Histological study of experimental murine amyloidosis

实验性小鼠淀粉样变性的组织学研究

• 批准号: 11671812
• 负责人: KUROIWA Mie
• 金额: $ 2.05万
• 依托单位: Showa University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11671812/
• 关键词: Amyloid; Amyloid Enhancing Factor; Mouse; Macrophage; Fibroblast; Oral Histology; Congo Red; Ultrastructure; AAアミロイド; ALアミロイド; アゾカゼイン; Amyloid Enhancing Factor

Amyloidosis is characterized by the deposition of amyloid fibrils in the extracellular matrix of various organs and oral tissues in animals as well as humans. In the experimental murine amyloidosis, the production of amyloidosis was examined by carrying out histological study.Female mice were given an intraperitoneal injection of 0.5 ml of amyloid enhancing factor (AEF) and a subcutaneously injection of 0.2 ml of an emulsion of M.Butyricum in Freund's complete adjuvant (FCA). The mice were sacrificed under ether anesthesia and blood was taken from the heart. Serum amyloid A (SAA) was measured using a Cytoscreen/Immunoassay kit for mouse SAA.The peritoneal macrophages were cultured with AEF and protein of the amyloid. Paraffin sections of the tissues fixed in paraformaldehyde were stained with Congo red, H-E, PAS and Berlin Blue and were immunostained with anti-amyloid P component, anti-macrophages F4/80. Subsequently, the ultrastructures of tissues fixed in Karnovsky's solution were examined with an electron microscope.The amyloidosis was induced in the perifollicular zone and marginal zone in spleen. SAA levels, amyloid P component and macrophage numbers of spleen and liver increased. The increased macrophages located in marginal zone and red pulp in spleen contained proteoglycan, ferric ion (Fe^<3+>) and amyloid P component. The deposition of amyloid is composed of the amyloid fibrils which are made up of amyloid P component, chondroitin sulfate proteoglycan, heparan sulfate proteoglycan and protein of the amyloid. The formation of amyloidosis is concerned with the macrophages and fibroblasts.

淀粉样变性的特征在于淀粉样纤维在动物以及人的各种器官和口腔组织的细胞外基质中的沉积。在实验性小鼠淀粉样变性中，通过进行组织学研究来检查淀粉样变性的产生，对雌性小鼠腹膜内注射0.5ml淀粉样增强因子（AEF）和皮下注射0.2ml丁酸分枝杆菌在弗氏完全佐剂（FCA）中的乳剂。在乙醚麻醉下处死小鼠，并从心脏取血。用小鼠血清淀粉样蛋白A（SAA）的Cytoscreen/Immunoassay试剂盒检测SAA。用多聚甲醛固定的组织石蜡切片用刚果红、HE、PAS和柏林蓝染色，并用抗淀粉样蛋白P成分、抗巨噬细胞F4/80进行免疫染色。电镜观察脾组织超微结构，发现脾滤泡周围带和边缘带发生淀粉样变性。脾、肝SAA含量、淀粉样蛋白P成分及巨噬细胞数增加。脾脏边缘区和红髓内巨噬细胞增多，含有蛋白多糖、铁离子（Fe^<3+>）和淀粉样蛋白P成分。淀粉样蛋白的沉积由淀粉样纤维组成，淀粉样纤维由淀粉样蛋白P成分、硫酸软骨素蛋白多糖、硫酸乙酰肝素蛋白多糖和淀粉样蛋白组成。淀粉样变性的形成与巨噬细胞和成纤维细胞有关。

项目成果

Sadayuki Inoue: "Basement membranes, microfibrils and β amyloid fibrillogenesis in Alzheimer's disease: high resolution ultrastructural findings."Brain Research Reviews. 29. 218-231 (1999)

Sadayuki Inoue：“阿尔茨海默病中的基底膜、微原纤维和 β 淀粉样原纤维形成：高分辨率超微结构发现。”《脑研究评论》29. 218-231 (1999)。

Sadayuki Inoue: "Basement membranes, microfibrils and β amyloid fibrillogenesis in Alzheimer's disease : high resolution ultrastructural findings,"Brain Research Reviews. 29. 218-231 (1999)

Sadayuki Inoue：“阿尔茨海默病中的基底膜、微原纤维和 β 淀粉样原纤维形成：高分辨率超微结构发现”，《大脑研究评论》29. 218-231 (1999)。

S.Inoue, M.Kuroiwa, R.Kisilevsky: "Basement membranes, microfibrils and β amyloid fibrillogenesis in Alzheimer's disease : high resolution ultrastructural findings."Brain Research Reviews. Vol.29. 218-231 (1999)

S.Inoue、M.Kuroiwa、R.Kisilevsky：“阿尔茨海默病中的基底膜、微原纤维和 β 淀粉样原纤维生成：高分辨率超微结构发现”。《脑研究评论》第 218-231 卷（1999 年）。