PATHOGENESIS OF PANCREATIC SOTONE : STUDIES ON THE ORGANIC MATRIX OF HUMAN PANCREATIC STONES.

我们在胰腺结石患者的胰腺结石取出后立即进行治疗，通过免疫组织化学方法来阐明有机基质的成分。将结石在4%缓冲甲醛中固定24小时，然后在室温下在含有0.1M EDTA的0.5N HCl中固定脱钙长达60分钟。所有结石均用气泡脱钙，获得坚固的海绵状基质。标准染色包括H-E、PAS和阿辛蓝。免疫组织化学染色与特定的纤连蛋白，层粘连蛋白，胶原蛋白IV，乳铁蛋白（DAKO），和PSP-reg抗体进行抗生物素蛋白-生物素复合物的方法。用非免疫血清的对照染色为阴性。标准染色显示，有机成分在整个结石中形成基质，基质质地不均匀，如网状、层状和透明蛋白样，包括改变的细胞成分。在所有7个基质中，有广泛的区域强烈染色的纤维连接蛋白和层粘连蛋白。所有标本的IV型胶原，乳铁蛋白和PSP-reg阳性区域，但强度的程度和分布的模式是不同的，在每个矩阵。本研究中的脱钙过程适合于免疫组化。在EDTA中不断搅拌脱钙时，基质发生破碎。在胰腺结石中发现了有机基质。如预期，基质中的乳铁蛋白和PSP-reg呈阳性。纤维连接蛋白、层粘连蛋白和IV型胶原在基质中呈阳性表达，说明细胞外基质和基底膜成分在胰腺结石的发病中起重要作用。

We treated pancreatic stones just after removal each from pancreatic lithiasis patients to clarify the components of the organic matrix by immunohistochemistry. Stones were fixed in 4% buffered formaldehyde for 24 hours, then decalcified stationary in 0.5N HCI with 0.1M EDTA at room temperature for up to 60 minutes. All stones were decalcified with bubble and firm sponge-like matrices were obtained. Standard staining included H-E,PAS,and alcian blue was made. Immunohistochemical staining with specific fibronectin, laminin, collagen IV,lactoferrin (DAKO), and PSP-reg antibodies were carried out by avidin-biotin complex methods. Control staining with non-immune serum was negative. Standard staining revealed that organic component formed matrix throughout the stone and matrix was heterogeneous in texture such as reticular, lamellar, and hyalin-like including altered cellular constituent. In all seven matrices, there were wide areas intensely stained for fibronectin and laminin. All specimens had positive areas for collagen IV,lactoferrin and PSP-reg, but the degree of intensity and the pattern of distribution were different in each matrix. Process of decalcification in this study is suitable for immunohistochemistry. Fragmentation of matrix occured when decalcification was carried out in EDTA with constant stirring. Organic matrix was found throughout the pancreatic stone. Lactoferrin and PSP-reg were positive in the matrix as were expected. Fibronectin, laminin and collagen IV were positive in the matrix, so extracellular matrix and basement membrane components play an important role in the pathogenesis of the pancreatic stones.