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观察羟乙葛根素对大鼠局灶性脑缺血再灌注损伤后TNF-α表达及NF-κB活性的影响。采用大鼠大脑中动脉内栓线阻断法(MCAO)建立大鼠脑缺血再灌注损伤模型,分别于缺血前30min及再灌注即刻由尾静脉注射羟乙葛根素(10,20及40mg.kg-1),缺血2h再灌注24h后取缺血侧脑组织,HE染色观察大鼠脑组织病理学变化并计数海马CA1区存活神经元数目,放射免疫分析测定脑组织匀浆中TNF-α含量,逆转录聚合酶链式反应(RT-PCR)测定脑组织中TNF-α mRNA表达情况,凝胶电泳迁移率实验(EMSA)观察NF-κB DNA结合活性改变,Western blotting检测观察IκBα蛋白表达情况。羟乙葛根素可明显改善大鼠海马CA1区损伤程度,升高锥体存活神经元数目,减少TNF-α蛋白及mRNA表达,抑制NF-κB DNA结合活性。羟乙葛根素可减轻大鼠脑缺血再灌注损伤后炎症反应,这可能是其发挥脑保护作用的机制之一。
To observe the effect of hydroxyethylpuerarin on TNF-α expression and NF-κB activity after focal cerebral ischemia-reperfusion injury in rats. The cerebral ischemic reperfusion injury model was established by middle cerebral artery occlusion (MCAO) in rats. The hydroxyethylpuerarin (10, 20 and 40 mg) was injected through the tail vein immediately 30 min before ischemia and immediately after reperfusion. .kg-1). Ischemic lateral cerebral tissue was taken after ischemia for 24 hours after reperfusion for 2 hours. HE staining was used to observe the pathological changes of rat brain and the number of surviving neurons in hippocampal CA1 area was counted. Radioimmunoassay was used to determine TNF in brain homogenate. -α content, reverse transcription polymerase chain reaction (RT-PCR) determination of TNF-α mRNA expression in brain tissue, electrophoretic mobility shift assay (EMSA) observed changes in NF-κB DNA binding activity, Western blotting detection of IκBα Protein expression. Hydroxy-ethylpuerarin can significantly improve the extent of injury in hippocampal CA1 region, increase the number of neurons in the pyramidal pyramidal neurons, reduce the expression of TNF-α protein and mRNA, and inhibit the DNA binding activity of NF-κB. Hydroxyethylpuerarin can reduce the inflammatory reaction after cerebral ischemia-reperfusion injury in rats, which may be one of the mechanisms that play a role in brain protection.