Department of Psychiatry, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi 710032, P.R. China.
Department of Anesthesiology, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi 710032, P.R. China.
Mol Med Rep. 2017 Nov;16(5):7305-7314. doi: 10.3892/mmr.2017.7547. Epub 2017 Sep 20.
Neonatal isoflurane exposure in rodents disrupts hippocampal cognitive functions, including learning and memory, and astrocytes may have an important role in this process. However, the molecular mechanisms underlying this disruption are not fully understood. The present study investigated the role of TWIK‑related K+ channel (TREK‑1) in isoflurane‑induced cognitive impairment. Lentiviruses were used to overexpress or knockdown TREK‑1 in astrocytes exposed to increasing concentrations of isoflurane or O2 for 2 h. Subsequently, the mRNA and protein expression of brain‑derived neurotrophic factor (BDNF), caspase‑3, Bcl‑2‑associated X (Bax) and TREK‑1 was measured by reverse transcription‑ quantitative polymerase chain reaction and western blot analysis, respectively. In addition, cell viability was assessed by a 2‑(4‑Iodophenyl)‑3‑(4‑nitrophenyl)‑5‑(2,4‑disulfophenyl)‑ 2H‑tetrazolium monosodium salt assay. The results demonstrated that, prior to manipulating TREK‑1, isoflurane significantly decreased the cell viability and BDNF expression, and increased Bax, caspase‑3 and TREK‑1 expression was observed. However, TREK‑1 overexpression in astrocytes significantly downregulated BDNF expression, and upregulated Bax and caspase‑3 expression. Furthermore, lentiviral‑mediated short hairpin RNA knockdown of TREK‑1 effectively inhibited the isoflurane‑induced changes in BDNF, Bax and caspase‑3 expression. Taken together, the results of the present study indicate that isoflurane‑induced cell damage in astrocytes may be associated with TREK‑1‑mediated inhibition of BDNF and provide a reference for the safe use of isoflurane anesthesia in infants and children.
新生鼠异氟烷暴露会破坏海马认知功能，包括学习和记忆，星形胶质细胞在此过程中可能发挥重要作用。然而，其破坏的分子机制尚不完全清楚。本研究探讨了 TWIK 相关钾通道（TREK-1）在异氟烷诱导认知障碍中的作用。使用慢病毒过表达或敲低暴露于不同浓度异氟烷或 O2 中 2 h 的星形胶质细胞中的 TREK-1。随后，通过逆转录定量聚合酶链反应和 Western blot 分析分别测量脑源性神经营养因子（BDNF）、半胱天冬酶-3、Bcl-2 相关 X（Bax）和 TREK-1 的 mRNA 和蛋白表达。此外，通过 2-（4-碘苯基）-3-（4-硝基苯基）-5-（2,4-二磺基苯基）-2H-四唑单钠盐测定法评估细胞活力。结果表明，在操纵 TREK-1 之前，异氟烷显著降低了细胞活力和 BDNF 表达，并且观察到 Bax、半胱天冬酶-3 和 TREK-1 的表达增加。然而，星形胶质细胞中 TREK-1 的过表达显著下调了 BDNF 的表达，并上调了 Bax 和半胱天冬酶-3 的表达。此外，慢病毒介导的 TREK-1 短发夹 RNA 敲低有效抑制了异氟烷诱导的 BDNF、Bax 和半胱天冬酶-3 表达的变化。综上所述，本研究结果表明，异氟烷诱导的星形胶质细胞损伤可能与 TREK-1 介导的 BDNF 抑制有关，为婴幼儿异氟烷麻醉的安全使用提供参考。
