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腰椎间盘突出症大鼠模型中尾段硬膜外脉冲射频后多个脊髓节段神经胶质细胞活性的变化

Changes in Neuroglial Activity in Multiple Spinal Segments after Caudal Epidural Pulsed Radiofrequency in a Rat Model of Lumbar Disc Herniation.

作者信息

Cho Hee Kyung, Kang Jae Hoon, Kim So-Yeon, Choi Mi-Jung, Hwang Se Jin, Cho Yun-Woo, Ahn Sang-Ho

机构信息

Department of Physical Medicine and Rehabilitation, College of Medicine, Catholic University, Daegu, Republic of Korea.

Goodtendon Rehabilitation Medicine Clinic, Busan, Republic of Korea.

出版信息

Pain Physician. 2016 Nov-Dec;19(8):E1197-E1209.

Abstract

BACKGROUND

Herniated lumbar discs can induce sciatica by mechanical compression and/or chemical irritation. It was recently reported that neuroglial cellular activity after pulsed radiofrequency (PRF) application to a single dorsal root ganglion (DRG) attenuated neuroglial activity at the corresponding spinal dorsal horn. Recently, caudal epidural PRF has been used to manage neuropathic pain, but evidence of molecular changes after the administration of caudal epidural PRF to attenuate neuropathic pain is lacking, and it has not been determined whether caudal epidural PRF affects neuroglial activity at different spinal levels.

OBJECTIVES

Using immunohistochemical methods in a rat model of lumbar disc herniation, the authors investigated the effects of caudal epidural PRF administration on pain-related behavior, on the activations of microglia and astrocytes in spinal cord, and on the expressions of calcitonin gene-related peptide (CGRP) and Transient receptor potential vanilloid 1(TRPV1) in the DRG at the L3, L4, L5, L6, and S1 levels.

STUDY DESIGN

Controlled animal trial.

SETTING

University hospital laboratory.

METHODS

Forty-five Sprague-Dawley rats were randomly assigned to a sham-operated group (n = 10) or a nucleus pulposus (NP)-exposed group (n = 35). Rats in the NP-exposed group were further subdivided into a NP-exposed with sham stimulation group (the NP-nonPRF group; n = 13) or a NP exposed with caudal epidural PRF stimulation group (the NP-PRF group; n = 22). Pulsed radiofrequency was administered on postoperative day 10 (POD 10) by placing an electrode in the caudal epidural space through the sacral hiatus and administering 5 Hz of PRF current for 600 seconds (maximum tip temperature 42°C). Rats were tested for mechanical allodynia on POD 10 and on days 7 and 14 after caudal epidural PRF administration (post-PRF). At 14 days post-PRF, sections of the spinal cord from L3, L4, L5, L6, and S1 were immunostained for ionized calcium-binding adapter molecule 1 (Iba1) and glial fibrillary acidic protein (GFAP), and DRGs from the same levels were immunostained for CGRP and TRPV1.

RESULTS

Mechanical withdrawal thresholds increased at 7 days post-PRF (P = 0.04), and the immunohistochemical expression of Iba1 in the L5 spinal dorsal horn and of CGRP in the L5 DRG were quantitatively reduced (P < 0.001) at 14 days post-PRF. Furthermore, the upregulations of Iba1 at L3, L4, L6, and S1 dorsal horns and CGRP at L6 DRG were also attenuated by caudal epidural PRF (P < 0.001).

LIMITATION

We examined molecular changes only in ipsilateral lumbar regions and at 14 days post-PRF.

CONCLUSION

Caudal epidural PRF reduced mechanical allodynia and downregulated microglia activity and CGRP expression at the lumbar disc herniated level and in adjacent lumbar spinal levels in a rat model of lumbar disc herniation.Key words: Caudal, pulsed radiofrequency, multisegmental, lumbar disc herniation, microglia, calcitonin gene-related peptide.

摘要

背景

腰椎间盘突出症可通过机械压迫和/或化学刺激诱发坐骨神经痛。最近有报道称,对单个背根神经节(DRG)进行脉冲射频(PRF)治疗后,神经胶质细胞活性会减弱相应脊髓背角处的神经胶质活性。近来,尾侧硬膜外PRF已被用于治疗神经性疼痛,但缺乏尾侧硬膜外PRF给药后减轻神经性疼痛的分子变化证据,且尚未确定尾侧硬膜外PRF是否会影响不同脊髓节段的神经胶质活性。

目的

作者采用免疫组化方法,在腰椎间盘突出症大鼠模型中,研究尾侧硬膜外PRF给药对疼痛相关行为、脊髓中小胶质细胞和星形胶质细胞的激活以及L3、L4、L5、L6和S1节段DRG中降钙素基因相关肽(CGRP)和瞬时受体电位香草酸亚型1(TRPV1)表达的影响。

研究设计

对照动物试验。

研究地点

大学医院实验室。

方法

45只Sprague-Dawley大鼠被随机分为假手术组(n = 10)或髓核(NP)暴露组(n = 35)。NP暴露组大鼠进一步细分为NP暴露加假刺激组(NP-非PRF组;n = 13)或NP暴露加尾侧硬膜外PRF刺激组(NP-PRF组;n = 22)。在术后第10天(POD 10)进行脉冲射频治疗,通过骶裂孔将电极置于尾侧硬膜外间隙,给予5 Hz的PRF电流600秒(最大尖端温度42°C)。在POD 10以及尾侧硬膜外PRF给药后(PRF后)第7天和第14天对大鼠进行机械性异常性疼痛测试。在PRF后第14天,对L3、L4、L5、L6和S1脊髓节段进行切片,免疫染色检测离子钙结合衔接分子1(Iba1)和胶质纤维酸性蛋白(GFAP),对相同节段的DRG进行免疫染色检测CGRP和TRPV1。

结果

PRF后第7天机械性撤针阈值升高(P = 0.04),PRF后第14天,L5脊髓背角Iba1的免疫组化表达以及L5 DRG中CGRP的表达定量降低(P < 0.001)。此外,尾侧硬膜外PRF也减弱了L3、L4、L6和S1背角Iba1以及L6 DRG中CGRP的上调(P < 0.001)。

局限性

我们仅检查了同侧腰部区域以及PRF后第14天的分子变化。

结论

在腰椎间盘突出症大鼠模型中,尾侧硬膜外PRF减轻了机械性异常性疼痛,并下调了椎间盘突出节段及相邻腰椎节段的小胶质细胞活性和CGRP表达。关键词:尾侧;脉冲射频;多节段;腰椎间盘突出症;小胶质细胞;降钙素基因相关肽

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