Demonstration of plasma proteinase inhibitors in beta 2-microglobulin amyloid deposits.
Demonstration of plasma proteinase inhibitors in beta 2-microglobulin amyloid deposits.
复制标题
β2-微球蛋白淀粉样沉积物中血浆蛋白酶抑制剂的演示。
DOI：
10.1038/ki.1992.368
复制
发表时间：
1992
影响因子：
19.6
通讯作者：
Skinner,M
中科院分区：
文献类型：
--
作者：
Campistol,JM;Shirahama,T;Abraham,CR;Rodgers,OG;Solé,M;Cohen,AS;Skinner,M
β2-微球蛋白淀粉样沉积物中血浆蛋白酶抑制剂的证明。β2-微球蛋白相关淀粉样变性（Aβ 2 M）是长期透析患者的常见并发症。虽然其发病机制尚未完全清楚，但已知淀粉样纤维通常由完整的β2-微球蛋白（β 2 m）分子组成。血浆蛋白酶抑制剂（PPI）是一个广泛的糖蛋白家族，具有消除丝氨酸蛋白酶不必要的蛋白水解的功能。它们在淀粉样蛋白形成中的作用已成为激烈讨论的主题，特别是自从最近在阿尔茨海默病的β-蛋白淀粉样沉积物中鉴定出α1-抗糜蛋白酶以来。我们用免疫化学和生物化学方法评价了淀粉样变性透析患者骨关节和内脏组织中Aβ 2 M沉积物中几种PPI（α1-蛋白酶抑制剂、α1-抗糜蛋白酶、抗凝血酶III、α2-巨球蛋白和组织抑制剂金属蛋白酶）和淀粉样蛋白P组分的存在和分布，以及非透析患者的两个腕管滑膜和一个阿尔茨海默病脑作为对照。免疫组化结果显示，除一种抗α1-抗糜蛋白酶抗体外，其余PPI抗体均呈不同程度的Aβ 2 M阳性反应。所有抗体（包括抗α1-抗糜蛋白酶抗体）也在一定程度上与其他非淀粉样内脏和结缔组织成分发生扩散性和/或选择性反应。其中只有抗淀粉样蛋白P成分的反应与Aβ 2 M沉积物有明显的特异性定位，刚果红染色和抗β 2 m免疫组化反应在相邻连续切片中均能识别。溶解的β 2 m-淀粉样蛋白原纤维的生化分析（SDS-PAGE和Western印迹分析）证实了免疫组织化学研究的结果，证明了与针对所有供试PPI的抗体的阳性反应。此外，α1-ACT的Western印迹分析表明存在一条分子量约为65 kDa的条带，该条带与α1-ACT标准品的条带一致。α1-ACT的免疫组化和生化分析结果之间的这种差异可以通过α1-ACT与淀粉样纤维的共纯化来解释。
Demonstration of plasma proteinase inhibitors in β2-microglobulin amyloid deposits. β2-microglobulin-related amyloidosis (Aβ2M) represents a frequent complication in long-term dialysis patients. Although the pathogenetic mechanism has yet to be fully understood, it is known that amyloid fibrils usually consist of intact molecules of β2-microglobulin (β2m). Plasma proteinase inhibitors (PPI) are a broad family of glycoproteins with the function of eliminating unwanted proteolysis of serine proteases. Their role in amyloidogenesis has become a subject of intense discussion, especially since the recent identification of α1-antichymotrypsin in the β-protein amyloid deposits of Alzheimer's disease. We evaluated immunohistochemically and biochemically the presence and distribution of several PPIs (α1-proteinase inhibitor, α1-antichymotrypsin, antithrombin III, α2-macroglobulin and tissue inhibitor metalloproteinase) and amyloid P component in Aβ2M deposits in osteo-articular and visceral tissues from dialysis patients with amyloidosis, as well as two carpal tunnel synovia from non-dialysis patients and one Alzheimer's brain as controls. The immunohistochemical study demonstrated that all but one (anti-α1-antichymotrypsin) of the PPI antibodies tested showed varying degrees of positive reaction against Aβ2M deposits. All the antibodies (including anti-α1-antichymotrypsin) also reacted to some extent with other non-amyloid visceral and connective tissue elements diffusely and/or selectively. Among them, only the reaction of anti-amyloid P component had significantly distinctive localization to Aβ2M deposits, which were identified in adjacent serial sections by Congo red staining and immunohistochemical reaction against anti-β2m. The biochemical analysis (SDS-PAGE and Western blot analysis) of the solubilized β2m-amyloid fibrils confirmed the results of the immunohistochemical study, demonstrating positive reactions with the antibodies directed against all of the tested PPIs. Furthermore, the Western blot analysis of α1-ACT demonstrated the presence of one band of approximate molecular weight of 65 kDa, which comigrated with the band of the α1-ACT standard sample. This discrepancy between the immunohistochemical and biochemical analyses findings on α1-ACT, could be explained by the co-purification of α1-ACT with the amyloid fibrils.