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

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

doi:10.4103/1673-5374.314322

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

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

Authors

Xiao-Min Zhang¹, Li-Ni Zeng², Wan-Yong Yang³, Lu Ding⁴, Kang-Zhen Chen¹, Wen-Jin Fu⁵, Si-Quan Zeng³, Yin-Ru Liang¹, Gan-Hai Chen⁶, Hong-Fu Wu¹

Affiliations

¹ Key Laboratory of Stem Cell and Regenerative Tissue Engineering, Guangdong Medical University, Dongguan, Guangdong Province, China.
² Key Laboratory of Stem Cell and Regenerative Tissue Engineering, Guangdong Medical University, Dongguan; Biology Research Group, Guangzheng Experimental School, Huizhou, Guangdong Province, China.
³ Geriatric Medicine Center, Dongguan Waterfront Zone Central Hospital, Dongguan, Guangdong Province, China.
⁴ Key Laboratory of Stem Cell and Regenerative Tissue Engineering, Guangdong Medical University, Dongguan; Scientific Research Center, the Seventh Affiliated Hospital, Sun Yat-sen University, Shenzhen, Guangdong Province, China.
⁵ Clinical Laboratory, Affiliated Houjie Hospital, Guangdong Medical University, Dongguan, Guangdong Province, China.
⁶ Department of Intensive Care Unit, Affiliated Houjie Hospital, Guangdong Medical University, Dongguan, Guangdong Province, China.

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.

Keywords: apoptosis; functional recovery; inflammation; long non-coding RNA ischemia related factor Vof-16; nerve regeneration; nerve repair; neurite extension; neuronal survival; proliferation; spinal cord injury.