Name: 著者スポットライト:電気鍼治療中のラットの固定を改善するための効率的なラットの服
Uploaded: 2023-08-18T14:05:00
Description: Watch this Scientific Journal Video about Efficient Rat Clothing for Improved Rat Fixation During Electroacupuncture at JoVE.com

著者らは、本論文の図2の作成に使用した資料を認証ライセンスID:URUUO5699eで提供してくれたHome for ResearchersのFigdraw科学研究図面プラットフォームに感謝の意を表します。この研究は、中国国家自然科学基金会(81873380、82074566年)、湖北省自然科学基金会(2022CFCO43)、および国家中医薬管理局の全国有名漢方薬遺産スタジオ建設プロジェクト([2022] No.5、[2022] No.75)の助成金から支援を受けています。

この研究でラットに実施されたすべての手順は、中華人民共和国によって正式に採択された実験動物福祉および倫理審査ガイドライン(2018)の国家基準および国立アカデミープレスの実験動物のケアと使用に関するガイド15に準拠していました。このプロトコルは、3つの主要なステップで構成されています。まず、ラットジャケットは、体重の異なるラットの個々の体型に基?...

ラットの電気鍼治療中の懸濁液固定を強化するラットジャケット

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T., Barbee, R. W., Garber, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Guide for the care and use of laboratory animals. 85 (3), 963-965 (2011).</li><li>Zheng, X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electroacupuncture+ameliorates+beta-amyloid+pathology+and+cognitive+impairment+in+Alzheimer+disease+via+a+novel+mechanism+involving+activation+of+TFEB+(transcription+factor+EB).">Electroacupuncture ameliorates beta-amyloid pathology and cognitive impairment in Alzheimer disease via a novel mechanism involving activation of TFEB (transcription factor EB).</a> Autophagy. 17 (11), 3833-3847 (2021).</li><li>Su, X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effectiveness+and+Safety+of+Acupuncture+for+Vascular+Cognitive+Impairment:+A+Systematic+Review+and+Meta-Analysis.">Effectiveness and Safety of Acupuncture for Vascular Cognitive Impairment: A Systematic Review and Meta-Analysis.</a> Frontiers In Aging Neuroscience. 13, 692508 (2021).</li><li>Wang, Q., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficacy+of+electroacupuncture+pretreatment+for+myocardial+injury+in+patients+undergoing+percutaneous+coronary+intervention:+A+randomized+clinical+trial+with+a+2-year+follow-up.">Efficacy of electroacupuncture pretreatment for myocardial injury in patients undergoing percutaneous coronary intervention: A randomized clinical trial with a 2-year follow-up.</a> International Journal of Cardiology. 194, 28-35 (2015).</li><li>Zhang, R., Lao, L., Ren, K., Berman, B. 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電気鍼治療は、認知障害16,17、心筋損傷18、炎症性疼痛19などのさまざまな疾患に良好な治療効果があることが示されています。その作用機序を理解するために、主にラットを用いた動物実験が行われています。正確な結果を得るためには、処置中の動物の固定が重要です。多くの研究が動物の麻酔後に電気鍼治?...

鍼治療は、病気の予防と治療のために特定のツボやその他の位置にニードルリングすることを特徴とする伝統的な治療法です。電気鍼治療は、糸状鍼と電気刺激1を組み合わせ、関連するパラメータを調整することで刺激量を正確に制御することができ、鍼治療の標準化を促進します2。近年、電気鍼治療のメカニズムに関する研究は拡大・深化を続けており3,4、電気鍼治療のメカニズムに着目した研究が年々増加している5。
電気鍼治療の基礎研究では、ラットが最も一般的に使用される動物です。電気鍼治療中、レシピエントはスムーズな手順を確実にするために静止している必要があります。ヒトを対象とした臨床試験とは異なり、動物実験で使用される小動物は、15〜30分間の電気鍼治療手順中に意識的にコマンドに従うことはできません。これは、安定した信頼性の高い固定を実現するという主要な課題を研究者に提示します。不適切な固定は、動物に不安、抑うつ、恐怖などのネガティブな感情をもたらす可能性があり6、ラットの怪我のリスクや実験担当者の咬まれる可能性を高めるだけでなく、研究結果の安定性と説得力にも影響を与えます。
以前の研究では、研究者は鍼治療または電気鍼治療の手術の前に、麻酔固定7、高プラットフォーム固定8、結合固定6,9、またはラットコート固定10をしばしば使用してきました。しかし、麻酔が身体11や脳機能12に及ぼす影響や、臨床例では意識のある患者に鍼治療が行われていることを考えると、麻酔は研究成果にある程度影響を与える可能性がある。したがって、麻酔固定と比較して、覚醒状態でのラットの快適な固定はより良い選択肢です。
しかしながら、麻酔をかけていない状態では、ラット10のラットコートの被覆によるツボ曝露の制限、手術13、調整不可能な固定具サイズ14、または強制位置6におけるラットのストレス状態など、様々な固定方法にもそれぞれの限界がある.これらの技術的問題に対処するために、ラットの体のほとんどの部分で鍼治療に適したラットジャケットと固定ラックを設計しました。これは、研究者がラットの鍼治療を効率的に行うのに役立ちます。
このプロトコルの理論的根拠は、まず、ネズミのジャケットの閉じたヘッドギアが、ネズミの暗闇や穴掘りに対する自然な傾向を利用し、活動範囲を制限することです。第二に、ラットの手足の関節の近くにある縛りロープは、物理的な圧迫拘束を提供し、ラットの動きに一定の制限を課します。しかし、ネズミはジャケットを着ていてもジャンプしたり、水平に移動したりすることができます。最後に、ラットは吊り下げられた状態では一定の力点がないと地面に触れることができないため、ラットジャケットにサスペンション固定装置を追加し、ラットの動きをさらに制限し、針の保持を確保しました。固定装置の構成を 図1に示す。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/65652/65652fig01.jpg"> 図1:ラットジャケットとサスペンション固定の模式図。(A)平面図でのラットジャケットの一般的な外観。(B)ラットジャケットの3次元図。(C)頭と腰の後ろのツボ付近に切り込みが入った特製仕立てのラットジャケット。(D)高架ラックに同時に吊り下げられた複数のネズミのイラスト。1:生地の全体像。2:前肢出口;3:後肢の出口;4:ロープを結ぶ;5:縫合後に形成された空洞;6:頭のツボを露出させるための切開。7:背中と腰のツボを露出させるための切開。8:縫合位置。<a href="https://www.jove.com/files/ftp_upload/65652/65652fig01large.jpg" target="_blank">この図の拡大版をご覧になるには、ここをクリックしてください。</a>

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	<tbody>
		<tr>
			<td>75% alcohol</td>
			<td>Shandong Lircon medical technology co., ltd</td>
			<td>2002-0059</td>
			<td>Disinfection</td>
		</tr>
		<tr>
			<td>acupuncture needles</td>
			<td>Beijing Luo ya shan chuan medical equipment co., Ltd.</td>
			<td></td>
			<td>needling</td>
		</tr>
		<tr>
			<td>Adult Sprague-Dawley (SD) rats</td>
			<td>Liaoning Changsheng biotechnology co., Ltd.</td>
			<td>SCXK 2020-0001</td>
			<td>The recipient&#160; subjected to fixation</td>
		</tr>
		<tr>
			<td>an elevated fixation rack&#160;</td>
			<td>Taizhou shiai instruments and equipments for scientific and teaching research&#160; Co., ltd</td>
			<td>/</td>
			<td>Fix rats</td>
		</tr>
		<tr>
			<td>Hans electronic acupuncture apparatus</td>
			<td>Beijing Huayun Ante Science and Technology Co. Ltd.&#160;</td>
			<td>HANS-200A</td>
			<td>electroacupuncture</td>
		</tr>
		<tr>
			<td>Medical cotton ball</td>
			<td>Jiangsu Misawa Medical Supplies Co., Ltd.</td>
			<td>20172142357</td>
			<td>Disinfection</td>
		</tr>
		<tr>
			<td>Pet electric clipper</td>
			<td>Fenghua Xikou Lubao Knives &#38; Scissors Electrical Appliance Factory</td>
			<td>/</td>
			<td>Shave the hair of rats</td>
		</tr>
		<tr>
			<td>Tweezers</td>
			<td>Qingdao Schultz Biotechnology Co., ltd</td>
			<td></td>
			<td>Pinch cotton balls</td>
		</tr>
	</tbody>
</table>

improved suspension fixation using rat jacket in rat acupuncture experiments

基礎的な実験鍼灸研究では、実験動物としてラットが一般的に用いられます。しかし、姿勢を固定することは困難です。電気鍼治療では、ラットの適切な固定が不可欠です。現在、ラット固定には、麻酔固定、高台固定、結合固定、自作ラットコートによる固定など、さまざまな方法が使用されています。しかし、これらの方法には限界があり、実験の効率や操作性にある程度影響する可能性があります。このプロトコルは、ラットの服を使用してラットを吊り下げて固定する方法を紹介します。まず、ネズミは体型に合ったネズミのジャケットを着せ、暗闇や穴を掘ることを好みます。ニードリング操作は、ラットがラットの服を着た後に実行できます。吊り下げられたとき、ネズミは手足を動かすことができないため、比較的静止しています。この固定方法は、経済的でユーザーフレンドリーな利点を提供するだけでなく、ラットを快適にリラックスした位置で安定して確実に固定することができます。また、時間消費、実験スペース、人的資源を効果的に最小限に抑えます。さらに、この方法では、ラットの鍼治療に使用されるほとんどの経穴の露出が可能になります。この記事では、主にデバイスの構成に焦点を当て、特別に設計されたラットジャケット、高架固定ラック、およびそれらの接続構造を網羅します。さらに、ラットの電気鍼治療手順におけるラットの衣服ベースの懸濁液固定法の適用を示すために、実例が提示されます。

Acupuncture is a traditional therapy characterized by needling at specific acupoints or other positions to prevent and treat diseases. Electroacupuncture combines filiform needles with electrical stimulation1, which allows for precise control over the amount of stimulation by adjusting relevant parameters, promoting the standardization of acupuncture treatment2. In recent years, research on the mechanisms of electroacupuncture has continued to expand and deepen3,4, which has led to an annual increase in studies focusing on the mechanisms of electroacupuncture5.
In basic research on electroacupuncture, rats are the most commonly used animals. During electroacupuncture treatment, the recipient must remain still to ensure a smooth procedure. Unlike in clinical trials with human subjects, small animals used in animal experiments cannot follow commands consciously during 15-30 min lengthy electroacupuncture procedures. This presents researchers with a primary challenge of achieving stable and reliable fixation. Improper fixation can lead to negative emotions such as anxiety, depression, and fear in animals6, which not only increases the risk of injury for rats and the chance of being bitten for the experimental personnel but also affects the stability and persuasiveness of research results.
In previous studies, researchers have often used anesthesia fixation7, high platform fixation8, binding fixation6,9, or rat coat fixation10 before acupuncture or electroacupuncture operations. However, given the effects of anesthesia on the body11 and brain function12, and considering that acupuncture is performed on conscious patients in clinical cases, anesthesia may impact research results to a certain extent. Therefore, compared with anesthesia fixation, comfortable fixation of rats in an awake state is a better option.
However, in the non-anesthetized state, various fixation methods also possess their respective limitations, including restricted acupoint exposure due to coverage of the rat coat on rats10, the necessity of at least two individuals to perform the operation13, non-adjustable fixator size14, or the stressed state of rats in a forced position6. To address these technical issues, we designed a rat jacket and a fixation rack suitable for acupuncture manipulation in most parts of the rat body. This can help researchers efficiently carry out acupuncture procedures with rats.
The rationale for this protocol is as follows: firstly, the closed headgear on the rat&#39;s jacket takes advantage of the rats&#39; natural inclination for darkness and burrowing, thus restricting their range of activities. Secondly, the binding ropes near the joints of the rats&#39; limbs provide physical compression restraint, imposing certain limitations on their movements. However, rats are still able to jump and move horizontally even while wearing the jacket. Lastly, since rats cannot touch the ground without a fixed force point when suspended, we added a suspension fixation device to the rat jacket, further restricting their movements and ensuring needle retention. The composition of the fixation device is shown in Figure 1.
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/65652/65652fig01.jpg" /> 
Figure 1: Schematic diagram of the rat jacket and suspension fixation. (A) The general appearance of the rat jacket in a flat view. (B) Three-dimensional view of the rat jacket. (C) Specially tailored rat jacket with an incision near the acupoints on the head and back waist. (D) Illustration of multiple rats simultaneously suspended on the elevated rack. 1: Fabric overall view; 2: Forelimb outlet; 3: Hind limb outlet; 4: Tying rope; 5: Cavity formed after suturing; 6: Incision for exposing head acupoints; 7: Incision for exposing back and waist acupoints. 8: Suture position. <a href="https://www.jove.com/files/ftp_upload/65652/65652fig01large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>

著者スポットライト:電気鍼治療中のラットの固定を改善するための効率的なラットの服 

ラット鍼治療実験におけるラットジャケットを用いた懸濁液固定の改善

We developed an improved method for suspending and fixing rats with rat clothes in an acupuncture experiment. This protocol aims to provide detailed descriptions of its procedural steps, including important precautions, offering a reference for researchers conducting fundamental experiments in the field of acupuncture and moxibustion. In previously reported studies, researchers have often used anesthesia fixation, high platform fixation, binding fixation, or various other self-made fixators while performing acupuncture or electroacupuncture operations on rats.Given the side effects of anesthesia on the body and the real clinical conditions, fixation in an awakened stage offers the better alternative. However, in the non-anesthetized stage, the mentioned fixations also possess limitations such as restricted acupoint exposure, requirement of multiple labels, non-adjustable fixator size, or forced posture. This method utilizes the rat's preference for burrowing to create rat jackets that fit their body shape.It offers several advantages over other techniques. It is cost-effective, user-friendly, manpower-and space-saving, accommodates rats of varying weights, and can also be used for needle operation on most acupoints. In subsequent studies, we will focus on investigating the impact of the restraints and suspensions of rat clothing on physiological indicators and behavioral aspects related to emotional cognition in rats.This will enable us to quantitatively assess its potential influence on research and refine the fixation method.

本実験では、合計12匹のラットを電気鍼治療用に選別し、固定法下での反応を観察した( 表2参照)。12匹のラットのうち、15分間の電気鍼保持期間中に激しい闘争を示したのは1匹のみであった。その激しい動きの結果、針は途中で外れ、ネズミの総数の8.33%を占めました。この結果は、一般的に使用される固定法が顕著な安定性と信頼性を示すことを示唆しています。1匹のラットが...

Watch this Scientific Journal Video about Efficient Rat Clothing for Improved Rat Fixation During Electroacupuncture at JoVE.com

ここでは、鍼治療実験中にラットを効果的に固定するために、ラットジャケットを用いた改良された懸濁液固定法を紹介します。さらに、この固定装置の構造と用途、ならびに電気鍼治療におけるその利用のための指示と評価が解明される。

In basic experimental acupuncture research, rats are commonly used as laboratory animals. However, it is difficult for them to maintain a fixed posture. ...

鍼灸実験において、ラットをラットの服を着せて吊るし固定する方法を改良した。このプロトコルは、重要な予防措置を含む手順の詳細な説明を提供することを目的としており、鍼治療および灸の分野で基礎実験を行う研究者の参考資料を提供します。以前に報告された研究では、研究者は、ラットに鍼治療または電気鍼治療の手術を行う際に、麻酔固定、高プラットフォーム固定、結合固定、またはその他のさまざまな自作固定具を使用することがよくあります。麻酔が身体に及ぼす副作用と実際の臨床状態を考えると、覚醒段階での固定はより良い選択肢を提供します。ただし、麻酔をかけていない段階では、前述の固定には、ツボへの露出の制限、複数のラベルの要件、調整不可能な固定具のサイズ、または強制的な姿勢などの制限もあります。この方法は、ネズミの穴掘りの好みを利用して、ネズミの体型に合ったネズミのジャケットを作成します。他の手法に比べていくつかの利点があります。費用対効果が高く、ユーザーフレンドリーで、人手とスペースを節約し、さまざまな体重のラットに対応し、ほとんどのツボの針操作にも使用できます。今後の研究では、ラットの衣服の拘束と停止が、ラットの情動認知に関連する生理学的指標と行動的側面に与える影響の調査に焦点を当てます。これにより、研究への影響を定量的に評価し、固定法の改良を図ることができます。

improved-suspension-fixation-using-rat-jacket-rat-acupuncture

Research

JoVE Journal

Medicine

Improved Suspension Fixation Using Rat Jacket in Rat Acupuncture Experiments

In basic experimental acupuncture research, rats are commonly used as laboratory animals. However, it is difficult for them to maintain a fixed posture. During electroacupuncture procedures, proper immobilization of rats is essential. Various methods of rat fixation are currently used, including anesthesia fixation, high-platform fixation, binding fixation, and fixation with a self-made rat coat. However, these methods have their limitations, which may affect the efficiency and operability of the experiment to some extent. This protocol introduces a method of suspending and fixing rats using rat clothes. Firstly, rats are clothed with rat jackets that match their body shape, taking advantage of their preference for darkness and burrowing. The needling operation can then be carried out after the rats have worn rat clothes. When suspended, the rats are relatively still, as their limbs cannot move. This fixation method offers not only economical and user-friendly benefits but also ensures a stable and reliable fixation of the rats in a comfortably relaxed position. It also effectively minimizes time consumption, experimental space, and manpower resources. Additionally, this method allows for the exposure of most acupoints used for acupuncture in rats. This article primarily concentrates on the device's composition, encompassing a specially designed rat jacket, an elevated fixation rack, and their connecting structures. Additionally, an illustrative example will be presented to demonstrate the application of the rat clothing-based suspension fixation method in rat electroacupuncture procedures.

Here, we present an improved suspension fixation method using a rat jacket to effectively immobilize rats during acupuncture experiments. Furthermore, the structure and application of this fixation apparatus, as well as instructions and evaluation for its utilization in electroacupuncture therapy, is elucidated.

医学

Securing the Rat in the Rat Jacket for Electroacupuncture

Wear a pair of gloves before proceeding to fit and secure the rat in the custom made rat jacket. Then using one hand hold the rat jacket's middle and back portion on the table. Using the other hand, gently hold the rat by its tail to pick it up from the feeding cage and place it into the jacket as the rat moves forward into the jacket and its head reaches the headgear point.Using both hands, carefully pull out its left and right upper limbs from the holes in the jacket. Securely wrap the tying ropes from the opening end of the jacket around the rat's trunk, and tie them tightly at the opposite end to adequately restrain its upper limbs. Tie the rope loose enough to prevent hypoxia or local limb ischemia, and tight enough to prevent the rat's escape.In a similar manner, secure its lower limbs.

電気鍼治療のためのラットジャケットにラットを固定する

Performing Electroacupuncture on Rat Immobilized Using Rat Jacket

Before beginning the procedure of electropuncture on the rat, dress the rat in the custom made rat jacket. Then disinfect the local skin of acupoints using alcohol swabs. After locating the acupoint accurately, insert the sterile disposable acupuncture needle into the spot at the appropriate depth.Gently suspend the animal from the horizontal iron rod with the help of the S-shaped hooks attached to the rat jacket. Wrap the electrode wire around the horizontal iron rod before securely attaching the small clip at the end of the electrode wire to the needle handle. Then after switching on the power, select the appropriate frequency, current, and time.Once the parameters are adjusted, commence the electroacupuncture treatment process. Using the S-shaped hooks, suspend the rats in the same group together on the iron rod for simultaneous electroacupuncture. When the treatment is over, remove the acupuncture needle along with the clamp from the animal.Then carefully take the needle out of the clamp and put it in the box. After removing the animal from the suspension hooks, loosen the straps, lift the head cover of the rat's jacket and position the rat above the cage. The rat will then be free to release itself from the jacket and enter the cage independently.While performing a 15 minute electroacupuncture experiment on 12 rats only one rat exhibited intense struggle during the needle retention period resulting in needle drop. While other rats maintained proper needle fixation throughout the treatment. When compared to the other fixation methods reported in the literature, the demonstrated method exhibited the combined benefits of stability and secure fixation, efficiency in terms of time and number of personnel required, affordability, as well as relative comfort for the rats.Additionally, in terms of the exposed acupoint range, the specially designed rat jackets in this protocol met the requirements for exposing most acupoints on the body.