Brody A R, George G, Hill L H
Lab Invest. 1983 Oct;49(4):468-75.
Chrysotile and crocidolite are commonly used forms of asbestos. Hemolysis has been widely used as a test of membrane injury, and it has been shown previously that chrysotile causes rapid breakdown of red blood cells (RBCs), whereas crocidolite is only weakly hemolytic. A reasonable hypothesis set forth to explain the cytotoxic effects of chrysotile maintains that positively charged chrysotile fibers bind to negatively charged sialic acid residues on RBC membranes causing clustering of membrane proteins and increased cell permeability to Na and K ions. Our studies presented here provide two lines of evidence in direct support of this hypothesis. (a) Morphologic--Ultrastructural techniques showed that both chrysotile and crocidolite asbestos bind to and distort more than 85% of RBCs treated for 15 minutes. The distorting effects of chrysotile, but not crocidolite, were almost totally ablated by pretreating the cells with neuraminidase. In addition, gold-conjugated wheat germ agglutinin was used to label the distribution of sialic acid groups on RBC membranes. Pretreatment of the RBCs with chrysotile, but not crocidolite, reduced the number of gold-conjugated wheat germ agglutinin-labeled sites to less than 30% of the control level. (b) Biochemical--The thiobarbituric acid assay was used to determine the percentage of sialic acid that remained with the cell pellet after neuraminidase and/or asbestos treatment. Asbestos treatment alone caused no release of sialic acid from the cells. Neuraminidase treatment for 3.5 hours removed more than 80% of the sialic acid from cell surfaces. Chrysotile, but not crocidolite, asbestos prevented neuraminidase-mediated removal of sialic acid from RBCs. In addition, x-ray energy spectrometry of freeze-dried cells showed that RBCs distorted by chrysotile, but not by crocidolite, exhibited significant alterations in intracellular Na:K ratios. The morphologic and biochemical data strongly support the hypothesis that chrysotile asbestos binds to sialic acid groups on RBC membranes. Consequently, the sialic acid residues are redistributed on the surfaces of distorted cells which then are unable to maintain a normal Na:K balance with the surrounding medium.
温石棉和青石棉是常用的石棉形式。溶血作用已被广泛用作膜损伤的检测方法,此前已有研究表明,温石棉会导致红细胞(RBC)迅速破裂,而青石棉的溶血作用较弱。为解释温石棉的细胞毒性作用而提出的一个合理假设认为,带正电荷的温石棉纤维与RBC膜上带负电荷的唾液酸残基结合,导致膜蛋白聚集,并增加细胞对Na和K离子的通透性。我们在此展示的研究提供了两条直接支持这一假设的证据。(a)形态学——超微结构技术显示,温石棉和青石棉都会与超过85%经15分钟处理的RBC结合并使其变形。用神经氨酸酶预处理细胞后,温石棉(而非青石棉)的变形作用几乎完全消除。此外,用金标记的麦胚凝集素用于标记RBC膜上唾液酸基团的分布。用温石棉(而非青石棉)预处理RBC后,金标记的麦胚凝集素标记位点的数量减少至对照水平的30%以下。(b)生物化学——硫代巴比妥酸测定法用于确定经神经氨酸酶和/或石棉处理后仍留在细胞沉淀中的唾液酸百分比。单独的石棉处理不会导致细胞释放唾液酸。神经氨酸酶处理3.5小时可去除细胞表面超过80%的唾液酸。温石棉(而非青石棉)可阻止神经氨酸酶介导的RBC唾液酸去除。此外,对冻干细胞的X射线能谱分析表明,因温石棉(而非青石棉)而变形的RBC细胞内Na:K比值发生了显著变化。形态学和生物化学数据有力地支持了温石棉与RBC膜上唾液酸基团结合的假设。因此,唾液酸残基在变形细胞表面重新分布,进而无法与周围介质维持正常的Na:K平衡。