川芎嗪对大鼠局灶性脑缺血再灌注损伤的保护作用及机制研究

目的探讨川芎嗪对大鼠局灶性脑缺血再灌注损伤的保护作用及机制。方法将64只健康雄性SD大鼠随机分为四组：（1）假手术组;（2）缺血再灌注组;（3）川芎嗪20 mg/kg处理组;（4）川芎嗪40 mg/kg处理组。每组各16只,8只用于脑梗死体积测定,8只用于mRNA和蛋白的测定。采用线栓法制备大鼠局灶性大脑中动脉栓塞模型,缺血2 h再灌注24 h。术后对大鼠神经功能进行评分;2,3,5-氯化三苯四氮唑染色观察脑梗死体积;酶联免疫吸附法测定脑组织中肿瘤坏死因子-α（TNF-α）、白介素-1β（IL-1β）和核因子-κBp65（NF-κBp65）水平;实时定量聚合酶链反应和蛋白质印迹法分别检测Toll样受体4（TLR4）mRNA和蛋白的表达。结果与假手术组比较,缺血再灌注组大鼠的神经功能缺陷评分和脑梗死体积明显增加;脑组织中TNF-α、IL-1β和NF-κBp65水平以及TLR4的表达也显著升高,差异均有统计学意义（P〈0.05）。川芎嗪处理组上述指标,与缺血再灌注组比较,差异有统计学意义（P〈0.05）;不同剂量川芎嗪处理组间比较,差异无统计学意义（P〉0.05）。结论川芎嗪可抑制脑缺血再灌注诱导的损伤,其机制与抑制TLR4/NF-κB信号通路,进而减少炎症因子的产生有关。

Protective effect and mechanism ofligustrazine on focal cerebral ischemia-reperfusion injury in rats

Objective To explore the protective effect and mechanism of ligustrazine on focal cerebral ischemia-reperfusion injury in rats. Methods Totally 64 healthy SD rats were divided into sham-operated group,ischemia-reperfusion group, 20 mg/kg ligustrazine group and 40 mg/kg ligustrazine group,16 cases in each group including 8 cases for determination of volume of cerebral infarction and 8 cases for determination of mRNA and protein. Focal cerebral ischemia-reperfusion model in rats was made by suture-occluded method through 24 h reperfusion after 2 h of ischemia. The neurological function was scored after opera-tion and the infarct volume was measured by 2,3,5-Triphenyltetrazoliumchloride staining. The levels of tumor necrosis factor-α（TNF-α）,interleukin-1β（IL-1β） and nuclear factor-κBp65（NF-κBp65） in brain tissue were measured by enzyme linked immunosorbent assay. The expression of Toll-likereceptor 4（TLR4） mRNA and protein was determined by real-time PCR and Western blotting,respectively. Results Compared with the sham-operated group,the neurological function score and infarct volume were significantly increased in ischemia-reperfusion group. The levels of TNF-α,IL-1β and NF-κBp65 in brain tissue and the expression of TLR4 were also markedly increased with statistically significant difference（P〈0.05）. However,the above effects induced by ischemia-reperfusion were significantly inhibited by ligustrazine. The difference between 20 mg/kg ligustrazine group and 40 mg/kg ligustrazine group had no statistical significance（P〉0.05）,while the difference between ligustrazine groups and ischemiareperfusion group was statistically significant（P〈0.05）. Conclusion Ligustrazine can inhibit cerebral ischemia-reperfusion injury, which might be related to decrease inflammatory factor production through inhibition of TLR4/ NF-κB signal pathway.