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SIRT1 通过 microRNA-494 抑制脊髓损伤的体内和体外模型中的细胞凋亡。

SIRT1 inhibits apoptosis in in vivo and in vitro models of spinal cord injury via microRNA-494.

机构信息

Department of Orthopaedics, Zhongshan Hospital of Dalian University, Dalian, Liaoning 116001, P.R. China.

出版信息

Int J Mol Med. 2019 Apr;43(4):1758-1768. doi: 10.3892/ijmm.2019.4106. Epub 2019 Feb 22.

Abstract

The aim of the present study was to investigate the function and mechanism of sirtuin 1 (SIRT1) in spinal cord injury (SCI). Reverse transcription‑quantitative polymerase chain reaction was used to measure the expression levels of microRNA (miR)‑494. MTT assay, lactate dehydrogenase activity assay and flow cytometry were used to analyze the effects of miR‑494 on cell growth and apoptosis in a model of SCI. The present study demonstrated that SIRT1 expression was reduced; whereas miR‑494 expression was increased in a rat model of SCI. Overexpression of miR‑494 suppressed the protein expression levels of SIRT1, and induced p53 protein expression. Conversely, knockdown of miR‑494 induced SIRT1 protein expression in an in vitro model of SCI. Furthermore, overexpression of miR‑494 promoted cell apoptosis and decreased cell growth in an in vitro model of SCI; however, miR‑494 knockdown enhanced cell growth and inhibited cell apoptosis. Administration of a SIRT1 agonist reduced the effects of miR‑494 overexpression on cell apoptosis in an SCI model, whereas treatment with a p53 agonist reduced the effects of miR‑494 knockdown on cell apoptosis in an SCI model. Together, these findings suggested that SIRT1 may inhibit apoptosis of SCI in vivo and in vitro through the p53 signaling pathway, whereas miR‑494 suppressed SIRT1 and induced apoptosis.

摘要

本研究旨在探讨沉默信息调节因子 1(SIRT1)在脊髓损伤(SCI)中的作用和机制。采用逆转录-定量聚合酶链反应(RT-qPCR)检测微小 RNA(miR)-494 的表达水平。MTT 检测、乳酸脱氢酶(LDH)活性检测和流式细胞术用于分析 miR-494 对 SCI 模型中细胞生长和凋亡的影响。本研究表明,在 SCI 大鼠模型中 SIRT1 表达降低,而 miR-494 表达升高。miR-494 的过表达抑制 SIRT1 蛋白表达,并诱导 p53 蛋白表达。相反,在 SCI 的体外模型中,miR-494 的敲低诱导 SIRT1 蛋白表达。此外,miR-494 的过表达促进 SCI 体外模型中的细胞凋亡并减少细胞生长;然而,miR-494 的敲低增强了细胞生长并抑制了细胞凋亡。SIRT1 激动剂的给药减轻了 miR-494 过表达对 SCI 模型中细胞凋亡的影响,而 p53 激动剂的治疗减轻了 miR-494 敲低对 SCI 模型中细胞凋亡的影响。综上所述,这些发现表明 SIRT1 可能通过 p53 信号通路抑制体内和体外 SCI 的细胞凋亡,而 miR-494 抑制 SIRT1 并诱导细胞凋亡。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4b4c/6414168/8a07d5f0a93c/IJMM-43-04-1758-g00.jpg

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