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抑制长链非编码RNA Vof-16的表达可促进脊髓损伤后的神经再生和功能恢复。

Inhibition of LncRNA Vof-16 expression promotes nerve regeneration and functional recovery after spinal cord injury.

作者信息

Zhang Xiao-Min, Zeng Li-Ni, Yang Wan-Yong, Ding Lu, Chen Kang-Zhen, Fu Wen-Jin, Zeng Si-Quan, Liang Yin-Ru, Chen Gan-Hai, Wu Hong-Fu

机构信息

Key Laboratory of Stem Cell and Regenerative Tissue Engineering, Guangdong Medical University, Dongguan, Guangdong Province, China.

Key Laboratory of Stem Cell and Regenerative Tissue Engineering, Guangdong Medical University, Dongguan; Biology Research Group, Guangzheng Experimental School, Huizhou, Guangdong Province, China.

出版信息

Neural Regen Res. 2022 Jan;17(1):217-227. doi: 10.4103/1673-5374.314322.

DOI:10.4103/1673-5374.314322
PMID:34100459
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8451561/
Abstract

Our previous RNA sequencing study showed that the long non-coding RNA ischemia-related factor Vof-16 (lncRNA Vof-16) was upregulated after spinal cord injury, but its precise role in spinal cord injury remains unclear. Bioinformatics predictions have indicated that lncRNA Vof-16 may participate in the pathophysiological processes of inflammation and apoptosis. PC12 cells were transfected with a pHBLV-U6-MCS-CMV-ZsGreen-PGK-PURO vector to express an lncRNA Vof-16 knockdown lentivirus and a pHLV-CMVIE-ZsGree-Puro vector to express an lncRNA Vof-16 overexpression lentivirus. The overexpression of lncRNA Vof-16 inhibited PC12 cell survival, proliferation, migration, and neurite extension, whereas lncRNA Vof-16 knockdown lentiviral vector resulted in the opposite effects in PC12 cells. Western blot assay results showed that the overexpression of lncRNA Vof-16 increased the protein expression levels of interleukin 6, tumor necrosis factor-α, and Caspase-3 and decreased Bcl-2 expression levels in PC12 cells. Furthermore, we established rat models of spinal cord injury using the complete transection at T10. Spinal cord injury model rats were injected with the lncRNA Vof-16 knockdown or overexpression lentiviral vectors immediately after injury. At 7 days after spinal cord injury, rats treated with lncRNA Vof-16 knockdown displayed increased neuronal survival and enhanced axonal extension. At 8 weeks after spinal cord injury, rats treated with the lncRNA Vof-16 knockdown lentiviral vector displayed improved neurological function in the hind limb. Notably, lncRNA Vof-16 knockdown injection increased Bcl-2 expression and decreased tumor necrosis factor-α and Caspase-3 expression in treated animals. Rats treated with the lncRNA Vof-16 overexpression lentiviral vector displayed opposite trends. These findings suggested that lncRNA Vof-16 is associated with the regulation of inflammation and apoptosis. The inhibition of lncRNA Vof-16 may be useful for promoting nerve regeneration and functional recovery after spinal cord injury. The experiments were approved by the Institutional Animal Care and Use Committee of Guangdong Medical University, China.

摘要

我们之前的RNA测序研究表明，长链非编码RNA缺血相关因子Vof-16（lncRNA Vof-16）在脊髓损伤后上调，但其在脊髓损伤中的具体作用仍不清楚。生物信息学预测表明，lncRNA Vof-16可能参与炎症和凋亡的病理生理过程。用pHBLV-U6-MCS-CMV-ZsGreen-PGK-PURO载体转染PC12细胞以表达lncRNA Vof-16敲低慢病毒，并用pHLV-CMVIE-ZsGree-Puro载体转染以表达lncRNA Vof-16过表达慢病毒。lncRNA Vof-16的过表达抑制了PC12细胞的存活、增殖、迁移和神经突延伸，而lncRNA Vof-16敲低慢病毒载体在PC12细胞中产生相反的效果。蛋白质印迹分析结果表明，lncRNA Vof-16的过表达增加了PC12细胞中白细胞介素6、肿瘤坏死因子-α和半胱天冬酶-3的蛋白表达水平，并降低了Bcl-2表达水平。此外，我们使用T10完全横断法建立了大鼠脊髓损伤模型。脊髓损伤模型大鼠在损伤后立即注射lncRNA Vof-16敲低或过表达慢病毒载体。脊髓损伤后7天，用lncRNA Vof-16敲低处理的大鼠神经元存活率增加，轴突延伸增强。脊髓损伤后8周，用lncRNA Vof-16敲低慢病毒载体处理的大鼠后肢神经功能改善。值得注意的是，lncRNA Vof-16敲低注射增加了治疗动物中Bcl-2的表达，并降低了肿瘤坏死因子-α和半胱天冬酶-3的表达。用lncRNA Vof-16过表达慢病毒载体处理的大鼠表现出相反的趋势。这些发现表明lncRNA Vof-16与炎症和凋亡的调节有关。抑制lncRNA Vof-16可能有助于促进脊髓损伤后的神经再生和功能恢复。实验获得中国广东医科大学实验动物管理与使用委员会的批准。

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