苦碟子注射液对大鼠脑缺血再灌注损伤炎症及TLR-4/NF-κB信号的影响

Effect of Kudiezi Injection on inflammation and TLR-4/NF-κB signal in cerebral ischemia reperfusion rats

目的考察苦碟子注射液对大鼠脑缺血再灌注损伤的保护作用,并从炎症角度探讨其保护机制。方法改良线栓法建立大鼠大脑中动脉栓塞(MCAO)模型,激光多普勒血流仪(LDF)监测大鼠脑血流变化。雄性SD大鼠随机分为假手术、模型组及苦碟子注射液高、低剂量组。缺血2 h再灌注24 h后进行神经功能缺损评分;断头取闹,称脑湿质量,计算脑指数;TTC染色法检测脑梗死面积;HE染色观察脑组织病理形态;并取血、脑组织,Elisa试剂盒检测炎症因子表达,Western Blotting技术检测TLR-4、NF-κB蛋白表达。结果与假手术组比较,模型组神经功能缺损较严重(P<0.01),脑指数和脑梗死面积也显著升高;给予苦碟子注射液后能改善神经功能缺损症状(P<0.01),显著降低模型组的脑指数和脑梗死面积,减少神经元坏死,同时苦碟子注射液还可显著降低脑匀浆和血清TNF-α水平(P<0.05),增加局部脑组织IL-10水平(P<0.05)。Western Blotting结果显示,苦碟子注射液可降低TLR-4、NF-κB蛋白的表达(P<0.05)。结论苦碟子注射液对大鼠脑缺血再灌注损伤有明确的保护作用,具有降低脑缺血再灌注后TNF-α,并升高IL-10水平的作用,其作用机制可能与其下调TLR-4/NF-κB信号通路有关。

Objective To investigate the protective effects ofKudiezi （KDZ） Injection on cerebral ischemia reperfusion injury in rats and to explore its protective mechanism. Methods The rat model of middle cerebral artery occlusion （MCAO） was established by modified suture method, and cerebral blood flow was monitored using laser Doppler flowmetry （LDF）. Male SD rats were randomly divided into control group, model group, Kudiezi Injection high and low dose groups. ARer ischemia-reperfusion for 24 11, the neurological scores were evaluated. After anesthesia, the blood samples and brain tissues were collected, and the expression of inflammatory was detected by Elisa. Western blotting was used to detect the expression of TLR-4 and NF-κB protein. Results Behavioral scores showed that neural function defect was serious in model group compared with control group （P 〈 0.01）. In model group, cerebral index and cerebral infarction area were significantly higher than those of the control group; After KDZ intervention, the symptoms of neurological deficit was alleviated （P 〈 0.01）, the cerebral index and cerebral infarction area of mode were decreased, and the neuronal necrosis was reduced. Kudiezi Injection could significantly reduce the cerebral homogenate and serum levels of TNF-α （P 〈 0.05） and increase IL-10 level （P 〈 0.05）. Westem blotting showed that Kudiezi Injection could reduce the expression of TLR-4 and NF-κB protein （P〈0.05）. Conclusion Kudiezi Injection has protective effect on cerebral ischemia reperfusion rats. After ischemia-reperfusion, Kudiezi Injection could reduce the levels of TNF-α and raise IL-10. Its mechanism may be associated to the down-regulation of TLR-4/NF-κB signaling pathway.