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--------------------------Page1------------------------------＼渊黜MRIMAGINGoFANTIBoDYTARGETINGPHENOTYPEVASCULARSMOOTHCELLSINVITROAThesisSubmittedtoSoutheastUniversityFortheAcademicofMasterofDegreeMedicineBYSHANGSong--anSupervisedbyProf．MAZhan--longMedicalSchoolSoutheastUniversity2014May万方数据--------------------------Page2------------------------------东南大学学位论文独创性声明本人声明所呈交的学位论文是我个人在导师指导下进行的研究工作及取得的研究成果。尽我所知，除了文中特别加以标注和致谢的地方外，论文中不包含其他人已经发表或撰写过的研究成果，也不包含为获得东南大学或其它教育机构的学位或***而使用过的材料。与我一同工作的同志对本研究所做的任何贡献均己在论文中作了明确的说明并表示了谢意。研究生签名：j纽立金L日期：盈瑙已生壮东南大学学位论文使用授权声明东南大学、中国科学技术信息研究所、国家图书馆有权保留本人所送交学位论文的复印件和电子文档，可以采用影印、缩印或其他复制手段保存论文。本人电子文档的内容和纸质论文的内容相一致。除在保密期内的保密论文外，允许论文被查阅和借阅，可以公布(包括以电子信息形式刊登)论文的全部内容或中、英文摘要等部分内容。论文的公布(包括以电子信息形式刊登)授权东南大学研究生院办理。研究生签名：、堑挫宴导师签名：日期：2班：互。29万方数据--------------------------Page3------------------------------中文摘要原肌球蛋白．4抗体靶向标记合成型血管平滑肌细胞MRI体外成像的实验研究硕士研究生：尚松安指导教师：马占龙东南大学医学院(210009)中文摘要血管平滑肌细胞(Vascularsmoothmusclecell，VSMC)来源于胚胎中胚层，是血管壁中膜层的重要组成成分，对血管张力、血流动力学等具有重要的调节作用。在血管内膜损伤、炎症因子刺激等因素作用下，VSMC去分化重新进入细胞循环，获得去分化表型，即合成型，伴随增殖、迁移、凋亡坏死等病理改变，最终导致动脉粥样硬化、血管再狭窄、主动脉夹层等血管性病变的发生。因此合成型VSMC是诊断及防治血管性疾病的研究重点。等合成型标记蛋白的确定为合成型VSMC的鉴定及研究提供了更为精确的靶点。分子影像学借助无创性的影像手段在活体内实时、重复的显示靶细胞及其分子变化的信息，并对其生物学行为进行定量和定性研究。随着血管性疾病的生物学发生机制以及防治方法研究的进展，分子影像学成为其早期诊断以及疗效评估的重要方法。研究主要聚焦于疾病形成的机制、体外活体特异性探测、预防治疗效果评价等，分子成像方式以核素、MIu两种分子成像技术为主。其中，M砌分子成像具有高空间分辨率、多序列成像等优点，能够在传统M砒技术的基础上，以特殊分子或细胞作为成像对象，把传统的物理成像转为特异性分子水平的分子成像，为VSMC的靶向性分子成像提供了广阔的应用空间。限制。本研究对颈动脉再狭窄模型MRj成像、再狭窄形成过程中合成型VSMC分布、高合成型VSMC体外分离及鉴定、靶向标记蛋白(TPM．4)表达变化等方面进行了系统研究，像学研究的可行性，为血管性疾病的早期诊断以及疗效评估提供更为精确科学的参照。本文主要分为以下3部分内容：第一部分：大鼠血管再狭窄形成中MRI成像及血管平滑肌细胞表型转化的实验研究T万方数据--------------------------Page4------------------------------东南大学硕士学位论文行7．0T测量结果行相关性分析；免疫组织化学染色分别鉴定收缩型、合成型VSMC的分布(SMA、及增殖活性改变。本部分旨在通过建立SD大鼠颈动脉RS模型，采用7．0TMIu动态观测再狭窄形成过程，研究VSMC在再狭窄形成过程中的表型转化，并探讨靶向合成型VSMC分子成像研究的可行性。第二部分：合成型血管平滑肌细胞分离、培养及鉴定的实验研究采用改良II型胶原酶消化法分离、体外培养SD大鼠胸主动脉VSMC，通过血小板衍生析细胞增殖活性与迁移活性；流式细胞周期测定细胞周期分布；倒置显微镜、结晶紫染色观察形态学改变并定量分析；免疫细胞化学染色鉴定VSMC并测定分析收缩型标记蛋白c【．讨以简便高效的方法获取VSMC，并分离、培养、鉴定处于高合成表型状态下VSMC，为血管性疾病的研究提供
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