我们感谢中国内蒙古医科大学蒙古医学院提供的仪器和实验室，本研究得到了必要的仪器和实验室设施的支持。本研究获得国家自然科学基金（Grant No. 81760762）和内蒙古医科大学项目（Grant No.YKD2022MS074）和内蒙古自治区高等教育科研项目（批准号）NJZY22661）内蒙古自治区中蒙医学重点实验室开放基金项目（批准号）MYX2023-K07）。

实验方案获得了内蒙古医科大学（YKD202301172）动物实验护理伦理委员会的批准，并遵守了美国国立卫生研究院的动物护理和伦理指南。我们动物中心的许可证号是NO.110324230102364187。获得 24 只雄性 Sprague-Dawley （SD） 大鼠，每只 8 周龄 （200 g ± 20 g），并将其饲养在温度为 22°C ± 2°C，湿度为 55% ± 15% 的受控环境中。用啮齿动物维持饲料喂养大鼠，用玉米芯喂养饲料和纯净水作为垫料。在实验前，将大鼠置于12小时/ 12小时的光/暗循环中1周。
1. CUMS大鼠模型的建立
<ol><li>分组<ol><li>将 24 只大鼠随机分为 2 组:对照 （CON） 组和模型 （MOD） 组，前者不会暴露于隔离或压力下。每组包含12只大鼠。</li>
		<li>将大鼠饲养在尺寸为55 cm x 40 cm x 20 cm的标准笼中，每个笼子6只大鼠。除非另有说明，否则在整个适应期间保留笼子分配。</li>
		<li>用新鲜的垫料填充每个育种笼，每周更换两次。</li>
		<li>进行为期 1 周的适应期。让大鼠不受限制地获取食物和水，除非在CUMS压力源应用期间。除非另有说明，否则保持恒定的环境，温度为 22°C ± 2°C，湿度为 55% ± 15%，从 08:00 到 20:00 为 12 小时的光/暗循环。</li>
		<li>在开始实验之前，每天处理大鼠，使它们适应研究人员，并尽量减少实验阶段的额外压力。</li>
	</ol></li>
	<li>与慢性不可预测的轻度应激隔离<ol><li>同时将 MOD 和 CON 组放置在不同的房间中。将MOD组大鼠单独饲养，同时将CON组大鼠保持在一起。保持所有其他条件不变。</li>
		<li>实施 28 天的压力源方案25.为了防止习惯化并确保压力源的不可预测性，每天随机管理一个压力源，避免连续几天使用相同的压力源。</li>
		<li>在不同的日子随机应用以下九个压力源26,27 之一:24 小时缺水、24 小时食物剥夺、湿填充物、笼子倾斜、明暗循环反转、4 °C 的冷暴露、45 °C 的热暴露、1 分钟的尾部夹紧或 15 分钟的 160 rpm 摇晃。具体设计见表1。</li>
		<li>在施加压力源期间，限制 MOD 组获得食物和水，直到压力结束，除非在明暗循环反转期间。CON组不需要限制水和饮食。</li>
	</ol></li>
	<li>压力方法<ol><li>通过对所有大鼠（不包括对照组）应用抑郁刺激并结合 28 天隔离来启动实验。将这些老鼠饲养在单独的笼子中。请参阅 表 2 了解与抑郁刺激相关的情况。</li>
		<li>为了执行尾夹方法，通过用标准回形针夹住尾巴，在距离大鼠尾巴根部1-2厘米的距离处，将大鼠的尾巴从MOD组中固定出来。测量 1 分钟的夹紧时间 （n = 12）。</li>
		<li>在禁水方法中，通过取出MOD组大鼠的水瓶来扣留水，并记录24小时。 注意:记录了缺水开始的时间，以便可以准确计算结束时间;观察大鼠在此期间的行为，包括活动、食欲和精神状态。</li>
		<li>在食物剥夺方法中，从MOD组大鼠中扣留食物并记录24小时。 注意:记录食物剥夺开始的时间，以便准确计算结束时间;确保大鼠在此期间获得足够的水分;并观察大鼠在此期间的行为，包括活动和精神状态等。</li>
		<li>对于4°C的冷刺激，将MOD组的大鼠置于一桶冷水中并记录5分钟。确保水温在整个测试过程中保持一致。在实验结束时，用鼓风机干燥大鼠并将其放回原来的笼子。 注意: 使用温度计和冰块将冷水温度保持在 4 °C，以在水温上升时调节水温。应定期更换水质，保证水质清洁，温度恒定。游泳时，除头部外，大鼠的所有肢体和躯干都应浸入冷水中。水的深度必须大于老鼠身体的长度，以防止老鼠因接触桶底而跳出水面。</li>
		<li>为了施行45°C热应激，将MOD组大鼠置于培养箱中并记录5分钟，确保温度在整个测试过程中保持稳定。</li>
		<li>在明暗循环的反转中，将笼子包裹在黑布中1小时，以模拟白天的黑暗。随后，在夜间照亮笼子12小时以模仿白天。记录大鼠的行为，食物和水的摄入量以及24小时的睡眠模式。</li>
		<li>对于湿填充物实验，将 200 mL 水引入含有 100 g 填充物的笼子中。将MOD组大鼠饲养在湿笼中，并记录大鼠在湿垫料压力下的行为，包括活动、食欲、饮水量等。观察是否有任何异常行为或不适反应，如大鼠的皮肤和毛发状况，这可能是由湿填充物引起的，并及时记录下来，以便后续分析24 h。测试后，用鼓风机干燥大鼠，然后将其放回装有新鲜木屑的笼子中。</li>
		<li>在笼子倾斜方法中，将MOD组大鼠置于以45°角倾斜的笼子中，靠墙并记录24小时。使用笼架结构调整角度并将笼子固定到位。 注:计算从开始到结束的时间，观察大鼠在倾斜笼子期间的行为，包括活动、食欲和精神状态，并确保倾斜笼子的角度设置正确并保持稳定，以确保实验的准确性和可重复性。</li>
		<li>对于高速振荡，将MOD组大鼠置于设置为160rpm的机械振荡器中，并记录大鼠15分钟。随后使用行为测试方法来评估模型的成功建立。</li>
		<li>施加压力源后，将 MOD 组笼从 CUMS 室移回饲养室。在为期 4 周的压力暴露期间，将 CON 组保持在位于住房房间的家庭笼子中。</li>
	</ol></li>
	<li>实验过程中的注意事项<ol><li>在CUMS室中施加压力源后，将MOD组的笼子转移回普通住房室。</li>
		<li>CUMS建模期间的动物监测<ol><li>在夹尾过程中，由于诱导刺激，动物可能会挣扎。在此期间，持续监控夹具。如果它脱落，暂停计时器，重新应用夹子，然后恢复计时器 1 分钟。</li>
			<li>不要同时施加缺水和湿床褥压力源。 注意:避免同时施加湿填充物和缺水有助于保持实验的完整性，减少混杂变量，并促进动物福利。</li>
			<li>在冷水游泳期间，动物的体温和环境室温可能会升高水温。因此，通过添加冰水或冰块进行调整，以保持恒定的水温。</li>
			<li>在施加压力源期间，以30分钟的间隔观察大鼠，昼夜反转期间除外。特别注意异常痛苦的迹象，例如颤抖、嗜睡或缺乏运动。如果观察到这些症状 - 特别是在4°C，冷水，游泳和潮湿的床上用品期间可能出现的体温过低 - 立即将大鼠从压力源中移开。 注意:当实验动物有健康问题时，如感染、严重创伤、攻击性行为、行动不便等，将实验动物从研究中移除的条件通常围绕着保护它们的健康和安全，同时确保实验结果的完整性。</li>
			<li>每天检查每只大鼠的伤口或其他身体或行为异常。如果观察到任何异常，请咨询实验室兽医，以确定是否应将大鼠排除在实验之外。</li>
			<li>每3天称重每只大鼠一次。如果动物失去超过其饲喂前基础体重的 20%，则必须将其排除在实验之外。</li>
		</ol></li>
	</ol></li>
</ol>2. 行为测试
<ol><li>首先，对所有大鼠（不包括对照组）进行抑郁刺激并隔离28天。将老鼠饲养在单独的笼子中。有关抑郁刺激条件的详细信息，请参阅 表 2 。</li>
	<li>对于旷场测试，将黑匣子分成 25 个面积相等的正方形部分。在箱内安装视频跟踪分析系统。将大鼠置于中央广场，并监测其水平和垂直活动5分钟。 注意: 盒子的尺寸为 500 mm x 500 mm x 300 mm。将使用视频跟踪系统收集活动数据，以评估啮齿动物在暴露于新环境中时与焦虑相关的行为。</li>
	<li>随后，使用所有爪子计算大鼠穿过的方格数，以量化水平活动。将站立和梳理的实例计为垂直活动的指标。每次测试后，使用 75% 的酒精对盒子进行消毒，以消除任何残留的老鼠气味，以便进行后续测试。</li>
	<li>接下来，通过蔗糖偏好测试评估快感缺乏。将两个瓶子放在笼盖上:瓶子 A 含有纯水，而瓶子 B 含有 1% 的蔗糖溶液。允许 随意 使用大鼠的两种解决方案。在食用前后称重瓶子，以计算第 0、7、14、21 和 28 天的 60 分钟蔗糖偏好率。计算公式如下: 蔗糖消耗量 = <img alt="Equation 1" src="/files/ftp_upload/65889/65889eq01.jpg" style="margin: auto;"> × 100%</li>
	<li>要测量空间记忆和学习能力，请采用莫里斯水迷宫测试。将池划分为四个象限，从 1 到 4 进行编号。在第三象限的水面以下 1 厘米处放置一个浸没式休息平台。</li>
	<li>将牛奶引入池中以增加水的不透明度，并在整个实验过程中保持约23°C的水温。</li>
	<li>将每只老鼠放在迷宫的各个象限中，让它们 120 秒找到隐藏的平台。老鼠必须依靠空间记忆和学习技能来记住平台的位置。一旦他们了解了平台的位置，他们就可以直接游到平台上。使用莫里斯水迷宫视频跟踪系统记录延迟时间。</li>
	<li>将大鼠放置在池中的固定位置。如果被摄体在 120 秒内未能找到隐藏的平台，则将延迟记录为 120 秒。</li>
	<li>最后，移除隐藏的平台，将老鼠放回水中，并记录 120 秒内的区域交叉次数。</li>
</ol>3. 统计分析
<ol><li>为了评估生化参数的显着差异，采用单因素方差分析 （ANOVA），然后进行 Duncan 事后检验。将数据显示为均值±标准误 （SE），并将小于 0.05 的 p 值视为具有统计显著性。</li>
</ol>

建立和验证慢性不可预测的轻度应激大鼠模型

作者没有要披露的利益冲突。

抑郁症是一种精神障碍，其特征是情绪低落、缺乏快乐和精力减少等症状30.在抑郁症研究领域，建立可靠的动物模型对于推进治疗干预至关重要。在各种动物模型中，CUMS模型因其高信度、效度以及与人类抑郁症31特征的一致性而特别值得关注。它非常适合在各种环境中模拟低水平压力源在长时间内的累积效应。在这项研究中，我们采用了步骤 1.2.3 和表 2</...

重度抑郁症 （MDD） 是一种普遍存在的精神疾病，是全球第三大残疾原因，影响着超过 3 亿人1,2,3.值得注意的是，据估计，至少有一半的受影响个体没有得到充分的治疗4.鉴于这一差距，动物模型可以作为调查抑郁症病因的重要工具。迄今为止，存在 20 多种不同的抑郁症动物模型5.其中，Paul Winer 于 1987 年改进的慢性不可预测的轻度压力 （CUMS） 模型是最常用的6.CUMS 模型的前提是，将啮齿动物暴露于各种社会环境压力源会导致类似于焦虑、紧张和抑郁的症状。该方法涉及在几周内将动物暴露于各种轻度压力源中，最终导致一系列行为改变，包括快感缺乏和抑郁样行为7,8.这些变化伴随着内分泌和神经递质谱的变化，例如 5-HT 的减少9,10.这些结果与在被诊断患有MDD的人类中观察到的结果非常相似，从而验证了该模型的实用性。CUMS 模型因其在评估抗抑郁药方面的有效性而受到特别重视，表现出高水平的表面、结构和预测有效性11,12.与其他模型不同，CUMS 对长期服用单胺能抗抑郁药的影响敏感。例如，选择性血清素再摄取抑制剂 （SSRIs），如西酞普兰、帕罗西汀和氟西汀，已被证明可以预防和逆转慢性压力条件下的快感缺乏12,13.此外，新的速效抗抑郁药，如氯胺酮，也证明了这种模型的疗效14,15.相比之下，其他测试，如强迫游泳测试（FST）和尾部悬挂测试（TST）在模拟长期行为变化方面不太可靠，通常反映出对急性压力的适应，而不是持久的抑郁症状16.这些特征强调了CUMS模型在抑郁症研究中的稳健有效性。CUMS模型最显着的特征之一是快感缺乏症，该模型因其在经典研究中的高可靠性而得到认可 - 无法在日常活动中体验快乐或兴趣17,18.这种现象通常使用蔗糖偏好测试进行评估，许多抗抑郁药已被证明可以逆转蔗糖消耗减少。CUMS 文献中也常用其他几个指标，包括开场测试 （OFT），它评估自愿运动行为、探索倾向和紧张程度，从而衡量抑郁症的严重程度19.其他测试，如高架加迷宫 （EPM） 评估类似焦虑的行为，莫里斯水迷宫测试 （MWM） 检查认知功能20，FST 评估对负面情绪和行为绝望的易感性20.此外，影响人类的大多数压力源本质上都是社会性的。社会关系欠佳的个体，其特征是社会活动、网络和支持有限，患各种疾病的风险更高21,22.这也与啮齿动物有关，啮齿动物是群居的群居动物。例如，隔离饲养的大鼠表现出所谓的隔离综合征的特征，这种特征会引起社会压力并加速抑郁症的发作23.
蒙古医学是中医的一个重要分支，它认为疾病的发作是内在因素和外部因素之间的复杂相互作用。这些外部因素被称为四个辅助条件，包括气候变化、饮食、生活方式和突发事件，如感染、令人震惊的事件和心理障碍。疾病过程被概念化为三个元素（称为三种类型的同源体）与七种身体成分与四种辅助条件之间的持续相互作用24.蒙古医学认为，人体作为一个整体运作，由三个同源之间的相对平衡维持。这种平衡的破坏被认为是疾病的前兆24.鉴于动物实验在连接传统医学和现代医学方面发挥着关键作用，开发与蒙古医学领域研究相关的动物模型至关重要。因此，我们采用了 28 天隔离方法与 CUMS 相结合来模拟这些生理和心理压力源。我们选择了九个特定的不可预测的应激源，并试图通过蒙古医学的三同源理论来支持这种建模方法。建立稳健的动物模型是推进蒙古医学基础研究的基础，并将对其基础研究做出重大贡献。

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		<tr>
			<td>High Speed Tissue Grinder</td>
			<td>service Biotechnology Co., Ltd</td>
			<td>KZ-<img alt="Equation 2" src="/files/ftp_upload/65889/65889eq02.jpg" style="margin: auto;" />-F</td>
			<td></td>
		</tr>
		<tr>
			<td>Horizontal Freezer</td>
			<td>Mellow Group Co., Ltd</td>
			<td>BCD-318AT</td>
			<td></td>
		</tr>
		<tr>
			<td>Laboratory Ultrapure Water Machine</td>
			<td>Jinan Aiken Environmental Protection Technology Co., Ltd&#160;&#160;</td>
			<td>AK-RO-C2</td>
			<td></td>
		</tr>
		<tr>
			<td>Morris water maze video trail analysing system&#160;</td>
			<td>Tai Meng Tech Co., Ltd</td>
			<td>WMT-200</td>
			<td></td>
		</tr>
		<tr>
			<td>Rat 5-HT ELISA Kit</td>
			<td>Lian Ke bio Co., Ltd,China</td>
			<td>96T/48T</td>
			<td></td>
		</tr>
		<tr>
			<td>SPF grade Sprague Dawley (SD) rats</td>
			<td>SPF (Beijing) Biotechnology Co&#160;</td>
			<td>SCXK(JING)2019-0010</td>
			<td></td>
		</tr>
		<tr>
			<td>Sprague Dawley rats</td>
			<td>Beijing Biotechnology Co., Ltd, China</td>
			<td>&#160;SCXK (JING) 2019-0010</td>
			<td></td>
		</tr>
		<tr>
			<td>Vertical Refrigerated Display Cabinet</td>
			<td>Xingx Group Co., Ltd</td>
			<td>LSC-316C</td>
			<td></td>
		</tr>
		<tr>
			<td>video tracking system</td>
			<td>Tai Meng Tech Co., Ltd</td>
			<td>ZH-ZFT</td>
			<td></td>
		</tr>
		<tr>
			<td>vortex mixer</td>
			<td>Servicebio technology Co., Ltd</td>
			<td>MV-100</td>
			<td></td>
		</tr>
	</tbody>
</table>

chronic unpredictable mild stress in rats based on the mongolian medicine

抑郁症是一种普遍存在的情感障碍，是导致全球残疾的主要原因。目前药物干预的局限性导致了这种疾病带来的巨大健康负担。迫切需要更深入地了解抑郁症的潜在机制，这使得具有转化潜力的临床前模型非常有价值。蒙古医学是传统医学的一个分支，认为疾病的发生与风、胆汁和痰液的平衡密切相关。在这项研究中，我们介绍了大鼠慢性不可预测的轻度应激（CUMS）方法的方案。在这个框架内，大鼠受到一系列波动的、温和的压力源的影响，以诱导出类似抑郁症的表型，模仿人类抑郁症的发病机制。该协议中采用的行为测定包括蔗糖偏好测试 （SPT），表明快感缺乏——抑郁症的核心症状;测量焦虑水平的旷场测试（OFT）;以及评估空间记忆和学习能力的莫里斯水迷宫测试（MWM）。CUMS 方法证明了诱导快感缺乏和导致长期行为缺陷的能力。此外，与其他旨在引发抑郁样行为的动物模型相比，该协议更符合蒙古医学理论。这种动物模型的开发和后续研究为蒙古医学领域未来的创新研究奠定了坚实的基础。

CUMS诱导的大鼠抑郁模型中的行为测试结果 为了证实 CUMS 程序在诱导抑郁样行为方面的功效，进行了操作检查。雄性Sprague-Dawley（SD）大鼠被随机分配到MOD或CON组，为期4周，如步骤1.2.3所述。随后，处死大鼠，并完全解剖其海马体，使用酶联免疫吸附测定（ELISA）28评估5-HT，这是一种与抑郁症病理生理学密切相关的神经递质10。
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Watch this Scientific Journal Video about Chronic Unpredictable Mild Stress in Rats based on the Mongolian medicine at JoVE.com

作者聚焦:建立啮齿动物模型以研究传统蒙古医学中的抑郁因素

该协议概述了基于蒙古医学理论的抑郁症慢性不可预测轻度压力 （CUMS） 模型，以及验证行为测试的方法。

基于蒙古药的大鼠慢性不可预测的轻度应激

Depression is a prevalent affective disorder and constitutes a leading cause of global disability. The limitations of current pharmacological ...

我们的研究范围是蒙古医学研究。在这项研究中，我们研究了导致蒙古医学抑郁症的因素，特别强调内部因素之间的相互作用，如风、生物膜和外部因素。解决了符合蒙古国传统医疗模式缺失的问题。该协议解决了基于蒙古医学理论的动物模型的建立，然后为蒙古医学的未来研究提供了一些建议。在抑郁症研究中，建立动物模型对于理解新药的病理机制和作用至关重要。与其他技术相比，该方案更符合蒙古医学中抑郁症的发病机制和治疗原则，有助于未来蒙古医学中抑郁症治疗的研究。该协议可以证明外部因素和内部因素相互作用产生疾病之间的关系，并符合蒙古医学的理论体系。本文将推进传统医学对抑郁症的基础研究，并为后续研究者研究、开发和利用传统医学提供一些建议。

chronic-unpredictable-mild-stress-rats-based-on-mongolian

Research

JoVE Journal

Medicine

Chronic Unpredictable Mild Stress in Rats based on the Mongolian medicine

681 次观看.  Inner Mongolia Medical University. 我们的研究范围是蒙古医学研究。在这项研究中，我们研究了导致蒙古医学抑郁症的因素，特别强调内部因素之间的相互作用，如风、生物膜和外部因素。解决了符合蒙古国传统医疗模式缺失的问题。该协议解决了基于蒙古医学理论的动物模型的建立，然后为蒙古医学的未来研究提供了一些建议。在抑郁症研究中，建立动物模型对于理解新药的病理机制和作用至关重要?...

视频: 作者聚焦:建立啮齿动物模型以研究传统蒙古医学中的抑郁因素

Acute and Chronic Tactile Sensory Testing after Spinal Cord Injury in Rats

Spinal cord injury (SCI) impairs sensory systems causing allodynia1-8. To identify cellular and molecular causes of allodynia, sensitive and valid sensory testing in rat SCI models is needed. However, until recently, no single testing approach had been validated for SCI so that standardized methods have not been implemented across labs. Additionally, available testing methods could not be implemented acutely or when severe motor impairments existed, preventing studies of the development of SCI-induced allodynia3. Here we present two validated sensory testing methods using von Frey Hair (VFH) monofilaments which quantify changes in tactile sensory thresholds after SCI4-5. One test is the well-established Up-Down test which demonstrates high sensitivity and specificity across different SCI severities when tested chronically5. The other test is a newly-developed dorsal VFH test that can be applied acutely after SCI when allodynia develops, prior to motor recovery4-5. Each VFH monofilament applies a calibrated force when touched to the skin of the hind paw until it bends. In the up-down method, alternating VFHs of higher or lower forces are used on the plantar L5 dermatome to delineate flexor withdrawal thresholds. Successively higher forces are applied until withdrawal occurs then lower force VFHs are used until withdrawal ceases. The tactile threshold reflects the force required to elicit withdrawal in 50% of the stimuli. For the new test, each VFH is applied to the dorsal L5 dermatome of the paw while the rat is supported by the examiner. The VFH stimulation occurs in ascending order of force until at least 2 of 3 applications at a given force produces paw withdrawal. Tactile sensory threshold is the lowest force to elicit withdrawal 66% of the time. Acclimation, testing and scoring procedures are described. Aberrant trials that require a retest and typical trials are defined. Animal use was approved by Ohio State University Animal Care and Use Committee.

We describe two tactile sensory testing methods for acute or chronic periods of spinal cord injury in rats. These validated procedures can detect the development and maintenance of allodynia-like sensations.

在大鼠脊髓损伤后的急性和慢性触觉测试

Spinal cord injury (SCI) impairs sensory systems causing allodynia1-8. To identify cellular and molecular causes of allodynia, sensitive and valid sensory ...

科研

医学

Chronic Constriction of the Sciatic Nerve and Pain Hypersensitivity Testing in Rats

Chronic neuropathic pain, resulting from damage to the central or peripheral nervous system, is a prevalent and debilitating condition, affecting 7-18% of the population1,2. Symptoms include spontaneous (tingling, burning, electric-shock like) pain, dysaesthesia, paraesthesia, allodynia (pain resulting from normally non-painful stimuli) and hyperalgesia (an increased response to painful stimuli). The sensory symptoms are co-morbid with behavioural disabilities, such as insomnia and depression. To study chronic neuropathic pain several animal models mimicking peripheral nerve injury have been developed, one of the most widely used is Bennett and Xie's (1988) unilateral sciatic nerve chronic constriction injury (CCI)3 (Figure 1). Here we present a method for performing CCI and testing pain hypersensitivity.
CCI is performed under anaesthesia, with the sciatic nerve on one side exposed by making a skin incision, and cutting through the connective tissue between the gluteus superficialis and biceps femoris muscles. Four chromic gut ligatures are tied loosely around the sciatic nerve at 1 mm intervals, to just occlude but not arrest epineural blood flow. The wound is closed with sutures in the muscle and staples in the skin. The animal is then allowed to recover from surgery for 24 hrs before pain hypersensitivity testing begins.
For behavioural testing, rats are placed into the testing apparatus and are allowed to habituate to the testing procedure. The area tested is the mid-plantar surface of the hindpaw (Figure 2), which falls within the sciatic nerve distribution. Mechanical withdrawal threshold is assessed by mechanically stimulating both injured and uninjured hindpaws using an electronic dynamic plantar von Frey aesthesiometer or manual von Frey hairs4. The mechanical withdrawal threshold is the maximum pressure exerted (in grams) that triggers paw withdrawal. For measurement of thermal withdrawal latency, first described by Hargreaves et al (1988), the hindpaw is exposed to a beam of radiant heat through a transparent glass surface using a plantar analgesia meter5,6. The withdrawal latency to the heat stimulus is recorded as the time for paw withdrawal in both injured and uninjured hindpaws. Following CCI, mechanical withdrawal threshold, as well as thermal withdrawal latency in the injured paw are both significantly reduced, compared to baseline measurements and the uninjured paw (Figure 3). The CCI model of peripheral nerve injury combined with pain hypersensitivity testing provides a model system to investigate the effectiveness of potential therapeutic agents to modify chronic neuropathic pain. In our laboratory, we utilise CCI alongside thermal and mechanical sensitivity of the hindpaws to investigate the role of neuro-immune interactions in the pathogenesis and treatment of neuropathic pain.

Due to the simplicity of surgery and the robust behavioural outcome, chronic constriction of the sciatic nerve is one of the pre-eminent animal models of neuropathic pain. Within 24 hrs following surgery, pain hypersensitivity is established and can be quantitatively measured using a von Frey aesthesiometer (mechanical test) and plantar analgesia meter (thermal test).

慢性缩窄大鼠坐骨神经疼痛过敏测试

Chronic neuropathic pain, resulting from damage to the central or peripheral nervous system, is a prevalent and debilitating condition, affecting 7-18% of ...

A Modified In vitro Invasion Assay to Determine the Potential Role of Hormones, Cytokines and/or Growth Factors in Mediating Cancer Cell Invasion

Blood serum serves as a chemoattractant towards which cancer cells migrate and invade, facilitating their intravasation into microvessels. However, the actual molecules towards which the cells migrate remain elusive. This modified invasion assay has been developed to identify targets which drive cell migration and invasion. This technique compares the invasion index under three conditions to determine whether a specific hormone, growth factor, or cytokine plays a role in mediating the invasive potential of a cancer cell. These conditions include i) normal fetal bovine serum (FBS), ii) charcoal-stripped FBS (CS-FBS), which removes hormones, growth factors, and cytokines and iii) CS-FBS + molecule (denoted &#8220;X&#8221;). A significant change in cell invasion with CS-FBS as compared to FBS, indicates the involvement of hormones, cytokines or growth factors in mediating the change. Individual molecules can then be added back to CS-FBS to assay their ability to reverse or rescue the invasion phenotype. Furthermore, two or more factors can be combined to evaluate the additive or synergistic effects of multiple molecules in driving or inhibiting invasion. Overall, this method enables the investigator to determine whether hormones, cytokines, and/or growth factors play a role in cell invasion by serving as chemoattractants or inhibitors of invasion for a particular type of cancer cell or a specific mutant. By identifying specific chemoattractants and inhibitors, this modified invasion assay may help to elucidate signaling pathways that direct cancer cell invasion.

A Modified In vitro Invasion Assay to Determine the Potential Role of Hormones, Cytokines and/or Growth Factors in Mediating Cancer Cell Invasion

This video article describes a modified in vitro method to examine the role of hormones, cytokines and/or growth factors in driving cancer cell invasion.

一种改进<em&gt;在体外</em&gt;入侵检测，以确定激素，在调节肿瘤细胞侵袭的细胞因子和/或生长因子的潜在作用

Blood serum serves as a chemoattractant towards which cancer cells migrate and invade, facilitating their intravasation into microvessels. However, the ...

A Novel Model of Mild Traumatic Brain Injury for Juvenile Rats

Despite growing evidence that childhood represents a major risk period for mild traumatic brain injury (mTBI) from sports-related concussions, motor vehicle accidents, and falls, a reliable animal model of mTBI had previously not been developed for this important aspect of development. The modified weight-drop technique employs a glancing impact to the head of a freely moving rodent transmitting acceleration, deceleration, and rotational forces upon the brain. When applied to juvenile rats, this modified weight-drop technique induced clinically relevant behavioural outcomes that were representative of post-concussion symptomology. The technique is a rapidly applied procedure with an extremely low mortality rate, rendering it ideal for high-throughput studies of therapeutics. In addition, because the procedure involves a mild injury to a closed head, it can easily be used for studies of repetitive brain injury. Owing to the simplistic nature of this technique, and the clinically relevant biomechanics of the injury pathophysiology, the modified weight-drop technique provides researchers with a reliable model of mTBI that can be used in a wide variety of behavioural, molecular, and genetic studies.

The modified weight-drop technique is an easy, cost-effective procedure used for the induction of mild traumatic brain injury in juvenile rats. This novel technique produces clinically relevant symptomology that will advance the study of mild traumatic brain injury (mTBI) and concussion.

轻度脑外伤的幼年大鼠一种新模式

Despite growing evidence that childhood represents a major risk period for mild traumatic brain injury (mTBI) from sports-related concussions, motor ...

Histological Quantification of Chronic Myocardial Infarct in Rats

Myocardial infarction is defined as cardiomyocyte death due to prolonged ischemia; an inflammatory response and scar formation (fibrosis) follow the ischemic injury. Following the initial acute phase, chronic remodeling of the left ventricle (LV) modifies the structure and function of the heart. Permanent coronary ligation in small animals has been widely used as a reference model for a chronic model of MI. Thinning of the infarcted wall progressively develops to transmural fibrosis. Histological assessment of infarct size is commonly performed; nevertheless, a standardization of the methods for quantification is missing. Indeed, important methodological aspects, such as the number of sections analyzed and the sampling and quantification methods, are usually not described and therefore preclude comparison across investigations. Too often, quantification is performed on a single section obtained at the level of the papillary muscles. Because novel strategies aimed at reducing infarct expansion and remodeling are under investigation, there is an important need for the standardization of accurate heart sampling protocols. We describe an accurate method to quantify the infarct size using a systematic sampling of harvested rat heart and image analyses of trichromatic stained histological sections obtained from base to apex. We also provide evidence that calculating the expansion index (EI) allowed for infarct size assessment, taking into account changes of the left ventricle throughout the remodeling.

The post-mortem assessment of myocardial infarction (MI) in rodents is based on quantification of the infarct on stained heart sections. We describe an accurate method to quantify the infarct size using systematic sampling of harvested rat hearts from base to apex and image analyses of trichrome-stained histological sections.

大鼠慢性心肌梗塞的组织学定量分析

Myocardial infarction is defined as cardiomyocyte death due to prolonged ischemia; an inflammatory response and scar formation (fibrosis) follow the ...

Ultrasonography in Experimental Reproductive Investigations on Rats

With the development of assisted reproductive technology and the ethical limitations of research on humans, rat animal models have been widely used in reproductive medicine. In the past, the study of reproductive system development in rodents has been based on one-time histological examination of excised tissues. Recently, with the development of high-resolution transabdominal ultrasound, high-quality sonography can now be performed to evaluate the reproductive organs of rats, allowing a new method for studying the reproductive system. Images were obtained using a high-resolution ultrasonographic system. Gynecological ultrasonography was performed on 28 eight-week-old non-pregnant rats and 5 pregnant Sprague-Dawley rats. We describe how to recognize organs of the reproductive system and associated structures in typical views during different phases of the estrus cycle. Color flow Doppler was used to measure uterine artery blood flow and evaluate uterine blood flow pattern changes during different stages of pregnancy. We have demonstrated that ultrasound exploration is a useful method for evaluating changes in internal reproductive organs. Its use raises the possibility of conducting additional experiments, including medical or surgical procedures, and provides the ability to monitor sonographic changes to internal organs without sacrificing animals.

This manuscript describes the utility of ultrasound performed on female rats to design experimental models for reproductive and gynecological investigation. A step-by-step explanation of how to perform ultrasonographic evaluation is shown.

大鼠实验性生殖研究中的超声检查

With the development of assisted reproductive technology and the ethical limitations of research on humans, rat animal models have been widely used in ...

Isolation of Intrapulmonary Artery and Smooth Muscle Cells to Investigate Vascular Responses

The intrapulmonary artery (IPA) and vascular smooth muscle cells (VSMCs) isolated from rat lungs can be used to study the underlying mechanisms of vasoconstriction and vasorelaxation. After isolating the IPA and VSMCs, the characteristics of vascular responses in physiological and pathological conditions can be assessed in the absence of extrinsic factors such as nerve signals, hormones, cytokines, etc. Thus, the IPA and VSMCs serve as excellent models for studying vascular physiology/pathophysiology, along with various experimental investigations, such as modulation by pharmacological agents, patch-clamp electrophysiological analysis, calcium imaging, etc. Here, we have used a technique for isolating the IPA to investigate vascular responses in an organ bath setup. IPA segments were mounted on the organ bath chamber via intraluminal wires and stimulated by various pharmacological agents. The changes in IPA vascular tone (i.e., vasoconstriction and vasorelaxation), were recorded using an isometric force transducer and physiological data analysis software program. We implemented several experimental protocols, which can be adapted to investigate the mechanisms of vasorelaxation/vasoconstriction for studying the pharmacological activities of phytochemical or synthetic drugs. The protocols can also be used to evaluate drugs&#39; roles in modulating various diseases, including pulmonary arterial hypertension. The IPA model allows us to investigate the concentration-response curve, which is crucial in assessing drugs&#39; pharmacodynamic parameters.

Vascular responses of arterial pulmonary circulation can be explored using intrapulmonary artery (IPA) and vascular smooth muscle cells (VSMCs). The present study describes the isolation of IPA in detail and the protocols used for investigating vasorelaxation in response to physiological stimuli.

分离肺内动脉和平滑肌细胞以研究血管反应

The intrapulmonary artery (IPA) and vascular smooth muscle cells (VSMCs) isolated from rat lungs can be used to study the underlying mechanisms of ...

Behavioral and Network Pharmacology-Based Analyses for the Traditional Mongolian Medicine Zadi-5 in a Rat Model of Depression

Zadi-5 is a traditional Mongolian medicine that is widely used for the treatment of depression and symptoms of irritation. Although the therapeutic effects of Zadi-5 against depression have been indicated in previously reported clinical studies, the identity and impact of the active pharmaceutical compounds present in the drug have not been fully elucidated.&#160;This study used network pharmacology to predict the drug composition and identify the therapeutically active compounds in Zadi-5 pills. Here, we established a rat model of chronic unpredicted mild stress (CUMS) and conducted an open field test (OFT), Morris water maze (MWM) analysis, and sucrose consumption test (SCT) to investigate the potential therapeutic efficacy of Zadi-5 in depression. This study aimed to demonstrate Zadi-5&#39;s therapeutic effects for depression and predict the critical pathway of the action of Zadi-5 against the disorder. The vertical and horizontal scores (OFT), SCT, and zone crossing numbers of the fluoxetine (positive control) and Zadi-5 groups were significantly higher (P &#60; 0.05) than those of the CUMS group rats without treatment. According to the results of network pharmacology analysis, the PI3K-AKT pathway was found to be essential for the antidepressant effect of Zadi-5.

The present protocol describes a method for the behavioral test validation and bioinformatical prediction of the therapeutic efficacy of Zadi-5, a traditional Mongolian medicine, in depression.

基于行为和网络药理学的蒙古医学Zadi-5在抑郁症大鼠模型中的分析

Zadi-5 is a traditional Mongolian medicine that is widely used for the treatment of depression and symptoms of irritation. Although the therapeutic ...

基于草药蛋糕治疗大鼠肾功能衰竭大鼠的艾灸平台

Using Herbal-Cake-Separated Moxibustion for the Treatment of Rats with Chronic Renal Failure

In the process of moxibustion in clinical practice, subjects need to be in a stable mood and comfortable posture to avoid problems such as moxa ash falling, scalding skin, and poor curative effect. Such problems also exist in the rat moxibustion experiment. To simulate clinical practice, it is necessary to introduce an experimental instrument in animal experiments, that is, a moxibustion device with fixed rats and moxibustion treatment synchronization, which can make experimental rats receive moxibustion treatment quietly and comfortably under non-anesthesia.
Our research group designed a rat moxibustion experimental platform. The device was framed by a wooden board with a supporting base plate, multiple fixed components, and partitioned components. The device can achieve the operation mode of moxibustion in rats without binding, avoiding anesthesia and scalding and simultaneously exposing multiple acupoints on the back. This operation can avoid physical and mental injury to rats and operators, which improves the research efficiency and further promotes the development and research of moxibustion animal experiments.
The device has a simple structure, is easy to operate and popularize, is comprehensively and innovatively designed, reusable, and is suitable for rat experiments mainly based on moxibustion. This article mainly introduces the structure of the experimental platform device for rat moxibustion, the basic procedure of herbal-cake-separated-moxibustion in experimental rats using the device and describes the establishment of a rat model of chronic renal failure (CRF) and representative experimental results.

作者聚焦:了解草药蛋糕分离艾灸对肾病大鼠的影响

The moxibustion experimental platform can fix the rats without restriction, and the rats can undergo multi-acupoint moxibustion in the awake state, ensuring the welfare of the rats, protecting the safety of the operators, ensuring therapeutic efficacy, and improving the efficiency of the experiment.

采用中药蛋糕分离艾灸治疗慢性肾功能衰竭大鼠

In the process of moxibustion in clinical practice, subjects need to be in a stable mood and comfortable posture to avoid problems such as moxa ash ...

Methods Article

基于蒙古药的大鼠慢性不可预测的轻度应激