阿尔茨海默病埋线针灸治疗的手术程序及注意事项
Summary
本研究提出了一种对阿尔茨海默病样大鼠进行线嵌入针灸治疗的方案。
Abstract
埋线疗法 （TEAT） 是一种通过将可生物降解的缝合线插入穴位来预防和管理疾病的治疗方法，提供持久的刺激。TEAT 是一种简单的方法，可以避免常规针灸的不适并提供持续的治疗效果。本文讨论了 TEAT 对具有阿尔茨海默病样症状的大鼠学习和记忆能力的潜在影响。由于化学诱导的大鼠神经元变性和认知障碍并不完全反映在阿尔茨海默病中观察到的真实病理变化。因此，我们的研究小组将这些表现指定为阿尔茨海默病样症状。已经制定了一份协议来概述穴位的选择、手术过程以及头部和下背部的必要预防措施。实验分为 3 组：对照组、模型组和 TEAT 组，每组 6 只大鼠。为了诱发阿尔茨海默病样症状，大鼠腹膜内注射 D-半乳糖 7 周 （49 天）。TEAT 组大鼠接受穴位肠埋伏治疗。干预期结束后，进行了莫里斯水迷宫 （MWM） 以评估大鼠的学习和记忆能力。随后，处死大鼠，检查它们的脑组织。进行组织学检查以了解 TEAT 对表现出阿尔茨海默病症状的大鼠病理的影响。这项研究表明，TEAT 可能会改善具有阿尔茨海默病样症状的大鼠的学习和记忆，这表明这种神经退行性疾病可能是一种有前途的新治疗方法。
Introduction
埋线针灸 （TEA） 涉及使用特殊针头将肠线或可吸收缝合线嵌入穴位。这种技术具有持久的效果，直到缝合线被吸收和降解。它为穴位提供持续和持续的刺激一周或更长时间1，无需每天重复手术即可达到可比的结果2。这减少了就医的患者数量，从而在一定程度上节省了医疗资源。近年来，TEAT 在中国的各个医学领域得到了越来越多的应用，包括内科、外科、皮肤病学、五官、妇科和儿科3。研究表明，它对某些慢性和复杂疾病，如癫痫4 （epilepsy）和面神经麻痹5（facial nerve paralysis））产生长期可靠的效果是有效的。具有创伤小、操作简单6.此外，据报道，TEAT 的治疗费用低于电针 （EA）7。因此，TEAT 在临床和科学研究中越来越受到重视。
阿尔茨海默病 （AD） 是一种随着时间的推移而进展的神经退行性疾病。它的病理生理过程在临床症状出现前大约 20 年开始积累，但其机制仍未完全清楚8。鉴于治疗 AD 的有效药物有限，研究越来越关注该疾病的前驱期和临床前阶段9。预防是解决 AD 发展和进展的关键策略 9,10。近年来，中医 （TCM） 取得了重大进展，有望为预防和治疗 AD 提供新的可能性11,12。作为针灸的一种亚型，TEAT 因其治疗范围广、创伤小、手术简单、治疗频率低和疗效持久而优于其他疗法6。新出现的文献表明，TEAT 可能对肥胖13,14、糖尿病15、失眠16、绝经后骨质疏松症17 和抑郁症18 的个体有益，所有这些都是 AD 的危险因素。它还通过调节线粒体自噬2 改善衰老大鼠的衰老，改善空间学习和记忆障碍，减轻海马体的病理损伤，并抑制血管性痴呆 （VD） 大鼠的炎症反应19。弄清楚 TEAT 是否也可以作用于 AD 期间的病理过程似乎很有吸引力。
目前的研究在 TEAT 操作的几个关键要素上缺乏统一性和标准化，包括嵌入方法、嵌入工具、线程和嵌入间隔周期。这种缺乏标准化影响了对治疗效果的评价和相关机制的探索，并限制了外部推广和交流。先前的研究 20,21 已经证明，腹膜内注射 D-半乳糖 （D-gal） 可诱导大鼠出现阿尔茨海默病样病理变化，包括神经元变性和认知障碍。此外，百会 （GV20） 和神枢 （BL23） 的电针已被证明可有效改善认知能力下降、减轻神经炎症和减少神经元损伤。本研究以阿尔茨海默病样模型大鼠为例，介绍了该模型下 TEAT 的穴位选择及操作过程和注意事项。它讨论了 TEAT 在实验研究中的关键操作要素，为未来的研究提供参考。
Protocol
所有实验均按照《美国国立卫生研究院实验动物护理和使用指南》进行，并经湖北省疾病预防控制中心实验动物管理和使用委员会批准。共 18 只 3 月龄雌性 Sprague-Dawley （SD） 大鼠，体重 220-250 g。所有大鼠均在 23 ± 1 °C 和 40%-50% 湿度下维持 12 小时的光照/黑暗循环，可自由获取食物和水。在为期一周的适应期之后，在此期间让大鼠适应实验室条件，开始实验。 1. 模型开?…
Representative Results
本文介绍了 TEAT 在阿尔茨海默病样大鼠中的具体操作方法。它讨论了 TEAT 在实验研究中的关键操作要素，为未来的研究提供参考。 TEAT 改善阿尔茨海默病样大鼠的学习如流程图（图 5）所示，在适应性训练期间，观察到每组中的大鼠在游泳时观察周围环境。游泳速度分析（图 6）显示各组之间没有显着差异，表明运?…
Discussion
干预时间
AD 的病因很复杂，其发病机制仍不确定。目前，该病尚无有效的治疗方法，其病程不可逆。因此，学术界已就关注 AD 预防和治疗的重要性达成共识。AD 的预防和治疗策略强调早期干预，遵循中医的理念，即在疾病发生之前进行治疗，强调在疾病发作、传播和复发之前进行治疗。对人类29,30 和啮齿动物31,32</s…
Divulgations
The authors have nothing to disclose.
Acknowledgements
这项工作得到了国家自然科学基金 （NSFC） 第 82374564 号（至 YJD）和第 82074566 号（至 YJD）和中国武汉世界针灸学会联合会孙国杰中医针灸传承基地（世界针灸学会联合会 [2019] 26 号）（至 YJD）和国家中医药管理局全国著名中医传承工作室建设项目的资金支持， 中国（国家中医药管理局 [2022] 5 号）（致 YJD）。
Materials
| 0.9% saline, 4% paraformaldehyde, Hematoxylin, Eosin Y (water-soluble), absolute ethanol, 0.01M PBS powder, xylene, hydrochloric acid, embedded paraffin, neutral gum | sinopharm (www.sinopharm.com) | ||
| 5-0 absorbable surgical suture | bodamedical (www.bodamedical.com) | ||
| 70% alcohol, iodophor, cotton ball | Used for disinfection. | ||
| chloral hydrate | sigma | C8383 | |
| coverslips, glass slides | Guangzhou Wuqiang Experimental Equipment Co., Ltd | ||
| Dehydrator | Wuhan Junjie JT-12J Computerized Biological Tissue Dehydrator | ||
| DELUXE THERMOSTAT HEAT MAT | Zhenhua Instruments | For maintaining the body temperature of rats. | |
| D-galactose | sigma | G0750 | |
| electric men's beard trimmer | Used for shaving rats. | ||
| Electronic scale for small animals (ZK-DST) | Henan Zhike Information Technology Co., Ltd | ||
| erythromycin ointment | frontpharm | ||
| glass dish | Used to place cut absorbable sutures. | ||
| High-Pressure Steam Sterilization Pot (YM100Z) | Shanghai Sanshen medical equipment factory | For sterilization of surgical instruments. | |
| Ice Maker (CMB40) | Shanghai Sheyan Instrument Co., Ltd. | ||
| Microscope (BX-53) | OLYMPUS | ||
| milk powder | Used for the MWM test. | ||
| Morris Water Maze Video Tracking System & Data Analysis Software (Version: Watermaze 2.0) | Chengdu Taimeng Technology Co., Ltd | ||
| No.6 thread-embedding needle | bodamedical (www.bodamedical.com) | ||
| ophthalmic scissors, trays, tweezers | |||
| Pathology Microtome (RM 2016) | LEICA, Germany | ||
| pentobarbital | Germany | P11011 | |
| permanent marker | Used to mark the acupoint. | ||
| Slicing knives (R35) | Feathers, Japanese | ||
| soap | Used to wash hands. | ||
| sterile gloves |
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You, M., Miao, J., Yang, M., Liu, Q., Liu, Y., Du, Y. Operation Procedure and Precautions of Thread-Embedding Acupuncture Therapy in Alzheimer. J. Vis. Exp. (207), e65895, doi:10.3791/65895 (2024).