Name: chronic post-ischemia pain model for complex regional pain syndrome type-i in rats
Uploaded: 2020-01-21T14:30:00
Description: Watch this Scientific Journal Video about Chronic Post-Ischemia Pain Model for Complex Regional Pain Syndrome Type-I in Rats at JoVE.com

该项目由中国国家自然科学基金（81873365和81603676）、浙江省杰出青年自然科学基金（LR17H270001）和浙江中医药大学（）资助。Q2019J01， 2018ZY37， 2018ZY19）.

动物规程经浙江医科大学动物伦理委员会批准。
1. 动物
<ol><li>从上海实验室动物中心获得雄性斯普拉格-道利（SD）大鼠（280~320克，8-10周龄）。在浙江中医药大学动物实验室动物中心饲养动物。请注意，繁殖条件应包括 12 h/ 2h 的光/暗周期，并保持温度稳定在 24°C。提供水和食物。请注意，本研究共使用了48只大鼠。有一点，在这个模型中，我们需要观察整个过程中的...

<ol>
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该协议描述了通过在大鼠后肢应用缺血/再灌注来建立大鼠CPIP模型的详细方法。它涉及评估后肢外观，水肿，机械/热过敏性，和急性角质行为，以回应辣椒素注射。
肢体缺血/再灌注是导致CRPS-I在人类患者12的常见因素。该协议描述了如何建立大鼠CPIP模型，这是一个常用的动物模型来概括人类CRPS-I6。在麻醉下，在大鼠的后肢诱导了缺血，在...

复杂的区域疼痛综合征 （CRPS） 引起骨折、创伤、手术、缺血或神经损伤1、2、3引起的复杂和慢性疼痛症状。CRPS分为2个子类别：CRPS类型I和II型（CRPS-I和CRPS-II）4。流行病学研究表明，CRPS的流行率约为1：20005。CRPS-I，显示没有明显的神经损伤，可能导致慢性疼痛，并极大地影响患者的生活质量。目前可用的治疗方法显示治疗效果不足。因此，CRPS-I 仍然是一个需要解决的重要且具有挑战性的临床问题。
建立模仿CRPS-I的临床前动物模型对于探索CRPS-I背后的机制至关重要。为了解决这个问题，Coderre等人设计了一个大鼠模型，将长期缺血和再灌注到后肢，以重述CRPS-I6。据了解，缺血/再灌注损伤是CRPS-I7的主要原因之一。大鼠CPIP模型表现出许多CRPS-I样的症状，包括后肢水肿和高血症在模型建立后的早期阶段，其次是持续热和机械高灵敏度6。在此模型的帮助下，建议中央疼痛敏化、外周TRPA1通道激活和活性氧物种生成等对CRPS-I8、9、10作出贡献。我们最近成功地建立了CPIP大鼠模型，并进行了背根结核（DRGs）的RNA测序，内骨对受影响的后爪11进行内脏测序。我们发现了一些潜在的机制，可能参与调解CRPS-I11的疼痛过度敏感。我们进一步确定DRG神经元中的瞬态受体电位万能1（TRPV1）通道是CRPS-I12的机械和热超灵敏度的重要原因。
在这项研究中，我们描述了建立CPIP的老鼠模型所涉及的详细程序。通过测量机械和热超敏感性及其对急性辣椒素挑战的反应能力，我们进一步评估了大鼠CPIP模型。提出大鼠CPIP模型是进一步研究CRPS-I中相关机制的可靠动物模型。

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	<tbody>
		<tr>
			<td>1.5 ml Eppendorf tube</td>
			<td>Eppendorf</td>
			<td>22431021</td>
			<td></td>
		</tr>
		<tr>
			<td>DMSO</td>
			<td>Sigma-Aldrich</td>
			<td>D1435</td>
			<td></td>
		</tr>
		<tr>
			<td>Capsaicin</td>
			<td>APEXBIO</td>
			<td>A3278</td>
			<td></td>
		</tr>
		<tr>
			<td>Digital caliper</td>
			<td>Meinaite</td>
			<td>NA</td>
			<td></td>
		</tr>
		<tr>
			<td>O-ring</td>
			<td>O-Rings West</td>
			<td>Nitrile 70 Durometer</td>
			<td>7/32 in. 
			internal diameter</td>
		</tr>
		<tr>
			<td>Plantar Test Apparatus</td>
			<td>UGO Basile, Italy</td>
			<td>37370</td>
			<td></td>
		</tr>
		<tr>
			<td>von Frey filaments</td>
			<td>UGO Basile, Italy</td>
			<td>NC12775</td>
			<td></td>
		</tr>
	</tbody>
</table>

chronic post-ischemia pain model for complex regional pain syndrome type-i in rats

复杂的区域疼痛综合征I型（CRPS-I）是一种神经系统疾病，在患者中引起剧烈疼痛，并且仍然是一个未解决的医疗状况。然而，CRPS-I的基本机制尚未透露。众所周知，缺血/再灌注是导致CRPS-I的主要因素之一。通过长期缺血和后肢再灌注，建立了大鼠慢性缺血后疼痛（CPIP）模型，以模仿CRPS-I。CPIP模型已成为研究CRPS-I机制的公认动物模型。该协议描述了建立CPIP大鼠模型所涉及的详细程序，包括麻醉，然后是后肢缺血/再灌注。通过测量后肢的机械和热超敏感性以及急性辣椒素注射的角质反应，进一步评价大鼠CPIP模型的特征。大鼠CPIP模型表现出几种CRPS-I样的表现，包括建立后早期后肢水肿和高血症，持续热和机械过敏，以及急性辣椒素注射增加的角质反应。这些特性使其成为进一步研究CRPS-I中涉及的机制的合适动物模型。

在脚踝上放置O环后，叶侧后爪皮肤出现青紫，指示组织缺氧（图1A）。切割O环后，叶侧后爪开始充满血液，并表现出强健的肿胀，这显示了高血症的强烈迹象（图1A）。爪子肿胀逐渐减少，并在缺血/再灌注程序后恢复正常的48小时（与Sidak的双向ANOVA，即位测试，图1B）。所有这些迹象都符合?...

Watch this Scientific Journal Video about Chronic Post-Ischemia Pain Model for Complex Regional Pain Syndrome Type-I in Rats at JoVE.com

Chronic Post-Ischemia Pain Model for Complex Regional Pain Syndrome Type-I in Rats (Video) | JoVE

此处提供了一个协议，详细说明了建立慢性缺血后疼痛（CPIP）动物模型的步骤。这是一个公认的模型，模仿人类复杂的区域疼痛综合征I型。进一步评价机械和热超敏感性，以及在CPIP大鼠模型中观察到的辣椒素引起的角感行为。

大鼠复杂区域疼痛综合征I型慢性缺血后疼痛模型

Complex regional pain syndrome type-I (CRPS-I) is a neurological disease that causes severe pain among patients and remains an unresolved medical ...

CRPS 背后的机制我仍然不清楚。建立一个临床前动物模型模仿CRPS I将是探索CRPS I的基础机制的关键。大鼠CPIP模型表现出许多CRPS我喜欢的simit，其中包括后肢水肿和高血症在早期阶段。模型建立后，然后是持续的热和机械超敏。首先用苯巴比他钠麻醉所有大鼠。用钳子捏住它们的后爪或尾巴，确保大鼠正确麻醉。把兽医软膏放在眼睛上在手术过程中，将大鼠放在37摄氏度的加热垫上。用甘油润滑右后爪和脚踝。然后将一个硝酸盐 70 测高仪 O 环滑入一点五毫升管的较大一侧，并切断卡扣盖。小心地将后爪插入空心管，直到其到达底部。逐渐将 O 环从管子滑到脚踝关节附近的右后肢。在那里离开三个小时对假老鼠群体应用同样的治疗方法，但不应诱发缺血的破O环除外。三个小时后，切断O环，监测大鼠，直到它恢复足够的意识，以保持胸骨的恢复。在老鼠完全从麻醉中恢复之前，不要将老鼠放在其他大鼠的陪伴下。将大鼠放在网状地板上的透明有机玻璃室中，并在任何行为测试前将大鼠与环境进行30分钟的习惯。使用冯弗雷灯丝测试大鼠的机械性肌酸。从中间灯丝开始测试，垂直地将其应用到后爪的中间植物表面。施加适当的力使灯丝弯曲长达五秒钟。爪子的突然缩回被认为是一种不费时的行为。第三天、第二天、第一天和每隔一天进行一次机械性阿洛迪尼亚试验，直到第13天。应用向上向下测试方法测试阈值，并采用迪克森方法计算 50% 的爪子提取阈值。要测试热超高血症，请使用哈格里夫斯方法，将50瓦灯泡发出的光束直接对瞄准后爪。测量爪子退出延迟，将 20 秒设置为截止阈值，以避免过度伤害。每只后爪在五分钟的间隔内重复每次测试三次，并以平均值作为爪子的退出延迟。在天三、第二天、第一天和每隔一天进行热超大病测试，直到第 13 天。为了测试辣椒素诱发急性中毒行为，用DMSO准备200毫摩尔辣椒素储存溶液，并在PBS中进一步稀释1至1000，用于注射。辣椒素是一种试剂，可引起皮肤、眼睛和呼吸的关节刺激。在准备辣椒素溶液时，戴上口罩和护目镜很重要。然后用连接到一毫升注射器的30针将50微升辣椒素或车辆注射到后爪中。在注射后 10 分钟内，使用摄像机记录注射爪子的舔、咬或退缩等无行为。通过测量模型建立后 15 分钟、24 小时、48 小时和 72 小时的爪子直径增加，评估后爪水肿。将 O 环放在脚踝上后，前爪皮肤出现青紫，这是组织缺氧的迹象。切割O环后，爪子开始充满血液，并表现出强烈的肿胀，表明强烈的高血症。肿胀逐渐减少，并在手术后48小时恢复正常。模型建立一天后，CPIP组的单体后爪表现出明显的机械性，持续了13天的观察时间框架。CPIP组的反向后爪也显示机械超高血症13天。在测量热高血痛时，CPIP大鼠的双边后爪与假组大鼠相比，对有毒热刺激的反应明显降低了戒断延迟。前爪的热超高热量一直持续到观察时间框架结束，而反面后爪的热超高热量持续七天。假和CPIP组被测试与辣椒素注射的无性行为。当车辆被注射时，假组表现出轻微的明显反应，CPIP组表现出明显较高的反应。更重要的是，CPIP大鼠对辣椒素注射的反应明显高于假组，这表明CPIP大鼠表现出与患有CRPS的人类患者相似的现象I.By这种程序，在模型建立之前和期间对大鼠进行麻醉。由缺血和人类CRPS建立的本协议中描述的CPIP大鼠模型我喜欢模拟。这些模型的家庭，更多的机制关于CRPS我可以探讨。

Research

JoVE Journal

Behavior

Chronic Post-Ischemia Pain Model for Complex Regional Pain Syndrome Type-I in Rats

Use of the Operant Orofacial Pain Assessment Device (OPAD) to Measure Changes in Nociceptive Behavior

Use of the Operant Orofacial Pain Assessment Device OPAD to Measure Changes in Nociceptive Behavior (Video) | JoVE

使用操作性口面疼痛评估设备（OPAD）测量伤害行为的变化

We present an operant  system for the detection of pain in awake, conscious rodents. The Orofacial  Pain Assessment Device (OPAD) assesses pain behaviors ...

科研

行为学

Determining Pain Detection and Tolerance Thresholds Using an Integrated, Multi-Modal Pain Task Battery

Determining Pain Detection and Tolerance Thresholds Using an Integrated, Multi-Modal Pain Task Battery (Video) | JoVE

确定疼痛的检测和容忍阈值使用集成的，多模式疼痛任务电池

Human pain models are useful in the assessing the analgesic effect of drugs, providing information about a drug&#39;s pharmacology and identify ...

A Protocol of Manual Tests to Measure Sensation and Pain in Humans

A Protocol of Manual Tests to Measure Sensation and Pain in Humans (Video) | JoVE

手动测试的协议来衡量感觉和疼痛人类

Numerous qualitative and quantitative techniques can be used to test sensory nerves and pain in both research and clinical settings. The current study ...

Psychophysically-anchored, Robust Thresholding in Studying Pain-related Lateralization of Oscillatory Prestimulus Activity

Psychophysically-anchored, Robust Thresholding in Studying Pain-related Lateralization of Oscillatory Prestimulus Activity (Video) | JoVE

在研究疼痛相关振荡Prestimulus活动偏侧Psychophysically锚，乐百氏阈值

In perceptual studies, it is often important to objectively assess the equality of delivered stimulation across participants or to quantify the ...

Treating Low Back Pain in Failed Back Surgery Patients with Multicolumn-lead Spinal Cord Stimulation

Treating Low Back Pain in Failed Back Surgery Patients with Multicolumn-lead Spinal Cord Stimulation (Video) | JoVE

多列铅脊髓刺激术后腰痛的治疗

Failed back surgery syndrome (FBSS) refers to persistent, chronic pain following spinal surgery. Spinal cord stimulation with dorsal epidural leads can be ...

Simultaneous Recordings of Cortical Local Field Potentials and Electrocorticograms in Response to Nociceptive Laser Stimuli from Freely Moving Rats

Simultaneous Recordings of Cortical Local Field Potentials and Electrocorticograms in Response to Nociceptive Laser Stimuli from Freely Moving Rats (Video) | JoVE

在自由运动大鼠的反应中, 皮质局部电位和电皮质图谱的同时记录

Electrocortical responses, elicited by laser heat pulses that selectively activate nociceptive free nerve endings, are widely used in many animal and ...

Multi-Modal Signals for Analyzing Pain Responses to Thermal and Electrical Stimuli

Multi-Modal Signals for Analyzing Pain Responses to Thermal and Electrical Stimuli (Video) | JoVE

分析疼痛对热和电刺激反应的多模态信号

The assessment of pain relies mostly on methods that require a person to communicate. However, for people with cognitive and verbal impairments, existing ...

Investigating Pain-Related Avoidance Behavior using a Robotic Arm-Reaching Paradigm

Investigating Pain-Related Avoidance Behavior using a Robotic Arm-Reaching Paradigm (Video) | JoVE

使用机械臂接触范式调查疼痛相关避免行为

Avoidance behavior is a key contributor to the transition from acute pain to chronic pain disability. Yet, there has been a lack of ecologically valid ...

A Complex Diving-For-Food Task to Investigate Social Organization and Interactions in Rats

A Complex Diving-For-Food Task to Investigate Social Organization and Interactions in Rats (Video) | JoVE

调查大鼠社会组织和互动的复杂潜水换食物任务

For many species, where status is a vital motivator that can affect health, social hierarchies influence behavior. Social hierarchies that include ...

Animal Models of Depression - Chronic Despair Model (CDM)

Animal Models of Depression - Chronic Despair Model CDM (Video) | JoVE

抑郁症的动物模型 - 慢性绝望模型（CDM）

Major depressive disorder is one of the most prevalent forms of mental illnesses and causes tremendous individual suffering and socioeconomic burden. ...