【摘要】： 本實驗旨在通過一種操作簡單、成功率高、耗資低廉的硬腦膜靜脈竇(Duralvenous sinus,DVS)栓塞動物模型,探討DVS栓塞后動物模型大腦的病理生理學及形態(tài)學改變,以期深入闡釋腦靜脈竇血栓形成(cerebral venous sinus thrombosis,CVST)等相關疾病的病理變化,并為臨床治療提供形態(tài)學方面的參考依據(jù)。實驗中選用成年健康SD大鼠58只(雌雄各半,體重200-300g),隨機分為栓塞組、假手術組及正常組(正常組6只,栓塞組及假手術組各26只,后兩組大鼠根據(jù)術后處死時間再次分為6h組7只、1d組7只、3d組6只和5d組6只)。栓塞組大鼠麻醉后,顱骨手術開窗,于上矢狀竇后部逆血流方向緩慢插入固體栓子并固定。假手術組僅開顱,正常組不手術。栓塞組與假手術組大鼠分別于術后6h、1d、3d、5d處死取腦,正常組大鼠隨機選擇處死時間。取栓塞段DVS旁腦皮質,采用干濕法測定腦含水量,并在石蠟切片后行HE染色及免疫組織化學染色,以觀察不同時間點腦含水量及凋亡相關蛋白Caspase-3、Bcl-2和Bax等指標的變化情況,并與大體標本及光鏡進行對照研究。以上實驗結果顯示:①栓塞組大鼠軟腦膜充血嚴重,腦組織表面呈現(xiàn)水腫改變;上矢狀竇內可見栓子前端附著有血栓形成;②栓塞組光鏡下可見明顯的腦水腫病理表現(xiàn),以栓塞后6h最為顯著;腦皮質內出現(xiàn)大量致密、深染的核固縮細胞,在栓塞6h組既可觀察到,而在栓塞1d時最明顯;假手術組中僅在術后短期內可觀察到輕微水腫及炎癥表現(xiàn),正常組中未見上述異常變化;③栓塞組大鼠術后6h腦含水量較假手術6h組及正常組顯著增加(P＜0.05),術后1d腦含水量仍高于正常組(P＜0.05);④栓塞組腦皮質中,Caspase-3、Bcl-2及Bax蛋白在術后一天內均有大量表達,且在1d組中表達最多,之后下降,而在所有時間點均與假手術組及正常組存在顯著性差異(P＜0.05);栓塞組Bcl-2/Bax值始終低于假手術組和正常組,并在手術1天后明顯下降。⑤栓塞組模型成功率為92.31%。由此可得出結論:①本實驗采用的造模方法簡單,手術耗時短,動物創(chuàng)傷小;栓子尾段留置在外的方法克服了栓子移位及再通的問題,造模成功率較高,該疾病動物模型對CVST的相關病理學以及臨床治療的深入研究都具有重要意義;②栓塞組大鼠腦水腫在手術6小時內達到高峰,之后逐漸恢復正常,說明僅造成上矢狀竇栓塞并不能長期影響大腦血液循環(huán),開放豐富的側支通路可能是此時機體調節(jié)顱內血流動力學的手段之一。③栓塞組中,栓塞段DVS旁腦皮質出現(xiàn)神經(jīng)細胞凋亡,提示細胞凋亡可能是DVS栓塞后引起繼發(fā)性神經(jīng)損害的機制之一。在此過程中,Caspase-3、Bcl-2及Bax等細胞凋亡相關的效應因子均有不同程度變化,因此在疾病早期采用針對凋亡相關基因的抗凋亡治療,可為降低CVST的死亡率提供新對策。
[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.
【學位授予單位】：天津醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2008
【分類號】：R322.81

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