神经损伤诱导的HDAC4细胞质滞留的调节促成大鼠神经性疼痛。

Modulation of Nerve Injury-induced HDAC4 Cytoplasmic Retention Contributes to Neuropathic Pain in Rats.

作者信息

Lin Tzer-Bin, Hsieh Ming-Chun, Lai Cheng-Yuan, Cheng Jen-Kun, Chau Yat-Pang, Ruan Ting, Chen Gin-Den, Peng Hsien-Yu

机构信息

From the Department of Physiology, School of Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan, Republic of China (T.-B.L.); Graduate Institute of Basic Medical Science, China Medical University, Taichung, Taiwan, Republic of China (T.-B.L.); Department of Biotechnology, Asia University, Taichung, Taiwan, Republic of China (T.-B.L.); Department of Medicine, Mackay Medical College, New Taipei, Taiwan, Republic of China (M.-C.H., C.-Y.L., Y.-P.C., H.-Y.P.); Department of Physiology, School of Medicine, College of Medicine, National Taiwan University, Taipei, Taiwan, Republic of China (M.-C.H.); Department of Veterinary Medicine, College of Veterinary Medicine, National Chung-Hsing University, Taichung, Taiwan, Republic of China (C.-Y.L.); Department of Anesthesiology, Mackay Memorial Hospital, New Taipei, Taiwan, Republic of China (J.-K.C.); School of Medicine, Fu-Jen Catholic University, New Taipei, Taiwan, Republic of China (T.R.); and Department of Obstetrics and Gynecology, Chung-Shan Medical University Hospital, Chung-Shan Medical University, Taichung, Taiwan, Republic of China (G.-D.C.).

出版信息

Anesthesiology. 2015 Jul;123(1):199-212. doi: 10.1097/ALN.0000000000000663.

DOI:10.1097/ALN.0000000000000663

PMID:25871743

Abstract

BACKGROUND

The histone deacetylases (HDACs) have been implicated in pain hypersensitivity. This study investigated the potential involvement of an HDAC4-related mechanism in the spinal nerve ligation (SNL)-induced nociceptive hypersensitivity.

METHODS

The left L5 to L6 spinal nerves of 627 adult male Sprague-Dawley rats were surgically ligated. The withdrawal threshold of hind paws and the abundances, cellular location, and interactions of proteins in the dorsal horn were assayed before and after surgery. The 14-3-3β-targeting small-interfering RNA, a serum- and glucocorticoid-inducible kinase 1 (SGK1) antagonist, or an HDAC inhibitor was spinally injected to elucidate the role of 14-3-3β, SGK1, and HDAC4.

RESULTS

Without affecting the HDAC4 level, SNL provoked SGK1 phosphorylation (mean ± SEM from 0.24 ± 0.02 to 0.78 ± 0.06 at day 7, n = 6), HDAC4 phosphorylation (from 0.38 ± 0.03 to 0.72 ± 0.06 at day 7, n = 6), 14-3-3β expression (from 0.53 ± 0.09 to 0.88 ± 0.09 at day 7, n = 6), cytoplasmic HDAC4 retention (from 1.18 ± 0.16 to 1.92 ± 0.11 at day 7, n = 6), and HDAC4-14-3-3β coupling (approximately 2.4-fold) in the ipsilateral dorsal horn in association with behavioral allodynia. Knockdown of spinal 14-3-3β expression prevented the SNL-provoked HDAC4 retention (from 1.89 ± 0.15 to 1.32 ± 0.08 at day 7, n = 6), HDAC4-14-3-3β coupling (approximately 0.6-fold above SNL 7D), and behavioral allodynia (from 0.16 ± 0.3 to 6 ± 1.78 at day 7, n = 7), but not SGK1 (from 0.78 ± 0.06 to 0.71 ± 0.04 at day 7, n = 6) or HDAC4 (from 0.75 ± 0.15 to 0.68 ± 0.11 at day 7, n = 6) phosphorylation.

CONCLUSION

Neuropathic pain maintenance involves the spinal SGK1 activation-dependent HDAC4 phosphorylation and its subsequent association with 14-3-3β that promotes cytoplasmic HDAC4 retention in dorsal horn neurons.

摘要

背景

组蛋白去乙酰化酶（HDACs）与痛觉过敏有关。本研究调查了HDAC4相关机制在脊髓神经结扎（SNL）诱导的伤害性超敏反应中的潜在作用。

方法

对627只成年雄性Sprague-Dawley大鼠的左侧L5至L6脊髓神经进行手术结扎。在手术前后测定后爪的退缩阈值以及背角中蛋白质的丰度、细胞定位和相互作用。脊髓注射靶向14-3-3β的小干扰RNA、血清和糖皮质激素诱导激酶1（SGK1）拮抗剂或HDAC抑制剂，以阐明14-3-3β、SGK1和HDAC4的作用。

结果

在不影响HDAC4水平的情况下，SNL引起同侧背角中SGK1磷酸化（第7天平均值±标准误从0.24±0.02增至0.78±0.06，n = 6）、HDAC4磷酸化（第7天从0.38±0.03增至0.72±0.06，n = 6）、14-3-3β表达（第7天从0.53±0.09增至0.88±0.09，n = 6）、HDAC4在细胞质中的滞留（第7天从1.18±0.16增至1.92±0.11，n = 6）以及HDAC4与14-3-3β的结合（约2.4倍），同时伴有行为性异常性疼痛。敲低脊髓14-3-3β表达可阻止SNL引起的HDAC4滞留（第7天从1.89±0.15降至1.32±0.08，n = 6）、HDAC4与14-3-3β的结合（比SNL第7天高约0.6倍）以及行为性异常性疼痛（第7天从0.16±0.3增至6±1.78，n = 7），但不影响SGK1（第7天从0.78±0.06降至0.71±0.04，n = 并6）或HDAC4（第7天从0.75±0.15降至0.68±0.11，n = 6）的磷酸化。