Laboratory of Clinical and Experimental Toxicology, and Poison Control Centre and National Toxicology Information Centre, Toxicology Unit, Istituti Clinici Scientifici Maugeri IRCCS, Via Maugeri 10, 27100, Pavia, Italy.
Department of Morphology, Surgery and Experimental Medicine, Section of Legal Medicine and LTTA Center, University of Ferrara, Ferrara, Italy.
Neurotox Res. 2021 Aug;39(4):1251-1273. doi: 10.1007/s12640-021-00369-3. Epub 2021 May 4.
Considering the consequences on human health, in general population and workplace, associated with the use of new psychoactive substances and their continuous placing on the market, novel in vitro models for neurotoxicology research, applying human-derived CNS cells, may provide a means to understand the mechanistic basis of molecular and cellular alterations in brain. Cytotoxic effects of MAM-2201, a potent-naphthoyl indole derivative-synthetic cannabinoid, have been evaluated applying a panel of human cell-based models of neurons and astrocytes, testing different concentrations (1-30 µM) and exposure times (3-24-48 h). MAM-2201 induced toxicity in primary neuron-like cells (hNLCs), obtained from transdifferentiation of mesenchymal stem cells derived from human umbilical cord. Effects occurred in a concentration- and time-dependent manner. The lowest concentration affecting cell viability, metabolic function, apoptosis, morphology, and neuronal markers (MAP-2, NSE) was 5 μM, and even 1 μM induced apoptosis. Effects appeared early (3 h) and persisted after 24 and 48 h. Similar behavior was evidenced for human D384-astrocytes treated with MAM-2201. Differently, human SH-SY5Y-neurons, both differentiated and undifferentiated, were not sensitive to MAM-2201. On D384, the different altered endpoints were reversed, attenuated, or not antagonized by AM251 indicating that CB1 receptors may partially mediate MAM-2201-induced cytotoxicity. While in hNLCs, all toxic effects caused by MAM-2201 were apparently unrelated to CB-receptors since they were not evidenced by immunofluorescence. The present in vitro findings demonstrate the cytotoxicity of MAM-2201 on human primary neurons (hNLCs) and astrocytes cell line (D384), and support the use of these cellular models as species-specific in vitro tools suitable to clarify the neurotoxicity mechanisms of synthetic cannabinoids.
考虑到与使用新精神活性物质及其不断投放市场相关的对人类健康的影响，特别是在一般人群和工作场所，应用源自中枢神经系统细胞的新型体外神经毒理学研究模型，可能有助于了解大脑中分子和细胞改变的机制基础。应用神经元和星形胶质细胞的人类细胞模型，评估了一种有效的萘酰吲哚衍生物合成大麻素 MAM-2201 的细胞毒性，测试了不同浓度（1-30µM）和暴露时间（3-24-48 小时）。MAM-2201 诱导源自人脐带间充质干细胞的神经元样细胞（hNLCs）的毒性。作用呈浓度和时间依赖性。影响细胞活力、代谢功能、凋亡、形态和神经元标志物（MAP-2、NSE）的最低浓度为 5µM，甚至 1µM 即可诱导凋亡。作用出现在早期（3 小时），并在 24 和 48 小时后持续存在。用 MAM-2201 处理的人 D384 星形胶质细胞也表现出类似的行为。不同的是，人 SH-SY5Y 神经元，无论是分化的还是未分化的，都对 MAM-2201 不敏感。在 D384 上，不同的改变终点被 AM251 逆转、减弱或不拮抗，表明 CB1 受体可能部分介导 MAM-2201 诱导的细胞毒性。而在 hNLCs 中，MAM-2201 引起的所有毒性作用显然与 CB 受体无关，因为它们没有通过免疫荧光法得到证实。本体外研究结果表明，MAM-2201 对人原代神经元（hNLCs）和星形胶质细胞系（D384）具有细胞毒性，并支持使用这些细胞模型作为种特异性体外工具，以阐明合成大麻素的神经毒性机制。
