Table_3_Transcriptome Analysis Reveals the Role of Cellular Calcium Disorder in Varicella Zoster Virus-Induced Post-Herpetic Neuralgia.XLSX

As a typical neuropathic pain, post-herpetic neuralgia (PHN) is a common complication of herpes zoster (HZ), which seriously affects the normal life and work of patients. The unclear pathogenesis and lack of effective drugs make the clinical efficacy of PHN unsatisfactory. Here, we obtained the transcriptome profile of neuroblastoma cells (SH-SY5Y) and DRG in rats infected with varicella zoster virus (VZV) by transcriptome sequencing (RNA-Seq) combined with publicly available gene array data sets. Next, the data processing of the transcriptome map was analyzed using bioinformatics methods, including the screening of differentially expressed genes (DEGs), Gene Ontology (GO), and the Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis. Finally, real-time fluorescent quantitative PCR (qRT-PCR) was used to detect the expression of calcium-related genes, and calcium fluorescent probes and calcium colorimetry were used to evaluate the distribution and content of calcium ions in cells after VZV infection. Transcriptome data analysis (GO and KEGG enrichment analysis) showed that calcium disorder played an important role in SH-SY5Y cells infected by VZV and dorsal root ganglion (DRG) of the PHN rat model. The results of qRT-PCR showed that the expression levels of calcium-related genes BHLHA15, CACNA1F, CACNG1, CHRNA9, and STC2 were significantly upregulated, while the expression levels of CHRNA10, HRC, and TNNT3 were significantly downregulated in SH-SY5Y cells infected with VZV. Our calcium fluorescent probe and calcium colorimetric test results showed that VZV could change the distribution of calcium ions in infected cells and significantly increase the intracellular calcium content. In conclusion, our results revealed that the persistence of calcium disorder caused by VZV in nerve cells might be a crucial cause of herpetic neuralgia, and a potential target for clinical diagnosis and treatment of PHN.

作为典型的神经性疼痛，带状疱疹后神经痛（post-herpetic neuralgia, PHN）是带状疱疹（herpes zoster, HZ）的常见并发症，严重影响患者的正常生活与工作。由于其发病机制尚不明确，且缺乏有效治疗药物，导致PHN的临床疗效欠佳。本研究通过转录组测序（transcriptome sequencing, RNA-Seq）结合公开基因芯片数据集，获取了水痘带状疱疹病毒（varicella zoster virus, VZV）感染的大鼠神经母细胞瘤细胞（SH-SY5Y）与背根神经节（dorsal root ganglion, DRG）的转录组谱。随后，采用生物信息学方法对转录组数据进行处理分析，包括差异表达基因（differentially expressed genes, DEGs）筛选、基因本体论（Gene Ontology, GO）分析及京都基因与基因组百科全书（Kyoto Encyclopedia of Genes and Genomes, KEGG）富集分析。最后，通过实时荧光定量PCR（real-time fluorescent quantitative PCR, qRT-PCR）检测钙相关基因的表达水平，并借助钙离子荧光探针与钙比色法，评估VZV感染后细胞内钙离子的分布与含量。转录组数据分析（GO与KEGG富集分析）结果显示，钙离子稳态紊乱在VZV感染的SH-SY5Y细胞及PHN大鼠模型的背根神经节中发挥了重要作用。qRT-PCR结果表明，在VZV感染的SH-SY5Y细胞中，钙相关基因BHLHA15、CACNA1F、CACNG1、CHRNA9及STC2的表达水平显著上调，而CHRNA10、HRC与TNNT3的表达水平显著下调。钙离子荧光探针与钙比色法的检测结果显示，VZV可改变感染细胞内钙离子的分布，并显著升高细胞内钙离子含量。综上，本研究结果揭示，VZV诱导神经细胞出现的持续性钙离子稳态紊乱，可能是疱疹性神经痛的关键致病原因之一，同时也可作为临床诊断与治疗PHN的潜在靶点。