Ketamine is a kind of anesthetic broadly applied in clinic. However, growing evidence has indicated that ketamine may induce neurotoxicity. Previous studies showed that mircoRNAs (miRNAs) participate in various aspects of biological regulations. In our work, we aimed to reveal the role of miR-429 in ketamine-induced neurotoxicity. The qRT-PCR was used to measure the miR-429 levels in ketamine-treated PC12 cells. TUNEL staining and caspase 3 activity detection assays were performed to assess cell apoptosis. A Cellular Reactive Oxygen Species Detection Assay Kit was utilized to detect ROS activity. A luciferase reporter assay was conducted in HEK-293T cells to test the binding between miR-429 and BAG5. Herein, we found that ketamine could induce the apoptosis and ROS activity in PC12 cells. The qRT-PCR results showed that miR-429 expression was downregulated by treatment of ketamine in a dose-dependent manner. Overexpression of miR-429 alleviated ketamine-induced neurotoxicity in PC12 cells. Mechanically, BAG5 was identified to be a target of miR-429 and negatively regulated by miR-429. Moreover, BAG5 expression was upregulated after ketamine treatment. Rescue assays revealed that overexpression of BAG5 reversed the suppressive effects of miR-429 upregulation on ketamine-induced neurotoxicity in PC12 cells. In summary, miR-429 attenuates ketamine-induced neurotoxicity in PC12 cells by the downregulation of BAG5.

中文翻译：

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MiRNA ‐429 通过靶向 BAG5 减轻氯胺酮诱导的神经毒性

氯胺酮是一种广泛应用于临床的麻醉剂。然而，越来越多的证据表明氯胺酮可能会引起神经毒性。先前的研究表明，miRNAs (miRNAs) 参与生物调控的各个方面。在我们的工作中，我们旨在揭示 miR-429 在氯胺酮诱导的神经毒性中的作用。qRT-PCR 用于测量氯胺酮处理的 PC12 细胞中的 miR-429 水平。进行 TUNEL 染色和半胱天冬酶 3 活性检测测定以评估细胞凋亡。使用细胞反应性氧物种检测分析试剂盒来检测 ROS 活性。在 HEK-293T 细胞中进行荧光素酶报告基因检测以测试 miR-429 和 BAG5 之间的结合。在此，我们发现氯胺酮可以诱导 PC12 细胞的凋亡和 ROS 活性。qRT-PCR 结果显示 miR-429 的表达被氯胺酮处理以剂量依赖性方式下调。miR-429 的过表达减轻了 PC12 细胞中氯胺酮诱导的神经毒性。在机械上，BAG5 被鉴定为 miR-429 的靶标并受 miR-429 负调控。此外，氯胺酮处理后 BAG5 表达上调。救援分析表明，BAG5 的过表达逆转了 miR-429 上调对 PC12 细胞中氯胺酮诱导的神经毒性的抑制作用。总之，miR-429 通过下调 BAG5 减弱 PC12 细胞中氯胺酮诱导的神经毒性。BAG5 被鉴定为 miR-429 的靶标并受 miR-429 负调控。此外，氯胺酮处理后 BAG5 表达上调。救援分析表明，BAG5 的过表达逆转了 miR-429 上调对 PC12 细胞中氯胺酮诱导的神经毒性的抑制作用。总之，miR-429 通过下调 BAG5 减弱 PC12 细胞中氯胺酮诱导的神经毒性。BAG5 被鉴定为 miR-429 的靶标并受 miR-429 负调控。此外，氯胺酮处理后 BAG5 表达上调。救援分析表明，BAG5 的过表达逆转了 miR-429 上调对 PC12 细胞中氯胺酮诱导的神经毒性的抑制作用。总之，miR-429 通过下调 BAG5 减弱 PC12 细胞中氯胺酮诱导的神经毒性。