【作者】 呼格吉乐图；

【导师】 佘锐萍；

【作者基本信息】 中国农业大学 ， 基础兽医学， 2004， 博士

【摘要】 本研究采用甲苯胺蓝染色法和免疫组织化学方法，对肥大细胞(Mast cell,MC)在家兔体内的分布定位进行了系统观察。结果表明，健康家兔肥大细胞分布于多种组织器官中，各种组织中肥大细胞多分布于浅层的上皮组织或靠近血管、腺管或神经等易表露于环境抗原的位点。分布于呼吸道的肥大细胞数量随兔年龄的增大而增多，在鼻腔粘膜显著高于喉、气管和肺。另外本研究还对健康家兔的圆小囊、空肠粘膜固有层内肥大细胞(mucosal mast cell,MMC)及粘膜下层结缔组织肥大细胞(connective tissue mast cell,CTMC)进行了透射电镜观察．未发现它们之间存在超微结构的显著差异。根据颗粒基质电子密度的差异，将家兔粘膜免疫器官肥大细胞的胞浆颗粒分为高电子密度基质和低电子密度基质两个类型。 本研究对比观察了常规甲苯胺蓝(Routine Toluidine Blue staining,RTB)染色及长时间甲苯胺蓝染色(LTB)对用Carnoy’s氏液或4％中性福尔马林(neutral buffered formalin,NBF)固定的家兔空肠及圆小囊肥大细胞的组织化学染色特性。结果表明，在Carnoy’s氏液或NBF固定的家兔组织均显示出了不同数量的甲苯胺蓝阳性细胞。但Carnoys氏液固定的家兔组织染色效果更佳，而中性福尔马林则明显地阻断肥大细胞的着染能力。Carnoy’s氏液或福尔马林固定的组织切片采用LTB染色均可不同程度的增加肥大细胞的着染能力。 本研究还对巴氏杆菌感染的家兔及未感染家兔的空肠、圆小囊粘膜肥大细胞(MMC)进行了定量观察，结果表明感染巴氏杆菌后，家兔空肠和圆小囊粘膜肥大细胞的数量显著增多。 为探讨家兔肥大细胞与脑肠肽的关系，本研究采用免疫组织化学方法，应用连续切片对应染色法，观察了P物质和血管活性肠肽(VIP)等脑肠肽类物质在肥大细胞内的表达。结果表明，家兔粘膜免疫器官内一部分肥大细胞的胞浆有P物质样物质表达，同时发现了某些肥大细胞的胞质内有VIP样物质。另外，为了从形态学角度验证肥大细胞与肽能神经“双响”调节的理论，采用免疫组织化学方法对家兔背部皮肤进行肥大细胞与肽能神经末梢关系的研究。结果表明家兔皮肤肥大细胞与P物质能神经末梢存在较为密切的形态学构筑关系，肥大细胞可与P物质能神经纤维紧密接触，两者间可能发生功能上的联系。另外，本研究还采用兔抗人溶菌酶多克隆抗体免疫组化技术对家兔空肠及圆小囊肥大细胞进行溶菌酶阳性结果的观察，但并未检测出溶菌酶阳性的肥大细胞。更多还原

【Abstract】 In this study, we make a systematic observation on localazation of mast cells in rabbit by toluidine blue staining method. The results showed that mast cell of health rabbit distributed in many tissues, but mainly located in the superficial layer of epithelial tissues or in a location easy to reveal to antigen near blood vascular, gland tube and neuron. Mast cell numbers in respiratory tract increase with age and the numbers per unit area in mucosa of nasal cavity were significant greater than that in larynx, trachea and lung. Besides, the lamina propria mast cells (MMC) of sacculus rotundus and jejunum and the submucosa connective tissue mast cell of jejunum(CTMC) in rabbits were observed under electron microscopy. No ultrastructural differences between those subpopulation of MC were detected. The cytoplasmic granules of MC in rabbit mucosa immune apparatus were divided into two sub types by their electron density: the high electron density homogeneous granules and the low electron density homogeneous granules.In this study, the histochemical characteristic of rabbit mast cell in the jejunum and sacculus rotundus fixed with Carnoys fluid and neutral buffered formalin(NBF) were studied by Routine Toluidine Blue(RTB) and long time Toluidine Blue(LTB) staining. Numerous toluidine blue stained mast cells were presented in All samples fixed with Carnoys fluid and NBF. Staining result of rabbit tissues that fixed with Carnoys fluid were excellent, But the staining characters of mast cells fixed with NBF were blocked by neutral buffered formalin. The staining affinity of MC were increased differently by LTB staining method in all sections whether fixed in Carnoys fluid or fixed in NBF.The jejunum and sacculus rotundus mucosal mast cells (MMC) of Pasteurella Cuniculum affected rabbits and health rabbits were fixed in Carnoys fluid and stained with Toluidine Blue (TB)and the tissue samples were counted under light microscope. It is shown that the mucosal mast cell numbers of jejunum and sacculus rotundus of Pasteurella Cuniculum affected rabbits were increased significantly.For the sake of investigation about relationship between mast cell and pepteds, we observed the expression of substance P and VIP in rabbit mast cell by histochemical staining method and corresponding staining method of serial section. It is show that The cytoplasm of some mast cells of rabbit mucosal immune apparatus were show substance P positive and were contain vasoactive intestinal peptide. In addition, in order to testing the double dimension regulative theory between mast cell and peptedergic nerve from a morphological angles, we studied the relationship between dorsal cutis mast cell and peptedergic nerve terminal by an immunohistochemistry method and discovered rather close morphological construction ties between them. Mast cells and substance P positive nerve fibers so g et in close touch with each others that may occurred functional relationship between them. In addsion, a histochemical study of rabbit mast cell in jejunum and sacculus rotundus were carried by an immunol histochemical technique using rabbit monoclonal antibody raised against human lysozyme. But no lysozyme active mast cells were discovered in the serial section by immunol histochemical technique.更多还原