4-氟甲基苯丙胺 (4-FMA) 是一种苯丙胺类精神活性物质，具有公认的致敏和兴奋作用，但毒理学机制尚不清楚。考虑到绝大多数 4-FMA 使用者通过口服途径消耗这种物质，预计肝脏会高度暴露。这项工作的目的是使用体外肝细胞模型确定 4-FMA 的肝毒性潜力：原代大鼠肝细胞 (PRH)、人肝癌细胞系 HepaRG 和 HepG2，并采用 37 μM 至 30 mM 的浓度，在24 小时暴露。欧共体50根据 MTT 活力测定数据估计的每个模型的值分别为 2.21 mM、5.59 mM 和 9.57 mM。然后将最敏感的模型 PRH 共同暴露于 4-FMA 和细胞色素 P450 (CYP) 抑制剂，以研究代谢对 4-FMA 毒性的影响。结果表明，CYP2E1、CYP3A4 和 CYP2D6 在 4-FMA 细胞毒性中起主要作用。抑制 CYP2D6 和 CYP3A4 导致 4-FMA 的浓度-反应曲线左移，暗示这些代谢酶对 4-FMA 的解毒作用，而 CYP2E1 抑制表明这种酶在 4-FMA 中的毒化作用生理相关浓度下的 FMA 代谢。该药物还破坏了线粒体膜电位并降低了 ATP 水平，增加了活性氧和氮物质的产生并损害了硫醇抗氧化防御。4-FMA 通过干扰细胞凋亡和坏死的机制，增加启动子和效应子半胱天冬酶的活性，并导致细胞膜完整性丧失，进一步影响 PRH 的完整性。还观察到了自噬的潜力。这项研究有助于提供越来越多的关于新型精神活性物质毒性的证据，特别是关于它们的肝毒性作用；代谢对所产生的 4-FMA 细胞毒性的明显影响表明，对于使用者的后果存在很大程度的不可预测性，可能与剂量无关。增加起始和效应半胱天冬酶的活性，并导致细胞膜完整性的丧失。还观察到了自噬的潜力。这项研究有助于提供越来越多的关于新型精神活性物质毒性的证据，特别是关于它们的肝毒性作用；代谢对所产生的 4-FMA 细胞毒性的明显影响表明，对于使用者的后果存在很大程度的不可预测性，可能与剂量无关。增加起始和效应半胱天冬酶的活性，并导致细胞膜完整性的丧失。还观察到了自噬的潜力。这项研究有助于提供越来越多的关于新型精神活性物质毒性的证据，特别是关于它们的肝毒性作用；代谢对所产生的 4-FMA 细胞毒性的明显影响表明，对于使用者的后果存在很大程度的不可预测性，可能与剂量无关。
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4-Fluoromethamphetamine (4-FMA) induces in vitro hepatotoxicity mediated by CYP2E1, CYP2D6, and CYP3A4 metabolism
4-Fluoromethamphetamine (4-FMA) is an amphetamine-like psychoactive substance with recognized entactogenic and stimulant effects, but hitherto unclear toxicological mechanisms. Taking into consideration that the vast majority of 4-FMA users consume this substance through oral route, the liver is expected to be highly exposed. The aim of this work was to determine the hepatotoxic potential of 4-FMA using in vitro hepatocellular models: primary rat hepatocytes (PRH), human hepatoma cell lines HepaRG and HepG2, and resorting to concentrations ranging from 37 μM to 30 mM, during a 24-h exposure. EC50 values, estimated from the MTT viability assay data, were 2.21 mM, 5.59 mM and 9.57 mM, for each model, respectively. The most sensitive model, PRH, was then co-exposed to 4-FMA and cytochrome P450 (CYP) inhibitors to investigate the influence of metabolism on the toxicity of 4-FMA. Results show that CYP2E1, CYP3A4 and CYP2D6 have major roles in 4-FMA cytotoxicity. Inhibition of CYP2D6 and CYP3A4 led to left-geared shifts in the concentration–response curves of 4-FMA, hinting at a role of these metabolic enzymes for detoxifying 4-FMA, while CYP2E1 inhibition pointed towards a toxifying role of this enzyme in 4-FMA metabolism at physiologically-relevant concentrations. The drug also destabilised mitochondrial membrane potential and decreased ATP levels, increased the production of reactive oxygen and nitrogen species and compromised thiol antioxidant defences. 4-FMA further affected PRH integrity by interfering with the machinery of apoptosis and necrosis, increasing the activity of initiator and effector caspases, and causing loss of cell membrane integrity. Potential for autophagy was also observed. This research contributes to the growing body of evidence regarding the toxicity of new psychoactive substances, in particular regarding their hepatotoxic effects; the apparent influence of metabolism over the resulting cytotoxicity of 4-FMA shows that there is a substantial degree of unpredictability of the consequences for users that could be independent of the dose.
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