丁苯酞对SD大鼠脑出血后血脑屏障的影响及其机制

目的：探讨SD大鼠脑出血后高尔基体和血脑屏障的变化及丁苯酞的干预作用和机制。方法：SD大鼠随机分为正常对照组、假手术组、脑出血组及丁苯酞组。造模48 h后测量各组的伊文思蓝含量,采用蛋白质印迹法及real-time PCR检测Cdc42,GM130,ZO-1和Occludin蛋白灰度比值及mRNA相对表达量,免疫荧光观察Cdc42和GM130是否存在共定位及脑出血后丁苯酞干预的荧光强度,免疫组织化学观察ZO-1和Occludin在血肿周围内皮细胞上的分布。结果：脑出血组的神经功能评分及伊文思蓝含量明显高于假手术组(P<0.05);丁苯酞组的神经功能评分及伊文思蓝含量较脑出血组明显降低(P<0.05)。 脑出血组Occludin,ZO-1,GM130及Cdc42蛋白灰度比值及mRNA相对表达量较假手术组降低(P<0.05);丁苯酞组中Occludin,ZO-1及Cdc42蛋白灰度比值及mRNA相对表达量较脑出血组升高(P<0.05),而GM130 mRNA差异无统计学意义(P>0.05)。免疫组织化学发现脑出血组中Occludin和ZO-1在血肿周围的表达较假手术组降低(P<0.05),而丁苯酞组则增加(P<0.05)。结论：脑出血后高尔基体形态发生改变,血脑屏障受到破坏;丁苯酞能减轻神经功能缺损及血脑屏障破坏程度,其机制与Cdc42的上调相关,而与GM130可能无关。

Effect of butylphthalide on blood-brain barrier after cerebral hemorrhage in SD rats and the mechanisms

Objective: To investigate the alteration in Golgi and blood-brain barrier aft er cerebral hemorrhagein SD rats, and to evaluate the eff ect of butylphthalide on blood-brain barrier.Methods: Sprague-Dawley rats were randomly distributed into 4 groups: a control group, a sham group, an intracerebral hemorrhage (ICH) group, and a butylphthalide group. Brain tissue was collected at 48 h after the blood brain barrier permeability was examined. Western blottingand real-time polymerase chain reaction (real-time PCR) were conducted to explore the changeof GM130, Cdc42 and tight junction protein and mRNA expression in rat brain after ICH.Immunohistochemistry (IHC) was performed to explore the distribution of ZO-1 and Occludin inthe cerebral vascular endothelial cells around the hematoma.Results: The Evans blue (EB) extravasation in the ICH group were much greater than that in thesham group (P<0.05). Butylphthalide treatment significantly decreased Evans blue extravasationcompared to the ICH group (P<0.05). Results of Western blotting and real-time PCR showed thatGM130, Cdc42, ZO-1/Occludin were decreased (P<0.05). The intervention of butylphthalidesignificantly upregulated the expressions of Cdc42 as well as ZO-1/Occludin (P<0.05), but exertedno effect on GM130 (P<0.05). Immunofluorescent staining showed that GM130 was co-localizedwith Cdc42 and administration of butylphthalide improved the expression of Cdc42 around thehematoma without affecting the expression of GM130. IHC showed that expressions of occludinand ZO-1 around the hematoma were significantly decreased in the ICH group (P<0.05), whereasbutylphthalide treatment elevated the expressions of ZO-1 and occludin around the hematomacompared with the ICH group (P<0.05).Conclusion: Morphology of Golgi apparatus is altered and the blood-brain barrier is destroyedafter ICH. The application of butylphthalide can alleviate neurological impairment and blood-brainbarrier disruption, which is related to the up-regulation of Cdc42, but not GM130.