高压氧干预后局灶性脑缺血再灌注损伤模型大鼠c-fos的表达(英文)

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背景:高压氧可增加氧弥散能力,从而改善脑水肿和脑组织缺氧,促进病灶区脑细胞的生理功能恢复、侧支循环的建立和脑细胞再生修复。目的:观察高压氧对大鼠急性局灶性脑缺血再灌注损伤后不同时间点c-fos癌基因表达的影响。设计:随机分组动物实验。单位:解放军第四军医大学唐都医院急诊科、西安高新医院检验中心、解放军空军总医院、解放军第四军医大学航空航天医学系高压氧治疗中心。材料:实验于2002-04在解放军第四军医大学航空航天医学系高压氧治疗中心完成,选用65只生后2个月健康雄性SD大鼠。方法:随机摸球法将大鼠分为4组,模型组(n=20):参照Koizumi等设计的方法,制备大鼠大脑中动脉缺血模型;正常对照组(n=5):手术方式同模型组,但不阻断动脉血流;纯氧治疗组(n=20):手术过程同模型组,动物缺血1h后抽出栓子,分别在插入栓子后2,9,21,45,69h将动物置于高压舱内,常压下吸100%纯氧。高压氧治疗组(n=20):手术过程同模型组,动物缺血1h后抽出栓子,分别在插入栓子后2,9,21,45,69h将动物置于高压氧舱内,0.25MPa吸纯氧1h。主要观察指标:利用免疫组织化学和病理组织学法观察各组大鼠脑缺血再灌注5,12,24,72h脑皮质、视前区与纹状体中性白细胞浸润及c-fos癌基因蛋白及阳性细胞表达的变化;计算脑缺血再灌注72h皮质、纹状体内侧区与视前区神经元坏死程度、大鼠左侧半球脑血管渗漏面积。结果:纳入大鼠65只均进入结果分析。①视前区c-fos阳性产物主要集中于中部。对侧皮质少见c-fos阳性反应,视前区有轻度表达,纹状体呈中等强度的反应。缺血12h,c-fos阳性产物在上述区域开始减弱,24h强度明显减弱!高压氧治疗组皮质和视前区较单纯缺血组c-fos阳性产物强度明显减弱!缺血再灌注12h,高压氧治疗组视前区、纹状体中性白细胞数明显低于模型组(P<0.05);缺血再灌注24h,脑皮质、视前区、纹状体中性白细胞低于模型组(P<0.05)。②高压氧治疗组血管渗漏面积与单纯模型组相比明显缩小(P<0.05);缺血再灌注72h,高压氧治疗组视交叉平面皮质、纹状体内侧区与视前区神经细胞损伤数目较模型组明显减轻(P<0.05),假手术组未见神经元损伤。结论:高压氧可明显缩小大鼠急性局灶性脑缺血再灌注损伤的血管渗漏面积,减轻神经系统症状,抑制中性白细胞浸润,抑制梗塞区c-fos癌基因表达,减少“半影区”神经元坏死。 BACKGROUND: Hyperbaric oxygen can increase the ability of oxygen diffusion, thereby improving brain edema and brain hypoxia, promote the recovery of physiological function of brain cells in focal area, establishment of collateral circulation and brain cell regeneration. Objective: To observe the effect of hyperbaric oxygen on c-fos oncogene expression at different time points after acute focal cerebral ischemia-reperfusion in rats. Design: Randomized animal experiments. Unit: Emergency Department of Tangdu Hospital, Fourth Military Medical University of PLA, Inspection Center of Xi’an High-tech Hospital, PLA General Hospital of Air Force, Hyperbaric Oxygen Therapy Center, Department of Aeronautics and Astronautics, Fourth Military Medical University of PLA. MATERIALS: Experiments were performed at the Hyperbaric Oxygen Therapy Center of the Department of Aeronautics and Astronautics, Fourth Military Medical University of PLA in 2002-04. Sixty-five healthy male Sprague Dawley rats aged 2 months and older were selected. Methods: The rats were randomly divided into 4 groups: model group (n = 20): model of middle cerebral artery occlusion (MCAO) was prepared by the method of Koizumi et al. The normal control group (n = 5) The same model group, but not block the arterial blood flow; pure oxygen treatment group (n = 20): the same surgical model group, the animals were removed after embolization 1h, respectively, after insertion of emboli 2,9,21,45 , 69h The animals were placed in a hyperbaric chamber, inhaled under normal pressure 100% pure oxygen. Hyperbaric oxygen treatment group (n = 20): The same procedure was performed in the model group. The animals were withdrawn after 1 h of ischemia and the animals were placed in hyperbaric oxygen chambers at 2, 9, 21, 45 and 69 h after insertion of the embolus, respectively MPa suction pure oxygen 1h. MAIN OUTCOME MEASURES: Immunohistochemistry and histopathology were used to observe the changes of neutrophil infiltration and c-fos oncogene in cerebral cortex, preoptic area and striatum at 5,12,24,72 h after cerebral ischemia-reperfusion in rats Protein and positive cells. The necrosis of neurons in medial area and preoptic area of ​​cortex, striatum at 72h after cerebral ischemia-reperfusion was calculated, and the area of ​​cerebral vascular leakage in the left hemisphere of rats was calculated. Results: 65 rats were included in the analysis of the results. ① preoptic area c-fos positive products are mainly concentrated in the middle. Contralateral cortical rare c-fos positive reaction, preoptic area mild expression, striatum showed moderate-intensity response. At 12h after ischemia, c-fos positive products began to weaken in the above-mentioned areas, and the intensity of 24h decreased significantly. The intensity of c-fos positive product in the cortex and the preoptic area was significantly weakened in hyperbaric oxygen treatment group 12h after ischemia-reperfusion The number of neutrophils in the preoptic area and striatum in the oxygen treatment group was significantly lower than that in the model group (P <0.05), while in the ischemic reperfusion group, the neutrophils in the cortex, preoptic area and striatum were lower than the model group <0.05). Compared with the model group, the area of ​​vascular leakage in the hyperbaric oxygen treatment group was significantly reduced (P <0.05); at 72h after ischemia-reperfusion, the neuronal cell damage in the cortical plane, medial striatum and preoptic area Compared with the model group, the number of neurons in the sham operation group was significantly reduced (P <0.05). CONCLUSION: Hyperbaric oxygen can significantly reduce the area of ​​vascular leakage in acute focal cerebral ischemia-reperfusion injury in rats, reduce the symptoms of nervous system, inhibit neutrophil infiltration, inhibit the expression of c-fos oncogene in infarct area, Film District "Neuronal necrosis.
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