-01
影响因子：
4.3
通讯作者：
Belli, Antonio
中科院分区：
文献类型：
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作者：
Di Pietro, Valentina;Amorini, Angela M.;Belli, Antonio
在这项研究中，我们调查的假设，轻度创伤性脑损伤（mTBI）触发一个受控的基因程序作为一种适应性反应完成神经保护，类似于在冬眠和缺血预处理。采用牵张损伤装置在大鼠海马脑片培养物中产生等双轴应变场，拉伸应变为10%，恒定应变率为20 s（-1）。损伤24 h后，分别进行碘化丙啶染色、代谢产物HPLC分析和cDNA微阵列分析，以评价细胞活力、细胞能量状态和基因表达。与对照培养物相比，10%牵张损伤的培养物的活力没有变化，但表现出低代谢状态（ATP、ATP/ADP和烟碱辅酶减少）和基因调节的特殊模式。后者的特点是下调的基因编码的蛋白质的复合物I，III和IV的线粒体电子传递链和ATP合酶;下调的转录和翻译基因;下调和上调的基因控制合成的谷氨酸和GABA受体，上调的钙调蛋白和钙调蛋白结合蛋白;适当的调制基因编码的促凋亡和抗凋亡蛋白。这些结果支持了以下假设，即在mTBI之后，在非冬眠物种中激活了冬眠型反应。与冬眠动物和缺血预处理不同，这种旨在实现最大神经保护的适应性基因程序不是由氧可用性降低触发的。在线粒体功能障碍的短暂时期，它似乎被激活以避免氧化/亚硝化应激和细胞凋亡的增加。
In this study, we investigated the hypothesis that mild traumatic brain injury (mTBI) triggers a controlled gene program as an adaptive response finalized to neuroprotection, similar to that found in hibernators and in ischemic preconditioning. A stretch injury device was used to produce an equi-biaxial strain field in rat organotypic hippocampal slice cultures at a specified Lagrangian strain of 10 % and a constant strain rate of 20 s(-1). After 24 h from injury, propidium iodide staining, HPLC analysis of metabolites and microarray analysis of cDNA were performed to evaluate cell viability, cell energy state and gene expression, respectively. Compared to control cultures, 10 % stretch injured cultures showed no change in viability, but demonstrated a hypometabolic state (decreased ATP, ATP/ADP, and nicotinic coenzymes) and a peculiar pattern of gene modulation. The latter was characterized by downregulation of genes encoding for proteins of complexes I, III, and IV of the mitochondrial electron transport chain and of ATP synthase; downregulation of transcriptional and translational genes; downregulation and upregulation of genes controlling the synthesis of glutamate and GABA receptors, upregulation of calmodulin and calmodulin-binding proteins; proper modulation of genes encoding for proapoptotic and antiapoptotic proteins. These results support the hypothesis that, following mTBI, a hibernation-type response is activated in non-hibernating species. Unlike in hibernators and ischemic preconditioning, this adaptive gene programme, aimed at achieving maximal neuroprotection, is not triggered by decrease in oxygen availability. It seems rather activated to avoid increase in oxidative/nitrosative stress and apoptosis during a transient period of mitochondrial malfunctioning.