TLR4, ATF-3 and IL8 inflammation mediator expression correlates with seizure frequency in human epileptic brain tissue
TLR4, ATF-3 and IL8 inflammation mediator expression correlates with seizure frequency in human epileptic brain tissue
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DOI：
10.1016/j.seizure.2013.04.023
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发表时间：
2013-10
期刊：
影响因子：
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通讯作者：
K. Pernhorst;S. Herms;P. Hoffmann;S. Cichon;H. Schulz;T. Sander;S. Schoch;A. Becker;A. Grote
中科院分区：
文献类型：
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作者：
K. Pernhorst;S. Herms;P. Hoffmann;S. Cichon;H. Schulz;T. Sander;S. Schoch;A. Becker;A. Grote
目的动物模型的数据已经很好地表明，中枢神经系统（CNS）的炎症过程可以调节癫痫发作频率。然而，在人类癫痫组织中，癫痫发作频率的调节和关键炎症因子的基因表达之间的潜在关系仍然没有得到解决。来自内侧颞叶癫痫（mTLE）耐药性患者的脑组织为临床-神经病理学相关性提供了独特的先决条件。在这里，我们集中在人类关键炎症介质，TLR 4，ATF-3和IL 8的基因表达，在相关的癫痫发作频率和其他临床参数在人类癫痫脑组织的耐药性mTLE患者。此外，我们的特点是细胞类型表达各自的蛋白质在epileptic campi.MethodsTotal RNA分离frommn = 26 campi的耐药性mTLE患者使用AllPrep DNA/RNA Mini Kit。使用Illumina直接杂交测定试剂盒将cRNA用于人HT-12 v3表达珠芯片上的杂交，并基于Illumina BeadStudio软件套件通过分位数标准化与背景扣除将所得基因表达数据标准化。相应的人类海马切片免疫组化探测与TLR 4，ATF-3，IL-8和胶质细胞酸性蛋白（GFAP），神经元核蛋白（NeuN）和小胶质细胞标志物HLA-DR的抗体。对于ATF-3，我们发现表达与癫痫发作频率呈负相关。IL 8低表达与高发作频率显著相关。此外，我们检测了TLR 4在耐药mTLE患者的神经元和GFAP阳性星形胶质细胞中的表达。只有人癫痫侧脑室的神经元表达ATF-3。IL 8在小胶质细胞和反应性星形胶质细胞中表达。ConclusionOur结果表明，癫痫患者的癫痫发作频率和关键炎症因子表达的差异相关性。这些过程的调节可能为治疗癫痫开辟新的治疗前景。
PurposeData from animal models has nicely shown that inflammatory processes in the central nervous system (CNS) can modulate seizure frequency. However, a potential relationship between the modulation of seizure frequency and gene expression of key inflammatory factors in human epileptic tissue is still unresolved. Brain tissue from pharmacoresistant patients with mesial temporal lobe epilepsy (mTLE) provides a unique prerequisite for clinico-neuropathological correlations. Here, we have concentrated on gene expression of the human key inflammatory mediators, TLR4, ATF-3 and IL8, in correlation to seizure frequency and additional clinical parameters in human epileptic brain tissue of pharmacoresistant mTLE patients. Furthermore, we characterized the cell types expressing the respective proteins in epileptic hippocampi.MethodsTotal RNAs were isolated fromn= 26 hippocampi of pharmacoresistant mTLE patients using AllPrep DNA/RNA Mini Kit. cRNA was used for hybridization on Human HT-12 v3 Expression BeadChips with Illumina Direct Hybridization Assay Kit and resulting gene expression data was normalized based on the Illumina BeadStudio software suite by means of quantile normalization with background subtraction. Corresponding human hippocampal sections for immunohistochemistry were probed with antibodies against TLR4, ATF-3, IL8 and glial fibrillary acidic protein (GFAP), neuronal nuclear protein (NeuN) and the microglial marker HLA-DR.ResultsWe observed abundantTLR4gene expression to relate to seizure frequency per month. ForATF-3, we found an inverse correlation of expression to seizure frequency. Lower expression ofIL8was significantly associated with high seizure frequency. Further, we detected TLR4 expression in neurons and GFAP-positive astrocytes of pharmacoresistant mTLE patients. Only neurons of human epileptic hippocampi express ATF-3. IL8 was expressed in microglia and reactive astrocytes.ConclusionOur results suggest a differential correlation of key inflammatory factor expression in epileptic hippocampi and seizure frequency in patients. The modulation of such processes may open new therapeutic perspectives for treating seizures.