Heuman R, Boeryd B, Bolin T, Magnusson K E, Sjödahl R, Tagesson C
Acta Chir Scand. 1980;146(3):195-201.
The subcellular organelles in diseased ileal mucosa of a patient with Crohn's disease have been separated by rate zonal density gradient centrifugation. Mucosal scrapings were obtained from a surgical specimen of diseased intestine and the cells disrupted by extrusion under controlled pressure. The homogenized cells were then centrifuged to prepare a cell extract and the extract fractionated in a single-step procedure by zonal centrifugation on a continuous sucrose gradient. The subcellular organelles (brush borders, basal-lateral membranes, lysosomes, mitochondria and endoplasmic reticulum) were located in the density gradient by assay of marker enzymes and their overall distribution compared with the distribution of organelles from a normal tissue. The centrifugal properties of individual organelles in the diseased tissue were largely unaffected but the lysosomes were more fragile than in the normal tissue. Since the fractionation technique used is on a large scale, each fraction of interest can be further analyzed for a variety of compounds. Therefore, the subcellular pathology and pathophysiology in this and other cases of Crohn's disease may eventually be studied and discussed in other than morphological and descriptive terms.
通过速率区带密度梯度离心法分离了一名克罗恩病患者病变回肠黏膜中的亚细胞器。从病变肠道的手术标本中获取黏膜刮片，并在控制压力下通过挤压使细胞破碎。然后将匀浆后的细胞离心以制备细胞提取物，并通过在连续蔗糖梯度上进行区带离心，以单步程序对提取物进行分级分离。通过测定标记酶确定亚细胞器（刷状缘、基底外侧膜、溶酶体、线粒体和内质网）在密度梯度中的位置，并将其总体分布与正常组织细胞器的分布进行比较。病变组织中各个细胞器的离心特性基本未受影响，但溶酶体比正常组织中的更脆弱。由于所使用的分级分离技术规模较大，每个感兴趣的级分都可以进一步分析各种化合物。因此，最终可能会以形态学和描述性术语以外的方式研究和讨论该病例及其他克罗恩病病例中的亚细胞病理学和病理生理学。
