艾灸对脑缺血再灌注损伤模型大鼠NF-κBp65蛋白表达的影响

［摘要］目的　观察艾灸对大脑皮质神经核因子κB p65(NF-κB p65)蛋白表达的影响，阐明艾灸治疗脑缺血再灌注损伤的作用机制。方法　雄性SD大鼠26只，分为假手术组(6只)、模型组(10只)和艾灸组(10只)。采用线栓法闭塞大脑中动脉2h后进行再灌注，制备局灶性脑缺血再灌注损伤大鼠模型。采用HE染色观察大鼠脑组织中炎细胞浸润情况；放射免疫分析法测定大鼠血清白介素-1β(IL-1β)和白介素-2(IL-2)的含量；Western blot技术检测大鼠大脑皮质NF-κB p65蛋白的表达。结果　与假手术组大鼠相比，模型组大鼠患侧脑组织中可见大量炎细胞浸润、血清IL-1β和IL-2含量增多、大脑皮质NF-κB p65蛋白表达量增加；而艾灸组大鼠患侧脑组织中炎细胞减少、血清IL-1β和IL-2含量降低、大脑皮质NF-κB p65蛋白表达减少。结论　减少大脑皮质NF-κB p65蛋白表达可能是艾灸治疗大鼠脑缺血再灌注损伤的作用机理之一。

［关键词］艾灸；脑缺血再灌注损伤；白介素-1β；白介素-2；神经核因子-κB

The effect of moxibustion on cortical expression of NF-κB p65 protein in rats with cerebral ischemia-reperfusion injury

［Abstract］Objective The study was to observe the effect of moxibustion on cortical expression of nuclear factor-κB(NF-κB)p65 protein in rats with cerebral ischemia-reperfusion injury.Methods Twenty-six male SD rats were randomly divided into three groups，sham group(n=6)，model group(n=10)and moxibustion group(n=10).Focal cerebral ischemia-reperfusion injury model was induced for 2h by middle cerebral artery occlusion(MCAO)followed by reperfusion.HE staining was used to examine inflammatory cells in the brain，radioimmunoassay was to calculate the concentration of interleukin-1βand interleukin-2 in theserum，and western blot was to detect the expression of NF-κB p65 protein in the cerebral cortex.Results Compared with sham group，a large number of inflammatory cells existed in the rat brain of model group，the serum concentrations of interleukin-1βand interleukin-2 were increased，and the cortical expression of NF-κB p65 protein was risen.Compared with model group，less inflammatory cells existed in the rat brain of moxibustion group，the serum concentrations of interleukin-1βand interleukin-2 were decreased，and the cortical expression of NF-κB p65 protein was reduced.Conclusion Reducing expression of NF-κB p65 protein might be one of mechanism of moxibustion attenuating focal cerebral ischemia-reperfusion injury.(www.xing528.com)

［Key words］moxibustion； cerebral ischemia-reperfusion injury； interleukin-1β； interleukin-2； nuclear factor-κB

脑血管疾病是危害人类生命和健康的主要疾病之一，缺血性脑血管病约占全部脑血管病患者的80%左右［1］，其中大多由大脑中动脉阻塞引起急性血流障碍，导致突发性局灶性脑功能障碍，引起感觉、运动功能丧失，甚至死亡。目前国际上通常采用溶栓疗法［2］以恢复脑组织血流供应，但再灌注本身有时也可加重组织的功能障碍和结构损伤，即引起脑缺血再灌注损伤。临床实践表明，艾灸治疗急性缺血性中风具有较好的疗效［3-4］。另外，前期实验研究发现，艾灸“大椎”穴能缩小大脑中动脉缺血再灌注损伤模型大鼠的脑梗死面积，减轻脑缺血再灌注损伤［5］。然而其发挥作用的机制还不明确。有研究认为，炎症反应参与了脑缺血再灌注损伤的病理过程［6］，而神经核因子-κB(nuclear factor-κB，NF-κB)在炎症反应中起着重要的调控作用，被认为是介导脑缺血后炎症级联反应的始动因素［7］。研究发现脑缺血再灌注后脑组织中NF-κB表达增加，加重脑组织损伤［8］。那么，艾灸是否通过调节大脑皮质NF-κB p65蛋白的表达而达到减轻大鼠脑缺血再灌注损伤?为此，本研究采用雄性SD大鼠，建立大脑中动脉栓塞(middle cerebral artery occlusion，MCAO)缺血再灌注损伤模型，采用Western blot技术观察艾灸对大鼠大脑皮质NF-κB p65蛋白表达的影响，以阐明艾灸治疗脑缺血再灌注损伤的机制。