长链非编码 RNA SNHG16 减轻氯胺酮诱导的人胚胎干细胞源性神经元毒性。
Long noncoding RNA SNHG16 reduced ketamine-induced neurotoxicity in human embryonic stem cell-derived neurons.
机构信息
Clinical laboratory, First Hospital of Jilin university, Changchun, 130021, China.
Department of Anesthesiology, China-Japan Union Hospital of Jilin University, Changchun 130033, China.
出版信息
J Chem Neuroanat. 2018 Dec;94:39-45. doi: 10.1016/j.jchemneu.2018.08.005. Epub 2018 Aug 29.
BACKGROUND
Clinical evidence demonstrates that prolonged exposure to ketamine may cause irreversible injury to immature human brains. In this study, we utilized an in vitro model to examine the function of long noncoding RNA (lncRNA) SNHG16 in ketamine-induced neurotoxicity in human embryonic stem cell (hESC)-derived neurons.
METHODS
HESCs were induced toward neuronsin vitro, and treated with ketamine, at various concentrations, for 48 h. Viability, apoptosis, caspase-3 activity and ROS activity were then examined among hESC-derived neurons. Ketamine-induced gene expression change of SNHG16 was assessed by qRT-PCR. SNHG16 was overexpressed in hESC-derived neurons, which were then treated with ketamine, followed by biochemical assays to assess the effects of SNHG16 upregulation on ketamine-induced neurotoxicity. Correlation between SNHG16 and NeuroD1 gene was assess by qRT-PCR. In SNHG16-upregulated hESC-derived neurons, they were double transfected with siRNA to knock down NeuroD1. The functions of NeuroD1 inhibition on SNHG16-associated neural protection on ketamine-induced neurotoxicity were further assessed.
RESULTS
48-h in vitro treatment of ketamine induced significant neurotoxicity, and downregulated SNHG16 among hESC-derived neurons. Conversely, SNHG16 upregulation reduced ketamine-induced neurotoxicity. NeuroD1 expression was downregulated by ketamine in hESC-derived neurons, and concomitantly upregulated by SNHG16 overexpression. SiRNA-mediated NeuroD1 inhibition reversed the protection of SNHG16 upregulation on ketamine-induced neurotoxicity.
CONCLUSIONS
SNHG16 is an important epigenetic factor which may functionally modulate ketamine-induced neurotoxicity through NeuroD1.
背景
临床证据表明，长时间接触氯胺酮可能会对人类未成熟大脑造成不可逆转的损伤。在这项研究中，我们利用体外模型研究了长链非编码 RNA（lncRNA）SNHG16 在氯胺酮诱导的人胚胎干细胞（hESC）衍生神经元神经毒性中的作用。
方法
体外诱导 hESC 向神经元分化，用不同浓度的氯胺酮处理 48 小时。然后检测 hESC 衍生神经元的活力、凋亡、caspase-3 活性和 ROS 活性。qRT-PCR 检测氯胺酮诱导的 SNHG16 基因表达变化。在 hESC 衍生神经元中转染 SNHG16 过表达质粒，然后用氯胺酮处理，进行生化测定，评估 SNHG16 上调对氯胺酮诱导的神经毒性的影响。qRT-PCR 检测 SNHG16 与 NeuroD1 基因的相关性。在 SNHG16 上调的 hESC 衍生神经元中，用 siRNA 双重转染敲低 NeuroD1。进一步评估 NeuroD1 抑制对 SNHG16 相关的氯胺酮诱导神经毒性的神经保护作用。
结果
体外 48 小时氯胺酮处理诱导 hESC 衍生神经元明显的神经毒性，并下调 SNHG16。相反，SNHG16 上调可减轻氯胺酮诱导的神经毒性。氯胺酮下调 hESC 衍生神经元中的 NeuroD1 表达，同时上调 SNHG16 表达。siRNA 介导的 NeuroD1 抑制逆转了 SNHG16 上调对氯胺酮诱导的神经毒性的保护作用。
结论
SNHG16 是一种重要的表观遗传因子，可能通过 NeuroD1 调节氯胺酮诱导的神经毒性。
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