利多卡因对兔蛛网膜下腔出血后早期脑损伤保护机制的研究

发布时间：2018-03-21 21:30

本文选题：蛛网膜下腔出血　切入点：早期脑损伤　出处：《贵州医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:探讨利多卡因(Lidocaine,LD)对兔蛛网膜下腔出血(subarachnoid hemorrhage,SAH)后早期脑损伤(early brain injury,EBI)的保护机制。方法:18只新西兰大白兔随机分为3组:假手术组(Sham);蛛网膜下腔出血组(SAH);利多卡因组(LD)。采用枕大池一次注血法复制兔蛛网膜下腔出血模型,各组动物称重后先予咪达唑仑肌肉注射做基础麻醉,然后进行耳缘静脉穿刺置管,经耳缘静脉缓慢注入咪达唑仑、枸橼酸舒芬太尼注射液及阿托品进行全麻诱导,待兔全麻生效后立即进行气管插管,气管插管成功后即可进行枕大池穿刺。触及枕骨下间隙,有突破感后见清亮脑脊液流出,然后采取兔耳中动脉血,蛛网膜下腔出血组及利多卡因组动物将兔自体动脉血1 mL/Kg经枕大池注入,假手术组则注入1 mL/Kg生理盐水,兔枕大池注射完毕后取30°头低位30 min,30 min后利多卡因组动物再次经枕大池注入2%利多卡因0.3 m L,蛛网膜下腔出血组、假手术组注入0.3 mL生理盐水,枕大池注液过程中给予呼吸机辅助呼吸,全部操作完毕后待兔自然苏醒,然后送回动物实验中心给予饲养观察。72 h后对兔进行摄食量和神经功能损害分级;采用实时荧光定量PCR(RT-PCR)方法及蛋白质免疫印迹(Western Blot)方法分别检测兔脑海马组织中含半胱氨酸的天冬氨酸蛋白水解酶3(cysteinyl aspartate specific proteinase 3,Caspase-3)、细胞色素C(Cytochrome C,Cyt-C)mRNA及蛋白表达变化;采用免疫组织化学染色,镜下观察兔脑海马CA1区Caspase-3、Cyt-C阳性表达情况。结果:对兔摄食量和神经功能损害分级发现,与Sham组比较,SAH、LD组动物摄食量减少,出现不同程度的神经功能损害(P0.05),与SAH组比较,LD组动物摄食量及神经功能损害无统计学差异(P0.05);RT-PCR、Western Blot统计结果显示,与Sham组比较,SAH、LD组Caspase-3、Cyt-C m RNA及蛋白表达增加(P0.05),与SAH组比较,LD组Caspase-3、Cyt-C m RNA及蛋白表达减少(P0.05);免疫组化检测Caspase-3、Cyt-C阳性细胞结果显示,与Sham组比较,SAH、LD组脑海马CA1区Caspase-3、Cyt-C阳性细胞数增加(P0.05),与SAH组比较,LD组Caspase-3、Cyt-C阳性细胞数减少(P0.05)。结论:利多卡因对兔蛛网膜下腔出血后早期脑损伤的保护机制可能与抑制线粒体通路激活有关。
[Abstract]:Objective: to investigate the protective mechanism of Lidocainedia lidocaine (LDD) on early brain injuryafter subarachnoid hemorrhage (SAH) in rabbits. Methods: eighteen New Zealand white rabbits were randomly divided into 3 groups: sham-operated group, subarachnoid hemorrhage group, Lidocaine group and lidocarcoma group. The rabbit model of subarachnoid hemorrhage was established by single blood injection into the cistern of occipital cistern. After weighing each group, midazolam was injected intramuscularly for basic anesthesia, then the ear margin vein was inserted into the tube, midazolam was injected slowly through the auricular vein, sufentanil citrate injection and atropine were induced by general anesthesia. Tracheal intubation was performed immediately after the rabbit general anesthesia took effect. After the intubation was successful, the cistern cistern was punctured. The suboccipital space was touched, the clear cerebrospinal fluid flowed out after a sense of breakthrough, and then the rabbit middle ear arterial blood was taken. Rabbits in subarachnoid hemorrhage group and lidocaine group were injected with 1 mL/Kg of autologous arterial blood through cistern magnum, while those in sham operation group were injected with 1 mL/Kg normal saline. After 30 掳cistern injection, the lidocaine group was injected with 2% lidocaine (0.3 mL) again through the cistern of the occipital cistern, subarachnoid hemorrhage group and sham operation group (0.3 mL normal saline). During the injection of fluid into the cistern of the occipital cistern, breathing machine was given to the rabbits. After all the operation was completed, the rabbits recovered naturally, and then sent back to the animal experimental center for feeding observation. 72 hours later, the feeding amount and the neurological damage were graded. The expression of 3(cysteinyl aspartate specific proteinase 3 caspase-3, cytochrome C(Cytochrome cyt mRNA and protein in rabbit brain tissues were detected by real-time fluorescence quantitative PCR assay and Western blot assay. Immunohistochemical staining was used to observe the positive expression of Caspase-3 Cyt-C in the CA1 region of rabbit hippocampus under microscope. Results: compared with the Sham group, the consumption of Caspase-3 Cyt-C was decreased in the Sham group. Compared with SAH group, there was no significant difference in food intake and nerve function damage between LD group and SAH group. The results of Western Blot analysis showed that there was no significant difference between LD group and LD group. Compared with Sham group, the expression of Caspase-3 Cyt-C m RNA and protein increased in SAH group, and the expression of Caspase-3 Cyt-C m RNA and protein decreased in LD group compared with SAH group. Compared with the Sham group, the number of Caspase-3 Cyt-C positive cells in the CA1 area of hippocampus increased in the Sham group, and the number of Caspase-3 Cyt-C positive cells decreased in the LD group compared with the SAH group. Conclusion: the protective mechanism of lidocaine on early brain injury after subarachnoid hemorrhage in rabbits may be related to the inhibitory line. Activation of granulocyte pathway is involved.
【学位授予单位】：贵州医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R743.35

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