偏头痛模型大鼠中影像学与组织学相关性的研究

发布时间：2018-03-22 09:37

本文选题：偏头痛　切入点：基于体素的形态学分析　出处：《中国人民解放军医学院》2017年博士论文　论文类型：学位论文

【摘要】：研究背景及目的偏头痛是一种原发性头痛,其发病机制尚不清楚。基于体素的形态学分析(VBM)可捕捉大脑结构的微细改变,对于参与偏头痛发病机制的大脑结构的揭示起着重要的作用。目前关于VBM显示结构改变的组织学机制尚不清楚,因此对于偏头痛患者VBM显示的改变结果需谨慎解释。本研究拟通过研究偏头痛模型大鼠大脑VBM显示结构改变与组织学改变的相关性明确VBM改变的组织学机制,同时进一步探索偏头痛的发病机制。方法1.通过炎症汤刺激清醒大鼠硬脑膜建立偏头痛模型将60只Sprague Dawley大鼠随机平均分配到10组:低频炎症汤刺激组(10μL炎症汤,每四天一次,刺激3周)、高频炎症汤刺激组(10μL炎症汤,每天一次,刺激3周)、低频炎症汤刺激恢复组(低频炎症汤刺激3周后停止刺激3周)、高频炎症汤刺激恢复组(高频炎症汤刺激3周后停止刺激3周)、长时间高频炎症汤刺激组(高频炎症汤刺激6周)以及相应对照组(将10μL炎症汤替换为10μL生理盐水)。2.行为学观察及机械痛阈值测定大鼠行为学在给药前录制15分钟,给药后录制30分钟,每4天录制一次。静止和休息行为时间通过Etho VisionXT动物跟踪软件获得。面部理毛时间通过人工计时记录。给药前用Von-Frey纤维丝测定大鼠给药同侧眶周皮肤机械痛阈值。低频炎症汤刺激组与其对照组每四天测一次,高频炎症汤刺激组与其对照组隔天一次。3.磁共振数据采集以及VBM分析给药3周、6周后使用7.0T超导磁共振机给予大鼠行T2核磁结构像扫描(每组6只)。使用spmratIHEP工具包对数据进行图像预处理及分析。4.组织学分析大鼠完成磁共振扫描之后,每组随机选取5只进行经心脏灌注取脑。采用免疫组化染色的方法标记感兴趣区域前额叶、中脑导水管周围灰质(PAG)、脑桥被盖部和三叉神经脊髓核尾部(Sp5C)的相关指标NeuN、GFAP、Ibal、MBP、PLP的表达情况。结果1.偏头痛模型大鼠炎症汤刺激组较相应对照组在给予药物刺激后大鼠出现更长的静止、休息时间和面部理毛时间,差异具有统计学意义(P0.05)。从给药后第三天开始,高频炎症汤刺激组大鼠眶周皮肤机械痛阈值较其对照组明显下降,具有统计学意义(P0.05)。2. VBM结果以及其与免疫组化染色结果的相关性与对照组相比,低频炎症汤刺激组在延髓和脑桥出现白质体积增加。高频炎症汤刺激组与对照组相比,白质体积增高表现在前额叶、前边缘叶、海马和运动皮层等区域。高频炎症汤刺激恢复组与对照组相比,白质体积增高表现在基底节、胼胝体以及海马等区域;丘脑、PAG、脑桥等区域出现灰质体积增加,海马区域灰质体积下降。与对照组相比,长时间高频炎症汤刺激组在前额叶、胼胝体、海马、丘脑等区域出现白质体积增高;灰质体积下降区域表现在胼胝体、基底节及感觉皮层,同时下丘脑灰质体积增加。白质体积的指标变化与髓鞘形成及继发性炎症改变如小胶质细胞增生和胶质细胞增生相关。PLP在反映髓鞘病变较MBP更敏感。结论1. VBM可发现传统T2加权像核磁不能发现的潜在病理改变结构。偏头痛模型大鼠中存在脑干尤其是三叉神经传导系统的结构异常,同时大鼠出现的痛觉过敏可能与前额叶的结构异常相关。2. VBM结构改变与髓鞘形成及继发性炎症改变相关。胶质细胞的活化和髓鞘异常与偏头痛相关性疼痛的发病机制相关。3.偏头痛患者VBM结构改变可能是头痛反复发作的结果,同时VBM显示的异常结构在偏头痛的慢性化过程中发挥作用。
[Abstract]:Background and objective: migraine is a primary headache, its pathogenesis is still not clear. Voxel based morphometry (VBM) can capture the fine structure of the brain changes, plays an important role in revealing the brain structures involved in the pathogenesis of migraine. At present the VBM display on the histological structure change mechanism not clear, so for migraine patients with VBM show a change in results should be interpreted with caution. In this study the brain VBM study of migraine model rats showed clear correlation is VBM changes of structure change and organizational changes to the organization, and to further explore the pathogenesis of migraine. Methods 1. Decoction by inflammatory stimulation of the dura mater in conscious rats to establish migraine model 60 Sprague Dawley rats were randomly assigned to 10 groups: low inflammation soup stimulation group (10 L inflammation soup, once every four days for 3 weeks, high frequency stimulation) Inflammation soup stimulation group (10 L inflammation soup, once every 3 weeks, low frequency stimulation) inflammation stimulation recovery group (low inflammation soup soup stimulation for 3 weeks after the cessation of stimulation for 3 weeks), high-frequency stimulation recovery group (high inflammation soup soup to stimulate inflammation 3 weeks after the cessation of stimulation for 3 weeks), long time with high frequency Zhengtang stimulation group (high frequency stimulation of inflammation Decoction for 6 weeks) and control group (10 L 10 L to replace the inflammatory soup saline).2. behavior observation and mechanical pain threshold determination of rat behavior before administration recorded 15 minutes, 30 minutes after administration of recording, recording every 4 rest and rest days. Time by Etho VisionXT animal tracking software. Facial grooming time by manual timing records. Before administration of Von-Frey fiber was administered to rats ipsilateral periorbital skin mechanical pain threshold. The low frequency stimulation of inflammation Decoction group and the control group were measured once every four days, the high frequency stimulation of inflammation soup group and The control group next time.3. magnetic resonance data acquisition and analysis of VBM treatment for 3 weeks, 6 weeks after the use of 7.0T superconducting magnetic resonance machine was given to rats for T2 magnetic structure imaging (n = 6). After using the spmratIHEP toolkit for image preprocessing and analysis of.4. histological analysis of rats performed MRI scans on the data in each group, 5 rats were randomly selected from perfusion through the heart and brain. The immunohistochemical staining method of marker region of interest in prefrontal cortex, midbrain periaqueductal gray (PAG), pons tegmentum and trigeminal spinal nucleus (Sp5C) tail correlation index NeuN, GFAP, Ibal, MBP, PLP expression. Compared with the control group in the presence of longer static rats were given drugs to stimulate the inflammation after the decoction of 1. migraine model rats were stimulated, rest time and facial hair, the difference was statistically significant (P0.05). From the third day after administration, high frequency Inflammation soup stimulated rats periorbital skin mechanical pain threshold decreased significantly compared to the control group, with statistical significance (P0.05) and the.2. VBM results and immunohistochemical staining of the relevance of the results compared with the control group, the low frequency of inflammatory stimulation group in the medulla oblongata and pons Decoction appear white matter volume increased. High frequency stimulation group and inflammatory soup compared with the control group, the white matter volume increased in the prefrontal cortex, anterior limbic lobe, hippocampus and motor cortex area. High frequency stimulation to restore inflammation Decoction group compared with the control group, the white matter volume increased in the basal ganglia, hippocampus and corpus callosum region; the thalamus, PAG, pons area increased gray matter volume, hippocampus regional gray matter volume decreased. Compared with the control group, long time high frequency stimulation group inflammatory soup in the prefrontal cortex and corpus callosum, hippocampus, thalamus and other regions appear white matter volume increased; a decrease in gray matter volume area now corpus callosum, basal And the sensory cortex, and hypothalamus increased gray matter volume. Changes of white matter volume and myelin formation and secondary inflammatory changes such as proliferation of microglia and glial cell proliferation related.PLP in reflecting the myelin lesions were more sensitive than MBP. Conclusion: 1. VBM can be found in traditional T2 weighted images can not be found in nuclear magnetic potential pathological structural changes. Especially the brainstem trigeminal nerve conduction system abnormalities in migraine rats, abnormal structure and hyperalgesia may also appear in the prefrontal cortex of rats.2. VBM structure change and myelin formation and secondary inflammatory changes. Change the structure of VBM associated with the pathogenesis of.3. by activation of glial cells and myelin abnormalities and migraine related pain the recurrent headache migraine may be the result of abnormal structure at the same time VBM display play a role in the progress of chronic migraine.

【学位授予单位】：中国人民解放军医学院
【学位级别】：博士
【学位授予年份】：2017
【分类号】：R747.2;R-332

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