Xiao Xingru, Deng Qingwen, Zeng Xiang, Lai Bi-Qin, Ma Yuan-Huan, Li Ge, Zeng Yuan-Shan, Ding Ying
Department of Histology and Embryology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, China.
Key Laboratory for Stem Cells and Tissue Engineering, Ministry of Education, Sun Yat-sen University, Guangzhou, China.
Neurospine. 2022 Sep;19(3):757-769. doi: 10.14245/ns.2244452.226. Epub 2022 Sep 30.
This study aimed to identify differentially expressed genes (DEGs) by transcriptome analysis to elucidate a potential mechanism by which governor vessel electroacupuncture (GV-EA) promotes neuronal survival, axonal regeneration, and functional recovery after complete transection spinal cord injury (SCI).
Sham, control, or GV-EA group adult female Sprague Dawley rats underwent a complete transection SCI protocol. SCI area RNA-seq investigated the DEGs of coding and noncoding RNAs 7 days post-SCI. Gene ontology (GO) and kyoto encyclopedia of genes and genomes (KEGG) enrichment analyses were used to classify DEGs functions, to explain a possible molecular mechanism. Immunofluorescence and BBB (Basso, Beattie, and Bresnahan) score were used to verify a GV-EA treatment effect following SCI.
GV-EA treatment could regulate the expression of 173 mRNA, 260 lncRNA, and 153 circRNA genes among these DEGs resulted by SCI. GO enrichment analysis showed that the DEGs were most enriched in membrane, actin binding, and regulation of Toll-like receptor signaling pathway. KEGG pathway analysis showed enriched pathways (e.g. , Toll-like receptors, MAPK, Hippo signaling). According to the ceRNA network, miR-144-3p played a regulatory role by interacting with lncRNA and circRNA. GV-EA also promoted the injured spinal cord neuron survival, axonal regeneration, and functional improvement of hind limb locomotion.
Results of our RNA-seq suggest that post-SCI GV-EA may regulate characteristic changes in transcriptome gene expression, potential critical genes, and signaling pathways, providing clear directions for further investigation into the mechanism of GV-EA in subacute SCI treatment. Moreover, we found that GV-EA promotes neuronal survival, nerve fiber extension, and motor function recovery in subacute SCI.
本研究旨在通过转录组分析鉴定差异表达基因（DEGs），以阐明督脉电针（GV-EA）促进完全性横断脊髓损伤（SCI）后神经元存活、轴突再生和功能恢复的潜在机制。
成年雌性Sprague Dawley大鼠分为假手术组、对照组或GV-EA组，进行完全性横断SCI实验。在SCI后7天，对SCI区域进行RNA测序，研究编码和非编码RNA的DEGs。采用基因本体论（GO）和京都基因与基因组百科全书（KEGG）富集分析对DEGs的功能进行分类，以解释可能的分子机制。采用免疫荧光和BBB（Basso、Beattie和Bresnahan）评分来验证SCI后GV-EA的治疗效果。
GV-EA治疗可调节SCI导致的这些DEGs中的173个mRNA、260个lncRNA和153个circRNA基因的表达。GO富集分析表明，DEGs在膜、肌动蛋白结合和Toll样受体信号通路调节方面富集程度最高。KEGG通路分析显示了富集的通路（如Toll样受体、MAPK、Hippo信号通路）。根据ceRNA网络，miR-144-3p通过与lncRNA和circRNA相互作用发挥调节作用。GV-EA还促进了损伤脊髓神经元的存活、轴突再生以及后肢运动功能的改善。
我们的RNA测序结果表明，SCI后GV-EA可能调节转录组基因表达、潜在关键基因和信号通路的特征性变化，为进一步研究GV-EA在亚急性SCI治疗中的机制提供了明确方向。此外，我们发现GV-EA可促进亚急性SCI中神经元的存活、神经纤维延伸和运动功能恢复。
