Calcitonin gene-related peptide regulates spinal microglial activation through the histone H3 lysine 27 trimethylation via enhancer of zeste homolog-2 in rats with neuropathic pain.
Calcitonin gene-related peptide regulates spinal microglial activation through the histone H3 lysine 27 trimethylation via enhancer of zeste homolog-2 in rats with neuropathic pain.
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降钙素基因相关肽通过 zeste 同源物 2 增强子通过组蛋白 H3 赖氨酸 27 三甲基化调节神经病理性疼痛大鼠脊髓小胶质细胞的活化
DOI：
10.1186/s12974-021-02168-1
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发表时间：
2021-05-21
影响因子：
9.3
通讯作者：
Wang Z
中科院分区：
文献类型：
--
作者：
An Q;Sun C;Li R;Chen S;Gu X;An S;Wang Z
背景降钙素基因相关肽（Calcitonin gene-related peptide，CGRP）在转录水平上作为小胶质细胞激活的介导因子，可能促进伤害性信息传递。通过zeste同源物增强子2（EZH 2）的H3赖氨酸27（H3 K27 me 3）的三甲基化是调节周围神经损伤后炎症相关基因表达的表观遗传标记。在这项研究中，我们探讨了CGRP和H3 K27 me 3在神经损伤后的小胶质细胞激活的关系，并阐明了慢性神经病理性pain.MethodsMicroglial细胞（BV 2）的发病机制，用CGRP处理和差异富集的H3 K27 me 3基因启动子使用ChIP-seq检查。采用慢性压迫性损伤（CCI）大鼠模型，研究CGRP对CCI诱导的神经病理性痛中小胶质细胞活化和EZH 2/H3 K27 me 3信号通路的影响。CGRP处理通过EZH 2诱导脊髓背角和培养的小胶质细胞（BV 2）中H3 K27 me 3表达增加。ChIP-seq数据表明，CGRP显著改变了CGRP处理后小胶质细胞中基因启动子上的H3 K27 me 3富集，其中173个获得H3 K27 me 3，75个失去该标记，其主要富集在细胞生长，吞噬体和炎症的调节中。qRT-PCR验证了代表性候选基因（TRAF 3 IP 2、BCL 2L 11、ITGAM、DAB 2、NLRP 12、WNT 3、ADAM 10）的表达，实时细胞分析（RTCA）验证了小胶质细胞增殖。此外，CGRP治疗和CCI增加了ITGAM、ADAM 10、MCP-1和CX 3CR 1的表达，这些是脊髓背角和培养的小胶质细胞中小胶质细胞活化的关键介质。这种增加的影响，由CCI引起的抑制CGRP拮抗剂和EZH 2抑制剂，这是同时与衰减的机械和热痛敏CCI ratus.ConclusionOur的研究结果高度表明，CGRP是牵连在神经病理性疼痛的发生，通过调节小胶质细胞激活EZH 2介导的H3 K27 me 3在脊髓背角。
BackgroundCalcitonin gene-related peptide (CGRP) as a mediator of microglial activation at the transcriptional level may facilitate nociceptive signaling. Trimethylation of H3 lysine 27 (H3K27me3) by enhancer of zeste homolog 2 (EZH2) is an epigenetic mark that regulates inflammatory-related gene expression after peripheral nerve injury. In this study, we explored the relationship between CGRP and H3K27me3 in microglial activation after nerve injury, and elucidated the underlying mechanisms in the pathogenesis of chronic neuropathic pain.MethodsMicroglial cells (BV2) were treated with CGRP and differentially enrichments of H3K27me3 on gene promoters were examined using ChIP-seq. A chronic constriction injury (CCI) rat model was used to evaluate the role of CGRP on microglial activation and EZH2/H3K27me3 signaling in CCI-induced neuropathic pain.ResultsOverexpressions of EZH2 and H3K27me3 were confirmed in spinal microglia of CCI rats by immunofluorescence. CGRP treatment induced the increased of H3K27me3 expression in the spinal dorsal horn and cultured microglial cells (BV2) through EZH2. ChIP-seq data indicated that CGRP significantly altered H3K27me3 enrichments on gene promoters in microglia following CGRP treatment, including 173 gaining H3K27me3 and 75 losing this mark, which mostly enriched in regulation of cell growth, phagosome, and inflammation. qRT-PCR verified expressions of representative candidate genes (TRAF3IP2, BCL2L11, ITGAM, DAB2, NLRP12, WNT3, ADAM10) and real-time cell analysis (RTCA) verified microglial proliferation. Additionally, CGRP treatment and CCI increased expressions of ITGAM, ADAM10, MCP-1, and CX3CR1, key mediators of microglial activation in spinal dorsal horn and cultured microglial cells. Such increased effects induced by CCI were suppressed by CGRP antagonist and EZH2 inhibitor, which were concurrently associated with the attenuated mechanical and thermal hyperalgesia in CCI rats.ConclusionOur findings highly indicate that CGRP is implicated in the genesis of neuropathic pain through regulating microglial activation via EZH2-mediated H3K27me3 in the spinal dorsal horn.
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影响因子：
15.1
作者：
Joseph M;Anglada-Huguet M;Paesler K;Mandelkow E;Mandelkow EM
通讯作者：
Mandelkow EM
影响因子：
5.3
作者：
Penas C;Navarro X
通讯作者：
Navarro X
影响因子：
8.2
作者：
Quan, Yi;Jiang, Chang-tao;Wang, Xian
通讯作者：
Wang, Xian
影响因子：
4
作者：
Gjorgjevski, Marko;Hannen, Ricarda;Nimsky, Christopher
通讯作者：
Nimsky, Christopher
雷公藤甲素通过抑制脊髓胶质细胞HDACs上调对骨癌疼痛大鼠的镇痛作用
影响因子：
9.3
作者：
Hu XF;He XT;Zhou KX;Zhang C;Zhao WJ;Zhang T;Li JL;Deng JP;Dong YL
通讯作者：
Dong YL