Department of Anesthesiology, The First People's Hospital of Lianyungang, Lianyungang 222000, China.
Department of Anesthesiology, Women's Hospital of Nanjing Medical University, Nanjing Maternity and Child Health Care Hospital, No. 123, Tianfeixiang, Mochou Road, Nanjing 210004, China.
Comput Math Methods Med. 2022 Jul 13;2022:2517463. doi: 10.1155/2022/2517463. eCollection 2022.
Isoflurane (ISO) is a type of anesthetic that might cause neurotoxicity in children. Although miR-424-5p is considerably downregulated in ISO-treated rat brain samples, its physiological role in ISO-induced neuronal injury in human embryonic stem cell-derived neurons remains unknown (hESC-derived neurons). miR-424-5p expression and fatty acid synthase (FASN) in ISO-treated hESC-derived neurons were tested via qRT-PCR. The amount of protein for Bax, Cleaved-caspase-8, Bcl-2, and FASN was investigated through western blot analysis. The viability and apoptosis of hESC-derived neurons were estimated through cell counting kit-8 assessment and TUNEL assay, accordingly. Superoxide dismutase, glutathione, and malondialdehyde levels were discovered via corresponding kits. The contents of inflammatory factors including interleukin-6 and tumor necrosis factor- were examined by enzyme-linked immunosorbent assays. The combination between FASN and miR-424-5p was resolute via dual-luciferase reporter assessment. After exposure to ISO, induced neurotoxicity and a decreased miR-424-5p production were identified in hESC-derived neurons. Upregulation of miR-424-5p repressed ISO-induced apoptosis and mitigated ISO-induced inflammatory response and oxidative stress in vitro. FASN expression levels were reduced by elevation of miR-424-5p and upregulated after ISO treatment. Mechanically, FASN was directly targeted by miR-424-5p in hESC-derived neurons. Of note, the miR-424-5p elevation-suppressed neuronal apoptosis, inflammatory response, and oxidative stress were countered by upregulation of FASN.
异氟醚（ISO）是一种可能导致儿童神经毒性的麻醉剂。尽管 miR-424-5p 在 ISO 处理的大鼠脑样本中显著下调，但它在人胚胎干细胞源性神经元中 ISO 诱导的神经元损伤中的生理作用尚不清楚（hESC 衍生神经元）。通过 qRT-PCR 测试 ISO 处理的 hESC 衍生神经元中的 miR-424-5p 表达和脂肪酸合酶（FASN）。通过 Western blot 分析研究 Bax、Cleaved-caspase-8、Bcl-2 和 FASN 的蛋白量。通过细胞计数试剂盒-8 评估和 TUNEL 测定分别评估 hESC 衍生神经元的活力和凋亡。通过相应的试剂盒发现超氧化物歧化酶、谷胱甘肽和丙二醛水平。通过酶联免疫吸附测定法检查包括白细胞介素-6 和肿瘤坏死因子-α 在内的炎症因子的含量。通过双荧光素酶报告基因评估确定 FASN 和 miR-424-5p 之间的结合。在 ISO 暴露后，在 hESC 衍生神经元中鉴定出诱导的神经毒性和 miR-424-5p 产生减少。miR-424-5p 的上调抑制了 ISO 诱导的凋亡，并减轻了 ISO 诱导的体外炎症反应和氧化应激。FASN 表达水平通过 miR-424-5p 的升高而降低，并且在 ISO 处理后上调。在机制上，FASN 是 hESC 衍生神经元中 miR-424-5p 的直接靶标。值得注意的是，FASN 的上调抑制了神经元凋亡、炎症反应和氧化应激，从而抑制了 miR-424-5p 的升高。
