Summary

小鼠或幼鼠的批量电针操作

Published: September 01, 2023
doi:

Summary

在这里,我们提出了一种使用三维(3D)打印支架在小鼠或年轻大鼠中进行高效和快速电针(EA)的方案。该技术允许在多只动物身上同时操作,节省时间并提高实验效率。

Abstract

电针(EA)被广泛用于治疗各种健康状况。然而,EA治疗的潜在机制尚不清楚,阻碍了其推广。机理研究需要小鼠或大鼠模型来解决这个问题。然而,这些动物不服从实验过程,这很耗时。为了解决这些问题,我们设计了3D打印的小动物身体散装固定器,以提高EA动物实验的效率。该视频详细介绍了如何使用固定器对小鼠或年轻大鼠进行批量 EA。对于穴位的选择,选取顶点颞部(MS6头部)和天枢点(ST25腹部)的前斜线,验证俯卧位和仰卧位固定装置的效果。使用3D打印的小动物支架可以同时固定和治疗多个啮齿动物,从而减少实验所需的时间和资源。这种技术可以通过3D打印不同尺寸的动物模型应用于其他动物模型,并有可能用于各种固定条件。该设备有利于促进EA的实验科学研究。

Introduction

电针疗法是一种独特的方法,将针灸针插入头皮并连接到电机以刺激特定穴位 1。与手动刺激不同,EA 通过稳定特定频率和波形来更好地控制刺激,以实现最佳治疗效果2。据调查,中国81.2%的医疗机构使用EA或手针灸治疗脑瘫、神经痛、面瘫等疾病。尽管很受欢迎,但EA的具体疗效机制仍然未知,这阻碍了其在神经系统疾病康复治疗中的推广和应用1,2,3。需要进一步的研究来充分了解EA的有效性机制,并促进其在神经系统疾病康复治疗中的应用。

随着针灸在世界范围内的影响力不断扩大,许多研究已经研究了对啮齿动物模型(例如大鼠或小鼠模型)进行 EA 的机制 1,2,3。在EA的啮齿动物实验中经常会遇到几个问题。首先是如何在没有麻醉的情况下固定啮齿动物,因为对清醒受试者进行的针灸更能反映临床实践。此外,一些实验要求动物保持清醒以观察治疗效果2,3。另一个挑战是准确定位小鼠或大鼠的穴位,这些穴位与人类的穴位相对应。目前,许多学者正在研究实验啮齿动物穴位的精确定位4,5。在该协议中,选择了MS6和ST25,它们已通过人体解剖定位的转换在啮齿动物中明确定义。MS6 通常用于治疗某些脑部疾病,例如帕金森病6。ST25 通常用于治疗胃肠道问题,例如腹泻7.选择这两个穴位主要是为了展示如何在仰卧位和俯卧位上有效地固定啮齿动物。此外,这些穴位已被广泛研究,并为研究目的提供了重要的见解 6,7

以前固定一只大鼠进行实验的方法不仅耗时,而且很难由一个人处理 8.此外,由于动物的不合作,在实践中成功率相对较低。因此,迫切需要创建一个易于建立的具有稳定特性的动物模型,以提高实验效率。在这篇文章中,介绍了一种用于小动物的3D打印支架,可以很容易地固定多只啮齿动物,导致运动受限。本文的目的是对一批年轻大鼠或小鼠进行EA治疗,重点研究小鼠批量限制、识别和刺激MS6和ST25穴位的策略。

Protocol

本研究的实验符合动物伦理学的“3R”原则,并已获得陆军医科大学实验动物福利和民族委员会(AMUWEC20234543)的批准。 1.啮齿动物身体固定装置的制备 基于中国实用新型专利9设计一种小动物支架。 通过3D打印技术生产设备。 准备一根 2 m 长的绳子,分成 6 根绳子。将这 6 根绳索穿过固定器上的孔。利用弹簧加载扣固定绳?…

Representative Results

我们设计的动物身体固定器的平面图如图1所示。此外,还提交了该固定器的 3D 图形模型,以提供设计的全面视图(补充文件 1)。这是一种允许 3 只啮齿动物固定并同时执行 EA 的设备。大鼠和小鼠在清醒状态下被限制在固定器内,俯卧位和仰卧位都可以牢固地固定,而不会对啮齿动物造成伤害(图2和图3</strong…

Discussion

电针 (EA) 是一种针灸形式,涉及在针灸针上使用电刺激11。该技术涉及使用特定强度和频率的微脉冲电流来刺激穴位并达到增强的治疗效果3.然而,EA的愈合机制是有限的,需要大量的基础研究来证明1,2,3。啮齿动物模型通常用于基本的 EA 研究 8,12,13<…

Declarações

The authors have nothing to disclose.

Acknowledgements

感谢陆军医科大学第二附属医院神经外科的现场支持。资助:本研究由中国自然科学基金(82104696)资助。

Materials

3D printing batch mice fixator MESH INVENT Custom made The fixator produced by 3D printer. The printing method is called fused deposition modeling (FDM), also known as fused filament fabrication (FFF).  
Electroacupuncture instrument Hwato, Suzhou Medical Appliance Factory SDZ-III www.Hwato-med.com
Disposable sterile acupuncture needle Suzhou Medical Appliance Factory N/A 0.25 mm x 13 mm

Referências

  1. Lee, S., Kim, S. N. The effect of acupuncture on modulating inflammatory cytokines in rodent animal models of respiratory disease: A systematic review and meta-analysis. Frontiers in Immunology. 13, 878463 (2022).
  2. Xin, Y. Y., Wang, J. X., Xu, A. J. Electroacupuncture ameliorates neuroinflammation in animal models. Acupuncture in Medicine: Journal of the British Medical Acupuncture Society. 40 (5), 474-483 (2022).
  3. Mengzhu, S., et al. Electroacupuncture at Tianshu (ST25) and Zusanli (ST36) alleviates stress-induced irritable bowel syndrome in mice by modulating gut microbiota and corticotropin-releasing factor. Journal of traditional Chinese medicine. 42 (5), 732-740 (2022).
  4. Xu, D. S., et al. A new attempt of re-mapping acupoint atlas in the rat. Acupuncture Research. 44 (1), 62-65 (2019).
  5. Yin, C. S., et al. A proposed transpositional acupoint system in a mouse and rat model. Research in Veterinary Science. 84 (2), 159-165 (2008).
  6. Cao, L., et al. The effectiveness of acupuncture for Parkinson’s disease: An overview of systematic reviews. Complementary Therapies in Medicine. 50, 102383 (2020).
  7. Yang, L., et al. Effect of warming moxibustion Tianshu (ST 25, bilateral) and Qihai (CV 6) for the treatment of diarrhea-dominant irritable bowel syndrome: a patient-blinded pilot trial with orthogonal design. Journal of Traditional Chinese Medicine. 37 (4), 538-545 (2017).
  8. Delcour, M., et al. Early movement restriction leads to maladaptive plasticity in the sensorimotor cortex and to movement disorders. Scientific Reports. 8 (1), 16328 (2018).
  9. Zhifeng, W. A fixing device for mice body. China Patent. , (2023).
  10. Ismail, K. I., Yap, T. C., Ahmed, R. 3D-printed fiber-reinforced polymer composites by fused deposition modelling (FDM): Fiber length and fiber implementation techniques. Polymers. 14 (21), 4659 (2022).
  11. Lu, C., et al. Efficacy of electroacupuncture with different frequencies in the treatment of chemotherapy-induced peripheral neuropathy: A study protocol for a randomized controlled trial. Frontiers in Neurology. 13, 843886 (2022).
  12. Zhang, L., et al. Modulation of colonic function in irritable bowel syndrome rats by electroacupuncture at ST25 and the neurobiological links between ST25 and the colon. Frontiers in Neuroscience. 16, 930489 (2022).
  13. Wang, J. H., et al. Anti-inflammation mechanism of electro-scalp acupuncture in treatment of ischemic stroke based on IL-12 mediated JAK/STAT signaling pathway. Chinese Acupuncture & Moxibustion. 42 (10), 1137-1144 (2022).
  14. Ren, D., et al. A novel design of a plate for posterolateral tibial plateau fractures through traditional anterolateral approach. Scientific Reports. 8 (1), 16418 (2018).
  15. Wu, L., Dong, Y., Zhu, C., Chen, Y. Effect and mechanism of acupuncture on Alzheimer’s disease: A review. Frontiers in Aging Neuroscience. 15, 1035376 (2023).
  16. Wu, L. Electroacupuncture for spinal cord injury: A scientific study of traditional medicine. Neurospine. 19 (3), 770-772 (2022).
  17. Gozzi, A., Zerbi, V. Modeling brain dysconnectivity in rodents. Biological Psychiatry. 93 (5), 419-429 (2023).
  18. Denic, A., Macura, S. I., Mishra, P., Gamez, J. D., Rodriguez, M., Pirko, I. MRI in rodent models of brain disorders. Neurotherapeutics. 8 (1), 3-18 (2011).
  19. Cong-hui, Q., et al. Efficacy observation of long-retaining scalp acupuncture plus interactive training for upper-extremity dysfunction after cerebral stroke. Journal of Acupuncture and Tuina Science. 19, 43-48 (2021).
  20. Kvist, L. J., Hall-Lord, M. L., Rydhstroem, H., Larsson, B. W. A randomised-controlled trial in Sweden of acupuncture and care interventions for the relief of inflammatory symptoms of the breast during lactation. Midwifery. 23 (2), 184-195 (2007).
  21. Boram, L., Chan-Young, K., Sun Haeng, L. Effectiveness and safety of auriculotherapy for breastfeeding: a systematic review. Journal of Traditional Chinese Medicine. 40 (5), 721-737 (2020).
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Yao, S., Wei, Y., Zhang, Y., Yang, X., Wu, Z. Bulk Electroacupuncture Operation for Mice or Young Rats. J. Vis. Exp. (199), e65648, doi:10.3791/65648 (2023).

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