Name: induction and assessment of exertional skeletal muscle damage in humans
Uploaded: 2016-12-11T13:00:00.000+00:00
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Description: Watch this Scientific Journal Video about Induction and Assessment of Exertional Skeletal Muscle Damage in Humans at JoVE.com

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以下程序均符合杨百翰大学机构审查委员会（IRB）的标准。 1.准备收缩协议注意:下面的协议说明基于所述Biodex Advantage软件。导航软件和如果使用不同的系统操作所述测力计将是不同的。 <ol><li>等速强度测试协议<ol><li>为了使等速协议，开测功机控制软件的计算机上，并选择"协议"选项卡，然后在屏幕顶部的工具栏上的"记录"选项卡。从下拉列表中选择"新建协议"。 </li><li>在"学习型"框中，选择"测试"选项。 </li><li>从下拉菜单中选择以下参数:在"模式"菜单中选择等速。在"联合"菜单中选择膝盖。在下面 ＆＃34;模式"菜单中选择扩展/屈选择两个伸屈运动心收缩选择。"销售代表"下的"同归"下拉菜单。 </li><li>在60°/秒，为离开那里，向收缩既有速度"单方面"标签计划1套3次重复。要做到这一点，输入"1"变成了"＃组"字段和"结束通过销售代表"行中输入"3"。然后在这两个"速度与千寻"和"速度走向"行键入60。 </li><li>使用90°为解剖参考点，并设置繁多的运动限制，以约10°，距离110°朝向（0°=全面推广，135° =完全屈曲）。为了做到这一点，选择在屏幕左侧的"设置光盘"按钮。校准杠杆臂，使得90°是平行于地面（如使用所提供的计算机屏幕上的杠杆臂角度指示器指示）。然后用90°作为所述解剖重ference点，向（0°=全面推广，135°=完全屈曲）的运动限制的范围内设定为约10°，距离110°。 注:此处给出的首选关节角度设置，但这些设置应根据具体情况逐案与固定屈曲畸形或其他限制对象的工作特别是当修改。 </li><li>在名称在屏幕左下方的"描述"字段的协议，并选择保存顶部菜单栏上的按钮。 </li></ol></li><li>等距强度测试协议<ol><li>重复步骤1.1.1开始制作等距协议。 </li><li>从"学习型"中的"测试"选项。 </li><li>从下拉菜单中选择以下参数:在"模式"菜单中选择"等距"。在"联合"菜单中选择"拐点"。在"模式"菜单中选择"扩展/屈"。下的"ContractioN的方向"菜单中选择"离开"，在"同归"菜单中选择"代表"。 </li><li>程序与1组3个等距收缩持续每5秒，在70°的关节角度进行测试。要做到这一点，输入"1"变成了"位置"字段中的"单方面"选项卡。在"结束由众议员"行中的位置＃1列中，键入"3"。在标有"角度"行选择"70"。 </li><li>选择90°为解剖参考点和远限制到70°。 </li><li>在名称在屏幕左下方的"描述"字段的协议，并选择保存顶部菜单栏上的按钮。 </li></ol></li><li>肌肉损坏了离心运动协议<ol><li>为了使离心运动的协议，开测功机控制软件的计算机上，并选择"协议"选项卡。然后选择屏幕顶部的工具栏上的"记录"选项卡。现在，点击"新建协议＆＃34;从下拉列表中。 </li><li>从"学习型"中的"运动"选项。 </li><li>从下拉菜单中选择以下参数:在"模式"菜单中选择"等速"。在"联合"菜单中选择"拐点"。在"模式"菜单中选择"扩展/屈"。在"收缩"菜单中选择"CON / ECC"。 </li><li>在"单方面"选项卡中，节目10套重复10次180°/秒，距离120°/秒速度向与集之间的60秒休息。 <ol><li>输入"10"变成了"＃设置"字段。 </li><li>在第一列（标示为＃1），标记该行中输入"10"，"通过重复次数结束"。然后，标有"速度与千寻"下一行中输入"180"。在标有"速度走向"下一行，键入"120"。在标有"扭矩"的行，键入200到600取决于S的预期强度ubject。 </li><li>通过＃10＃套2重复步骤1.3.4.2。 </li><li> 60类中的"休息时间秒"字段。 </li></ol></li><li>的90°到"解剖参考"字段。 </li><li>选择在屏幕的左侧向（0°=完全伸展，135°=完全屈曲）"设置的ROM"按钮的运动限制到40°，距离110°设置范围。这些设置提供运动，通过它的主题必须提供抵抗测功机的臂的范围内。为安全起见，测功机将停止，如果这个问题停止这种指定的运动范围内提供移动性。 <ol><li>设定的运动范围中，选择在屏幕左侧的"设定ROM"按钮。校准杠杆臂，使得90°是平行于地面（如使用所提供的计算机屏幕上的杠杆臂角度指示器指示）。然后用90°作为解剖参考点，S等运动限制到40°，距离110°朝向范围（0°=全面推广，135°=完全屈曲）。 注:这些关节的角度设置是一个典型的健康主题的建议。这些应该进行修改，以适应能力，从一个人到另一个。 </li></ol></li><li>名称中的"描述"字段的协议，并在顶部工具栏中选择"保存"按钮。 </li></ol></li></ol> 2.测量基线<ol><li>测量酸痛<ol><li>在页面上绘制水平100毫米线做一个视觉模拟评分。 </li><li>上线的左侧表示"否疼痛"，并在该行的右手侧表示"极端疼痛"（ 图1）。此方法通常用于评估延迟性肌肉酸痛1,10,11。 </li><li>指示主题做两体重蹲。执行武器举行straigh都蹲在肩膀和双脚与肩同宽，前面T出。当上腿与地板平行返回到开始位置之前达到适当的深度。 </li><li>让拍摄对象表示他或她在股四头肌觉得通过对视觉模拟评分适当的位置画一条垂直线的股四头肌在深蹲肌肉酸痛的力度。 </li><li>通过测量受试者的印记从视觉模拟评分的无酸痛结束毫米的距离量化酸痛的主题的水平。 </li></ol></li><li>血清肌酸激酶 注意:处理血液之前戴防护手套，并在锐器容器按医院或大学政策使用后立即针的处理。 <ol><li>遵循标准化放血过程从禁食条件下受验者的手臂的肘前区域撤出血液以获得5至6毫升血液12。 </li><li>离心机在15 XG血液样本在3 15分钟 °C到从血细胞中分离血清。 </li><li>用移液管，转移血清样品放入适当标记的主题和时间点离心管。 </li><li>储存在-80℃以备后用的样品。 </li><li>测定血清样品用于使用根据制造商的协议市售的试剂盒的肌酸激酶活性。可替代地，血清样品可以运输到当地的医疗诊断实验室进行分析。 </li></ol></li><li>主题准备<ol><li>前主体的访问四十八小时，指示他们要避免剧烈的或不熟悉的运动，非甾体类抗炎药和酒精。 </li><li>当拍摄对象到达时，收集体重，身高，性别和之前安装的腿主导地位的信息。腿霸主地位通过询问用哪条腿，他们会选择踢球的主题被确定。 </li><li>有subjec在50进行的强度试验上前t升温到150瓦用于在自行车测功计10分钟。 </li><li>热身后，问这个问题的测力计的椅子上坐下。调整倾斜，高度，深度，斜倚，并且座椅和轴臂的旋转，以适应受试者。使用股骨外侧髁作为参考并对齐膝的旋转轴与该测力计轴臂的。调整轴臂长度，使得它仅仅是接近内踝，并用带固定。主题应能舒适地跖屈全运动范围。 </li><li>离开座位的前边缘和腿部的后面之间有足够的空间（25毫米），以允许膝关节的最大屈曲，而不会挤压或不适。 </li><li>受稳定与肩，髋，踝关节和大腿绑带。以分离膝伸肌和避免辅助肌受累，至关重要的是，该主题被牢固地稳定。 </li><li>有主题演出3-5实践分机之前开始收缩协议可能存在的，并根据需要调整膝关节/屈曲机动，运动的监控范围和不适的迹象。 </li><li>现在，测功机的可调部件正确安装到这个问题，录制在以后的访问重复设置的座椅位置设置和轴手臂的位置。 注:重要的是，受试者是安全和舒适。测功机配备有用于在紧急的情况下，主体和操作都紧急停止按钮。 </li></ol></li><li>力测量<ol><li>这个问题是在座位上正确定向和等距强度试验加载协议之后，指导如何执行测试的主题。 <ol><li>告诉他或她将被收缩对杠杆臂，将留固定的主题。请告诉主体按照计算机屏幕做3宫缩持续5秒EAC的提示H。 </li><li>告诉主体维护每个收缩的最大努力。告诉主体在宫缩时呼吸。让拍摄对象在测试过程中交叉于胸前交叉双臂。 </li><li>启动等距协议，允许受实践与次最大努力的考验。这种做法审判将在熟悉受制于测试，以及提供特定的测试热身。允许受试者练试验2至3倍。 </li></ol></li><li>一旦受试者是准备好了，开始测试和口头鼓励受试者以提供每个收缩的最大努力。 </li><li>记录扭矩，功率，和工作。该软件还计算变异系数（基于性能重复性试验有效性的统计表示，较低的值表示较高的再现性）。如果变异系数超过15％，重复测试。重复测试高达4倍。如果主体是无法生成系数Ø˚F方差后4尝试小于15％使用从产生方差值的最低系数的集合的数据。 </li><li>允许受到两到三分钟休息，然后加载等速协议来获取等速强度数据。 </li><li>这个问题被固定在座位上后，指导如何进行等速测试。 <ol><li>告诉他们会做对杠杆臂，将在一个固定的速度动议三项屈伸收缩的主题。给该主题的目标是在整个每个收缩运动的整个范围内为有力地收缩。 </li><li>告诉他们，当他们到达运动范围的结束杠杆臂将停止。让拍摄对象在测试过程中交叉于胸前交叉双臂。提醒受不宫缩时屏住呼吸 注意:当从收缩的屈曲部件收集的数据可能并不感兴趣的研究者，主体谁为屈伸演习提供最大的努力学分倾向于延长演习中，以便获得更好的力量的一致性。 </li></ol></li><li>开始测试和口头鼓励受提供最大的努力。 </li><li>记录扭矩，功率，和工作。该软件还计算变异系数（基于性能重复性试验有效性的统计表示，较低的值表示较高的再现性）。如果变异系数超过15％，重复测试。 <ol><li>重复测试高达4倍。如果主体是无法产生偏差小于15％的系数后4次尝试，从产生偏差值系数最低的一组使用的数据。 </li></ol></li></ol></li></ol> 3.感应损坏<ol><li>一到第2部分的基线测量三天后，让主体回归到实验室进行的损伤诱导部协议</li><li>对一个周期测功计在50受试者热身10分钟，以150瓦</li><li>经过热身，东方和使用记录在2.3.7步调整设置固定在测功机的主题，加载肌肉损伤离心运动的协议和指导如何正确地执行演习的主题。 <ol><li>告诉它们将收缩对杠杆臂，以启动轴臂的移动对每个收缩的主题。告诉主体，由于轴臂以固定利率向他们移动，他们的目标是尽可能有力地收缩反对。 </li><li>允许受试者把握位于座椅的任一侧的手柄。 </li><li>告诉主体在宫缩时呼吸;在偏心阶段呼气。 </li></ol></li><li>当拍摄对象准备就绪后，按下屏幕上的启动按钮启动第一套收缩。 </li><li>口头鼓励的主题，以保持最大的EF堡垒每个收缩。 </li><li>按照指导的主题，通过10套10重复与集之间有1分钟的休息时间在屏幕上的提示进行操作。 </li><li>一旦10套完成允许受试者1至5分钟，以其余重复协议之前。 注:套偏心协议下可用的最大数量是10然而这可以根据所使用的测力计和软件变化。其余部分提供时间如有必要，运营商重新加载协议。 </li><li>重复该过程两次，总共300主要离心收缩。 </li></ol> 4.肌肉损伤评估<ol><li>重复两等速和作为有害运动后立即在后24，48，72和96小时在2.4中描述等角强度试验。 </li><li>重复测量疼痛于24，48，72部分2.1所述，破坏运动后96小时。一定要使用新的视觉模拟评分对每个评估使主体不能看到他们以前的响应。 </li><li>重复抽血破坏运动后48和120小时为肌酸激酶检测部分2.2所描述的，或由实验方案规定的时间点。 </li><li>绘制关于对时间的曲线图上的x轴的y轴的肌酸激酶，强度和酸痛数据。 </li><li>观察这些变量是如何偏离，并在运动后的日子里向基线值回归，说明损伤修复，分别为。 </li></ol>

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Physiol. 294 (5), R1628-R1637 (2008).</li><li>Stupka, N., Tarnopolsky, M. A., Yardley, N. J., Phillips, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cellular+adaptation+to+repeated+eccentric+exercise-induced+muscle+damage.">Cellular adaptation to repeated eccentric exercise-induced muscle damage.</a> J. Appl. Physiol. 91 (4), 1669-1678 (2001).</li><li>Smith, L. L., 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=Changes+in+serum+cytokines+after+repeated+bouts+of+downhill+running.">Changes in serum cytokines after repeated bouts of downhill running.</a> Appl. Physiol. Nutr. Metab. 32 (2), 233-240 (2007).</li><li>Marqueste, T., Giannesini, B., Fur, Y. L., Cozzone, P. J., Bendahan, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparative+MRI+analysis+of+T2+changes+associated+with+single+and+repeated+bouts+of+downhill+running+leading+to+eccentric-induced+muscle+damage.">Comparative MRI analysis of T2 changes associated with single and repeated bouts of downhill running leading to eccentric-induced muscle damage.</a> J. Appl. Physiol. 105 (1), 299-307 (2008).</li><li>Crameri, R. M., 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=Myofibre+damage+in+human+skeletal+muscle:+effects+of+electrical+stimulation+versus+voluntary+contraction.">Myofibre damage in human skeletal muscle: effects of electrical stimulation versus voluntary contraction.</a> J. Physiol. 583 (Pt 1), 365-380 (2007).</li><li>Yu, J. G., Malm, C., Thornell, L. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Eccentric+contractions+leading+to+DOMS+do+not+cause+loss+of+desmin+nor+fibre+necrosis+in+human+muscle.">Eccentric contractions leading to DOMS do not cause loss of desmin nor fibre necrosis in human muscle.</a> Histochem. Cell Biol. 118 (1), 29-34 (2002).</li><li>Jamurtas, A. Z., 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=Comparison+between+leg+and+arm+eccentric+exercises+of+the+same+relative+intensity+on+indices+of+muscle+damage.">Comparison between leg and arm eccentric exercises of the same relative intensity on indices of muscle damage.</a> Eur. J. Appl. Physiol. 95 (2-3), 179-185 (2005).</li></ol>

The authors have nothing to disclose.

有几个步骤是获得本协议的预期效果的关键。首先，受试者必须充分熟悉到收缩的协议，特别是测力。要确保拍摄理解他们应该做的，给他们练习之前，数据采集强度测试的机会到底是什么。谁不熟悉充分使用这些程序的主体会显示损伤诱导下过天的学习曲线。这可以是一个混杂变量呈现强度测量无效。 图5从谁可能没有被正确熟悉的个人资料显示。本课题显示，由于越来越多的实力，也许，尽管破坏性锻炼学习在实验的过程中。第二，该受试者群体的选择也可以是该协议的关键。劳力肌肉损伤可能取决于许多f起受受试者变化很大行动者，包括个人的训练的状态。习惯了离心运动的个人将比较不习惯的人表现出很大的损伤小。选择的受试者群体时，这可能是一个重要的考虑因素。如果研究者有兴趣观察大损伤反应，人口不习惯离心运动将是最有可能提供这一结果。最后，这是至关重要的受试者一致地定位在测力计上的反复访问。研究者应该记录在测功机每个科目的每个可调节部件的位置设置，并相应地调整它的每个后续测试。这将最大限度地减少可变性因体位的不同。 许多不同dynameters可用。而这里所提供的方法是特定于Biodex测力计和控制软件，这些方法可占操作系后适于与其他测力计使用摹分歧。使用如这里介绍的测力计的提供对用于诱导人类肌肉损伤一些其它方法的优点。这些优势包括精确量化在演习期间所做的总功，控制角速度和精确测量的力量和强度损失。然而，损伤诱导的其它方法可以在一个测力计，不提供或不优选的情况下使用。胡巴尔和同事13所使用的重复椅子坐和上升的方法来有效地诱导的肌肉损伤。在这项研究中，受试者进行降低到椅子延长收缩。斯图普卡和同事14使用期间，受试者在同心一个重复最多的120％进行收缩的延长部件传统伸膝锻炼。下坡行驶也是造成肌肉损伤15,16的有效刺激。如果研究人员是损伤诱导的这些其他方法可以是优选感兴趣观察从一个更真实的或特定的运动刺激，或从一个封闭的动能上链机芯的损害。这个协议，研究人员可以考虑另一种修饰是采血的肌酸激酶测定频率。为了得到血清肌酸激酶运动后的最翔实的图片，研究人员可能选择采取血液样本，每24小时的损害后5〜6天。这将确保高峰期血清肌酸激酶值不会错过。也这将提供作为到的变化在血清肌酸激酶，如曲线是否遵循单或双相形状的性质更好的信息。 此协议的一个限制是，强度损失，血清肌酸激酶和疼痛，是损害的间接标记。而损伤的直接证据可以通过获得肌肉样品并用电子显微镜观察到的，在啮齿动物研究中使用的其他组织学方法不检测VOLU在人体肌肉17,18 ntary收缩诱导的损伤。考虑到能够直接检测在人类自主收缩引起的损伤情况下肌肉损伤的方法有限，损坏的间接指标是最好的选择10名。 二者的数据收集和分析的各个方面，可能需要排除故障。在肌肉损伤协议中，受试者必须触发对每个离心收缩轴臂的运动。主题，直到阈力值满足触发轴运动和离心收缩在扩展膝盖位置对承包固定臂做到这一点。阈值力值是程序性进入收缩协议（步骤1.3.4.2）的扭矩值的10％。主体可能满足在伸展膝盖开始位置的门槛难度力，已建成了多套特之后。研究人员可以帮助主体到达THReshold通过拉动轴臂以引发收缩。可替代地，能够减少程序性到协议（1.3.4.2）的每个收缩的转矩值，使得该阈值是较低的。然而，扭矩值不能设置得太低，因为如果这个问题超出了扭矩值轴手臂将停止收缩将被中断。在肌酸激酶和酸痛数据的性质也可以为研究者一些麻烦。这些数据集常常不是正态分布。两个变量往往显示方差不均匀性，使得增加的变异性较高的平均值发生。由于这些变量的性质，对数变换往往是适当的归一化分布和均匀化之前的统计分析的方差。可替代地，可以使用非参数检验。 在本文所描述的运动引起的肌肉损伤协议具有广阔的应用前景。主要海峡使用的协议的ength是研究支配肌肉的修复，再生，和适应直接在人受试者的细胞和分子过程的潜力。此外，延迟性肌肉酸痛和临时实力的损失往往是剧烈的，或新的锻炼不良的后果。利用如一个在这里描述的基于运动损伤协议，提供了研究人员验证的模型可以测试营养，营养食品，或药物干预或为了防止延迟性肌肉酸痛以下锻炼其他治疗的有效性。 总之，本手稿描述了一种安全可靠的方法以引起和量化在人类收缩诱导的骨骼肌损伤。它使用由一测力计控制以诱导损伤膝伸肌的离心收缩。肌肉损伤无创性与延迟性肌肉酸痛评估，血清肌酸激酶和强度损失。而该协议对损害诱导和力的测量的特定测力计写入，该协议可以适于一起用于与其他测力计以及收缩诱导损伤其它模式中使用。

The overall goal of this procedure is to induce exertional damage to the quadriceps femoris muscles using voluntary lengthening (eccentric) contractions in human subjects.
Contraction-induced skeletal muscle damage is a common consequence of exercise that is marked by delayed onset muscle soreness1, transient strength loss, and elevated muscle-specific enzymes in the blood2. Exertional muscle damage is most pronounced following exercise to which the subject is unaccustomed, particularly when eccentric contractions are involved3. Exertional muscle damage is typically benign. Soreness subsides, and both serum proteins and strength typically return to pre-damage levels within a few days to weeks after the damaging insult. In extreme cases, exertional muscle damage can lead to a life-threatening syndrome know as rhabdomyolysis. However, exertional muscle damage is usually insufficient to cause clinical rhabdomyolysis in healthy individuals4 in the absence of compounding factors including heat stress, dehydration5, infection6 or rare genetic predispositions7. 
Contraction-induced muscle damage is typically less severe than toxin-induced or freezing-induced injury, methods often used in rodent studies8,9. Yet, contraction-induced injury provides a useful method to study the muscle damage response with notable advantages. First, it is a safe and ethical method for use with human subjects1-3. Thus, interspecies translation of the results is not needed as data can be obtained directly from human subjects. Moreover, translating data obtained from rodent studies is very difficult given that the severity of injury seen in the rodent injury models exceeds the level of damage that would be ethical to induce in human subjects. Second, contraction-induced damage is commonly experienced and a natural process of exercise. Therefore, this mode of damage induction is useful for studying muscle damage in the context of exercise, adaptation to exercise as well as overt muscle injury. Here we describe a safe and reliable method to induce and evaluate skeletal muscle damage using lengthening contractions in humans.

<table><tbody><tr><td>Biodex Dynomometer</td><td>Biodex Medical Systems</td><td>850-000</td><td>Other models are available and should produce similar results</td></tr><tr><td>Creatine Kinase kit</td><td>Sigma-Aldrich&amp;nbsp;</td><td>MAK116</td><td></td></tr><tr><td>Serum Vacutainers</td><td>BD Bioscience</td><td>367812</td><td></td></tr><tr><td>Winged safety push button blood collection set</td><td>BD Bioscience</td><td>367338</td><td></td></tr><tr><td>Cryogenic vials</td><td>Sigma-Aldrich</td><td>V5007</td><td>We use the 2&amp;nbsp;ml vials to store serum aliquots</td></tr></tbody></table>

induction and assessment of exertional skeletal muscle damage in humans

通过自愿偏心（延长）收缩收缩引起的肌肉损伤提供了研究人类肌肉的适应和恢复的优秀典范。在这里我们讨论的离心运动协议的设计诱导四头肌肌肉，在力量，疼痛，和血浆肌酸激酶水平的变化标志着损害。因为它是在人参加演出，并消除了结果的种间翻译此法简单，伦理和适用广泛。科目的速度进行伸膝肌群的最大300离心收缩 120°/上等速测力秒。损害的程度是衡量使用强度的损失，疼痛，和血浆肌酸激酶水平的相对非侵入等速和等距的措施在几天后的锻炼。因此，它的应用，可向特定人群，以试图确定肌肉机制适应和再生。

使用这里介绍的方法，基线酸胀，血清肌酸激酶活性和强度（等距和等速）测量是在7未受过训练的年轻男子。第二天，受试者接受了肌肉损伤上述离心收缩协议。为了提供肌肉损伤的指标，跟进的实力，疼痛和血清肌酸激酶活性的评估进行的。强度运动后以及24，48，72，和96小时后，立即进行测定。疼痛测定24，48，72，和运动后96小时。血清肌酸激酶在48和120小时的运动后测得的。使用方差用Dunnett多重比较测试的单向重复测量分析，以运动后的值进行比较，以在运动前值这些数据进行分析。相比运动前，等距和等速力分别减少到24和48小时后的运动，以及<html>返回类似运动前的值此后（ 图2）。酸痛24,48被显著增加，运动后72小时（ 图3）。血清肌酸激酶升高显著运动后48小时（ 图4）。 图5示出在天以下破坏性锻炼非典型等距力曲线。 <img alt="图1" src="/files/ftp_upload/54859/54859fig1.jpg" /> 图1: 用于量化股四头肌的肌肉酸痛的视觉模拟量表 股四头肌 肌肉。受试者被指示做两体重深蹲，然后在该行表示酸胀，他们在四头肌肌肉下蹲动作时感觉到的程度。研究者通过测量从规模的无酸痛结束毫米大关的距离量化这一点。"https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54859/54859fig1large.jpg"目标="_空白"&gt;点击此处查看该图的放大版本。 <img alt="图2" src="/files/ftp_upload/54859/54859fig2.jpg" /> 图2: 长力（A）和60°/ s的等速膝伸肌的力量（B）前（PRE），后（IMM POST），24，48，72，和一个回合后96小时1天300最大努力延长收缩。数据被表示为运动前力值的百分比（平均值±SD）。与一个Dunnett检验多重比较方差单向重复测量分析数据进行分析。 *表示从预（P &lt;0.05）显著差异。这个数字是改编自参考3。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54859/54859fig2large.jpg" target="_blank">请点击此处查看大图已经</a>这个数字rsion。 <img alt="图3" src="/files/ftp_upload/54859/54859fig3.jpg" /> 图3: 运动前（预）的疼痛响应曲线有一天，当以及在24，48，72，和96小时以下300最大省力延长收缩。这些数据是在对数刻度10表示（平均值±SD）。 *表示从运动前（P &lt;0.05）显著差异。这个数字是改编自参考3。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54859/54859fig3large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图4" src="/files/ftp_upload/54859/54859fig4.jpg" /> 图4: 血清肌酸激酶（CK）活性（平均值±SD）24小时前（PRE），48后最大的300省力延长续120小时ractions（LC）。数据显示为运动前值上的日志10的范围的百分比。 *表示从预（P &lt;0.05），NS表示无显著差异显著差异。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54859/54859fig4large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图5" src="/files/ftp_upload/54859/54859fig5.jpg" /> 图5: 一个主题的膝伸肌谁发现非典型响应等长收缩力。力测量后立即（IMM POST）24，48，300最大努力延长收缩回合后72和96小时相比，在这个问题的运动前（PRE）值并没有减少。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54859/54859fig5large.jpg" target="_blank">请点击此处查看该图的放大版本。</a></a>

Watch this Scientific Journal Video about Induction and Assessment of Exertional Skeletal Muscle Damage in Humans at JoVE.com

Induction and Assessment of Exertional Skeletal Muscle Damage in Humans

This article describes a safe and reliable method to induce and quantify exertional skeletal muscle damage in human subjects.

诱导和人类劳力骨骼肌损伤评估

Contraction-induced muscle damage via voluntary eccentric (lengthening) contractions offers an excellent model for studying muscle adaptation and recovery ...

induction-assessment-exertional-skeletal-muscle-damage

Nanyang Technological University

Research

JoVE Journal

Medicine

7.4K 次观看.  Brigham Young University. This article describes a safe and reliable method to induce and quantify exertional skeletal muscle damage in human subjects.

视频: Induction and Assessment of Exertional Skeletal Muscle Damage in Humans

In Vivo Canine Muscle Function Assay

We describe a minimally-invasive and reproducible method to measure canine pelvic limb muscle strength and muscle response to repeated eccentric 
contractions. The pelvic limb of an anesthetized dog is immobilized in a stereotactic frame to align the tibia at a right angle to the femur. Adhesive wrap affixes the 
paw to a pedal mounted on the shaft of a servomotor to measure torque. Percutaneous nerve stimulation activates pelvic limb muscles of the paw to either push (extend) or 
pull (flex) against the pedal to generate isometric torque. Percutaneous tibial nerve stimulation activates tibiotarsal extensor muscles. Repeated eccentric (lengthening) 
contractions are induced in the tibiotarsal flexor muscles by percutaneous peroneal nerve stimulation. The eccentric protocol consists of an initial isometric contraction 
followed by a forced stretch imposed by the servomotor. The rotation effectively lengthens the muscle while it contracts, e.g., an eccentric contraction. During 
stimulation flexor muscles are subjected to an 800 msec isometric and 200 msec eccentric contraction. This procedure is repeated every 5 sec. To avoid fatigue, 4 min rest 
follows every 10 contractions with a total of 30 contractions performed.

In Vivo Canine Muscle Function Assay

We describe a minimally-invasive and painless method to measure canine hindlimb muscle strength and muscle response to repeated eccentric contractions.

在犬体内的肌肉功能测定

We describe a minimally-invasive and reproducible method to measure canine pelvic limb muscle strength and muscle response to repeated eccentric 
...

科研

医学

Manual Muscle Testing:  A Method of Measuring Extremity Muscle Strength Applied to Critically Ill Patients 

Survivors of acute respiratory distress syndrome (ARDS) and other causes of critical illness often have generalized weakness, reduced exercise tolerance, and persistent nerve and muscle impairments after hospital discharge.1-6 Using an explicit protocol with a structured approach to training and quality assurance of research staff, manual muscle testing (MMT) is a highly reliable method for assessing strength, using a standardized clinical examination, for patients following ARDS, and can be completed with mechanically ventilated patients who can tolerate sitting upright in bed and are able to follow two-step commands. 7, 8 
This video demonstrates a protocol for MMT, which has been taught to &ge;43 research staff who have performed &gt;800 assessments on &gt;280 ARDS survivors. Modifications for the bedridden patient are included. Each muscle is tested with specific techniques for positioning, stabilization, resistance, and palpation for each score of the 6-point ordinal Medical Research Council scale.7,9-11 Three upper and three lower extremity muscles are graded in this protocol: shoulder abduction, elbow flexion, wrist extension, hip flexion, knee extension, and ankle dorsiflexion. These muscles were chosen based on the standard approach for evaluating patients for ICU-acquired weakness used in prior publications. 1,2.

Manual Muscle Testing:  A Method of Measuring Extremity Muscle Strength Applied to Critically Ill Patients

Survivors of acute respiratory distress syndrome (ARDS) and critical illness frequently develop long-lasting muscle weakness. Manual muscle testing (MMT) is a standardized clinical examination commonly used to measure strength of peripheral skeletal muscle groups. This video demonstrates MMT using the 6-point Medical Research Council scale.

危重病人中的应用手册肌力测试：下肢肌力测量方法

Survivors of acute respiratory distress syndrome (ARDS) and other causes of critical illness often have generalized weakness, reduced exercise tolerance, ...

An in vivo Rodent Model of Contraction-induced Injury and Non-invasive Monitoring of Recovery

Muscle strains are one of the most common complaints treated by physicians. A muscle injury is typically diagnosed from the patient history and physical exam alone, however the clinical presentation can vary greatly depending on the extent of injury, the patient's pain tolerance, etc. In patients with muscle injury or muscle disease, assessment of muscle damage is typically limited to clinical signs, such as tenderness, strength, range of motion, and more recently, imaging studies. Biological markers, such as serum creatine kinase levels, are typically elevated with muscle injury, but their levels do not always correlate with the loss of force production. This is even true of histological findings from animals, which provide a "direct measure" of damage, but do not account for all the loss of function. Some have argued that the most comprehensive measure of the overall health of the muscle in contractile force. Because muscle injury is a random event that occurs under a variety of biomechanical conditions, it is difficult to study. Here, we describe an in vivo animal model to measure torque and to produce a reliable muscle injury. We also describe our model for measurement of force from an isolated muscle in situ. Furthermore, we describe our small animal MRI procedure.

An in vivo Rodent Model of Contraction-induced Injury and Non-invasive Monitoring of Recovery

An in vivo animal model of injury is described. The method takes advantage of the subcutaneous position of the fibular nerve. Velocity, timing of muscle activation, and arc of motion are all pre-determined and synchronized using commercial software. Post injury changes are monitored in vivo using MR imaging/spectroscopy.

一个在体内收缩引起的损伤，非侵入性的监测恢复啮齿动物模型

Muscle strains are one of the most  common complaints treated by physicians. A muscle injury is typically diagnosed from the patient history and  physical ...

The Use of Thermal Infra-Red Imaging to Detect Delayed Onset Muscle Soreness

Delayed onset muscle soreness (DOMS), also known as exercise induced muscle damage (EIMD), is commonly experienced in individuals who have been physically inactive for prolonged periods of time, and begin with an unexpected bout of exercise1-4, but can also occur in athletes who exercise beyond their normal limits of training5. The symptoms associated with this painful phenomenon can range from slight muscle tenderness, to severe debilitating pain1,3,5. The intensity of these symptoms and the related discomfort increases within the first 24 hours following the termination of the exercise, and peaks between 24 to 72 hours post exercise1,3. For this reason, DOMS is one of the most common recurrent forms of sports injury that can affect an individual&#x2019;s performance, and become intimidating for many1,4.
For the last 3 decades, the DOMS phenomenon has gained a considerable amount of interest amongst researchers and specialists in exercise physiology, sports, and rehabilitation fields6. There has been a variety of published studies investigating this painful occurrence in regards to its underlying mechanisms, treatment interventions, and preventive strategies1-5,7-12. However, it is evident from the literature that DOMS is not an easy pathology to quantify, as there is a wide amount of variability between the measurement tools and methods used to quantify this condition6. It is obvious that no agreement has been made on one best evaluation measure for DOMS, which makes it difficult to verify whether a specific intervention really helps in decreasing the symptoms associated with this type of soreness or not. Thus, DOMS can be seen as somewhat ambiguous, because many studies depend on measuring soreness using a visual analog scale (VAS)10,13-15, which is a subjective rather than an objective measure. Even though needle biopsies of the muscle, and blood levels of myofibre proteins might be considered a gold standard to some6, large variations in some of these blood proteins have been documented 6,16, in addition to the high risks sometimes associated with invasive techniques. 
Therefore, in the current investigation, we tested a thermal infra-red (IR) imaging technique of the skin above the exercised muscle to detect the associated muscle soreness. Infra-red thermography has been used, and found to be successful in detecting different types of diseases and infections since the 1950&#x2019;s17. But surprisingly, near to nothing has been done on DOMS and changes in skin temperature. The main purpose of this investigation was to examine changes in DOMS using this safe and non-invasive technique.

The purpose of this investigation was to assess whether using an infra-red thermal camera is a valid tool for detecting and quantifying the muscle soreness after exercising.

热红外成像来检测迟发性肌肉酸痛

Delayed onset muscle soreness (DOMS), also known as exercise induced muscle damage (EIMD), is commonly experienced in individuals who have been physically ...

Induction of Acute Skeletal Muscle Regeneration by Cardiotoxin Injection

Skeletal muscle regeneration is a physiological process that occurs in adult skeletal muscles in response to injury or disease. Acute injury-induced skeletal muscle regeneration is a widely used, powerful model system to study the events involved in muscle regeneration as well as the mechanisms and different players. Indeed, a detailed knowledge of this process is essential for a better understanding of the pathological conditions that lead to skeletal muscle degeneration, and it aids in identifying new targeted therapeutic strategies. The present work describes a detailed and reproducible protocol to induce acute skeletal muscle regeneration in mice through a single intramuscular injection of cardiotoxin (CTX). CTX belongs to the family of snake venom toxins and causes myolysis of myofibers, which eventually triggers the regeneration events. The dynamics of skeletal muscle regeneration is evaluated by histological analysis of muscle sections. The protocol also illustrates the experimental procedures for dissecting, freezing, and cutting the Tibialis Anterior muscle, as well as the routine Hematoxylin &#38; Eosin staining that is widely used for subsequent morphological and morphometric analysis.

This manuscript describes a detailed protocol to induce acute skeletal muscle regeneration in adult mice and subsequent manipulations of the muscles, such as dissection, freezing, cutting, routine staining, and myofiber cross-sectional area analysis.

通过注射心脏毒素急性骨骼肌再生的诱导

Skeletal muscle regeneration is a physiological process that occurs in adult skeletal muscles in response to injury or disease. Acute injury-induced ...

发育生物学

Human Skeletal Muscle Biopsy Procedures Using the Modified Bergstr&#246;m Technique

The percutaneous biopsy technique enables researchers and clinicians to collect skeletal muscle tissue samples. The technique is safe and highly effective. This video describes the percutaneous biopsy technique using a modified Bergstr&#246;m needle to obtain skeletal muscle tissue samples from the vastus lateralis of human subjects. The Bergstr&#246;m needle consists of an outer cannula with a small opening (&#8216;window&#8217;) at the side of the tip and an inner trocar with a cutting blade at the distal end. Under local anesthesia and aseptic conditions, the needle is advanced into the skeletal muscle through an incision in the skin, subcutaneous tissue, and fascia. Next, suction is applied to the inner trocar, the outer trocar is pulled back, skeletal muscle tissue is drawn into the window of the outer cannula by the suction, and the inner trocar is rapidly closed, thus cutting or clipping the skeletal muscle tissue sample. The needle is rotated 90&#176;&#160;and another cut is made. This process may be repeated three more times. This multiple cutting technique typically produces a sample of 100-200 mg or more in healthy subjects and can be done immediately before, during, and after a bout of exercise or other intervention. Following post-biopsy dressing of the incision site, subjects typically resume their activities of daily living right away and can fully participate in vigorous physical activity within 48-72 hr. Subjects should avoid heavy resistance exercise for 48 hr to reduce the risk of herniation of the muscle through the incision in the fascia.

The purpose of this video is to present the modified Bergstr&#246;m skeletal muscle biopsy technique on human subjects.

人骨骼肌活检手术使用修改伯格斯坦技术

The percutaneous biopsy technique enables researchers and clinicians to collect skeletal muscle tissue samples. The technique is safe and highly ...

Ex Vivo Assessment of Contractility, Fatigability and Alternans in Isolated Skeletal Muscles

Described here is a method to measure contractility of isolated skeletal muscles. Parameters such as muscle force, muscle power, contractile kinetics, fatigability, and recovery after fatigue can be obtained to assess specific aspects of the excitation-contraction coupling (ECC) process such as excitability, contractile machinery and Ca2+ handling ability. This method removes the nerve and blood supply and focuses on the isolated skeletal muscle itself. We routinely use this method to identify genetic components that alter the contractile property of skeletal muscle though modulating Ca2+ signaling pathways. Here, we describe a newly identified skeletal muscle phenotype, i.e., mechanic alternans, as an example of the various and rich information that can be obtained using the in vitro muscle contractility assay. Combination of this assay with single cell assays, genetic approaches and biochemistry assays can provide important insights into the mechanisms of ECC in skeletal muscle.

Ex Vivo Assessment of Contractility, Fatigability and Alternans in Isolated Skeletal Muscles

We describe a method to directly measure muscle force, muscle power, contractile kinetics and fatigability of isolated skeletal muscles in an in vitro system using field stimulation. Valuable information on Ca2+ handling properties and contractile machinery of the muscle can be obtained using different stimulating protocols.

体外评估对离体骨骼肌的收缩，疲劳性和交替“

 
Described here is a method to measure contractility of isolated skeletal muscles. Parameters such as muscle force, muscle power, contractile kinetics, ...

生物学

Isometric and Eccentric Force Generation Assessment of Skeletal Muscles Isolated from Murine Models of Muscular Dystrophies

Critical to the evaluation of potential therapeutics for muscular disease are sensitive and reproducible physiological assessments of muscle function. Because many pre-clinical trials rely on mouse models for these diseases, isolated muscle function has become one of the standards for Go/NoGo decisions in moving drug candidates forward into patients. We will demonstrate the preparation of the extensor digitorum longus (EDL) and diaphragm muscles for functional testing, which are the predominant muscles utilized for these studies. The EDL muscle geometry is ideal for isolated muscle preparations, with two easily accessible tendons, and a small size that can be supported by superfusion in a bath. The diaphragm exhibits profound progressive pathology in dystrophic animals, and can serve as a platform for evaluating many potential therapies countering fibrosis, and promoting myofiber stability. Protocols for routine testing, including isometric and eccentric contractions, will be shown. Isometric force provides assessment of strength, and eccentric contractions help to evaluate sarcolemma stability, which is disrupted in many types of muscular dystrophies. Comparisons of the expected results between muscles from wildtype and dystrophic muscles will also be provided. These measures can complement morphological and biochemical measurements of tissue homeostasis, as well as whole animal assessments of muscle function.

Muscle function measurements contribute to the evaluation of potential therapeutics for muscle pathology, as well as to the determination of mechanisms underlying physiology of this tissue. We will demonstrate the preparation of the extensor digitorum longus and diaphragm muscles for functional testing. Protocols for isometric and eccentric contractions will be shown, as well as differences in results between dystrophic muscles, representing a pathological state, and wildtype muscles.

等距和骨骼肌产生偏心力评估隔离肌营养不良症的小鼠模型

Critical to the evaluation of potential  therapeutics for muscular disease are sensitive and reproducible physiological  assessments of muscle function. ...

Skeletal Muscle Neurovascular Coupling, Oxidative Capacity, and Microvascular Function with 'One Stop Shop' Near-infrared Spectroscopy

Exercise represents a major hemodynamic stress that demands a highly coordinated neurovascular response in order to match oxygen delivery to metabolic demand. Reactive hyperemia (in response to a brief period of tissue ischemia) is an independent predictor of cardiovascular events and provides important insight into vascular health and vasodilatory capacity. Skeletal muscle oxidative capacity is equally important in health and disease, as it determines the energy supply for myocellular processes. Here, we describe a simple, non-invasive approach using near-infrared spectroscopy to assess each of these major clinical endpoints (reactive hyperemia, neurovascular coupling, and muscle oxidative capacity) during a single clinic or laboratory visit. Unlike Doppler ultrasound, magnetic resonance images/spectroscopy, or invasive catheter-based flow measurements or muscle biopsies, our approach is less operator-dependent, low-cost, and completely non-invasive. Representative data from our lab taken together with summary data from previously published literature illustrate the utility of each of these end-points. Once this technique is mastered, application to clinical populations will provide important mechanistic insight into exercise intolerance and cardiovascular dysfunction.

Here, we describe a simple, non-invasive approach using near-infrared spectroscopy to assess reactive hyperemia, neurovascular coupling and skeletal muscle oxidative capacity in a single clinic or laboratory visit.

"一站式" 近红外光谱对骨骼肌神经血管的耦合、氧化能力及微血管功能的作用

Exercise represents a major hemodynamic stress that demands a highly coordinated neurovascular response in order to match oxygen delivery to metabolic ...

Non-invasive Assessment of Dorsiflexor Muscle Function in Mice

Assessment of skeletal muscle contractile function is an important measurement for both clinical and research purposes. Numerous conditions can negatively affect skeletal muscle. This can result in a loss of muscle mass (atrophy) and/or loss of muscle quality (reduced force per unit of muscle mass), both of which are prevalent in chronic disease, muscle-specific disease, immobilization, and aging (sarcopenia). Skeletal muscle function in animals can be evaluated by a range of different tests. All tests have limitations related to the physiological testing environment, and the selection of a specific test often depends on the nature of the experiments. Here, we describe an in vivo, non-invasive technique involving a helpful and easy assessment of force frequency-curve (FFC) in mice that can be performed on the same animal over time. This permits monitoring of disease progression and/or efficacy&#160;of a potential therapeutic treatment.

Measurement of rodent skeletal muscle contractile function is a useful tool that can be used to track disease progression as well as efficacy of therapeutic intervention. We describe here the non-invasive, in vivo assessment of the dorsiflexor muscles that can be repeated over time in the same mouse.

小鼠肌功能的无创评价

Assessment of skeletal muscle contractile function is an important measurement for both clinical and research purposes. Numerous conditions can negatively ...