羟基红花黄色素A、芍药苷单用及联合对脑缺血再灌注损伤大鼠脑组织NF-κB及炎症因子的影响

Protective effect of hydroxylsafflower yellow A and paeonifolrin on cerebral NF-κB expression and inflammation factors of rats with cerebral ischemia reperfusion injury

目的:探讨羟基红花黄色素A、芍药苷单用及联合用药对脑缺血再灌注损伤的保护作用及其机制。方法:120只SD雄性大鼠随机分为假手术组、模型组、银杏内酯5mg/kg组、羟基红花黄色素A 5mg/kg组、芍药苷5mg/kg组和羟基红花黄色素A与芍药苷联合用药组(5mg/kg+5mg/kg)。采用线栓法建立大鼠局部脑缺血再灌注模型(MCAO)。脑缺血1h再灌注6h后进行神经功能缺失评分及尾静脉注射给药;给药7天后,再次进行神经功能缺失评分;氯化三苯基四氮唑(TTC)染色检测梗死面积;酶联免疫法(ELISA)测定脑组织中IL-1β和TNF-α含量;免疫组化法(ICH)检测缺血脑组织中核转录因子NF-κB/p65的表达情况。结果:与假手术组比较,模型组大鼠治疗前神经功能评分均明显高于假手术组;用药7天后,与模型组比较,银杏内酯组(5mg/kg)、羟基红花黄色素A组(5mg/kg)、芍药苷组(5mg/kg)和合用组(5mg/kg+5mg/kg)的神经功能评分均降低,脑梗死面积显著减少,脑组织IL-1β和TNF-α含量明显减少,海马组织NF-κB p65表达明显减弱;且合用组对炎症因子表达的抑制效应更加明显。结论:羟基红花黄色素A与芍药苷联合用药对脑缺血再灌注大鼠的脑损伤具有协同保护作用,且显著优于单独使用,协同作用与下调脑组织中NF-κB的表达,减少炎性因子IL-1β和TNF-α的生成,从而减轻脑缺血后脑组织的继发性炎症反应,发挥对脑组织的保护作用有关。

Objective： To observe the protection and mechanism of hydroxysafflower yellow A and paeoniflorin in acute focal cerebral reperfosion injury in rats. Methods： 120 male SD rats were randomly divided into six groups,including the sham group,the model group,the gingko lactone group（5 mg/kg）,hydroxysafflower yellow A group（5 mg/kg）,paeoniflorin group（5 mg/kg） and hydroxysafflower yellow A combined with paeoniflorin group（5 mg/kg ＋ 5 mg/kg）. The model of rat with middle cerebral artery occlusion（MCAO） was replicated by modified thread embolism method. Nerve function loss was scored and drugs were administered by intravenous injection after 1 h cerebral ischemia and6 h reperfusion. Neurological deficit was revalued after 7 days＇ treatment. The area of cerebral infarction was detected by triphenyl tetrazolium chloride（TTC） staining. Contents of IL-1β and TNF-α in cerebral homogenate were examined by ELISA. The expression of NF-κB was examined by using immunohistochemistry. Results： Compared with the sham group,scores of pre-treatment rats in operation groups were significantly higher than those in the sham group. Compared with the model group,neurological deficits,cerebral infarction area,levels of IL-1βand TNF-α and the expression of NF-κB were all attenuated in four drug-treated groups. Meanwhile,compared with the combination of hydroxysafflor yellow A and paeoniflorin group,neurological deficits,cerebral infarction area,levels of IL-1β and TNF-α and the expression of NF-κB were increased in other three groups. Conclusion： Hydroxysafflower yellow A combined with paeoniflorin displays a synergistic protection against cerebral ischemia and reperfusion injury,which is better than that in single drug group. It may be associated with the downregulation of the expression of NF-κB,which leads to the decrease of inflammatory factor expression and the response in the cerebral tissue.