Departamento de Toxicología y Farmacología, Facultad de Veterinaria, Universidad Complutense of Madrid, 28040 Madrid, Spain.
Departamento de Toxicología y Farmacología, Facultad de Veterinaria, Universidad Complutense of Madrid, 28040 Madrid, Spain.
Toxicol Lett. 2014 Mar 3;225(2):252-63. doi: 10.1016/j.toxlet.2013.12.016. Epub 2013 Dec 27.
4-Aminopyridine (4-AP) is an orphan drug indicated for the treatment of neuromuscular disorders. There is a great controversy around the use of this drug because of its narrow safety index and because a large number of adverse effects have been reported. Moreover, it was shown to induce cell death in different cell lines, being reported mainly apoptosis and necrosis as the principal pathways of cell death mediated by blockage of K channels or the Na, K-ATPase, but until now it was not described in vivo cell death induced by 4-aminipyridine. To provide new subchronic toxicity data and specifically, evaluate if 4-AP is able to induce in vivo cell death process and the main pathways related to it, a repeated dose (28 days) oral toxicity study, at therapeutic range of doses, was conducted in rats. The anatomical pathology, the biochemical and hematological parameters were analyzed and a real-time PCR array analysis was developed with an Ingenuity Pathway Analysis (IPA). The leucocytes number, the lactate dehydrogenase (LDH) and aspartate aminotransferase (AST) enzymatic activity were increased at all dose but the erythrocytes number, the hemoglobin concentration, the alkaline phosphatase (FAL) and alanine aminotransferase (ALT) enzymatic activity were increased only at highest dose studied. However, glucose levels decreased at all doses. The biochemical results are indicative of hepatic damage. The anatomy pathology studies showed cell death only on liver and kidney, and the real-time PCR array on liver tissue expressed a gene expression profile of necrotic and apoptotic induced cell death. The present work shows for the first time in vivo cell death on liver and kidney with features of apoptosis and necrosis induced by 4-AP and the gene expression profile shows that the cell death is mediated by necrotic and apoptotic pathways that support this finding.
4-氨基吡啶（4-AP）是一种用于治疗神经肌肉疾病的孤儿药。由于其安全性指数狭窄，并且已经报道了大量不良反应，因此围绕该药物的使用存在很大争议。此外，它已被证明可在不同的细胞系中诱导细胞死亡，主要报道的是凋亡和坏死作为阻断 K 通道或 Na，K-ATP 酶介导的细胞死亡的主要途径，但直到现在，尚未在体内描述过 4-氨基吡啶诱导的细胞死亡。为了提供新的亚慢性毒性数据，并特别评估 4-AP 是否能够诱导体内细胞死亡过程及其相关的主要途径，在大鼠中进行了重复剂量（28 天）口服毒性研究，剂量处于治疗范围内。分析了解剖病理学、生化和血液学参数，并使用实时 PCR 阵列分析和 IPA 进行了分析。白细胞数、乳酸脱氢酶（LDH）和天冬氨酸转氨酶（AST）酶活性在所有剂量下均增加，但红细胞数、血红蛋白浓度、碱性磷酸酶（FAL）和丙氨酸转氨酶（ALT）酶活性仅在研究的最高剂量下增加。然而，所有剂量下的血糖水平均降低。生化结果表明存在肝损伤。解剖病理学研究仅在肝脏和肾脏中显示出细胞死亡，实时 PCR 阵列在肝脏组织中表达了诱导细胞坏死和凋亡的基因表达谱。本工作首次在体内显示了 4-AP 诱导的肝脏和肾脏细胞凋亡和坏死的特征性细胞死亡，基因表达谱表明细胞死亡是由坏死和凋亡途径介导的，这支持了这一发现。
