Identification of key regulatory genes involved in myelination after spinal cord injury by GSEA analysis

IF 4.6 2区医学 Q1 NEUROSCIENCES

Experimental Neurology Pub Date : 2024-09-24 DOI:10.1016/j.expneurol.2024.114966

Yehua Lv , Lingyun Ji , Hui Dai, Shanru Qiu, Yu Wang, Cheng Teng, Bin Yu, Daguo Mi, Chun Yao

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引用次数: 0

Abstract

Multilayer dense myelin tissue provides insulating space and nutritional support for axons in healthy spinal cord tissue. Oligodendrocyte precursor cells (OPCs) are the main glial cells that complement myelin loss in the central nervous system and play an important role in the repair of spinal cord injury (SCI). However, the regulation of axonal remyelination after SCI is still insufficient. In this study, we focused on the changes in genes related to myelin repair after rat hemisection SCI by gene set enrichment analysis (GSEA). Key genes proteolipid protein 1 (Plp1), hexosaminidase subunit alpha (Hexa), and hexosaminidase subunit beta (Hexb) during remyelination after SCI were found. Through quantitative real-time polymerase chain reaction (qPCR) experiments, we confirmed that within 28 days after rat hemisection SCI, the mRNA expression of gene Plp1 gradually decreased, while the expressions of gene Hexa and Hexb gradually increased, which was consistent with RNA sequencing results. In vitro, we performed EdU proliferation assays on OPC cell line OLN-93 and primary rat OPCs. We found that interference of Plp1 promoted OPC proliferation, while interference of Hexa and Hexb inhibited OPC proliferation. In addition, we performed in vitro differentiation experiments on primary rat OPCs. By measuring myelin sheath branch outgrowth and the fluorescence intensity of the mature myelin sheath marker myelin basic protein (MBP), we found that interference of Hexa or Hexb promoted OPC differentiation and maturation, but interference of Plp1 inhibited this process. Finally, we injected Hexb siRNA in vivo and found that interfering Hexb could improve motor movements and myelin regeneration after SCI in rats. Our results provide new target genes that can selectively regulate the proliferation and differentiation of endogenous OPCs, providing new ideas for promoting remyelination and functional recovery after SCI.

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通过 GSEA 分析鉴定脊髓损伤后髓鞘形成过程中的关键调控基因

在健康的脊髓组织中，多层致密的髓鞘组织为轴突提供绝缘空间和营养支持。少突胶质细胞前体细胞（OPCs）是补充中枢神经系统髓鞘损失的主要胶质细胞，在脊髓损伤（SCI）的修复中发挥着重要作用。然而，目前对 SCI 后轴突再髓鞘化的调控仍不充分。本研究通过基因组富集分析（GSEA），重点研究了大鼠半切脊髓损伤后髓鞘修复相关基因的变化。研究发现了髓鞘损伤后再髓鞘化过程中的关键基因蛋白脂质蛋白1（Plp1）、己糖胺酸酶亚基α（Hexa）和己糖胺酸酶亚基β（Hexb）。通过实时定量聚合酶链式反应（qPCR）实验，我们证实了在大鼠断肢后28天内，基因Plp1的mRNA表达量逐渐减少，而基因Hexa和Hexb的表达量逐渐增加，这与RNA测序结果一致。在体外，我们对 OPC 细胞系 OLN-93 和原代大鼠 OPC 进行了 EdU 增殖实验。我们发现，干扰 Plp1 会促进 OPC 增殖，而干扰 Hexa 和 Hexb 则会抑制 OPC 增殖。此外，我们还对原代大鼠 OPC 进行了体外分化实验。通过测量髓鞘分支的生长和成熟髓鞘标志物髓鞘碱性蛋白（MBP）的荧光强度，我们发现干扰Hexa或Hexb会促进OPC的分化和成熟，而干扰Plp1则会抑制这一过程。最后，我们在体内注射了Hexb siRNA，发现干扰Hexb可以改善大鼠的运动能力和脊髓损伤后的髓鞘再生。我们的研究结果提供了可选择性调控内源性OPC增殖和分化的新靶基因，为促进损伤后髓鞘再形成和功能恢复提供了新思路。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Experimental Neurology 医学-神经科学

CiteScore

10.10

自引率

3.80%

发文量

258

审稿时长

42 days

期刊介绍： Experimental Neurology, a Journal of Neuroscience Research, publishes original research in neuroscience with a particular emphasis on novel findings in neural development, regeneration, plasticity and transplantation. The journal has focused on research concerning basic mechanisms underlying neurological disorders.