目的:本研究旨在探索A型肉毒杆菌毒素(BoNT/A)通过P2X7受体(P2X7R)抑制小胶质细胞炎症激活的潜在机制。
背景:BoNT/A是一种很有前途的镇痛药物,和先前的研究已经确定它通过抑制小胶质细胞炎症激活来减轻神经性疼痛(NP)。这项研究检查了BoNT/A通过介导小胶质细胞P2X7R和分析小鼠BV-2小胶质细胞的转录组测序数据来缓解神经性疼痛的生物标志物和潜在机制。
目的:P2X7R激动剂Bz-ATP用于诱导小胶质细胞炎症激活,而RNAseq技术用于探索BoNT/A抑制小胶质细胞炎症的生物标志物和潜在机制。
方法:对三个用P2X7R特异性激活剂(Bz-ATP)处理的BV-2细胞样品和三个用BoNT/A预处理的BV-2细胞样品进行RNA测序。只有成功通过质量控制措施的数据才包括在随后的分析中。最初,从BoNT/A和对照样品中鉴定出差异表达基因(DEGs),其次是基因本体论(GO)和京都基因和基因组百科全书(KEGG)富集分析。然后通过构建蛋白质-蛋白质相互作用(PPI)网络并利用Cytoscape软件中的CytoHubba插件来鉴定生物标志物。最后,进行了富集分析和调节网络分析,以阐明BoNT/A治疗NP的潜在机制。
结果:确定了与“细胞成分大小调节”GO术语相关并富集在“轴突指导”KEGG途径中的93DEGG。随后,确定了6种生物标志物,即PTPRF,CHDH,CKM,Ky,Sema3b,和Sema3f,富含与生物合成和代谢相关的途径,疾病进展,信号转导,和细胞器功能,包括“核糖体”和“Wnt信号通路”。“最后,从6个mRNA构建竞争内源性RNA(CERNA)网络,66miRNAs,和31个lncRNAs,形成复杂的关系网络。
结论:六个基因(PTPRF,Sema3b,Sema3f,CHDH,CKM,和Ky)被鉴定为BoNT/A治疗后小胶质细胞炎症激活的生物标志物。这一发现可为神经病理性疼痛的缓解和治疗提供有价值的参考。
OBJECTIVE: This study aims to explore the potential mechanism by which Botulinum toxin type A (BoNT/ A) inhibits microglial inflammatory activation through P2X7 receptors (P2X7R).
BACKGROUND: BoNT/A is a promising analgesic drug, and previous studies have established that it alleviates Neuropathic Pain (NP) by inhibiting microglial inflammatory activation. This study examined the biomarkers and potential mechanisms by which BoNT/A relieves neuropathic pain by mediating microglial P2X7R and analyzing transcriptome sequencing data from mouse BV-2 microglial cells.
OBJECTIVE: The P2X7R agonist Bz-ATP was used to induce microglial inflammatory activation, whilst RNAseq technology was used to explore the biomarkers and potential mechanisms through which BoNT/A suppresses microglial inflammation.
METHODS: RNA sequencing was performed on three BV-2 cell samples treated with a P2X7R specific activator (Bz-ATP) and three BV-2 cell samples pre-treated with BoNT/A. Only data that successfully passed quality control measures were included in subsequent analysis. Initially, Differentially Expressed Genes (DEGs) were identified from BoNT/A and control samples, followed by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses. Biomarkers were then identified by constructing a Protein- Protein Interaction (PPI) network and utilizing the CytoHubba plug-in in Cytoscape software. Lastly, enrichment analysis and regulatory network analysis were performed to elucidate the potential mechanism of BoNT/A in the treatment of NP.
RESULTS: 93 DEGs related to the \"cell component size regulation\" GO term and enriched in the \"axon guidance\" KEGG pathway were identified. Subsequently, 6 biomarkers were identified, namely PTPRF, CHDH, CKM, Ky, Sema3b, and Sema3f, which were enriched in pathways related to biosynthesis and metabolism, disease progression, signal transduction, and organelle function, including the \"ribosome\" and \"Wnt signaling pathway.\" Finally, a competing endogenous RNA (ceRNAs) network was constructed from 6 mRNAs, 66 miRNAs, and 31 lncRNAs, forming a complex relationship network.
CONCLUSIONS: Six genes (PTPRF, Sema3b, Sema3f, CHDH, CKM, and Ky) were identified as biomarkers of microglial inflammatory activation following BoNT/A treatment. This finding may provide a valuable reference for the relief and treatment of neuropathic pain.