Casartelli A, Bonato M, Cristofori P, Crivellente F, Dal Negro G, Masotto I, Mutinelli C, Valko K, Bonfante V
Safety Assessment Department, GlaxoSmithKline Research Centre, Verona, Italy.
Cell Biol Toxicol. 2003 Jun;19(3):161-76. doi: 10.1023/a:1024778329320.
Phospholipidosis is a term commonly used to indicate a phospholipid storage disorder; in affected cells, phospholipids accumulate in lysosomes that acquire a multilamellar morphological appearance. Cationic amphiphilic drugs (CADs) are suggested to induce phospholipidosis by direct interaction of xenobiotics with intracellular phospholipids or by the action of xenobiotics on the synthesis and metabolism of phospholipids. To date, electron microscopy (EM) represents the most reliable and the preferred method for the demonstration of phospholipidotic cell damage. Nevertheless, EM has a low throughput, it is expensive, and it is not suitable for screening purposes. We discuss here the assessment of the the phospholipidogenic potential of drugs using a cell culture-based model. In this test, intracellular phospholipids of treated U-937 cells (a human monocyte-derived cell line) were measured using the fluorescent probe Nile red. Eleven CADs reported to induce phospholipidosis in vivo and eight nonphospholipidogenic drugs were tested. Results obtained with the U-937 model confirmed the phospholipidogenic potential of drugs tested as described in the literature. Results have also been correlated with data obtained with a physical-chemical model (chromatographic hydrophobicity index measurement). Good correlation was obtained, confirming that the physical-chemical properties of CADs play a crucial role in the development of phospholipidosis. This work demonstrates that the U-937 model is a rapid and sensitive method for the determination of phospholipidosis-mediated cell damage. The specificity and the predictive potency observed make this method suitable for screening purposes in pharmaceutical development.
磷脂沉积症是一个常用于表示磷脂储存障碍的术语；在受影响的细胞中，磷脂在溶酶体中积累，溶酶体呈现出多层形态外观。阳离子两亲性药物（CADs）被认为可通过异生物质与细胞内磷脂的直接相互作用或异生物质对磷脂合成和代谢的作用来诱导磷脂沉积症。迄今为止，电子显微镜（EM）是证明磷脂沉积性细胞损伤最可靠且首选的方法。然而，EM通量低、成本高，且不适用于筛选目的。我们在此讨论使用基于细胞培养的模型评估药物的磷脂生成潜力。在该试验中，使用荧光探针尼罗红测量经处理的U - 937细胞（一种人单核细胞衍生细胞系）的细胞内磷脂。测试了11种据报道在体内可诱导磷脂沉积症的CADs和8种非磷脂生成药物。用U - 937模型获得的结果证实了文献中所述测试药物的磷脂生成潜力。结果还与通过物理化学模型（色谱疏水性指数测量）获得的数据相关。获得了良好的相关性，证实了CADs的物理化学性质在磷脂沉积症的发生中起关键作用。这项工作表明，U - 937模型是一种快速且灵敏的测定磷脂沉积症介导的细胞损伤的方法。所观察到的特异性和预测效力使该方法适用于药物研发中的筛选目的。
