TRESK Regulates Gm11874 to Induce Apoptosis of Spinal Cord Neurons via ATP5i Mediated Oxidative Stress and DNA Damage
TRESK Regulates Gm11874 to Induce Apoptosis of Spinal Cord Neurons via ATP5i Mediated Oxidative Stress and DNA Damage
复制标题
TRESK 通过 ATP5i 介导的氧化应激和 DNA 损伤调节 Gm11874 诱导脊髓神经元凋亡
DOI：
10.1007/s11064-021-03318-w
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发表时间：
2021-05-10
影响因子：
4.4
通讯作者：
Zhou, Jun
中科院分区：
文献类型：
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作者：
Liu, Pei;Cheng, Ye;Zhou, Jun
据报道，脊髓神经元中twik相关的脊髓K+(TRESK)缺乏与神经性疼痛（NP）的机制正相关。然而，TRESK对脊髓神经元的确切影响仍然难以捉摸。在本研究中，我们研究了TRESK沉默对脊髓神经元的影响，以进一步阐明TRESK的下游机制。本实验以脊髓背侧神经元为细胞模型进行研究。评估细胞凋亡、氧化应激和DNA损伤相关蛋白。此外，还进行了流式细胞术、微阵列分析、实时聚合酶链反应（PCR）、western blotting、荧光原位杂交（FISH）、免疫荧光和酶联免疫吸附试验（ELISA）。在培养神经元中，下调TRESK mRNA表达可诱导背脊髓神经元凋亡。利用real-time PCR和western blotting技术，证实从基因芯片中筛选出的LncRNA Gm11874 （Gm11874）和ATP5i表达上调。沉默TRESK后，氧化应激和DNA损伤指标γ-H2AX、聚[adp -核糖]聚合酶1 （PARP-1）、FoxO1、FoxO3、MitoSOX、丙二醛（MDA）和8-羟基-2′-脱氧鸟苷（8-OHdG）的表达水平显著升高。此外，ATP诱导的氧化应激、DNA损伤和细胞凋亡被ATP5i siRNA抑制。最后，在FISH实验中，Gm11874和ATP5i在脊髓神经元中共表达，Gm11874正向调节ATP5i的表达。这些结果表明，ATP5i诱导氧化应激和DNA损伤，导致神经元凋亡，Gm11874被证实作用于ATP5i的上游。我们的研究发现，TRESK沉默上调Gm11874诱导脊髓神经元凋亡，导致ATP5i促进氧化应激和DNA损伤。这些发现可以突出tresk介导的NP机制。
Reportedly, TWIK-related spinal cord K+(TRESK) deficiency in spinal cord neurons positively correlates with the mechanism underlying neuropathic pain (NP). However, the precise effects of TRESK on neurons of the spinal cord remain elusive. In the present study, we investigated the impact of TRESK silencing on spinal cord neurons to further elucidate the downstream mechanisms of TRESK. Herein, neurons of the dorsal spinal cord were cultured as a cell model for investigations. Apoptosis, oxidative stress, and DNA damage-related proteins were evaluated. Additionally, flow cytometry, microarray profiling, real-time polymerase chain reaction (PCR), western blotting, fluorescence in situ hybridization (FISH), immunofluorescence, and enzyme-linked immunosorbent assay (ELISA) were performed. In cultured neurons, the downregulation of TRESK mRNA expression induced apoptosis of dorsal spinal cord neurons. Using real-time PCR and western blotting, the upregulation of LncRNA Gm11874 (Gm11874) and ATP5i, screened from the gene chip, was confirmed. On silencing TRESK, expression levels of γ-H2AX, poly [ADP-ribose] polymerase 1 (PARP-1), FoxO1, FoxO3, MitoSOX, malondialdehyde (MDA), and 8-hydroxy-2′ -deoxyguanosine (8-OHdG), which are known indices of oxidative stress and DNA damage, were significantly elevated. Moreover, ATP induced oxidative stress, DNA damage, and apoptosis were reduced by ATP5i siRNA. Finally, Gm11874 and ATP5i were co-expressed in spinal cord neurons in a FISH experiment, and the expression of ATP5i was positively regulated by Gm11874. These results implied that ATP5i induced oxidative stress and DNA damage, resulting in neuronal apoptosis, and Gm11874 was confirmed to act upstream of ATP5i. Our study revealed that TRESK silencing upregulated Gm11874 to induce apoptosis of spinal cord neurons, which resulted in ATP5i promoting oxidative stress and DNA damage. These findings could highlight the TRESK-mediated NP mechanism.