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背景:目前,有很多关于预处理诱导脑缺血耐受及其可能机制的报道,如脑缺血预处理、吸氧、针灸、异氟醚预处理等,国外一些研究也已证实肿瘤坏死因子-α(tumornecrosisfactor-α,TNF-α)能诱导脑缺血,但机制尚未阐明。迄今,大鼠脑缺血再灌注损伤中TNF-α预处理的影响作用及其机制尚不十分清楚。目的:探讨TNF-α预处理对大鼠脑缺血再灌注损伤的影响及可能机制。设计:完全随机设计,对照实验研究。地点和材料:实验地点为哈尔滨医科大学附属二院动物实验中心。材料:健康雄性Wistar大鼠60只,体质量200～300g,完全随机分为TNF-α0.05μg组、0.5μg组、1.0μg组及对照组,每组15只大鼠。实验动物由哈尔滨医科大学附属二院动物实验中心提供,显微器械、25μL微量进样器及微动脉夹均购自上海仪欣医疗器械公司,日本产钓鱼线(直径0.17mm),恒温箱,数码相机,北航图像分析系统,PBS液,重组大鼠肿瘤坏死因子-α(购自美国Sigma公司),CD54,GFAP试剂盒、相应二抗及DAB染色试剂盒皆购自武汉博士德生物工程有限公司,10g/L四氮唑红(TTC)及实验室常规使用材料等。干预:采用线栓法制作大鼠局灶性脑缺血模型(阻断大鼠大脑中动脉血流),并予以改良。在脑缺血前48h,给大鼠小脑延髓池内注射不同剂量的TNF-α(0.05μg,0.5μg,1.0μg)或PBS液
BACKGROUND: At present, there are many reports on pretreatment induced cerebral ischemic tolerance and its possible mechanisms, such as cerebral ischemic preconditioning, oxygen inhalation, acupuncture and isoflurane preconditioning. Some foreign studies have also confirmed that tumor necrosis factor -α (tumor necrosis factor-α, TNF-α) can induce cerebral ischemia, but the mechanism has not yet been clarified. Up to now, the effect of TNF-α preconditioning in rats with cerebral ischemia-reperfusion injury and its mechanism is not yet clear. Objective: To investigate the effect of TNF-α preconditioning on cerebral ischemia-reperfusion injury in rats and its possible mechanism. Design: Completely randomized design, controlled experimental study. Venue and material: The experimental site is the Animal Experimental Center of the Second Affiliated Hospital of Harbin Medical University. MATERIALS: Sixty healthy male Wistar rats weighing 200-300g were randomly divided into TNF-α 0.05μg group, 0.5μg group, 1.0μg group and control group with 15 rats in each group. Experimental animals were provided by Animal Experimental Center, Second Affiliated Hospital of Harbin Medical University. Microsurgery, 25μL microinjector and arteriolar clips were purchased from Shanghai Yixin Medical Device Co., Ltd., fishing lines (diameter 0.17mm) made in Japan, incubators, Digital camera, Beihang image analysis system, PBS solution, recombinant rat tumor necrosis factor-alpha (purchased from Sigma, USA), CD54, GFAP kit, corresponding secondary antibody and DAB staining kit were purchased from Wuhan Boster Bioengineering Limited Company, 10g / L tetrazolium red (TTC) and laboratory routine use of materials. Intervention: A rat model of focal cerebral ischemia (blocking middle cerebral artery blood flow) was made by the method of thread embolization, and was improved. 48h before cerebral ischemia, the rats were injected with different doses of TNF-α (0.05μg, 0.5μg, 1.0μg) or PBS solution