补阳还五汤类方对大鼠脑缺血再灌注损伤炎症的保护作用

Protective effect of Buyang Huanwu Decoction categorized formula on cerebral ischemia-reperfusion injury in rats

目的:探究补阳还五汤类方对大鼠脑缺血再灌注损伤(CIRI)炎症的保护作用。方法:建立大鼠大脑中动脉栓塞(MCAO)模型,随机将大鼠分为假手术组、模型组、补阳还五汤组(4.11g·kg^(-1)·d^(-1))、养阴通脑颗粒组(3.25g·kg^(-1)·d^(-1))和脑心通胶囊组(0.15g·kg^(-1)·d^(-1)),造模结束后灌胃给药,24h后进行神经功能评分;TTC染色测定大鼠脑梗死体积;HE染色观察大鼠海马CA1区病理状态改变;ELISA试剂盒检测肿瘤坏死因子(TNF-α)和白细胞介素1β(IL-1β)的含量变化;Western blot和免疫组化法检测TLR4/NF-κB信号通路中关键蛋白、TLR4、IκBa、NF-κB p65的表达。结果:与模型组比较,补阳还五汤类方均可改善大鼠的神经功能缺损现象(P<0.05)和海马CA1区的病理状态,减小大鼠脑梗死百分比(P<0.01),抑制小胶质细胞的激活(P<0.01),升高IκBα(P<0.01),降低TLR4、NF-κB p65等关键蛋白的表达量和炎症因子TNF-α、IL-1β的释放(P<0.01)。3种中药复方对比发现,脑心通胶囊对大鼠炎症的改善效果较为明显。结论:补阳还五汤类方均可改善CIRI大鼠的炎症反应,其中脑心通胶囊在大鼠炎症急性期炎症损伤的药物治疗效果相对显著,其机制可能与TLR4/NF-κB信号通路有关。

Objective: To investigate the protective effect of Buyang Huanwu Decoction categorized formula on cerebral ischemia-reperfusion injury(CIRI) inflammation in rats. Methods: The rat middle cerebral artery occlusion(MCAO) model was establised. The rats were randomly divived into sham operation group, model group, Buyang Huanwu Decoction group(4.11 g·kg^(-1)·d^(-1)). Yangyin Tongnao Granules group(0.15 g·kg^(-1)·d^(-1)) and Naoxintong Capsule group(3.25 g·kg^(-1)·d^(-1)), Rats were administrated by gavage. Score neurological deficit was evaluated after 24 h;The cerebral infarction volume was determined by TTC staining;HE staining was used to detect the pathological changes of rat hippocampal CA1 area;The changes of tumor necrosis factor(TNF-α)and interleukin-1β(IL-1β) were detected by enzyme-linked immunosorbent assay(ELISA);Western blot and immunohistochemistry were used to detect the expression of key proteins in TLR4/NF-κB signaling pathway(TLR4, IκBα, NF-κB p65). Results: Compared to the model group, Buyang Huanwu Decoction categorized formula could improve the neurological deficit of rats(P<0.05) and the pathological state of hippocampal CA1 area, reduce the infarct volume of rats(P<0.01), inhibit the activation of microglia(P<0.01), improve the expression of IκBα(P<0.01), reduce the expression of TLR4, NF-κB p65 and other key proteins and the release of inflammatory factors TNF-α and IL-1β(P<0.01). The comparison of the three traditional Chinese medicine compounds found that the Naoxintong Capsule was more significant for cerebral ischemia reperfusion injury in rats. Conclusion: Buyang Huanwu Decoction categorized formula can improve the inflammation in rats with cerebral ischemia-reperfusion injury and the drug treatment effect of Naoxintong Capsule in the acute phase of inflammation in rats is relatively significant, its mechanism may be related to TLR4/NF-κB signaling pathway.