麝香酮联合灯盏花素对大鼠脑缺血再灌注损伤后血脑屏障的保护作用

Protective effects of muscone combined with breviscapine on blood-brain barrier in rats after cerebral ischemia-reperfusion injury

目的探讨麝香酮联合灯盏花素对大鼠脑缺血再灌注损伤后MMP-2和MMP-9的表达及血脑屏障通透性的影响。方法选择SD大鼠504只,按照简单随机化方法分为假手术组、模型组、麝香酮组(1 mg/kg)、灯盏花素组(200 mg/kg)和麝香酮(1 mg/kg)联合灯盏花素高、中、低剂量组(400、200、100 mg/kg),每组72只。采用线栓法制作大鼠大脑中动脉栓塞(middle cerebral artery occlusion,MCAO)模型,再灌注伤后3、6、12、24、48、72 h,Western blot测定MMP-2和MMP-9蛋白的表达水平;同时测定脑组织伊文思蓝(evans blue,EB)的含量观察血脑屏障通透性。结果与模型组相比,灯盏花素组和麝香酮联合高、中、低剂量灯盏花素组可明显降低大鼠缺血再灌注损伤后MMP-2和MMP-9的表达(P<0.01),降低脑组织中EB的含量(P<0.01),而麝香酮组与模型组差异无统计学意义(P>0.05);对所有组进行析因分析,表明1 mg/kg麝香酮和高、中、低剂量灯盏花素配伍具有叠加的交互作用(P<0.01)。结论麝香酮联合灯盏花素比单用灯盏花素对大鼠脑缺血再灌注损伤后血脑屏障具有更好的保护作用。

Objective To determine the effect of combination of muscone and breviscapine on the expression of MMP-2 and MMP-9 and the permeability of blood-brain barrier in rats after cerebral ischemia-reperfusion injury. Methods A total of 504 rats were randomly divided into sham operation group, model group, muscone group （1 mg/kg）, breviscapine group （200 mg/kg） and combination groups （1 mg/kg muscone plus 400, 200 and 100 mg/kg breviscapine, respectively）, 72 animals in each group. Middle cerebral artery occlusion model rats were established by suture-occluded method. After reperfusion for 3, 6, 12, 24, 48 and 72 h, the levels of MMP-2 and MMP-9 in the brain tissue were evaluated by Western blotting, and the permeability of blood-brain barrier was evaluated by evaluating the contents of Evans Blue in brain tissue. Results Compared with model group, the expression levels of MMP-2 and MMP-9 and the content of EB in brain tissue were decreased significantly in the breviscapine group and the combination groups （P〈0.01）. But there was no significant difference in the expression levels and the content between the model group and the muscone group（P〉0.05）. Factorial analysis showed that muscone of 1 mg/kg combined with breviscapine of high, middle and low doses exerted synergetic effects （P〈0.01）. Conclusion Muscone combined with breviscapine shows better protective effects than breviscapine alone on the blood-brain barrier in rats after cerebral ischemia-reperfusion injury.