硬脑膜静脉窦栓塞的应用解剖学研究
发布时间：2018-08-24 19:12
【摘要】： 本实验旨在通过一种操作简单、成功率高、耗资低廉的硬脑膜静脉窦(Duralvenous sinus,DVS)栓塞动物模型,探讨DVS栓塞后动物模型大脑的病理生理学及形态学改变,以期深入阐释脑静脉窦血栓形成(cerebral venous sinus thrombosis,CVST)等相关疾病的病理变化,并为临床治疗提供形态学方面的参考依据。实验中选用成年健康SD大鼠58只(雌雄各半,体重200-300g),随机分为栓塞组、假手术组及正常组(正常组6只,栓塞组及假手术组各26只,后两组大鼠根据术后处死时间再次分为6h组7只、1d组7只、3d组6只和5d组6只)。栓塞组大鼠麻醉后,颅骨手术开窗,于上矢状窦后部逆血流方向缓慢插入固体栓子并固定。假手术组仅开颅,正常组不手术。栓塞组与假手术组大鼠分别于术后6h、1d、3d、5d处死取脑,正常组大鼠随机选择处死时间。取栓塞段DVS旁脑皮质,采用干湿法测定脑含水量,并在石蜡切片后行HE染色及免疫组织化学染色,以观察不同时间点脑含水量及凋亡相关蛋白Caspase-3、Bcl-2和Bax等指标的变化情况,并与大体标本及光镜进行对照研究。以上实验结果显示:①栓塞组大鼠软脑膜充血严重,脑组织表面呈现水肿改变;上矢状窦内可见栓子前端附着有血栓形成;②栓塞组光镜下可见明显的脑水肿病理表现,以栓塞后6h最为显著;脑皮质内出现大量致密、深染的核固缩细胞,在栓塞6h组既可观察到,而在栓塞1d时最明显;假手术组中仅在术后短期内可观察到轻微水肿及炎症表现,正常组中未见上述异常变化;③栓塞组大鼠术后6h脑含水量较假手术6h组及正常组显著增加(P＜0.05),术后1d脑含水量仍高于正常组(P＜0.05);④栓塞组脑皮质中,Caspase-3、Bcl-2及Bax蛋白在术后一天内均有大量表达,且在1d组中表达最多,之后下降,而在所有时间点均与假手术组及正常组存在显著性差异(P＜0.05);栓塞组Bcl-2/Bax值始终低于假手术组和正常组,并在手术1天后明显下降。⑤栓塞组模型成功率为92.31%。由此可得出结论:①本实验采用的造模方法简单,手术耗时短,动物创伤小;栓子尾段留置在外的方法克服了栓子移位及再通的问题,造模成功率较高,该疾病动物模型对CVST的相关病理学以及临床治疗的深入研究都具有重要意义;②栓塞组大鼠脑水肿在手术6小时内达到高峰,之后逐渐恢复正常,说明仅造成上矢状窦栓塞并不能长期影响大脑血液循环,开放丰富的侧支通路可能是此时机体调节颅内血流动力学的手段之一。③栓塞组中,栓塞段DVS旁脑皮质出现神经细胞凋亡,提示细胞凋亡可能是DVS栓塞后引起继发性神经损害的机制之一。在此过程中,Caspase-3、Bcl-2及Bax等细胞凋亡相关的效应因子均有不同程度变化,因此在疾病早期采用针对凋亡相关基因的抗凋亡治疗,可为降低CVST的死亡率提供新对策。
[Abstract]:The purpose of this study was to investigate the pathophysiological and morphological changes of Duralvenous sinus,DVS model after DVS embolization by a simple, high success rate and low cost dural venous sinus (Duralvenous sinus,DVS) embolization model. In order to explain the pathological changes of cerebral venous sinus thrombosis (cerebral venous sinus thrombosis,CVST) and provide the morphological basis for clinical treatment. 58 healthy adult SD rats were randomly divided into three groups: embolization group (n = 6), sham operation group (n = 6), embolization group (n = 26) and sham operation group (n = 26). According to the time of death, the latter two groups were divided into 6 h group (7 rats), 7 rats in the 1st day group (6 rats in the 3rd day group) and 6 rats in the 5 day group (6 rats in the 5th day group). After anaesthesia, the rats in the embolization group underwent craniotomy and inserted the solid embolus slowly in the posterior part of the superior sagittal sinus. Sham operation group only craniotomy, normal group does not operate. The rats in embolization group and sham operation group were killed at 6 hours and 3 days after operation, and the normal rats were killed at random. The cerebral water content was measured by dry-wet method. After paraffin section, HE staining and immunohistochemical staining were performed to observe the changes of cerebral water content and apoptosis-related proteins (Caspase-3,Bcl-2 and Bax) at different time points. The results were compared with gross specimen and light microscope. The results showed that the pia meningeal congestion was severe and the surface of brain tissue showed edema in the embolization group, and the pathological manifestations of cerebral edema could be seen under light microscope in the embolization group with thrombus attached to the anterior end of the embolus in the superior sagittal sinus. A large number of dense and deeply stained nuclear pyknotic cells were observed in the cerebral cortex at 6 h after embolization, and at 1 day after embolization, and mild edema and inflammation were observed in the sham operation group only within a short period of time after embolization. No abnormal changes were found in the normal group. 3The cerebral water content in the embolization group was significantly higher than that in the sham operation group at 6 h after operation (P < 0. 05). The cerebral water content in the embolization group was still higher than that in the normal control group (P < 0. 05). The expression of Caspase-3, Bcl-2 and Bax protein in the cerebral cortex of the embolization group was significantly higher than that in the control group at 1 day after operation (P < 0. 05). The expression of Bcl-2/Bax in embolization group was lower than that in sham operation group and normal group (P < 0. 05), and the expression of Bcl-2/Bax in embolization group was lower than that in sham operation group and normal group at all time points (P < 0. 05), and the expression of Bcl-2/Bax in embolization group was lower than that in sham operation group and normal group. The success rate of the embolization group was 92.31%. It can be concluded that the method used in this experiment is simple, the operation time is short, the animal trauma is small, the method of leaving the tail segment of embolus out overcomes the problems of embolus displacement and recanalization, and the success rate of making model is high. The animal model of the disease has important significance for the further study of the pathology and clinical treatment of CVST. The cerebral edema in the embolization group reached the peak within 6 hours of operation, and then gradually returned to normal. It is suggested that only superior sagittal sinus embolism can not affect the cerebral blood circulation for a long time, and opening the abundant collateral pathway may be one of the methods of regulating intracranial hemodynamics in the embolization group. Neuronal apoptosis in the paracentral cortex of DVS suggests that apoptosis may be one of the mechanisms of secondary nerve damage after embolization of DVS. In this process, the apoptosis-related effectors such as Caspase-3, Bcl-2 and Bax all changed to varying degrees. Therefore, antiapoptotic therapy aimed at apoptosis-related genes in the early stage of disease may provide a new strategy for reducing the mortality of CVST.
【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R322.81
本文编号：2201767
[Abstract]:The purpose of this study was to investigate the pathophysiological and morphological changes of Duralvenous sinus,DVS model after DVS embolization by a simple, high success rate and low cost dural venous sinus (Duralvenous sinus,DVS) embolization model. In order to explain the pathological changes of cerebral venous sinus thrombosis (cerebral venous sinus thrombosis,CVST) and provide the morphological basis for clinical treatment. 58 healthy adult SD rats were randomly divided into three groups: embolization group (n = 6), sham operation group (n = 6), embolization group (n = 26) and sham operation group (n = 26). According to the time of death, the latter two groups were divided into 6 h group (7 rats), 7 rats in the 1st day group (6 rats in the 3rd day group) and 6 rats in the 5 day group (6 rats in the 5th day group). After anaesthesia, the rats in the embolization group underwent craniotomy and inserted the solid embolus slowly in the posterior part of the superior sagittal sinus. Sham operation group only craniotomy, normal group does not operate. The rats in embolization group and sham operation group were killed at 6 hours and 3 days after operation, and the normal rats were killed at random. The cerebral water content was measured by dry-wet method. After paraffin section, HE staining and immunohistochemical staining were performed to observe the changes of cerebral water content and apoptosis-related proteins (Caspase-3,Bcl-2 and Bax) at different time points. The results were compared with gross specimen and light microscope. The results showed that the pia meningeal congestion was severe and the surface of brain tissue showed edema in the embolization group, and the pathological manifestations of cerebral edema could be seen under light microscope in the embolization group with thrombus attached to the anterior end of the embolus in the superior sagittal sinus. A large number of dense and deeply stained nuclear pyknotic cells were observed in the cerebral cortex at 6 h after embolization, and at 1 day after embolization, and mild edema and inflammation were observed in the sham operation group only within a short period of time after embolization. No abnormal changes were found in the normal group. 3The cerebral water content in the embolization group was significantly higher than that in the sham operation group at 6 h after operation (P < 0. 05). The cerebral water content in the embolization group was still higher than that in the normal control group (P < 0. 05). The expression of Caspase-3, Bcl-2 and Bax protein in the cerebral cortex of the embolization group was significantly higher than that in the control group at 1 day after operation (P < 0. 05). The expression of Bcl-2/Bax in embolization group was lower than that in sham operation group and normal group (P < 0. 05), and the expression of Bcl-2/Bax in embolization group was lower than that in sham operation group and normal group at all time points (P < 0. 05), and the expression of Bcl-2/Bax in embolization group was lower than that in sham operation group and normal group. The success rate of the embolization group was 92.31%. It can be concluded that the method used in this experiment is simple, the operation time is short, the animal trauma is small, the method of leaving the tail segment of embolus out overcomes the problems of embolus displacement and recanalization, and the success rate of making model is high. The animal model of the disease has important significance for the further study of the pathology and clinical treatment of CVST. The cerebral edema in the embolization group reached the peak within 6 hours of operation, and then gradually returned to normal. It is suggested that only superior sagittal sinus embolism can not affect the cerebral blood circulation for a long time, and opening the abundant collateral pathway may be one of the methods of regulating intracranial hemodynamics in the embolization group. Neuronal apoptosis in the paracentral cortex of DVS suggests that apoptosis may be one of the mechanisms of secondary nerve damage after embolization of DVS. In this process, the apoptosis-related effectors such as Caspase-3, Bcl-2 and Bax all changed to varying degrees. Therefore, antiapoptotic therapy aimed at apoptosis-related genes in the early stage of disease may provide a new strategy for reducing the mortality of CVST.
【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R322.81
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