亚甲基蓝对脑缺血再灌注大鼠血脑屏障的保护作用  

Protective effect of methylene blue on blood-brain barrier in rats with cerebral ischemia-reperfusion

作　　者：程方圆 路凯[1] 刘捷[2] 白宁[1] 徐瑞芬[1] 霍苗[1] 陈方[1] CHENG Fang-yuan;LU KAI;LIU Jie;BAI Ning;XU Rui-fen;HUO Miao;CHEN Fang(Anesthesiology Department,Shaanxi People′s Hospital,Xi′an 710068,China;Operating Room,Shaanxi People′s Hospital,Xi′an 710068,China)

机构地区：[1]陕西省人民医院麻醉科,陕西西安710068 [2]陕西省人民医院手术室,陕西西安710068

出　　处：《实用药物与临床》2021年第4期289-294,共6页Practical Pharmacy and Clinical Remedies

基　　金：陕西省科技厅2019年自然科学基础研究计划项目(2019JQ-975)。

摘　　要：目的探讨亚甲基蓝对大鼠脑缺血再灌注损伤的保护作用及机制。方法SD大鼠随机分为假手术组、模型组、亚甲基蓝低剂量组(1 mg/kg)、亚甲基蓝中剂量组(2 mg/kg)、亚甲基蓝高剂量组(4 mg/kg),每组12只。线栓法制备大鼠左侧颈动脉栓塞2 h再灌注24 h模型。亚甲基蓝组于术前1 d,再灌注时腹腔注射相应剂量亚甲基蓝溶液。假手术组与模型组给予生理盐水。行为学测试检测神经功能,TTC染色检测大脑梗死面积,伊文思蓝染色检测血脑屏障完整性,试剂盒与Western blot检测氧化应激水平、炎症反应与血脑屏障相关蛋白表达。结果亚甲基蓝能够显著改善神经功能,减少梗死面积并且维持血脑屏障结构完整。此外,脑缺血再灌注造成脑中氧化应激与炎症反应发生,抑制occludin与claudin-5蛋白表达,然而,亚甲基蓝治疗能够显著改善脑中氧化应激水平与炎症反应,并升高occludin与claudin-5蛋白表达。结论亚甲基蓝可能通过抑制脑中氧化应激水平与炎症反应,升高occludin与claudin-5蛋白表达,维持血脑屏障结构,减少脑中梗死面积,改善神经功能。Objective To investigate the protective effect and mechanism of methylene blue on cerebral ischemia-reperfusion injury in rats.Methods SD rats were randomly divided into sham group,model group,methylene blue low-dose group(1 mg/kg),methylene blue middle-dose group(2 mg/kg)and methylene blue high-dose group(4 mg/kg),with 12 rats in each group.The model of left carotid artery embolization was performed for 2 hours and reperfusion for 24 hours.At one day before surgery and the moment of reperfusion,the methylene blue group was intraperitoneally injected with the corresponding dose of methylene blue solution.Sham operation and model group were given saline.Behavioral tests was used to detect neurological function,TTC staining was used to detect cerebral infarct size,Evans blue staining was used to detect the integrity of blood-brain barrier,and kits and Western blot was used to detect oxidative stress levels,inflammatory response and blood-brain barrier-associated protein expression.Results Methylene blue could significantly improve the nerve function,reduce infarct size and maintaine the integrity of blood-brain barrier.In addition,cerebral ischemia-reperfusion caused oxidative stress and inflammatory reaction in brain,inhibited the expression of occludin and claudin-5 protein.However,methylene blue treatment could significantly improve the oxidative stress and inflammation in brain,and increase the expression of occludin and claudin-5 protein.Conclusion Methylene blue can maintain the blood-brain barrier structure,reduce the infarct area in the brain,and improve the nerve function by inhibiting the oxidative stress level and inflammatory reaction in the brain and increasing the expression of occludin and claudin-5 protein.

关 键 词：亚甲基蓝 脑缺血再灌注 血脑屏障 

分 类 号：R743.3[医药卫生—神经病学与精神病学]