迷走神经刺激对兔脑爆炸伤神经保护作用机制的实验研究

发布时间：2018-05-10 23:20

本文选题：神经调控 + 兔　；参考：《厦门大学》2014年硕士论文

【摘要】：目的：实验的第一阶段通过利用爆炸冲击波与铁屑碎片结合,建立了一种与实际情况接近、操作简单、质控标准化、稳定性高、可重复性好的兔脑爆炸弹片伤模型,并观察实验兔的伤情变化。实验的第二、三阶段通过利用第一阶段建立的简易兔脑爆炸弹片伤模型的方法制作动物模型,并分别观察及探讨了迷走神经电刺激对脑爆炸弹片伤后脑组织病理、水肿变化的影响,以及对脑及全身炎症反应和自主神经系统功能紊乱的调节作用。方法：实验的第一阶段选取雄性新西兰大白兔24只,随机分成预实验组(4只)、空白对照组(4只)、假手术组(6只)、爆炸弹片伤组(10只),采用自行设计的爆炸装置,爆炸经过开颅处理后的兔脑右侧顶叶皮质。观测伤后兔生命体征、头颅CT表现、脑组织病理。通过对简易兔脑爆炸弹片伤模型质控性、稳定性高和可重复性的研究,建立合适的动物模型,实验进入第二、三阶段。在实验的第二阶段中,选取另外24只雄性新西兰大白兔按随机数字表法分为假手术组(6只)、爆炸弹片伤组(10只)和爆炸弹片伤+迷走神经刺激组(8只),爆炸弹片伤组和爆炸弹片伤+迷走神经刺激组按照以上方法建立兔脑爆炸弹片伤模型,爆炸弹片伤+迷走神经刺激组爆炸伤后接受右侧颈迷走神经电刺激(10V,5HZ,5ms,20min)。对各组实验兔进行血清.(伤后6h)和脑组织匀浆(伤后24h)TNF-a、IL-1β和IL-10浓度检测及脑组织含水量测定,并观察伤后24h实验兔脑组织病理变化。在实验的第三阶段中,另选32只雄性新西兰大白兔按随机数字表法分为假手术组(8只)、爆炸伤组(12只)和爆炸弹片伤+迷走神经刺激组(12只),爆炸伤组和爆炸弹片伤+迷走神经刺激组制作兔脑爆炸伤模型,爆炸弹片伤+迷走神经刺激组爆炸伤后接受右侧颈迷走神经电刺激(10V,5HZ,5ms,20min)。对各组兔进行血清(伤后8h及24h)和脑脊液(伤后24h) TNF-a和IL-10浓度检测及脑组织含水量测定,并记录各组实验兔伤前、伤后6h-8h和22h-24h三个时间长度为2h的长城心电图。结果：实验兔脑爆炸弹片伤后头颅CT检查可见明显的脑挫伤、脑出血,并有颅内积气和铁屑残留等开放性颅脑损伤的表现；脑组织切片呈现神经元变性坏死、脑组织水肿、出血等病理改变。在脑爆炸伤早期(伤后6h-8h),爆炸弹片伤组兔血清和脑内促炎因子TNF-a和IL-1p浓度及脑组织含水量显著高于假手术组和迷走神经刺激组(P0.01),而抗炎因子IL-10浓度介于假手术组和迷走神经刺激组之间(P0.05)。在脑爆炸伤后期(24h),爆炸弹片伤组兔血清和脑内促炎因子TNF-a浓度及脑组织含水量显著高于假手术组和迷走神经刺激组(P0.05),抗炎因子IL-10浓度显著低于假手术组和迷走神经刺激组(P0.05)；而假手术组与迷走神经刺激组之间抗炎因子IL-10浓度差异不明显(P0.10)。与假手术组相比,在脑爆炸弹片伤早期,爆炸弹片伤组和爆炸弹片伤+迷走神经刺激组反映自主神经系统功能平衡的指标低频/高频(LF/HF)比值显著升高(P0.01)；在脑爆炸弹片伤后期,爆炸弹片伤组LF/HF比值明显降低(P=0.01),而爆炸弹片伤+迷走神经刺激组无显著变化(P=0.320)。结论：本实验建立的兔脑爆炸弹片伤模型符合颅脑爆炸伤模型的影像学和病理学等特点,并且简便易行、质控标准,可应用于基层颅脑爆炸伤的基础研究。迷走神经电刺激可以减轻脑爆炸弹片伤兔脑组织水肿程度,降低血清及脑组织内TNF-a和IL-1β的浓度,升高IL-10的浓度,改善脑内及全身炎症反应程度从而发挥对脑爆炸弹片伤兔的脑保护作用。迷走神经电刺激可以改善和调控自主神经系统功能平衡,调节呼吸循环系统功能,发挥促进脑爆炸弹片伤兔良好预后的作用。
[Abstract]:Objective: in the first stage of the experiment, by combining the blast shock wave with the iron debris, a rabbit model of explosive fragment injury was established, which was close to the actual situation, the operation was simple, the quality control was standardized, the stability was high and the repeatability was good, and the injury changes of the experimental rabbits were observed. The second, third stage of the test was established by the first stage. The animal model was made by the model of rabbit brain exploding bullet wound, and the effects of electric stimulation of the vagus nerve on brain tissue pathology, edema change and the regulation of brain and systemic inflammatory response and autonomic nervous system dysfunction were observed and discussed respectively.
Methods: the first stage of the experiment was to select 24 male New Zealand white rabbits, randomly divided into pre experimental group (4), blank control group (4), sham operation group (6), explosive fragment injury group (10), using self designed explosive device to explode the right parietal cortex of rabbit brain after craniotomy. The signs of life, CT and brain of the rabbit after injury were observed. Through the study of the quality control, high stability and repeatability of the simple rabbit brain explosion bullet wound model, a suitable animal model was established and the experiment entered the second, third stage. In the second stage of the experiment, the other 24 New Zealand white rabbits were divided into the sham group (6) and the explosive fragment group (10) according to the random number table method. The explosive fragment injury group and the vagus nerve stimulation group (8), the explosion bullet wound group and the explosive fragment injury + vagus nerve stimulation group established the rabbit model of the explosion bullet wound according to the above methods, and the explosive bullet wound + vagus nerve stimulation group received the right cervical vagus nerve stimulation (10V, 5HZ, 5ms, 20min) after the blast injury. After 6h) and brain tissue homogenate (post injury 24h) TNF-a, IL-1 beta and IL-10 concentration detection and determination of brain tissue water content, and observe the pathological changes of brain tissue in 24h experimental rabbits after injury. In the third stage of the experiment, 32 New Zealand white rabbits were divided into sham group (8) according to random number table method, explosion injury group (12) and explosive fragment injury + fan walk The nerve stimulation group (12), the explosion injury group and the explosive fragment injury + vagus nerve stimulation group made the rabbit brain explosion injury model, the explosion bullet wound + vagus nerve stimulation group received the right cervical vagus nerve stimulation (10V, 5HZ, 5ms, 20min). The serum (8h and 24h after injury) and the cerebrospinal fluid (24h) TNF-a and IL-10 concentrations were detected in each group of rabbits. And the water content of brain tissue was measured. The electrocardiogram of the the Great Wall with three time 2h of 6h-8h and 22h-24h after injury was recorded.
Results: the head CT examination of the rabbit brain explosion shrapnel showed obvious brain contusion, cerebral hemorrhage, and the expression of open craniocerebral injury, such as intracranial gas and iron residue, and the pathological changes of neuron degeneration and necrosis, brain edema, bleeding and other pathological changes. In the early stage of brain explosion injury (6h-8h after injury), rabbit blood was injured by explosive fragment. The concentration of TNF-a and IL-1p in the brain and brain water content in the brain and brain tissue were significantly higher than that of the sham operation group and the vagus nerve stimulation group (P0.01), but the concentration of anti inflammatory factor IL-10 was between the sham operation group and the vagus nerve stimulation group (P0.05). In the late stage of the brain explosion injury (24h), the serum and brain proinflammatory factor TNF-a concentration and brain group in the rabbits in the explosive fragment injury group were in the brain group and brain group. The water content of the fabric was significantly higher than that of the sham operation group and the vagus nerve stimulation group (P0.05), the IL-10 concentration of the anti inflammatory factor was significantly lower than that of the sham operation group and the vagus nerve stimulation group (P0.05), but the difference of the concentration of anti inflammatory factors IL-10 between the sham operation group and the vagus nerve stimulation group was not significant (P0.10). The ratio of low frequency / high frequency (LF/HF) to the functional balance of the autonomic nervous system was significantly increased (P0.01), and the LF/HF ratio of the explosive fragment was significantly decreased (P=0.01) in the late stage of the explosion of the brain exploded fragment (P=0.01), but there was no significant change in the explosive fragment and the vagus nerve stimulation group (P=0.320).
Conclusion: the model of rabbit brain explosion bullet wound in this experiment is conformed to the imaging and pathological features of the model of craniocerebral explosive injury, and it is simple and easy to use. The standard of quality control can be applied to the basic study of brain explosion injury at the base level. The electric stimulation of vagus nerve can reduce the degree of brain edema in rabbits injured by the brain explosions and reduce the serum and brain tissue. The concentration of TNF-a and IL-1 beta, increase the concentration of IL-10 and improve the degree of inflammation in the brain and the whole body, thus exerting protective effect on the brain of rabbits injured by brain explosions. Electrical stimulation of the vagus nerve can improve and regulate the function balance of the autonomic nervous system, regulate the function of the respiratory circulatory system, and promote the good prognosis of the rabbit with the brain explosion flick.

【学位授予单位】：厦门大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R651.15

【参考文献】