亚甲基蓝对大鼠局灶性脑缺血再灌注后血脑屏障的保护作用

发布时间：2018-10-26 17:18

【摘要】：目的探讨亚甲基蓝对大鼠局灶性脑缺血再灌注引起血脑屏障破坏的保护作用。方法雄性Sprague-Dawley大鼠18只随机分为假手术组(n=6)、模型组(n=6)和亚甲基蓝组(n=6)。用线栓法制备大鼠左侧大脑中动脉栓塞1 h再灌注模型。亚甲基蓝组于再灌注即刻及再灌注2 h分别静脉给予亚甲基蓝3 mg/kg、1.5 mg/kg;模型组给予相同体积的生理盐水;假手术组手术操作与模型组相同,不插入线栓,不静脉注射药物。再灌注47 h后HE染色观察大鼠脑缺血周边皮层组织学结构,白蛋白免疫组化染色法检测血脑屏障通透性的变化,免疫组化染色法和免疫荧光双标染色法检测胶质原纤维酸性蛋白(GFAP)和水通道蛋白-4(AQP-4)的表达。结果 HE染色显示,模型组缺血周边大脑皮层的细胞和血管形态不完整,而亚甲基蓝组病理变化减轻;模型组白蛋白、GFAP和AQP-4表达均明显高于假手术组(P0.01),亚甲基蓝组白蛋白、GFAP、AQP-4表达均低于模型组(P0.05)。免疫荧光双标染色显示,AQP-4与GFAP在星形胶质细胞上共定位。结论亚甲基蓝可能是通过减轻胶质细胞增生及下调AQP-4表达来改善局灶性脑缺血再灌注损伤引起的血脑屏障破坏。
[Abstract]:Objective to investigate the protective effect of methylene blue on blood-brain barrier damage induced by focal cerebral ischemia reperfusion in rats. Methods Eighteen male Sprague-Dawley rats were randomly divided into three groups: sham operation group (n = 6), model group (n = 6) and methylene blue group (n = 6). The left middle cerebral artery (MCAE) reperfusion model was established by thread embolization for 1 h. Methylene blue 3 mg/kg,1.5 mg/kg; model group was given the same volume of normal saline respectively immediately after reperfusion and 2 hours after reperfusion. The sham-operation group had the same operation procedure as the model group, without insertion of thread embolus and intravenous injection of drugs. The histological structure of cerebral ischemic cortex was observed by HE staining and the changes of blood-brain barrier permeability were detected by albumin immunohistochemical staining after 47 h reperfusion. The expression of glial fibrillary acidic protein (GFAP) and aquaporin-4 (AQP-4) were detected by immunohistochemistry and double immunofluorescence staining. Results HE staining showed that the morphology of cells and blood vessels in the cerebral cortex around ischemia in the model group was incomplete, while the pathological changes in the methylene blue group were alleviated. The expression of albumin, GFAP and AQP-4 in model group was significantly higher than that in sham operation group (P0.01), and the expression of albumin and GFAP,AQP-4 in methylene blue group was lower than that in model group (P0.05). Double immunofluorescence staining showed that AQP-4 and GFAP were co-located on astrocytes. Conclusion methylene blue may ameliorate the blood-brain barrier damage induced by focal cerebral ischemia-reperfusion injury by reducing glial cell proliferation and down-regulation of AQP-4 expression.
【作者单位】：首都医科大学北京市神经外科研究所病理生理室首都医科大学附属北京天坛医院;北德克萨斯大学医学中心神经与药理系;
【基金】：国家自然科学基金项目(No.81271286,No.81228009)
【分类号】：R743.3

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