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小干扰 LncRNA-TUG1(siTUG1)可降低氯胺酮诱导的大鼠海马神经元神经毒性。

Small interfering LncRNA-TUG1 (siTUG1) decreases ketamine-induced neurotoxicity in rat hippocampal neurons.

机构信息

Department of Hyperbaric Oxygen Therapy, Yantai Yuhuangding Hospital , Yantai , China.

Department of Neurology, Yantai Yuhuangding Hospital , Yantai , China.

出版信息

Int J Neurosci. 2019 Oct;129(10):937-944. doi: 10.1080/00207454.2019.1594805. Epub 2019 May 17.

Abstract

Ketamine (KET) is known to be used in pediatric anaesthetic, whereas many evidences revealed that KET generated neurotoxicity. In our study, we found KET decreased the level of LncRNA taurine-upregulated gene 1 (TUG1). Hence, the aim of our study is to investigate whether small interfering TUG1 (siTUG1) has effect on KET-induced rat hippocampal neurons. siTUG1 and KET alone or in combination was used to treat with neurons for 12 h. MTT assays was used to detect hippocampal neurons viability. Cell apoptosis and reactive oxygen species (ROS) level were analysed by flow cytometry assay. TUG1 level was determined via reverse transcription-quantitative polymerase chain reaction (RT-qPCR) assay. The levels of Bax, Bcl-2, cleaved-caspase-3, p38 and p-p38 were measured by western blot (WB) assay. Neurons treated by siTUG1 and KET had a higher optical density (OD) value, a lower apoptosis rate and lower ROS level. Neurons treated by the combination of siTUG1 and KET had lower levels of TUG1, cleaved-caspase-3 and p-p38 than those under KET treatment. Besides, siTUG1 + KET group (siTUG1 and KET treated neurons) had a higher Bcl-2 level than KET group. We found that siTUG1 decreased KET-induced hippocampal neurons apoptosis and ROS level, and increased hippocampal neurons viability. siTUG1 decreased neurons apoptosis via signaling pathways of Bax/Bcl-2 and Caspases, and increased neurons viability by signaling pathway of p38 MAPK. Our results indicate that KET-induced neurotoxicity might be reduced by inhibiting TUG1.

摘要

氯胺酮(KET)已知可用于儿科麻醉,而许多证据表明 KET 会产生神经毒性。在我们的研究中,我们发现 KET 降低了长链非编码 RNA 牛磺酸上调基因 1(TUG1)的水平。因此,我们的研究旨在探讨小干扰 TUG1(siTUG1)是否对 KET 诱导的大鼠海马神经元有影响。用 siTUG1 和 KET 单独或联合处理神经元 12 小时。MTT 法检测海马神经元活力。通过流式细胞术检测细胞凋亡和活性氧(ROS)水平。采用逆转录-定量聚合酶链反应(RT-qPCR)检测 TUG1 水平。Western blot(WB)检测 Bax、Bcl-2、cleaved-caspase-3、p38 和 p-p38 的水平。用 siTUG1 和 KET 处理的神经元具有更高的光密度(OD)值、更低的凋亡率和更低的 ROS 水平。与 KET 处理相比,siTUG1 和 KET 联合处理的神经元 TUG1、cleaved-caspase-3 和 p-p38 水平更低。此外,siTUG1+KET 组(siTUG1 和 KET 处理的神经元)的 Bcl-2 水平高于 KET 组。我们发现,siTUG1 降低了 KET 诱导的海马神经元凋亡和 ROS 水平,增加了海马神经元活力。siTUG1 通过 Bax/Bcl-2 和 Caspases 信号通路降低神经元凋亡,通过 p38 MAPK 信号通路增加神经元活力。我们的结果表明,通过抑制 TUG1,可能减轻 KET 诱导的神经毒性。

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