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长链非编码 RNA-LIN-微小 RNA-9-DRD2 调控网络对吸入麻醉剂七氟醚后神经元系统发育的影响。

Effect of the LncRNA-LIN-miRNA-9-DRD2 regulatory network on the development of the neuronal system after inhalation of the anesthetic sevoflurane.

机构信息

Department of Anesthesiology, Jiangxi Medical College, The Third Affiliated Hospital of Nanchang University, Nanchang University, Jiangxi, China.

Department of Anesthesiology, The First Hospital of Nanchang City, Jiangxi, China.

出版信息

Exp Brain Res. 2024 Nov 29;243(1):6. doi: 10.1007/s00221-024-06960-2.

Abstract

Animal studies have shown that exposure of newborns to general anesthesia drugs can lead to neurodegenerative diseases and subsequent decline in learning and memory abilities. The neurotoxicity of general anesthesia drugs can also occur in the fetus. Therefore, in order to investigate the effect of the Long non-coding RNA(LncRNA)-LIN-microRNA(miRNA)-9-Dopamine receptor D2(DRD2) regulatory network on the development of the neuronal system after the inhalation of the anesthetic sevoflurane, RT-qPCR was used to detect the mRNA levels of LncRNA-LIN, miRNA-9, and DRD2. A dual-luciferase reporter system was used to detect the relationship between LncRNA-LIN and miRNA-9, and miRNA-9 and DRD2. Western blotting and immunofluorescence staining was employed to detect the protein levels of DRD2 and cleaved caspase-3. Flow cytometry was carried out to detect the number of apoptotic cells. The escape latency, swimming distance, and platform crossing times were analyzed using the Morris water maze. The results showed that, after treatment with sevoflurane, the mRNA levels of LncRNA-LIN and DRD2, the expression levels of the DRD2 protein, and the number of neuronal levels of DRD2 were significantly decreased, whereas the expression levels of miRNA-9 and the cleaved caspase-3 protein and neuronal apoptosis were significantly increased. miR-9 knockdown revealed that miRNA-9 regulated DRD2 expression and affected the function of mouse neuronal cells. In turn, LncRNA-LIN overexpression indicated that LncRNA-LIN regulated miR-9 and affected the function of mouse neuronal cells. The present results demonstrated that the LncRNA-LIN-miRNA-9-DRD2 regulatory network is involved in the effects of the inhalation anesthetic sevoflurane on neuronal system development.

摘要

动物研究表明,新生暴露于全身麻醉药物可导致神经退行性疾病和随后学习和记忆能力下降。全身麻醉药物的神经毒性也可能发生在胎儿中。因此,为了研究长链非编码 RNA(LncRNA)-LIN-微小 RNA(miRNA)-9-多巴胺受体 D2(DRD2)调节网络对吸入麻醉药七氟醚后神经元系统发育的影响,采用 RT-qPCR 检测 LncRNA-LIN、miRNA-9 和 DRD2 的 mRNA 水平。双荧光素酶报告系统用于检测 LncRNA-LIN 与 miRNA-9 之间的关系,以及 miRNA-9 与 DRD2 之间的关系。Western blotting 和免疫荧光染色用于检测 DRD2 和 cleaved caspase-3 的蛋白水平。流式细胞术用于检测凋亡细胞数量。采用 Morris 水迷宫分析逃避潜伏期、游泳距离和平台穿越次数。结果表明,七氟醚处理后,LncRNA-LIN 和 DRD2 的 mRNA 水平、DRD2 蛋白表达水平以及神经元水平的 DRD2 数量均显著降低,而 miRNA-9 的表达水平和 cleaved caspase-3 蛋白以及神经元凋亡显著增加。miR-9 敲低表明 miRNA-9 调节 DRD2 表达并影响小鼠神经元细胞的功能。反过来,LncRNA-LIN 过表达表明 LncRNA-LIN 调节 miR-9 并影响小鼠神经元细胞的功能。本研究结果表明,LncRNA-LIN-miRNA-9-DRD2 调节网络参与了吸入麻醉药七氟醚对神经元系统发育的影响。

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