靶向动脉粥样硬化斑块内Tenascin-C表达的磁共振体内成像实验研究

发布时间：2017-12-31 18:42

本文关键词：靶向动脉粥样硬化斑块内Tenascin-C表达的磁共振体内成像实验研究　出处：《南京医科大学》2015年硕士论文　论文类型：学位论文

【摘要】：第一部分ApoE~(-/-)小鼠腹主动脉粥样硬化MR成像及Tenascin-C、CD68表达的实验研究目的探讨小鼠腹主动脉粥样硬化模型7.0TMR成像的可行性,检测动脉粥样硬化斑块中Tenascin-C(TN-C)的表达,并探讨其与CD68的相关性,为进一步研究其在动脉粥样硬化发生发展中靶向成像的可行性奠定试验研究基础。方法高脂饲养法建立实验组ApoE~(-/-)小鼠(n=20)和对照组野生型C57小鼠腹主动脉AS模型(n=10),14周后行MRI检查,后处死动物取腹主动脉,行HE染色观察血管病理学变化,并分别使用油红O、anti-CD68及TN-C抗体染色观察斑块脂质分布情况及CD68、TN-C的表达。结果建模14周后,MRI见实验组腹主动脉血管壁增厚,在T1WI、PDWI呈高信号,T2WI呈稍低信号;病理学检查见血管内膜明显增厚,管腔不规则狭窄;TN-C及CD68在斑块内均高表达,且二者表达区域相一致;对照组腹主动脉MRI未见管强狭窄,病理检查均未见内膜增厚。结论高脂饲养法可快速建立ApoE~(-/-)小鼠AS模型,7.0MR可成功检测之;TN-C在AS斑块中高表达且与CD68关系密切,二者可能通过相互协同,在AS发生发展中共同起作用。检测TN-C的表达,将有助于动脉粥样硬化病变的进一步研究。第二部分靶向动脉粥样硬化斑块内Tenascin-C表达的MR体内成像实验研究目的检测TN-C在动脉粥样硬化斑块中的表达并探讨MR抗体靶向TN-C体内成像的可行性。方法采用高脂饲养法建立ApoE~(-/-)小鼠主动脉粥样硬化模型,14周后使用7.0micro-MR观察主动脉斑块情况;HE及油红O染色观察血管壁病理学改变,免疫组织化学检测TN-C在斑块内的表达;化学偶联法合成抗体靶向TN-C的MR成像探针(anti-TN-C-SPIO探针),并通过尾静脉注人小鼠体内,以单纯USPIO为对照组,于8小时后进行MR扫描。成像后立即取标本,行普鲁士蓝染色。结果建模后14周,MRI见小鼠腹主动脉壁增厚,T1WI、PDWI为稍高信号,T2WI为稍低信号;HE染色见动脉内膜明显增厚。油红O染色证实为脂质沉积,免疫组化显示TN-C在AS斑块内高表达;实验组小鼠注射anti-TNC-SPIO探针8小时后,见增厚的血管壁于T2WI信号较前降低,与注射前相比,差异有统计学意义(P0.05)。相应的普鲁士蓝染色见斑块内蓝色颗粒明显沉积;对照组MRI未见明显信号改变,普鲁士蓝染色仅见极少量蓝色颗粒沉积。结论Anti-TNC-SPIO可以显示AS斑块内信号的改变,可为进一步开展斑块的MR分子影像学研究奠定实验基础。
[Abstract]:Part I Mr imaging and Tenascin-C of abdominal aorta atherosclerosis in mice. Objective to investigate the feasibility of 7.0T Mr imaging of abdominal aortic atherosclerosis model in mice. The expression of tenascin-Con TN-C in atherosclerotic plaques was detected and its correlation with CD68 was investigated. In order to further study the feasibility of target imaging in atherosclerosis. Methods the experimental group ApoEN-/-/-mice were established by high fat feeding method. And the wild type C57 mice as model of abdominal aorta in the control group (n = 10). After 14 weeks MRI examination was performed and the abdominal aorta was taken from the animals. The pathological changes of the vessels were observed by HE staining and oil red O was used respectively. The distribution of plaque lipid and the expression of CD68 TN-C were observed by anti-CD68 and TN-C antibody staining. The signal intensity was slightly low on T _ 1WI and T _ 2WI. Pathological examination showed obvious thickening of the intima and irregular stenosis of the lumen. Both TN-C and CD68 were highly expressed in plaque, and their expression regions were consistent. In the control group, there was no strong stenosis in abdominal aorta MRI and no intimal thickening was observed in pathological examination. Conclusion High fat feeding method can be used to quickly establish as model in mice. TN-C is highly expressed in atherosclerotic plaques and is closely related to CD68. They may play a role in the pathogenesis and development of as by cooperating with each other. The expression of TN-C is detected. In vivo Mr imaging of Tenascin-C expression targeting atherosclerotic plaques objective to detect TN-C in atherosclerotic plaques. Expression of TN-C in sclerosing plaques and the feasibility of Mr antibody targeted imaging in vivo. Methods High fat feeding method was used to establish ApoEG (ApoEN). Rat aortic atherosclerosis model. The plaque of aorta was observed with 7.0micro-Mr after 14 weeks. The pathological changes of vascular wall were observed by HE and oil red O staining, and the expression of TN-C in plaque was detected by immunohistochemistry. Anti-TN-C-SPIO probes targeting TN-C were synthesized by chemical coupling method, and then injected into mice via tail vein. USPIO was used as the control group. Mr scanning was performed 8 hours later. The specimens were taken immediately after imaging and stained with Prussian blue. Results at 14 weeks after modeling, the thickening of abdominal aorta wall in mice was seen as slightly hyperintense on T1WII-PDWI. The signal intensity was slightly low on T2WI. The thickening of intima was observed by HE staining. Oil red O staining confirmed lipid deposition and high expression of TN-C in atherosclerotic plaques by immunohistochemistry. Eight hours after injection of the anti-TNC-SPIO probe, the thickened vascular wall in the experimental group showed a decrease in signal intensity on T2WI, which was compared with that before injection. The difference was statistically significant (P 0.05). The blue particles in the plaques were obviously deposited in the corresponding Prussian blue staining. In control group, there was no obvious signal change in MRI, but only a few blue particles were deposited in Prussian blue staining. Conclusion Anti-TNC-SPIO can show the change of signal in atherosclerotic plaques. It can lay an experimental foundation for the further study of Mr molecular imaging of plaque.
【学位授予单位】：南京医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R543.5;R445.2

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