推拿对大鼠模型的镇痛作用，压迫背根神经节疼痛
1Department of Tuina,Yueyang Hospital of Integrated Traditional Chinese and Western Medicine Affiliated to Shanghai University of Traditional Chinese Medicine, 2Rehabilitation Medicine Department,Yueyang Hospital of Integrated Traditional Chinese and Western Medicine Affiliated to Shanghai University of Traditional Chinese Medicine, 3Department of Tuina,Shuguang Hospital of Shanghai University of Traditional Chinese Medicine
Summary
本文介绍了一种使用推拿疗法治疗大鼠背根神经节慢性压迫的操作方法，以及一种基于疼痛行为和组织病理学结果评估其有效性的方法。
Abstract
神经性疼痛是一种普遍存在的疾病，影响 6.9%-10% 的人口，由各种病因引起的神经损伤引起，例如腰椎间盘突出症、椎管狭窄和椎间孔狭窄。虽然推拿作为一种传统的中国手法疗法，在临床上显示出治疗神经性疼痛的镇痛作用，但其潜在的神经生物学机制尚不清楚。动物模型对于阐明推拿的基本原理至关重要。在这项研究中，我们提出了一种标准化的推拿方案，用于压迫背根神经节 （DRG） 的大鼠，其中包括通过将不锈钢棒插入椎间孔来诱导 DRG 压迫，在受控环境中以特定的位置、强度和频率参数进行推拿操作，并评估推拿治疗的行为和组织病理学结果。本文还讨论了该研究的潜在临床意义和局限性，并提出了未来推拿研究的方向。
Introduction
在临床环境中，由于各种原因，观察到神经根受压引起的神经系统病理性疼痛是很常见的。这种神经性疼痛最典型的形式是腰椎间盘突出症 （LDH），它通常是持续的、反复发作的且难以治愈。全球约有 9% 的人口受到乳酸脱氢酶的影响，导致严重的社会和经济负担1.由于人类生产和生活方式的改变，这种类型的神经性疼痛的发病率逐年增加，患者有年轻化的趋势2.尽管使用非甾体类止痛药，但患者的症状仍不能完全缓解。因此，用于治疗乳酸脱氢酶引起的疼痛的替代疗法（如推拿）越来越受到关注。
推拿疗法是乳酸脱氢酶的一种保守治疗形式，在世界范围内的各种临床实践指南中被广泛推荐用于预防和治疗腰痛 3,4。研究表明，推拿可显著降低乳酸脱氢酶患者的血清IL-6和肿瘤坏死因子-α（TNF-α）等炎症因子，同时改善患者的疼痛和腰椎功能损害5。然而，推拿疗法缓解疼痛效果背后的具体机制尚不清楚。
动物模型是研究 LDH6 引起的神经性疼痛的宝贵工具。它们允许进行行为测量以评估推拿疗法的有效性，并提供乳酸脱氢酶的病理生理学样本。例如，可以从大腿背根神经节采集样本来验证背根神经节细胞的变化。背根神经节 （CCD） 模型的慢性压迫通常用于评估 LDH 的病理生理学，因为它对背根神经节细胞的形态造成损害，这与椎间盘突出引起的神经压迫临床病例中观察到的病理变化一致7.
许多学者对穴位按压镇痛进行了几次动物实验8,9,10。然而，在动物模型上实施穴位按摩操作时，他们经常模仿人类穴位按摩。穴位按摩的治疗效果受大小、频率和施加力的方向等因素的影响11,12,13。如果实验缺乏统一的穴位按摩标准，如手术的力度、频率、持续时间等，可能会导致实验结果出现一定偏差。本文介绍了一套基于CCD大鼠特点的穴位按压治疗方案，并促进了动物模型中标准化穴位按压操作的发展。
Protocol
这项工作是在复旦大学神经生物学研究所疼痛实验室进行的。这些实验已获得批准，并严格遵守国际疼痛研究协会（LASP）为所有外科手术和动物处理制定的实验动物保护指南。本研究使用清洁级 Sprague-Dawley （SD） 大鼠，由 32 只 40-50 日龄的雄性组成，平均体重为 220 ± 1.38 g。这些大鼠购自中国科学院上海生命科学研究院实验动物中心。动物得到适当的照顾，并被安置在一个有独立通风、调节温度?…
Representative Results
推拿疗法可以帮助降低CCD建模引起的大鼠机械和热刺激阈值推拿治疗17天后，接受推拿治疗的CCD大鼠与未治疗的CCD组之间的PWT阈值存在显着差异（P = 0.021，<0.05）（图6 和 表1）。 CCD组接受推拿治疗的大鼠从治疗开始疼痛阈值就有所改善，取模后第14天CCD组与接受推拿治疗组的热痛阈值差异有统计学意义（P = 0.0047?…
Discussion
课题组对早期推拿操作的参数进行了相关研究。首先，设置推拿操作的力强度很重要。在临床推拿中，从业者根据自己的经验和患者的主观感受调整用力强度，通过沟通达到最佳的推拿效果。然而，这在动物实验中是不可行的。在动物实验中，“反应阈值”用于定义推拿操作的强度。这些根据动物的本能反应来判断施加的推拿力是否合适。Randall17 使用这种方法来研究大鼠炎症模…
Disclosures
The authors have nothing to disclose.
Acknowledgements
这项工作得到了上海市临床重症专科项目（批准号：Shslczdzk04001）的支持;上海市科委航海项目（批准号：22YF1444300）;上海中医药大学预算内项目（批准号：2021LK091）.
Materials
| "L" stainless steel rod (4 mm long and 0.4 mm in diameter) | hand-made | / | For CCD models making |
| ALMEMO admeasuring apparatus | ahlborn | 2450-1 | Mechanical Withdrawal Threshold test |
| Constant temperature slicer CM-1900 | Leica | 1491950C1US | For specimen production |
| Disinfectant (iodine) 100 mL/bottle | LIRCON/Shandong Lilkang | / | For disinfection |
| Disposable sterile syringe 5 mL | Shanghai Misha Wa Medical Industry | / | For injection |
| Electron microscope CX-31 | Olympus, Japan | BJ002318 | For specimen observation |
| Finger pressure recordings | Suzhou Changxian Optoelectronic Technology | CX1003w | For Tuina manipulation |
| Foam board (35 cm x 20 cm) | hand-made | / | It is our homemade apparatus for fixing rats |
| MERSILK W2512 | Johnson & Johnson | / | For tissue suture |
| Neutral balsam | Sinopharm Chemical Reagent | 10004160 | For specimen production |
| paraformaldehyde | China National Chemical Reagent | / | For specimen production |
| Pentobarbital sodium | Sigma-Aldrich | P3761 | For anesthesia of rat |
| Plantar Test Apparatus (Hargreaves Method) for Mice and Rats | IITC Life Science | / | Paw Withdrawal Latency |
| Precision electronic scale for experiment JY3002 | Shanghai Precision Scientific Instrument | / | Weighing of rat |
| Rat hair clipper | Philips | HP6341/00 | Shaving of rat fur |
| Restrainer for rats | Tongji University (self-made) | / | It is a homemade apparatus made by Tongji University, which can effectively immobilize the rats and fully expose their hind limbs. |
| Tissue-Tek O.C.T. Compound | SAKURA | 4583 | For specimen production |
| Uratan | China National Chemical Reagent | / | For anesthesia of rat |
| X-ray detector XR-600 | Dongguan Kaso Electronic Technology | / | Examination of CCD models |
| xylene | Shanghai Sinopharm Group | 100092 | For specimen production |
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Cite This Article
Meng, F., Xing, H., Su, X., Xu, W., Song, P., Gong, L. Analgesic Effect of Tuina on Rat Models with Compression of the Dorsal Root Ganglion Pain. J. Vis. Exp. (197), e65535, doi:10.3791/65535 (2023).