Isoflurane, one of the commonly used inhalation anesthetics worldwide in clinical practice, may generate substantial risks of neurotoxicity in the developing brains. The present study aimed to illustrate the effects and underlying mechanisms of miR-214 on isoflurane-induced neurotoxicity in human neuroblastoma cell line SH-SY5Y. SH-SY5Y cells were transfected with miR-214 or miR-con alone or in combination with pcDNA empty vector or pcDNA-PTEN in the presence of 3% isoflurane and incubated for 48 h. Cell viability, lactate dehydrogenase (LDH) release, apoptosis, and caspase-3/7 activity were evaluated using CCK-8, LDH release assay, flow cytometry analysis, and caspase-3/7 activity assay, respectively. The superoxide dismutase (SOD), glutathione (GSH), and malondialdehyde (MDA) activities were measured using commercial kits. miR-214 expression and alterations of the phosphatase and tensin homolog (PTEN)/phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt) pathway were detected by qRT-PCR and Western blot, respectively. The interaction between miR-214 and PTEN was explored by luciferase reporter assay. We found that isoflurane exposure induced neurotoxicity in SH-SY5Y cells, as evidenced by the reduced cell viability, increased LDH release, apoptotic rate, caspase-3/7 activity, and oxidative stress levels. Moreover, isoflurane exposure decreased the expression of miR-214 and affected the PTEN/PI3K/Akt pathway in SH-SY5Y cells. miR-214 overexpression significantly suppressed isoflurane-induced viability reduction, LDH release, apoptosis and oxidative stress, as well as inactivation of the PI3K/Akt pathway in SH-SY5Y cells. Interestingly, PTEN was identified as a target of miR-214. Moreover, PTEN upregulation blocked the effects of miR-214 on isoflurane-induced neurotoxicity in SH-SY5Y cells. In conclusion, miR-214 protected against isoflurane-induced neurotoxicity in SH-SY5Y cells via regulation of PI3K/Akt pathway by targeting PTEN, contributing to better understanding the underlying mechanisms of anesthetics-induce neurotoxicity.

异氟烷是临床实践中全球范围内最常用的吸入麻醉药之一，可能会在发育中的大脑中产生很大的神经毒性风险。本研究旨在阐明miR-214对异氟烷诱导的人神经母细胞瘤细胞系SH-SY5Y产生神经毒性的作用及其潜在机制。在3％异氟烷存在下，单独用miR-214或miR-con或与pcDNA空载体或pcDNA-PTEN组合转染SH-SY5Y细胞，并孵育48小时。分别使用CCK-8，LDH释放测定，流式细胞仪分析和caspase-3 / 7活性测定来评估细胞活力，乳酸脱氢酶（LDH）释放，凋亡和caspase-3 / 7活性。使用商业试剂盒测量了超氧化物歧化酶（SOD），谷胱甘肽（GSH）和丙二醛（MDA）的活性。分别通过qRT-PCR和Western blot检测miR-214的表达以及磷酸酶和张力蛋白同源物（PTEN）/磷酸肌醇3-激酶（PI3K）/蛋白激酶B（Akt）途径的变化。miR-214和PTEN之间的相互作用是通过萤光素酶报告基因检测来进行的。我们发现异氟烷暴露可诱导SH-SY5Y细胞发生神经毒性，这可通过降低细胞活力，增加LDH释放，凋亡率，caspase-3 / 7活性和氧化应激水平来证明。此外，异氟烷暴露降低了SH-SY5Y细胞中miR-214的表达并影响了PTEN / PI3K / Akt途径。miR-214过表达显着抑制了异氟烷诱导的活力降低，LDH释放，细胞凋亡和氧化应激以及SH-SY5Y细胞中PI3K / Akt途径的失活。有趣的是，PTEN被确定为miR-214的靶标。此外，PTEN上调阻断了miR-214对异氟烷诱导的SH-SY5Y细胞神经毒性的作用。总之，miR-214可通过靶向PTEN来调节PI3K / Akt通路，从而防止SH-SY5Y细胞中异氟烷诱导的神经毒性，有助于更好地理解麻醉剂的潜在机制，从而诱导神经毒性。