Department of Orthopedics, The 981st Hospital of the Chinese People's Liberation Army, Chengde 067000, Hebei Province, P.R. China.
Department of Orthopedics, Tianjin Medical University General Hospital, Tianjin 300052, P.R. China.
Int J Biol Sci. 2019 Jun 2;15(7):1557-1570. doi: 10.7150/ijbs.31904. eCollection 2019.
Sensory dysfunction post spinal cord injury causes patients great distress. Sciatic nerve conditioning injury (SNCI) has been shown to restore sensory function after spinal cord dorsal column injury (SDCL); however, the underlying mechanism of this recovery remains unclear. We performed a microarray assay to determine the associated miRNAs that might regulate the process of SNCI promoting SDCL repair. In total, 13 miRNAs were identified according to our inclusion criteria, and RT-qPCR was used to verify the microarray results. Among the 13 miRNAs, the miR-155-5p levels were decreased at 9 h, 3 d, 7 d, 14 d, 28 d, 2 m and 3 m timepoints in the SDCL group, while the SNCI group had a smaller decrease. Thus, miR-155-5p was chosen for further study after a literature review and an analysis with the TargetScan online tool. Specifically, miR-155-5p targets PKI-α, and the expression pattern of PKI-α was opposite that of miR-155-5p in both the SDCL and SNCI groups. Interestingly, miR-155-5p could promote dorsal root ganglion (DRG) neuron axon growth via the cAMP/PKA pathway and in a TNF-α, IL-1β or MAG inhibitory microenvironment in vitro. Furthermore, miR-155-5p could regulate the cAMP/PKA pathway and promote sensory conduction function recovery post dorsal column injury as detected by NF-200 immunohistochemistry, somatosensory-evoked potentials, BBB scale and tape removal test. Collectively, our results demonstrated that miR-155-5p participates in the molecular mechanism by which SNCI promotes the repair of SDCL and that upregulated miR-155-5p can repair SDCL by enhancing DRG neuron axon growth via the cAMP/PKA pathway. These findings suggest a novel treatment target for spinal cord injury.
脊髓损伤后的感觉功能障碍给患者带来极大的痛苦。坐骨神经条件损伤(SNCI)已被证明可在脊髓背柱损伤(SDCL)后恢复感觉功能;然而,这种恢复的潜在机制尚不清楚。我们进行了微阵列分析,以确定可能调节 SNCI 促进 SDCL 修复过程的相关 miRNA。根据我们的纳入标准,共鉴定出 13 个 miRNA,并用 RT-qPCR 验证了微阵列结果。在这 13 个 miRNA 中,miR-155-5p 的水平在 SDCL 组的 9 h、3 d、7 d、14 d、28 d、2 m 和 3 m 时间点下降,而 SNCI 组下降幅度较小。因此,在文献回顾和使用 TargetScan 在线工具进行分析后,选择 miR-155-5p 进行进一步研究。具体而言,miR-155-5p 的靶标是 PKI-α,PKI-α 的表达模式与 SDCL 和 SNCI 组中的 miR-155-5p 相反。有趣的是,miR-155-5p 可以通过 cAMP/PKA 通路促进背根神经节(DRG)神经元轴突生长,并在体外 TNF-α、IL-1β 或 MAG 抑制微环境中发挥作用。此外,miR-155-5p 可以通过 NF-200 免疫组织化学、体感诱发电位、BBB 量表和胶带去除试验来调节 cAMP/PKA 通路,并促进背柱损伤后的感觉传导功能恢复。综上所述,我们的结果表明,miR-155-5p 参与了 SNCI 促进 SDCL 修复的分子机制,上调的 miR-155-5p 可以通过增强 DRG 神经元轴突生长来修复 SDCL,该生长是通过 cAMP/PKA 通路实现的。这些发现为脊髓损伤提供了一种新的治疗靶点。