Department of Veterinary Clinical Science, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, 210095, P. R. China.
ACS Chem Neurosci. 2020 Dec 16;11(24):4329-4335. doi: 10.1021/acschemneuro.0c00586. Epub 2020 Nov 24.
Emulsified isoflurane (EISO) is an intravenous anesthetic. However, researchers have not clearly determined how emulsified isoflurane affects the central nervous system during the process of anesthesia. The aim of this study was to explore changes in the gamma-aminobutyric acid type A receptor subunit (GABA), 61 kD isoform of striatal-enriched protein phosphatase (STEP61) signaling pathway, and epigenetic regulation in cortical neurons after treatment with emulsified isoflurane. After immunological identification, the isolated neurons were randomly divided into three groups: the blank group (Con), intralipid treatment group (FE), and emulsified isoflurane treatment group (EISO). Neuron viability was assayed using cell counting kit-8 (CCK-8). The expression levels of target nucleic acids, proteins, and corresponding ligands were detected. Using real-time polymerase chain reaction (PCR) to assess the promoter methylation of ion channel proteins in the cerebral cortex of rats anesthetized with EISO, we observed changes in promoter methylation of the genes encoding gamma-aminobutyric acid type A receptor α1 subunit (GABA), -methyl-d-aspartate receptor subunit 1 (NMDAR1), and mu opioid receptor 1 (OPRM1), accompanied by changes in the levels of their messenger ribonucleic acids (mRNAs) and proteins. The levels of ligands for these receptors were also altered. EISO altered the methylation rate of the promoter region of channel protein-coding genes involved in the GABA/STEP61 signaling pathway in cerebral cortical neurons to regulate gene expression. The ligands for the receptors were also changed.
乳化异氟醚（EISO）是一种静脉麻醉剂。然而，研究人员尚未明确确定乳化异氟醚在麻醉过程中如何影响中枢神经系统。本研究旨在探讨皮质神经元在接受乳化异氟醚处理后，γ-氨基丁酸 A 型受体亚基（GABA）、富含纹状体的 61kD 同工型蛋白磷酸酶（STEP61）信号通路和表观遗传调控的变化。经过免疫鉴定后，分离的神经元被随机分为三组：空白组（Con）、中链脂肪乳剂处理组（FE）和乳化异氟醚处理组（EISO）。使用细胞计数试剂盒-8（CCK-8）测定神经元活力。检测目标核酸、蛋白质和相应配体的表达水平。通过实时聚合酶链反应（PCR）评估 EISO 麻醉大鼠大脑中离子通道蛋白启动子的甲基化，观察编码γ-氨基丁酸 A 型受体α1 亚基（GABA）、-甲基-d-天冬氨酸受体亚基 1（NMDAR1）和μ阿片受体 1（OPRM1）的基因启动子甲基化的变化，以及它们的信使核糖核酸（mRNA）和蛋白质水平的变化。这些受体的配体水平也发生了改变。EISO 改变了大脑皮质神经元中 GABA/STEP61 信号通路中参与通道蛋白编码基因的启动子区域的甲基化率，以调节基因表达。受体的配体也发生了改变。
