Chen Shuhui, Gu Xinpei, Li Ruidi, An Shuhong, Wang Zhaojin
Department of Human Anatomy, Shandong First Medical University & Shandong Academy of Medical Sciences, 2 Ying Sheng Dong Lu, Taian, 271000, China.
Neurochem Res. 2023 Jun;48(6):1945-1957. doi: 10.1007/s11064-023-03879-y. Epub 2023 Feb 10.
The histone H3 lysine 27 trimethylation (H3K27me3) is one of the most important chromatin modifications, which is associated with injury-activated gene expression in Schwann cells (SCs). However, the alteration of genome-wide H3K27me3 enrichments in the development of neuropathic pain is still unknown. Here, we applied the chromatin immunoprecipitation sequencing (ChIP-seq) approach to identify the alteration of differential enrichments of H3K27me3 in chronic constriction injury (CCI) sciatic nerve of rats and potential molecular mechanisms underlying the development of neuropathic pain. Our results indicated that CCI increased the numbers of SCs displaying H3K27 methyltransferase enhancer of zeste homolog 2 (EZH2) and H3K27me3 in the sciatic nerve. ChIP-seq data showed that CCI significantly changed H3K27me3 enrichments on gene promoters in the sciatic nerve. Bioinformatics analyses exhibited that genes gaining H3K27me3 were mostly associated with regulation of cell proliferation, response to stress and oxidation-reduction process. Genes losing this mark were enriched in neuronal generation, and MAPK, cAMP as well as ERBB signaling pathways. Importantly, IL1A, CCL2, NOS2, S100A8, BDNF, GDNF, ERBB3 and C3 were identified as key genes in neuropathic pain. CCI led to significant upregulation of key genes in the sciatic nerve. EZH2 inhibitor reversed CCI-induced increases of H3K27me3 and key gene protein levels, which were accompanied by relieved mechanical allodynia and thermal hyperalgesia in CCI rats. These results indicate that genes with differential enrichments of H3K27me3 in SCs function in various cellular processes and pathways, and many are linked to neuropathic pain after peripheral nerve injury.
组蛋白H3赖氨酸27三甲基化(H3K27me3)是最重要的染色质修饰之一,它与雪旺细胞(SCs)中损伤激活的基因表达相关。然而,在神经性疼痛发展过程中全基因组H3K27me3富集的变化仍不清楚。在此,我们应用染色质免疫沉淀测序(ChIP-seq)方法来鉴定大鼠慢性压迫性损伤(CCI)坐骨神经中H3K27me3差异富集的变化以及神经性疼痛发展潜在的分子机制。我们的结果表明,CCI增加了坐骨神经中显示H3K27甲基转移酶增强子同源物2(EZH2)和H3K27me3的雪旺细胞数量。ChIP-seq数据显示,CCI显著改变了坐骨神经中基因启动子上的H3K27me3富集。生物信息学分析表明,获得H3K27me3的基因大多与细胞增殖调控、应激反应和氧化还原过程相关。失去该标记的基因富集于神经元生成以及MAPK、cAMP和ERBB信号通路。重要的是,IL1A、CCL2、NOS2、S100A8、BDNF、GDNF、ERBB3和C3被鉴定为神经性疼痛中的关键基因。CCI导致坐骨神经中关键基因显著上调。EZH2抑制剂逆转了CCI诱导的H3K27me3和关键基因蛋白水平的增加,同时CCI大鼠的机械性异常性疼痛和热痛觉过敏得到缓解。这些结果表明,雪旺细胞中H3K27me3差异富集的基因在各种细胞过程和通路中发挥作用,并且许多与周围神经损伤后的神经性疼痛相关。