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环加氧酶-2在大鼠原代皮质神经细胞缺氧损伤中的作用

Role of cyclooxygenase-2 in injury induced by hypoxia/reoxygenation in cultured rat cortical neurons

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【作者】 张艳桥陈仁武徐长庆Graham H Steven

【Author】 ZHANG Yan-qiao1, CHEN Ren-wu2, XU Chang-qing3, Graham H Steven2 (1Department of Geriatrics, The Second Hospital of Harbin Medical University, Harbin 150086, China; ?2?Department of Neurology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15213, USA; ?3?Department of Pathophysiology, Harbin Medical University, Harbin 150086, China)

【机构】 哈尔滨医科大学附属第二医院老年病科美国匹兹堡大学神经病科美国匹兹堡大学神经病科 黑龙江哈尔滨150086匹兹堡15213黑龙江哈尔滨150086匹兹堡15213

【摘要】 目的观察环加氧酶-2(cyclooxygenase-2,COX-2)在原代培养的大鼠皮质神经细胞缺氧-再给氧损伤中的作用以及COX-2特异性抑制剂NS398的保护作用。方法原代培养大鼠皮质神经细胞,随机分为对照组(未处理组)、缺氧再给氧组、COX-2特异性抑制剂NS398+缺氧再给氧组。用噻唑蓝(MTT)比色法测定细胞生存率,用Western印迹法检测COX-2蛋白质的表达,用DNA凝胶电泳法检测细胞凋亡情况。结果培养的大鼠原代皮质神经细胞经缺氧再给氧处理后,COX-2蛋白质表达水平明显高于对照组及缺氧再给氧+NS398组(P<005);缺氧再给氧+NS398予处理组神经细胞生存率明显高于单独缺氧再给氧组(P<005和P<001),DNA凝胶电泳显示DNA断裂显著减轻。结论COX-2参与缺氧再给氧后神经细胞凋亡的病理过程,COX-2特异性抑制剂明显减轻缺氧再给氧后神经细胞的损伤,保护神经细胞免遭缺氧诱导的细胞凋亡。

【Abstract】 AIM: To observe the role of cyclooxygenase-2 (COX-2) in injury induced by hypoxia and reoxygenation in cultured rat cortical neurons and protective effects of COX-2 specific inhibitor NS398.METHODS: Primary rat cortical neuronal cells were cultured. Experiments were divided into control group, hypoxia/reoxygenation group and hypoxia/reoxygenation with COX-2 inhibitor group. Cell viability was measured by MTT assay. COX-2 protein expression was examined by Western blotting. Apoptosis was measured by DNA agarose electrophoresis.RESULTS: The expression levels of COX-2 increased significantly after neurons were treated with hypoxia and reoxygenation, compared with control group and hypoxia/reoxygenation with COX-2 inhibitor group (P<0.05). COX-2 specific inhibitor NS398 protected neurons from death (P<0.05 and P<0.01), DNA fragmentation analysis showed DNA fragmentation was inhibited significantly by NS398.CONCLUSION: COX-2 is involved in the pathogenesis of neuron apoptosis induced by hypoxia/reoxygenation. COX-2 specific inhibitor significantly protects cortical neurons against hypoxia/reoxygenation injury and inhibits apoptosis induced by hypoxia. [

【基金】 黑龙江省教育厅海外学人科研资助项目(1054HQ016)
  • 【文献出处】 中国病理生理杂志 ,Chinese Journal of Pathophysiology , 编辑部邮箱 ,2004年12期
  • 【分类号】R363
  • 【被引频次】6
  • 【下载频次】107
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