Department of Biochemistry and Biophysics, Stockholm University, Stockholm, Sweden.
Unit of Toxicology Sciences, Swedish Toxicology Sciences Research Center (Swetox), Karolinska Institutet, Södertälje, Sweden.
Sci Rep. 2020 Oct 7;10(1):16714. doi: 10.1038/s41598-020-73698-6.
Acrylamide (ACR) is a known neurotoxicant which crosses the blood-brain barrier, passes the placenta and has been detected in breast milk. Hence, early-life exposure to ACR could lead to developmental neurotoxicity. The aim of this study was to elucidate if non-cytotoxic concentrations of ACR alter neuronal differentiation by studying gene expression of markers significant for neurodevelopment in the human neuroblastoma SH-SY5Y cell model. Firstly, by using RNASeq we identified two relevant pathways that are activated during 9 days of retinoic acid (RA) induced differentiation i.e. RA receptor (RAR) activation and the cAMP response element-binding protein (CREB) signalling pathways. Next, by qPCR we showed that 1 and 70 µM ACR after 9 days exposure alter the expression of 13 out of 36 genes in the RAR activation pathway and 18 out of 47 in the CREB signalling pathway. Furthermore, the expression of established neuronal markers i.e. BDNF, STXBP2, STX3, TGFB1 and CHAT were down-regulated. Decreased protein expression of BDNF and altered ratio of phosphorylated CREB to total CREB were confirmed by western blot. Our results reveal that micromolar concentrations of ACR sustain proliferation, decrease neurite outgrowth and interfere with signalling pathways involved in neuronal differentiation in the SH-SY5Y cell model.
丙烯酰胺 (ACR) 是一种已知的神经毒素，它可以穿过血脑屏障，通过胎盘，并在母乳中检测到。因此，生命早期接触 ACR 可能导致发育性神经毒性。本研究旨在通过研究对神经发育有重要意义的标志物在人神经母细胞瘤 SH-SY5Y 细胞模型中的基因表达，阐明非细胞毒性浓度的 ACR 是否会改变神经元分化。首先，通过使用 RNASeq，我们鉴定了两条在视黄酸 (RA) 诱导分化的 9 天内激活的相关途径，即 RA 受体 (RAR) 激活和 cAMP 反应元件结合蛋白 (CREB) 信号通路。接下来，通过 qPCR，我们发现 9 天暴露于 1 和 70 μM ACR 后，RAR 激活途径中的 36 个基因中有 13 个和 CREB 信号通路中的 47 个基因中的 18 个表达发生改变。此外，已建立的神经元标志物 BDNF、STXBP2、STX3、TGFB1 和 CHAT 的表达下调。Western blot 进一步证实 BDNF 的蛋白表达减少和磷酸化 CREB 与总 CREB 的比值改变。我们的结果表明，微摩尔浓度的 ACR 维持增殖，减少突起生长，并干扰 SH-SY5Y 细胞模型中参与神经元分化的信号通路。
