【作者】 车彦军；

【导师】 苏芳忠；

【作者基本信息】 郑州大学 ， 神经外科， 2004， 硕士

【摘要】 目的： 脑外伤(Traumatic Brain Injury，TBI)是临床常见的一种创伤性疾病，大量实验和临床研究证实，外伤造成的脑损伤并不仅仅是在伤后瞬时完成的，伤后逐渐发展演化形成的继发脑损伤才是主要病理过程。近年来国内外研究结果表明，外伤所导致的局部炎症反应在继发性脑损伤方面起着重要作用。局部神经组织变性坏死、血脑屏障(Blood Brain Barrier，BBB)的破坏和局部脑水肿都与炎症反应有着密切的关系。炎症反应的一个重要特点是白细胞浸润，而细胞间粘附分子-1(ICAM-1)和单核细胞趋化因子-1(MCP-1)是使白细胞游出血管壁并激活和引导白细胞向炎症区浸润的重要因子，其表达的程度与炎症反应的强弱有着密切关系。 高压氧(Hyperbaric Oxygen，HBO)治疗颅脑损伤，在促醒、促进神经功能的恢复和减少后遗症方面的效果已被多数学者所接受。但其确切的治疗机理目前尚不十分清楚。有学者在对肌瓣移植的缺血再灌注损伤进行实验研究时发现：HBO能够减少白细胞与血管内皮细胞粘附，并减少白细胞的浸润，减轻局部的炎症反应，从而减轻肌瓣的缺血再灌注损伤。但HBO对TBI后脑组织中白细胞浸润和ICAM-1、MCP-1的表达是否有影响，目前国内外尚未见报道。本实验研究目的在于通过对大鼠TBI后HBO的治疗，观察其对外伤脑组织内的ICAM-1和MCP-1表达的影响，进而了解HBO对TBI后脑组织中自细胞浸润的影响，为HBO治疗脑外伤提供理论依据。娜州大拳2004月卜.士布呀，七息毕J七今全文高压反对大双实脸性肺栩伤后肺姐旗1〔胡~1和峨P一]表达的影响方法: 采用250一3009成年健康封闭群SD雄性大鼠，随机分为A、B两组，A、B两组又分别分为6个亚组。所有大鼠均采用Freeney自由落体脑损伤模型的制作方法，制造出中度的脑损伤模型。A组于外伤后不施加任何处理因素，B组于外伤后30分钟内送入高压氧舱进行HBO治疗。将各亚组分别于伤后3小时，6小时、12小时、24小时、72小时、7天断头处死，处死后立即采集标本，每个标本的一部分送电镜中心进行透射电镜观察，另一部分进行ICAM一1、MCP一1免疫组化染色，染色后阳性细胞用图像分析系统进行测定。测定结果在计算机上用SPSS软件包(10.0版)进行统计学处理。计量资料均以x士s表示，采用成组样本t检验，以。=0 .05为差异显著性的检验标准。结果:1.透射电镜下超微结构的变化 A组(对照组)于伤后3h即有明显的血管周围水肿、细胞内及线粒体肿胀。伤后6h血管周围水肿达高峰，线粒体呈球形，有的线粒体空泡化甚至碎裂，线粒体峭消失;有的神经细胞胞质内线粒体及其他细胞器已不存在。伤后12h血管周围水肿及细胞内肿胀均减轻。B组(HBO组)于伤后3h和6h均亦出现血管周围水肿，但明显较对照组为轻，无线粒体空泡化和破裂。伤后24h两组己无明显差别。2.ICAM一1的表达 A组在伤后3h即有ICAM一1的表达，伤后6h表达至高峰，伤后24h开始下降，至伤后第7天仍有少量表达，B组在伤后3h、6h及伤后12h的ICAM一l的表达与A组相比有明显降低(P<0 .05)，但两组在伤后24小时及以后，ICAM一1的表达则无明显差异(P>0.05)。3.MCP一1的表达 两组均于伤后3小时即有MCP一1的表达，伤后12小时达高峰，伤后24小时仍有较高表达，但B组MCP一l表达于伤后3小时、6小时、12小时和24小时明显低于(P>0 .05)。A组(P(0.两组于伤后05)。两组于伤后72小时毗P一1的表达即无明显差异7天MCP一1均仍有表达。娜州大学2004年硕士研免生早业论文高压孰对大民实脸性肺扭伤后脸组织IC侧-1和式P一1表达的影响结论: 1.炎症反应的一个重要特点是白细胞浸润，而白细胞浸润是TBI后的关键事件，BSB的破坏、局部脑组织水肿与白细胞浸润密切相关;且ICAMeel和MCP一1是使白细胞穿越血管内皮细胞向炎症部位游走和浸润的重要因子;本实验证实了工CAMeel的表达高峰与脑水肿的高峰相一致，成P一1的表达的高峰与单核细胞、淋巴细胞的迟发浸润相一致。 2.HBO能明显减轻大鼠脑外伤ICAM一l和MCP一1在伤后3h、6h和 12h的表达，因而减少了白细胞向血管外游出和向外伤灶的游走和浸润，缓解了局部脑组织的炎症反应和脑水肿，减轻了由炎症反应引起的继发性脑损伤，为HBO治疗TBI提供理论依据。 3.本实验表明，TBI后3小时即有ICAMeel和MCP一l的表达，因此，HBO治疗TBI应尽早进行，以有效降低工CAM一1和MCP一1的表达。 4.由于治疗组与对照组ICAM一1和MCP一1的表达分别于伤后24小时和72小时已无明显差异，提示应最迟于TBI后24小时内行第二次HBO治疗，进一步减少IcAM一1和MCP一1的迟发表达和白细胞的浸润，减轻迟发性炎症反应，改善脑功能。更多还原

【Abstract】 ObjectivesTraumatic brain injury (TBI) is very common in neurosurgery. Many experimental and clinical researches have conformed that the secondary brain injury is an important pathological process after TBI. Recently, the results of oversea and domestic researches have indicated that inflammation after TBI play a critical role in the secondary brain injury. Inflammation contributes closely to brain tissue degeneration and necrosis, blood brain barrier (BBB) damage and local brain tissue edema after TBI. Leukocyte infiltration is a main characteristic of inflammation. Intercellular adhesion molecule-1 (1CAM-1) andmonocyte chemoattractant protein- 1 (MCP-1) are the key factors which activate and direct leukocytes moving out of capillary vessels and gathering in the inflammatory area. Therefore, ICAM-1 and MCP-1 are closely related to the inflammatory course after TBI.Many researches indicate that hyperbaric oxygen(HBO) treatment can effectively alleviate TBI in recovering consciousness, improving nervous function and improving prognosis. But the accurate mechanisms are not known. In the course of experimental study on re-perfusion of ischemic skeletal-muscle graft, Zamboni found that HBO could reduce leukocyte adhering to endothelia and infiltrating. Whether HBO affects leukocyte infiltration and the expression of ICAM-1 and MCP-1 in brain tissue has been not reported. The objectives of present experimental research are to explore the HBO influence on the expression of ICAM-1 and MCP-1 after TBI, which may provide the theoretical evidences for the effect of HBO in treating TBI.MethodsAdult healthy male SD rats are chosen. The range of their-5-weight was 250 - 300g. All rats were divided into two groups: group A and group B, then each group was divided into 6 sub-group at random. According to Freeney’s experimental TBI model, moderate damage was done on all rats. No administration was done on group A and group B were sent to HBO treatment in 30 minutes after TBI. Then the rats of sub-group 1,2,3,4,5,6 were killed at 3h, 6h, 12h, 24h, 72h and on day 7 respectively after TBI. One part of each specimen was sent to electron microscope center, the rest was stained with immunohistochemical method of ICAM-1 and MCP-1. The positive cells were measured by map analysis system(MAS). The results were analysized by SPSS on computer. Data were expressed as x s with a =0.05 considered statistically significant.Results1, The change of brain tissue ultrastructure under electromicroscopeIn group A, vascular periphery edema and mitochondria swollen were obviously observed at 3h after TBI. At 6h, vascular periphery edema was the most apparent and mitochondria was ball-like, even blebbing and fragmentary. Mitochondria matrixes vanished. At 12h, the conditions were alleviated. Vascular periphery edema in every sub-group ofgroup B was slighter than that of group A respectively. The blibbing and fragmentary of mitochondria wasn’t exist in group B. At 24h, both group A and B showed no significant difference .2, The expression of ICAM-1In group A, the expression of ICAM-1 began to up-regulate at 3h and the expression reached the highest level at 6h and began to reduce after 24h. The expression of ICAM-1 became minor on 7th day after TBI. The expression of ICAM-1 in group B was dramatically lower than that in group A at 3h, 6h and 12h respectively (P<0.05). However, the conditions in both group A and B had not difference at 24h after TBI ( P>0.05 ) . 3, The expression of MCP-1In both group A and B, the expression of MCP-1 began to be observed at 3h after TBI, and the expression reached the highest level at 12h, and the expression was high at 24h after TBI. But, the expression of MCP-1 in each sub-group of group B was lower than of group A at 3h, 6h, 12h and 24h respectively after TBI (P<0.05). The expression of MCP-1 in both group A and B showed no significant difference (P>0.05) . The expression of MCP-1 was faint on 7 day after TBI.-7-Conclusion1, A critical characteris更多还原