Laboratory Animal Research Center, Ajou University School of Medicine, Suwon 16499, Korea.
Department of Pharmacy, College of Pharmacy and Institute of Pharmaceutical Sciences, CHA University, Pocheon 11160, Korea.
Int J Mol Sci. 2022 Jun 30;23(13):7294. doi: 10.3390/ijms23137294.
Excessive use of alcohol can induce neurobiological and neuropathological alterations in the brain, including the hippocampus and forebrain, through changes in neurotransmitter systems, hormonal systems, and neuroimmune processes. We aimed to investigate the effects of ethanol on the expression of coding and noncoding RNAs in a brain-derived cell line exposed to ethanol. After exposing Neuro2a cells, a neuroblastoma cell line, to ethanol for 24 and 72 h, we observed cell proliferation and analyzed up- and downregulated mRNAs and long noncoding RNAs (lncRNAs) using total RNA-Seq technology. We validated the differential expression of some mRNAs and lncRNAs by RT-qPCR and analyzed the expression of and through knock-down of . Cell proliferation was significantly reduced in cells exposed to 100 mM ethanol for 72 h, with 1773 transcripts up- or downregulated by greater than three-fold in ethanol-treated cells compared to controls. Of these, 514 were identified as lncRNAs. Differentially expressed mRNAs and lncRNAs were mainly observed in cells exposed to ethanol for 72 h, in which and decreased, but , , and increased. On the other hand, lncRNAs , , and were changed by ethanol, and in particular was greatly increased by chronic ethanol treatment through inhibition of . Our results increase the understanding of cellular and molecular mechanisms related to coding and noncoding RNAs in an in vitro model of acute and chronic exposure to ethanol.
过度饮酒会通过改变神经递质系统、激素系统和神经免疫过程,在大脑中引起神经生物学和神经病理学的改变,包括海马体和前脑。我们旨在研究乙醇对暴露于乙醇的脑源性细胞系中编码和非编码 RNA 表达的影响。在将神经母细胞瘤细胞系 Neuro2a 细胞暴露于乙醇 24 和 72 小时后,我们观察到细胞增殖,并使用总 RNA-Seq 技术分析上调和下调的 mRNA 和长非编码 RNA(lncRNA)。我们通过 RT-qPCR 验证了一些 mRNA 和 lncRNA 的差异表达,并通过 敲低分析了 和 的表达。与对照组相比,暴露于 100mM 乙醇 72 小时的细胞中细胞增殖显著降低,乙醇处理的细胞中 1773 个转录物的表达上调或下调超过三倍。其中 514 个被鉴定为 lncRNA。差异表达的 mRNA 和 lncRNA 主要在暴露于乙醇 72 小时的细胞中观察到,其中 和 降低,但 、 、 和 增加。另一方面,乙醇改变了 lncRNA 、 、 和 ,特别是通过抑制 ,慢性乙醇处理大大增加了 lncRNA 。我们的结果增加了对体外急性和慢性乙醇暴露模型中编码和非编码 RNA 相关的细胞和分子机制的理解。