Department of Anesthesiology, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Center for Specialty Strategy Research of Shanghai Jiao Tong University China Hospital Development Institute, Shanghai, China.
PLoS One. 2020 Dec 9;15(12):e0243644. doi: 10.1371/journal.pone.0243644. eCollection 2020.
Exposure to anesthetics during brain development may impair neurological function, however, the mechanisms underlying anesthetic neurotoxicity are unclear. Recent studies indicate that long non-coding RNAs (lncRNAs) are crucial for regulating the functional brain development during neurogenesis. This study aimed to determine the regulatory effects and potential mechanisms of lncRNA Peg13 (Peg13) on sevoflurane exposure-related neurotoxicity against neural stem cells (NSCs).
Mouse embryotic NSCs were isolated and their self-renewal and differentiation were characterized by immunofluorescence. NSCs were exposed to 4.1% sevoflurane 2 h daily for three consecutive days. The potential toxicities of sevoflurane against NSCs were evaluated by neurosphere formation, 5-ethynyl-2'-deoxyuridine (EdU) incorporation and flow cytometry assays. The Peg13, miR-128-3p and Sox13 expression in NSCs were quantified. The potential interactions among Peg13, miR-128-3p and Sox13 were analyzed by luciferase reporter assay. The effects of Peg13 and/or miR-128-3p over-expression on the sevoflurane-related neurotoxicity and Sox13 expression were determined in NSCs.
The isolated mouse embryotic NSCs displayed potent self-renewal ability and differentiated into neurons, astrocytes and oligodendrocytes in vitro, which were significantly inhibited by sevoflurane exposure. Sevoflurane exposure significantly down-regulated Peg13 and Sox13, but enhanced miR-128-3p expression in NSCs. Transfection with miR-128-3p mimics, but not the control, significantly mitigated the Peg13 or Sox13-regulated luciferase expression in 293T cells. Peg13 over-expression significantly reduced the sevoflurane-related neurotoxicity and increased Sox13 expression in NSCs, which were mitigated by miR-128-3p transfection.
Such data indicated that Peg13 mitigated the sevoflurane-related neurotoxicity by sponging miR-128-3p to preserve Sox13 expression in NSCs.
在大脑发育过程中接触麻醉剂可能会损害神经功能，但麻醉剂神经毒性的机制尚不清楚。最近的研究表明，长链非编码 RNA（lncRNA）对于调控神经发生过程中的功能性大脑发育至关重要。本研究旨在确定 lncRNA Peg13（Peg13）对七氟醚暴露相关神经毒性对神经干细胞（NSCs）的调节作用及其潜在机制。
分离小鼠胚胎 NSCs 并通过免疫荧光法对其自我更新和分化进行鉴定。将 NSCs 暴露于 4.1%七氟醚中，每天 2 小时，连续 3 天。通过神经球形成、5-乙炔基-2'-脱氧尿苷（EdU）掺入和流式细胞术检测评估七氟醚对 NSCs 的潜在毒性。定量检测 NSCs 中 Peg13、miR-128-3p 和 Sox13 的表达。通过荧光素酶报告基因检测分析 Peg13、miR-128-3p 和 Sox13 之间的潜在相互作用。在 NSCs 中检测 Peg13 和/或 miR-128-3p 过表达对七氟醚相关神经毒性和 Sox13 表达的影响。
分离的小鼠胚胎 NSCs 具有强大的自我更新能力，并在体外分化为神经元、星形胶质细胞和少突胶质细胞，而七氟醚暴露显著抑制了这一过程。七氟醚暴露显著下调 Peg13 和 Sox13，同时增强 NSCs 中 miR-128-3p 的表达。转染 miR-128-3p 模拟物而非对照可显著减轻 293T 细胞中 Peg13 或 Sox13 调节的荧光素酶表达。Peg13 过表达显著减轻 NSCs 中七氟醚相关的神经毒性并增加 Sox13 的表达，而 miR-128-3p 的转染可减轻这一作用。
这些数据表明，Peg13 通过海绵 miR-128-3p 减轻七氟醚相关的神经毒性，从而维持 NSCs 中的 Sox13 表达。
