Amphetamine (AMPH) is a systemic stimulant used to treat a variety of diseases including Attention Deficit Hyperactive Disorder, narcolepsy and obesity. Previous data showed that by binding to catecholamine transporters, AMPH prevents the reuptake of the neurotransmitters dopamine (DA) and norepinephrine (NE). Because AMPH, either used therapeutically at final concentrations of 1-10 µM or abused as recreational drug (50-200 µM), is taken over long periods of time, we investigated the prolonged effects of this drug on the uptake of DA. We found that, in LLC-PK1 cells stably expressing the human DA transporter (hDAT), pretreatments with 1 or 50 µM AMPH caused significant reduction in DA uptake right after the 15-h pretreatment. Remarkably, after 50 but not 1 µM AMPH pretreatment, we observed a significant reduction in DA uptake also after one, two or three cell divisions. To test whether these long-term effects induced by AMPH where conserved in a model comparable to primordial neuronal cells and native neurons, we used the human neuroblastoma cell line SH-SY5Y cells, which were reported to endogenously express both hDAT and the NE transporter. Pretreatments with 50 µM AMPH caused a significant reduction of DA uptake both right after 15 h and 3 cell divisions followed by neuro-differentiation with retinoic acid (RA) for 5 days. Under these same conditions, AMPH did not change the intracellular concentrations of ATP, ROS and cell viability suggesting, therefore, that the reduction in DA uptake was not cause by AMPH-induced toxicity. Interestingly, while 1 µM AMPH did not cause long-term effects in the LLC-PK1 cells, in the SH-SY5Y cells, it decreased the DA uptake after one, two, but not three, cell divisions and 5-day RA differentiation. These data show that besides the well-known acute effects, AMPH can also produce long-term effects in vitro that are maintained during cell division and transmitted to the daughter cells.

中文翻译：

---

长期的苯丙胺治疗会导致培养细胞中多巴胺的长期吸收减少。

苯丙胺（AMPH）是一种全身性兴奋剂，用于治疗多种疾病，包括注意力缺陷多动障碍，发作性睡病和肥胖症。先前的数据显示，AMPH通过结合儿茶酚胺转运蛋白，阻止了神经递质多巴胺（DA）和去甲肾上腺素（NE）的再摄取。由于AMPH的治疗时间长于1-10 µM的最终浓度，或者被滥用为休闲药（50-200 µM），因此需要长期服用，因此我们研究了该药对DA吸收的延长作用。我们发现，在稳定表达人DA转运蛋白（hDAT）的LLC-PK1细胞中，用15或1μMAMPH进行预处理可在15 h预处理后立即引起DA摄取的显着降低。值得注意的是，经过50次但不是1 µM AMPH预处理后，我们观察到在进行1次AMPH预处理后，DA吸收也显着降低，两个或三个单元格。为了测试由AMPH诱导的这些长期效应在与原始神经元细胞和天然神经元相当的模型中是否保守，我们使用了人类神经母细胞瘤细胞系SH-SY5Y细胞，据报道其内源性表达hDAT和NE转运蛋白。用15 µM AMPH预处理可在15 h和3个细胞分裂后立即显着降低DA摄取，然后用视黄酸（RA）进行神经分化5天。在这些相同条件下，AMPH不会改变细胞内ATP，ROS的浓度和细胞活力，因此，DA摄取的减少不是由AMPH诱导的毒性引起的。有趣的是，虽然1 µM AMPH不会对LLC-PK1细胞造成长期影响，但在SH-SY5Y细胞中，在一，两，但不是三个，分别是细胞分裂和5天RA分化。这些数据表明，除了众所周知的急性作用外，AMPH还可以在体外产生长期作用，这种作用在细胞分裂期间得以维持并传递给子细胞。