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犬颅脑枪弹伤后局部脑组织BDNF蛋白表达及意义的实验研究

Expression of BDNF in Canines with Craniocerebral Gunshot Wounds

【作者】 杨江河

【导师】 章翔;

【作者基本信息】 第四军医大学 , 外科学, 2004, 硕士

【摘要】 颅脑火器伤具有死亡率、致残率高、救治难度大等特点,其死亡率历来居各部位火器伤之首。颅脑战伤的发生率在第一次世界大战未用钢盔时为25%,以后为9%~15%或10%~17%。自二战以来,其死亡率一直保持在30%~40%,伤死率为10%~20%,伤残率为40%,在火器伤致死的总人数中,颅脑伤几乎占50%。颅脑火器伤不仅多见于战时,平时也时有发生,平时枪击伤的死亡率超过60%。在国内、外颅脑火器伤研究中,存在一个共同问题是动物模型制作困难,体现在动物伤后存活时间过短,无法进入更深入的研究。本实验采用低速德国小口径步枪射击犬头颅,建立犬脑贯通伤(penetrating craniocerebral injury,PCI)和脑切线伤模型,研究犬颅脑火器伤后局部脑组织脑源性神经营养因子(brain-derived neurotrophic factor,BDNF)蛋白的表达与意义。 一、犬脑枪弹伤模型建立及相关生理、病理指标观察:目的建立犬脑枪弹伤致伤模型,检验该模型用于颅脑枪弹伤进一步研究的可行性。方法 杂种犬16只,采用低速德国小口径步枪子弹致伤。测定弹丸致伤参数,观察实验犬伤后相关生理与病理指标改变。结果 脑切线伤组和PCI组的弹丸撞击速度与撞击能量基第四军医大学硕士学位论文本一致;脑切线伤组的组织对弹丸吸收能量较大(P<0.05),且实验犬伤后存活时间也较长(P<0.05)。两组均在伤后数分钟至lh内出现明显的心率减慢、血压下降、颈动脉血流量减少、呼吸暂停或减慢、颅内压(运让acranial pressure,ICP)逐渐升高。两组挫伤区病理表现为脑血管扩张、充血、血管周围环形出血、神经细胞轻度缺血、坏死及脑水肿(b rain edema,BE)。结论上述模型均具有弹道伤恒定、重复性好、伤后动物存活时间较长等特点,尤以脑切线伤模型更为理想。 二、犬颅脑枪弹伤后局部脑组织BDNF蛋白的表达:目的探讨犬颅脑火器伤后脑组织内BDNF的变化规律及其对颅脑火器创伤神经元的保护作用。方法建立犬颅脑枪弹伤模型,制备不同部位不同时间点脑组织切片,免疫组化法染色,检测BDNF的表达。与设立的对照组相比较,并进行组间比较。结果损伤组脑组织BDNF含量明显高于对照组,动物的伤情相对愈重,表达愈明显。在2一4h为表达高峰期,不同组织间亦存在明显差异(P<0 .01),以海马及挫伤区含量高。结论颅脑枪弹伤后不同部位脑组织BDNF随时间的变化出现不同表达,提示BDNF在颅脑枪弹伤的早期修复中发挥作用。 综上所述,采用低速德国小口径步枪致伤犬颅脑、建立PCI和脑切线伤模型,两模型具有存活时间长、重复好等特点,尤以脑切线伤模型存活时间更长,为颅脑火器伤的进一步研究提供了较为理想的动物模型。通过对枪弹伤后脑组织神经元BDNF蛋白的表达变化的研究,从分子水平阐述了颅脑火器伤后BDNF在早期修复中所发挥的作用。

【Abstract】 The craniocerebral missile wounds have the characteristic of a high death rate and mutilation rate and are particularly difficult to be managed. They are the most common cause of death in missile injure victims. Since world war two, the craniocerebral missile wounds contributed to 30%~40% of death rate under shield, 10%~20% of casualty rate and 40% of mayhem rate. Up to 50% of deaths caused by missile injury are penetrating missile injuries of the head. It happened not only in wartime, but also in peacetime. The mortality of gunshot wound is more than 60% during times of peace. The common problem of research for craniocerebral missile wounds is difficult to make the animal models because the animals will survive a short time after injury. The aim of this study was to make the canine models of penetrating craniocerebral injury (PCI) and tangent brain injury by the bullets of the small-calibre rifle made in German and test the BDNF expression of cerebral neuronal cells after craniocerebral missile wounds and high energy missile wounds in the extremity of canines. 1. Model establishment and observation of related pbysiopatholog-ical parameters in canines with craniocerebral gunshot woundsAIM To set up the experimental model of canine craniocerebral gunshot wound and examine its feasibility, for further study in craniocerebral gunshot wound. METHODS Sixteen canines, whichwere wounded by the bullets of the small-calibre rifle made in German, were made into penetrating craniocerebral injury (PCI) group (n=10) and tangent brain injury group (n=6). The wounded parameters of bullets were measured, the physiopathological changes of canines were observed. RESULTS The impact velocity and the impact energy of the two groups were recorded. There was no significant difference between two groups. The absorbed energy in the TBI group was more than that in the PCI group (P<0.05). The average survival time of the canines in the tangent brain injury group was longer than that of the PCI group (P<0.05). The heart rate, respiratory frequency, blood pressure and carotid blood flow were reduced, and the intracranial pressure (ICP) was increased obviously in both groups at a few minutes to 1 hour post-wound. There were cerebral vascular dilation, hyperemia, perivascular hemorrhage, ischemia necrosis of the neunons and perivascular edema in the specimen of two groups. CONCLUSION The experimental model showed invariable wound trajectory. The reiteration of the model was good and the survival times were long, especially in the model of the tangent brain injury. 2. Expression brain-derived neurotrophic factor (BDNF) in canines with craniocerebral gunshot woundsAIM To observe the expression of BDNF in cerebral cortex, hippocampus CA2 and brain stem in canines with cranicerebral gunshot wounds. METHODS Twenty canines were divided into normal control and injury group. Using the model of gunshot-injured canine cerebrum made by the bullets of small-caliber rifles made in German, BDNF positive neurons were observed in the contusion trajectory area, the concussion trajectory area, hippocampus, and brain stem at 30 min,l, 2, 4, 6, 8 and 12 h after trauma by immunohistochemical method. The results of the control group and injury group were compared. RESULTS The Expression of BDNF in the injury group was obviously higher than that of the control group. The expression appeared 2-4 hour after trauma (p<0.01).CONCLUSION BDNF plays a positive role in repair of brain injury.In summary, we conclude that using the bullets of the small-calibre rifle made in German, we established the canine models of PCI and tangent brain injury. The canines with PCI or tangent brain injury, especially the tangent brain injury, will survive long time and the model is easy to repeat. Therefore, the models can serve better for the research of craniocerebral missile wounds. The study that BDNF expressed in cerebral neuronal cells after gunshot injury explained the effect of craniocerebral missile wound and the principle of the cerebral secondary

  • 【分类号】R651.15
  • 【下载频次】49
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