Damodaran Tirupapuliyur V, Greenfield Stephen T, Patel Anand G, Dressman Holly K, Lin Siomon K, Abou-Donia Mohamed B
Department of Pharmacology and Cancer Biology, Duke University Medical Center, P.O. Box 3813, Durham, NC, 27710, USA.
Neurochem Res. 2006 Mar;31(3):367-81. doi: 10.1007/s11064-005-9023-5. Epub 2006 May 13.
We have studied sarin-induced global gene expression patterns at an early time point (2 h: 0.5 x LD50) using Affymetrix Rat Neurobiology U34 chips and male Sprague-Dawley rats. A total of 46 genes showed statistically significant alterations from control levels. Three gene categories contained more of the altered genes than any other groups: ion channel (8 genes) and calcium channel and binding proteins (6 genes). Alterations were also found in the following gene groups: ATPases and ATP-based transporters (4), growth factors (4), G-protein-coupled receptor pathway-related molecules (3), neurotransmission and neurotransmitter transporters (3), cytoskeletal and cell adhesion molecules (2), hormones (2), mitochondria-associated proteins (2), myelin proteins (2), stress-activated molecules (2), cytokine (1), caspase (1), GABAnergic (1), glutamergic (1), immediate early gene (1), prostaglandin (1), transcription factor (1), and tyrosine phosphorylation molecule (1). Persistent alteration of the following genes also were noted: Arrb1, CaMKIIa, CaMKIId, Clcn5, IL-10, c-Kit, and Plp1, suggesting altered GPCR, kinase, channel, and cytokine pathways. Selected genes from the microarray data were further validated using relative RT-PCR. Some of those genes (GFAP, NF-H, CaMKIIa, Calm, and MBP) have been shown by other laboratories and ours, to be involved in the pathogenesis of sarin-induced pathology and organophosphate-induced delayed neurotoxicity (OPIDN). Induction of both proapoptotic (Bcl2l11, Casp6) and antiapoptotic (Bcl-X) genes, besides suppression of p21, suggest complex cell death/protection-related mechanisms operating early on. Principal component analysis (PCA) of the expression data confirmed that the changes in gene expression are a function of sarin exposure, since the control and treatment groups separated clearly. Our model (based on current and previous studies) indicates that both degenerative and regenerative pathways are activated early and contribute to the level of neurodegeneration at a later time, leading to neuro-pathological alterations.
我们使用Affymetrix大鼠神经生物学U34芯片和雄性Sprague-Dawley大鼠，在早期时间点（2小时：0.5倍半数致死剂量）研究了沙林诱导的全基因组表达模式。共有46个基因表现出与对照水平相比具有统计学意义的改变。三个基因类别包含的改变基因比其他任何组都多：离子通道（8个基因）以及钙通道和结合蛋白（6个基因）。在以下基因组中也发现了改变：ATP酶和基于ATP的转运蛋白（4个）、生长因子（4个）、G蛋白偶联受体途径相关分子（3个）、神经传递和神经递质转运蛋白（3个）、细胞骨架和细胞粘附分子（2个）、激素（2个）、线粒体相关蛋白（2个）、髓磷脂蛋白（2个）、应激激活分子（2个）、细胞因子（1个）、半胱天冬酶（1个）、γ-氨基丁酸能（1个）、谷氨酸能（1个）、即早基因（1个）、前列腺素（1个）、转录因子（1个）以及酪氨酸磷酸化分子（1个）。还注意到以下基因的持续改变：Arrb1、CaMKIIa、CaMKIId、Clcn5、IL-10、c-Kit和Plp1，提示GPCR、激酶、通道和细胞因子途径发生改变。使用相对逆转录聚合酶链反应（RT-PCR）对微阵列数据中选定的基因进行了进一步验证。其他实验室和我们的实验室已表明，其中一些基因（胶质纤维酸性蛋白、神经丝重链、CaMKIIa、钙调蛋白和髓磷脂碱性蛋白）参与沙林诱导的病理和有机磷酸酯诱导的迟发性神经毒性（OPIDN）的发病机制。除了p21受到抑制外，促凋亡基因（Bcl2l11、Casp6）和抗凋亡基因（Bcl-X）的诱导表明早期存在复杂的细胞死亡/保护相关机制。对表达数据进行主成分分析（PCA）证实，基因表达的变化是沙林暴露的函数，因为对照组和治疗组明显分开。我们的模型（基于当前和先前的研究）表明，退化和再生途径在早期均被激活，并在后期对神经退行性变的程度产生影响，导致神经病理改变。
