由11-易位甲基胞嘧啶双加氧酶1和3介导的DNA羟甲基化在慢性炎症性疼痛模型中调节伤害性敏感化。

DNA Hydroxymethylation by Ten-eleven Translocation Methylcytosine Dioxygenase 1 and 3 Regulates Nociceptive Sensitization in a Chronic Inflammatory Pain Model.

作者信息

Pan Zhiqiang, Xue Zhou-Ya, Li Guo-Fang, Sun Meng-Lan, Zhang Ming, Hao Ling-Yun, Tang Qian-Qian, Zhu Li-Jiao, Cao Jun-Li

机构信息

From Jiangsu Province Key Laboratory of Anesthesiology (Z.P., Z.-Y.X., G.-F.L., M.-L.S., M.Z., L.-Y.H., Q.-Q.T., L.-J.Z., J.-L.C.) and Jiangsu Province Key Laboratory of Anesthesia and Analgesia Application Technology (Z.P., Z.-Y.X., G.-F.L., M.-L.S., M.Z., L.-Y.H., Q.-Q.T., L.-J.Z., J.-L.C.), Xuzhou Medical University, Xuzhou; and Department of Anesthesiology, The Affiliated Hospital of Xuzhou Medical University, Xuzhou (J.-L.C.), China.

出版信息

Anesthesiology. 2017 Jul;127(1):147-163. doi: 10.1097/ALN.0000000000001632.

DOI:10.1097/ALN.0000000000001632

PMID:28437360

Abstract

BACKGROUND

Ten-eleven translocation methylcytosine dioxygenase converts 5-methylcytosine in DNA to 5-hydroxymethylcytosine, which plays an important role in gene transcription. Although 5-hydroxymethylcytosine is enriched in mammalian neurons, its regulatory function in nociceptive information processing is unknown.

METHODS

The global levels of 5-hydroxymethylcytosine and ten-eleven translocation methylcytosine dioxygenase were measured in spinal cords in mice treated with complete Freund's adjuvant. Immunoblotting, immunohistochemistry, and behavioral tests were used to explore the downstream ten-eleven translocation methylcytosine dioxygenase-dependent signaling pathway.

RESULTS

Complete Freund's adjuvant-induced nociception increased the mean levels (± SD) of spinal 5-hydroxymethylcytosine (178 ± 34 vs. 100 ± 21; P = 0.0019), ten-eleven translocation methylcytosine dioxygenase-1 (0.52 ± 0.11 vs. 0.36 ± 0.064; P = 0.0088), and ten-eleven translocation methylcytosine dioxygenase-3 (0.61 ± 0.13 vs. 0.39 ± 0.08; P = 0.0083) compared with levels in control mice (n = 6/group). The knockdown of ten-eleven translocation methylcytosine dioxygenase-1 or ten-eleven translocation methylcytosine dioxygenase-3 alleviated thermal hyperalgesia and mechanical allodynia, whereas overexpression cytosinethem in naïve mice (n = 6/group). Down-regulation of spinal ten-eleven translocation methylcytosine dioxygenase-1 and ten-eleven translocation methylcytosine dioxygenase-3 also reversed the increases in Fos expression (123 ± 26 vs. 294 ± 6; P = 0.0031; and 140 ± 21 vs. 294 ± 60; P = 0.0043, respectively; n = 6/group), 5-hydroxymethylcytosine levels in the Stat3 promoter (75 ± 16.1 vs. 156 ± 28.9; P = 0.0043; and 91 ± 19.1 vs. 156 ± 28.9; P = 0.0066, respectively; n = 5/group), and consequent Stat3 expression (93 ± 19.6 vs. 137 ± 27.5; P = 0.035; and 72 ± 15.2 vs. 137 ± 27.5; P = 0.0028, respectively; n = 5/group) in complete Freund's adjuvant-treated mice.

CONCLUSIONS

This study reveals a novel epigenetic mechanism for ten-eleven translocation methylcytosine dioxygenase-1 and ten-eleven translocation methylcytosine dioxygenase-3 in the modulation of spinal nociceptive information via targeting of Stat3.

摘要

背景

10-11易位甲基胞嘧啶双加氧酶将DNA中的5-甲基胞嘧啶转化为5-羟甲基胞嘧啶，其在基因转录中发挥重要作用。虽然5-羟甲基胞嘧啶在哺乳动物神经元中含量丰富，但其在伤害性信息处理中的调节功能尚不清楚。

方法

在完全弗氏佐剂处理的小鼠脊髓中测量5-羟甲基胞嘧啶和10-11易位甲基胞嘧啶双加氧酶的整体水平。采用免疫印迹、免疫组织化学和行为测试来探索下游依赖10-11易位甲基胞嘧啶双加氧酶的信号通路。

结果

与对照小鼠（每组n = 6）相比，完全弗氏佐剂诱导的伤害感受增加了脊髓5-羟甲基胞嘧啶（178±34 vs. 100±21；P = 0.0019）、10-11易位甲基胞嘧啶双加氧酶-1（0.52±0.11 vs. 0.36±0.064；P = 0.0088）和10-11易位甲基胞嘧啶双加氧酶-3（0.61±0.13 vs. 0.39±0.08；P = 0.0083）的平均水平（±标准差）。敲低10-11易位甲基胞嘧啶双加氧酶-1或10-11易位甲基胞嘧啶双加氧酶-3可减轻热痛觉过敏和机械性异常性疼痛，而在未处理的小鼠（每组n = 6）中过表达胞嘧啶则无此作用。脊髓中10-11易位甲基胞嘧啶双加氧酶-1和10-11易位甲基胞嘧啶双加氧酶-3的下调也逆转了完全弗氏佐剂处理小鼠中Fos表达的增加（分别为123±26 vs. 294±6；P = 0.0031；和140±21 vs. 294±60；P = 0.0043；每组n = 6）、Stat3启动子中5-羟甲基胞嘧啶水平的增加（分别为75±16.1 vs. 156±28.9；P = 0.0043；和91±19.1 vs. 156±28.9；P = 0.0066；每组n = 5）以及随之而来的Stat3表达的增加（分别为93±19.6 vs. 137±27.5；P = 0.035；和72±15.2 vs. 137±27.5；P = 0.0028；每组n = 5）。