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背景:任何生理状况的改变和表型的变化,最初的基础激发点是基因表达的改变,基因芯片技术是利用碱基配对的原理来快速、大量筛选目标基因的新方法,能整体宏观地研究生物体基因的表达及功能。目的:应用基因芯片技术研究大鼠脑出血早期差异表达基因,为探讨脑出血的病理机制奠定理论基础。设计:随机对照实验。单位:川北医学院附属医院神经内科。材料:实验于2002-10/2003-12在川北医学院附属医院完成。选取20只无特殊病原体级Wistar大鼠,雌雄各半,体质量220～260g,由重庆医科大学实验动物中心提供,将全部大鼠随机分成对照组和脑出血组,每组10只。方法:采用Ⅶ型胶原酶立体定位法制备大鼠急性脑出血动物模型,模型建立后4h取血肿周围组织和相同部位的正常脑组织进行基因芯片对照检测,用扫描仪扫描芯片荧光信号并进行计算机分析,用反转录-聚合酶链反应来考证基因表达谱的结果。主要观察指标:大鼠脑组织基因芯片检测结果及基因反转录-聚合酶链反应的计算结果。结果:大鼠急性脑出血后4h有差异表达基因129个,上调基因114个,下调基因15个,这些基因主要涉及到应激和免疫应答、细胞凋亡、能量代谢及信号转导。有关炎性损害的基因上调最为明显。反转录-聚合酶链反应计算结果显示,基因表示水平与芯片检测结果相符,说明基因芯片所建立的基因表达谱具有相当的可靠性。结论:脑出血早期存在多个差异表达基因,这些差异表达的基因可能在出血性脑损伤中发挥重要作用。
Background: Physiological changes and changes in phenotype, the initial basis of the trigger point is the change of gene expression, gene chip technology is the use of base pairing principle to quickly and massively screen the target gene of the new method, the overall macro-graduate students Object gene expression and function. OBJECTIVE: To study differentially expressed genes in early stage of ICH in rats by gene chip technique, and to lay a theoretical foundation for exploring the pathological mechanism of ICH. Design: Randomized controlled experiment. SETTING: Department of Neurology, Affiliated Hospital of North Sichuan Medical College. Materials: The experiment was performed at Affiliated Hospital of North Sichuan Medical College from October 2002 to December 2003. Twenty non-specific pathogen-grade Wistar rats were randomly divided into control group and intracerebral hemorrhage group, with 10 animals in each group. The body weight was 220-260g and provided by Experimental Animal Center of Chongqing Medical University. All rats were randomly divided into control group and intracerebral hemorrhage group. Methods: Rat model of acute intracerebral hemorrhage was established by Ⅶ collagenase stereotactic method. After establishment of the model, the surrounding tissues of hematoma and normal brain tissue at the same site were taken for 4h to detect the gene chip. The fluorescence signal of the chip was scanned with a scanner Analysis, reverse transcription - polymerase chain reaction to test the results of gene expression profiling. MAIN OUTCOME MEASURES: The results of gene chip detection in rat brain and the results of gene reverse transcription - polymerase chain reaction. Results: There were 129 differentially expressed genes, 114 up-regulated genes and 15 down-regulated genes at 4h after acute cerebral hemorrhage in rats. These genes mainly involved in stress and immune response, apoptosis, energy metabolism and signal transduction. The genes that cause inflammatory damage are most noticeably up-regulated. Reverse transcriptase-polymerase chain reaction results showed that the level of gene expression was in good agreement with the chip test results, indicating that the gene expression profile established by the gene chip is of considerable reliability. Conclusion: There are many differentially expressed genes in early stage of ICH. The differentially expressed genes may play an important role in hemorrhagic brain injury.