Zuo Shanhuai, Shi Guidong, Fan Jianchao, Fan Baoyou, Zhang Xiaolei, Liu Shen, Hao Yan, Wei Zhijian, Zhou Xianhu, Feng Shiqing
Department of Radiology, Tianjin Medical University General Hospital, Heping, Tianjin 300052, P.R. China.
Department of Orthopedics, Tianjin Medical University General Hospital, Heping, Tianjin 300052, P.R. China.
Exp Ther Med. 2021 Jan;21(1):48. doi: 10.3892/etm.2020.9479. Epub 2020 Nov 18.
Schwann cells are unique glial cells in the peripheral nervous system. These cells provide a range of cytokines and nutritional factors to maintain axons and support axonal regeneration. However, little is known concerning adhesion-associated epigenetic changes that occur in Schwann cells after peripheral nerve injury (PNI). In the present study, adhesion-associated DNA methylation biomarkers were assessed between normal and injury peripheral nerve. Specifically, normal Schwann cells (NSCs) and activated Schwann cells (ASCs) were obtained from adult Wistar rats. After the Schwann cells were identified, proliferation and adhesion assays were used to assess differences between NSCs and ASCs. Methylated DNA immunoprecipitation-sequencing and bioinformatics analysis were used to identify and analyze the differentially methylated genes. Reverse transcription-quantitative PCR was performed to assess the expression levels of adhesion-associated genes. In the present study, the proliferation and adhesion assays demonstrated that ASCs had a more robust proliferative activity and adhesion compared with NSCs. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses were performed to identify methylation-associated biological processes and signaling pathways. Protein-protein interaction network analysis revealed that Fyn, Efna1, Jak2, Vav3, Flt4, Epha7, Crk, Kitlg, Ctnnb1 and Ptpn11 were potential markers for Schwann cell adhesion. The expression levels of several adhesion-associated genes, such as vinculin, BCAR1 scaffold protein, collagen type XVIII α1 chain and integrin subunit β6, in ASCs were altered compared with those in NSCs. The current study analyzed adhesion-associated DNA methylation patterns of Schwann cells and identified candidate genes that may potentially regulate Schwann cell adhesion in Wistar rats before and after PNI.
施万细胞是周围神经系统中独特的神经胶质细胞。这些细胞提供一系列细胞因子和营养因子以维持轴突并支持轴突再生。然而，关于周围神经损伤（PNI）后施万细胞中发生的与黏附相关的表观遗传变化，我们所知甚少。在本研究中，评估了正常和损伤的周围神经之间与黏附相关的DNA甲基化生物标志物。具体而言，从成年Wistar大鼠中获取正常施万细胞（NSCs）和活化施万细胞（ASCs）。在鉴定出施万细胞后，使用增殖和黏附试验来评估NSCs和ASCs之间的差异。采用甲基化DNA免疫沉淀测序和生物信息学分析来鉴定和分析差异甲基化基因。进行逆转录定量PCR以评估与黏附相关基因的表达水平。在本研究中，增殖和黏附试验表明，与NSCs相比，ASCs具有更强的增殖活性和黏附能力。进行基因本体论和京都基因与基因组百科全书富集分析以鉴定与甲基化相关的生物学过程和信号通路。蛋白质-蛋白质相互作用网络分析显示，Fyn、Efna1、Jak2、Vav3、Flt4、Epha7、Crk、Kitlg、Ctnnb1和Ptpn11是施万细胞黏附的潜在标志物。与NSCs相比，ASCs中几种与黏附相关基因的表达水平发生了改变，如纽蛋白、BCAR1支架蛋白、 XVIII型胶原α1链和整合素亚基β6。本研究分析了施万细胞与黏附相关的DNA甲基化模式，并鉴定了可能在PNI前后调节Wistar大鼠施万细胞黏附的候选基因。
