这项工作得到了国家中医药临床研究基地（JDZX2015127，依托安徽省中医院）第二批专项科研项目的支持。

所有动物实验均经南京中医药大学动物护理与使用专业委员会批准（许可号：202209A033）。选取SPF等级、8周龄、体重20±4g的健康雄性C57BL/6J小鼠。将小鼠饲养在20-22°C的温度下进行12小时的光/暗循环，并保持45%-50%的相对湿度。动物们可以自由地进食和饮水。
1. T2DM小鼠模型的建立
<ol><li>首先以规律的饮食喂养小鼠1周，让小鼠适应动物中心的新环境。1 周后，提供高脂肪饮食（饮食成分：20.0% 猪油、10.0% 蔗糖、2.5% 胆固醇、1.0% 胆酸盐、66.5% 常规饮食），为期 4 周。</li>
	<li>从第5周的第1天开始，腹膜内注射小鼠链脲佐菌素溶液，每天35mg/kg，连续3天。用 10 mg 链脲佐菌素粉末制备 1 mL 链脲佐菌素溶液，以 0.1 M/L 柠檬酸钠缓冲液为溶剂。 注意：注射应在溶液制备后30分钟内完成，使用过程中溶液应避光放置。</li>
	<li>在首次注射链脲佐菌素溶液后第 5 天和第 7 天随机测试血糖。两次随机血糖水平高于 16.7 mM/L 的动物被认为是成功的 T2DM 模型。</li>
	<li>在第一次注射链脲佐菌素溶液后第 8 天开始训练干预。</li>
</ol>2. 小鼠的分组和处理
<ol><li>使用随机数表法将T2DM小鼠分为模型组，训练组和二甲双胍组，每组6只小鼠。</li>
	<li>对模型组不执行任何干预。</li>
	<li>将二甲双胍片研磨成粉末并溶解在纯水中。通过管饲法给予二甲双胍组，剂量为 200 mg/kg，每天一次，持续 3 周。</li>
	<li>让训练组做 30 分钟的静态力量训练，每天一次，每周 5 天，持续 3 周。培训每周一至周五下午举行。</li>
	<li>随机分配 6 只非 T2DM 小鼠作为对照组。这些小鼠不进行建模和干预。</li>
</ol>3.静态力量训练器的制造
<ol><li>准备一块 5 毫米厚的透明亚克力板，尺寸为 20 厘米 x 20 厘米。使用透明板观察小鼠的训练。</li>
	<li>获取直径为 3 毫米的木棍和一卷宽度为 1 厘米的胶带。用切纸刀切两根 4 厘米长的棍子和四根 1 厘米长的棍子。</li>
	<li>使用胶带将两根长度为 1 厘米的棍子连接到长度为 4 厘米的棍子的末端（图 1A）。短棍应与长棍固定在同一侧。</li>
	<li>将透明板放在平坦的表面上，然后使用热胶枪将两组 1 厘米的短棍固定在板上。确保 4 厘米的木棍不与木板接触。在长棍和板之间留出 2 厘米的间隙，以将鼠标固定到位（图 1B）。让两根长棍对齐并平行，它们之间的距离为 6 厘米（图 1C）。</li>
</ol><img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/65556/65556fig01.jpg"> 图 1：将木棍组装并固定到透明板上。 （A） 将 1 厘米的木棍粘在 4 厘米木棍的两端。（B） 使用热熔胶将 1 厘米长的棒子和透明板连接起来，缝隙长度为 2 厘米。 （C） 两根 4 厘米的棒子，间隔 6 厘米。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/65556/65556fig01large.jpg" target="_blank">请点击这里查看此图的较大版本.</a>
4. 小鼠的静态力量训练
<ol><li>剪下两根 15 厘米长的羊毛纱线。使用滑结在小鼠的每个上脚踝上系一根羊毛线（图2A）。 注意：弹性绳不能固定老鼠的姿势，麻绳会磨损老鼠的脚踝，所以这里选择无弹性但柔软的羊毛纱线。</li>
	<li>将板子水平放在桌子上，用棍子朝上。小心地将鼠标按在两根长摇杆之间，将其头部和尾部定位以与摇杆下方的间隙对齐。</li>
	<li>将纱线穿过尾部末端的缝隙。然后，调整纱线，直到鼠标的脚踝紧贴棍子的边缘。</li>
	<li>用胶带固定羊毛纱线（图2B）。羊毛纱的末端应固定好，否则老鼠会爬上掉落的羊毛。将羊毛拉得尽可能紧，否则鼠标将很容易挣脱。</li>
	<li>将透明板倒置。在透明板的两端取 2 个相同高度（15-20 厘米高）的盒子。将板水平放置在一定高度。盒子位于鼠标头部和尾部的两端。如果它太高，实验者将无法轻松处理它;如果温度太低，鼠标可能会接触桌面。</li>
	<li>一旦板子被掀翻，鼠标就会倒挂。由于扶正反射，小鼠卷曲腹部并伸展前肢以抓住后肢或棍子。在这个阶段，将一根 20 厘米的棍子放在老鼠面前，并小心地引导它用前肢抓住棍子。重复该过程，直到鼠标熟练掌握手机摇杆。</li>
	<li>使用棍子将鼠标的前肢移动到透明板上的另一根 4 厘米的棍子上。不断调整棍子的角度，使小鼠能够主动抓住板上静止的4厘米棍子（图2C）。</li>
	<li>重复上一步，鼠标松开前爪，直到鼠标耗尽。30分钟后，大多数老鼠无法抬起上半身并用前肢抓住棍子。 注意：在实施干预之前，有必要对小鼠进行为期一周的习惯训练。最初的训练时间为每天 10 分钟，然后应每天增加 5 分钟，直到达到每天 30 分钟。</li>
	<li>训练 30 分钟后，立即释放鼠标并解开羊毛，以防止因长时间捆绑而导致脚红肿。<ol><li>如果训练后小鼠的脚踝上出现皮肤病变，请停止训练小鼠并寻求兽医专业护理，直到脚踝健康为止。</li>
		<li>一些老鼠可能偶尔会在训练过程中通过激烈的斗争来摆脱自己。当老鼠的后腿没有牢固固定时，它们会挣扎。为避免因挣扎而受伤，请在整个训练过程中观察小鼠。当发现后肢没有牢固固定或小鼠开始挣扎时，松开小鼠并重新固定。 注意：先前的实验表明，每周 5 天，每天倒挂 30 分钟不会导致脚踝磨损，只要它们在适当的时间内释放10。小鼠可以在 1-3 个疗程中学习并习惯训练。</li>
	</ol></li>
</ol><img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/65556/65556fig02.jpg"> 图 2：小鼠的固定方法。 （A） 用滑结系住脚踝顶部。（B） 将绳子的末端穿过缝隙并拉紧，然后用胶带固定。（C）小鼠的静态力量训练。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/65556/65556fig02large.jpg" target="_blank">请点击这里查看此图的较大版本.</a>

探索 T2DM 运动干预：小鼠静态训练装置

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静态力量训练可以减少脂肪堆积，帮助减肥，增加新陈代谢8.此外，它还增强了骨骼肌细胞中 PGC-1α 和线粒体生物发生的表达，从而改善了 2 型糖尿病小鼠的葡萄糖代谢，从而降低了血糖水平11。为了确认静态训练对T2DM的影响和机制，需要开发合适的设备对实验动物进行静态训练。
该协议引入了一种用于小鼠静态训练的设备。所需的设备，如亚克力板、木棒、毛纱、热胶枪等一应俱全，生产方法简单易行。这样可以有效降低实验的成本，提高实验的可行性和可重复性。在设计静态训练设备时，在训练期间保持小鼠的固定位置可能具有挑战性。在该协议中，可以通过收紧柔软的羊毛将鼠标的脚固定在小棍子的前部。扶正反射使鼠标的上半身向上弯曲，从而使其能够用前爪抓住前面的杆子。杆很短，限制了前爪的运动范围。经过两到三次的适应性训练后，小鼠能够保持稳定的姿势。由于前爪的力量有限，老鼠经常失去对横杆的抓握。实验人员有必要全程监测小鼠，以帮助它们用前肢抓住横杆并防止小鼠意外受伤。通过训练，小鼠可以一次保持固定姿势1-2分钟，重复几次，耗尽后约30分钟。当筋疲力尽时，老鼠不再滚动腹部或抬起前腿。当老鼠在棍子的引导下用前爪抓住时，老鼠可以抓住，但上半身无法向上转动。放下老鼠后尽快解开结，可以有效避免脚踝水肿和磨损。
初步实验表明，经过一段时间的静态训练后，与模型小鼠对照组相比，T2DM小鼠的血糖水平明显下降。骨骼肌是外周葡萄糖代谢的关键组成部分，对于维持血糖稳态至关重要12。电子显微镜分析表明，T2DM小鼠的腓肠肌细胞发生了严重的变性，而静态训练可以减轻肌肉组织变性。这些发现表明，静态训练可能与骨骼肌功能的调节有关。研究表明，维持骨骼肌功能受到线粒体动力学的显着影响13。我们观察到，与野生小鼠和静态运动小鼠相比，T2DM小鼠模型组的腓肠肌细胞中线粒体数量显着减少，线粒体活性缺乏。这些发现表明，静态训练可以通过增强骨骼肌细胞中的线粒体功能来促进骨骼肌功能。根据上述实验结果，可以确定静态训练可以显著提高T2DM小鼠的血糖和胰岛素代谢。干预机制可能与静态训练对骨骼肌线粒体功能的调节有关。
该协议有一些限制。最初，一个人单独捆绑小鼠的后肢是很困难的，因为它需要一个人抓住后腿，而另一个人进行捆绑。随着实验者的一些调整和技能的增加，小鼠变得更加容易适应。这允许后腿独立捆绑。此外，柔软的羊毛可以减轻小鼠脚踝的瘀伤。
总之，该协议提供了一种为小鼠制作静态训练设备的简单方法。同样，可以通过增加木板和木棍的尺寸来制作简单的老鼠静态训练设备。该装置可以保持小鼠四肢肌肉的等长收缩，从而验证中医传统运动对T2DM的干预效果，为T2DM的临床治疗提供新的视角。

2 型糖尿病 （T2DM） 是一种以胰岛素抵抗和β细胞功能障碍为特征的慢性疾病，对全球健康构成重大威胁1.运动是管理2型糖尿病的重要干预措施。大量研究表明，太极拳和八段拳等中国传统锻炼方法可显着改善 T2DM2、3、4、5 患者的血糖水平和生活质量。为了执行这些动作，教练必须在一段时间内保持稳定的身体和关节位置。静态姿势是通过进行静态肌肉收缩来维持的，这通常被称为静态力量6。
然而，静态力量训练干预在T2DM中的机制尚未明确。要回答这个问题，动物实验是必不可少的。在等长运动中，肌肉被激活，保持恒定的长度，并安全地达到最大张力7。在静态力量训练的实验中，实验动物需要进行等长肌肉收缩并保持这种肌肉收缩状态。如何对小鼠、大鼠等实验动物进行静态力量训练，成为研究中的一大难题。首先，动物难以服从命令并根据需要收缩肌肉。其次，动物在阻力下难以保持稳定的位置，无法达到等长肌肉收缩的目的。在让动物根据需要进行训练的同时，重要的是要解决与动物福利相关的问题，例如缓解压力和焦虑、尽量减少疼痛和改善整体条件。该协议涉及大鼠8,9的静态训练模型，这里我们介绍一种用于小鼠静态训练的简单设备。当小鼠的后腿被抬起时，它们的腹肌由于扶正反射而收缩，前爪抓住前面的横杆，然后前后肢体在重力的作用下收缩。老鼠在抓住短杆后无法移动，导致它们的肌肉处于等长收缩状态。

<table><tbody><tr><td>Acrylic boards</td><td></td><td></td><td>Transparent acrylic boards with 5mm thickness. The size should be larger than 20cm&amp;times;20cm</td></tr><tr><td>Boxes</td><td></td><td></td><td>Two boxes of the same height (15~20cm)</td></tr><tr><td>ELISA KIT</td><td></td><td>H203-1-2</td><td>Nanjing Jiancheng Bioengineering Institute</td></tr><tr><td>Hot melt glue gun</td><td></td><td></td><td>Avoid touching the gun head to cause burns</td></tr><tr><td>Knives</td><td></td><td></td><td>No special requirement</td></tr><tr><td>Metformin tablets</td><td></td><td>1396309</td><td>Sigma</td></tr><tr><td>scissors</td><td></td><td></td><td>No special requirement</td></tr><tr><td>Sticks</td><td></td><td></td><td>Several wooden sticks with a diameter of 3mm</td></tr><tr><td>Streptozotocin</td><td></td><td>S0130</td><td>Sigma</td></tr><tr><td>Tape</td><td></td><td></td><td>No special requirement</td></tr><tr><td>Transmission Electron Microscope (TEM)</td><td></td><td>HT7700</td><td>HITACHI&amp;nbsp;</td></tr></tbody></table>

static strength training method for type 2 diabetic mice

2型糖尿病（T2DM）的治疗是改善患者健康的一大难点。运动是T2DM的主要干预措施之一。静态力量训练是中国传统体育运动的重要形式之一。研究表明，静态力量训练是T2DM干预的一种有效的临床方法，但目前还没有适合小鼠静态训练的实验设备。从临床研究转向基础研究的困难之一是设计合适的实验设备。为了进一步研究静态训练干预在T2DM中的机制，本文介绍了一种简单的小鼠静态训练装置制作方法。该装置具有操作简单、材料便宜、可行性高等优点。先前根据该协议进行的研究表明，静态训练可以有效降低血糖水平并改善 T2DM 小鼠骨骼肌细胞的线粒体功能。引进该装置的目的是促进传统运动干预T2DM作用机制的研究，为运动的定量干预奠定基础。

按照上述方案，将小鼠的后肢固定，前肢自主抓住前杆。狭窄的运动范围使鼠标保持在相对固定的位置。可以通过触摸腹部和腿部的肌肉来确认小鼠的肌肉收缩。这与静态力量训练中对等长肌肉收缩状态的需求是一致的。按照协议训练小鼠，随着训练时间的增加，将帮助小鼠适应训练，同时减少它们挣扎的欲望。可以避免逃跑的斗争。
对T2DM小鼠血糖和胰岛素水平的影响 共24只C57BL/6J小鼠随机分为对照组（n=6）和T2DM模型组（n=18）。然后将T2DM小鼠随机分为模型组、训练组和二甲双胍组。对照组未给予建模或干预，模型组小鼠未给予干预。训练组的小鼠每天接受一次30分钟的静态力量训练，每周5天，持续3周。二甲双胍组小鼠给予二甲双胍200mg/kg，每日一次，持续3周。干预 3 周后的空腹血糖 （FBG） 结果如图 3A 所示。从图中可以看出，T2DM模型小鼠的FBG水平明显高于对照组小鼠。与模型组相比，在静态训练中训练的小鼠表现出显着降低的 FBG 水平，表明静态训练可有效减少 T2DM 小鼠的 FBG。模型组的空腹血清胰岛素（FINS）水平明显低于对照组， 如图3B所示。在训练组和二甲双胍组中，与模型组相比，FINS水平有所增加。 在图3C中，模型组小鼠的胰岛素抵抗指数（HOMA-IR）明显高于对照组，而静态训练组和二甲双胍组的HOMA-IR明显低于模型组，证明了它们在缓解T2DM小鼠胰岛素抵抗状态方面的功效。这些结果表明，该方案提供的静态力量训练策略在调节T2DM小鼠血糖水平方面与二甲双胍具有相似的效果。
<img alt="Figure 3" class="xfigimg" src="/files/ftp_upload/65556/65556fig03.jpg"> 图 3：血糖和胰岛素水平。 （A） 干预 3 周后 T2DM 小鼠空腹血糖水平的比较。* p <0.05 与对照组相比，# p <0.05 与模型组相比。（B） 干预 3 周后 T2DM 小鼠空腹血清胰岛素水平的比较。* p <0.05 与对照组相比，# p <0.05 与模型组相比。（C） 干预 3 周后 T2DM 小鼠胰岛素抵抗指数的比较。* p <0.05 与对照组相比，# p <0.05 与模型组相比。采用单因素方差分析（ANOVA）进行统计分析。定量数据表示为SEM±平均值（n=6）。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/65556/65556fig03large.jpg" target="_blank">请点击这里查看此图的较大版本.</a>
对T2DM小鼠骨骼肌的影响 通过透射电子显微镜观察小鼠的腓肠肌。与对照小鼠相比，T2DM小鼠的骨骼肌肌细胞退化，肌原纤维结构疏松，肌节排列不规则（图4A，B）。在协议中描述的静态训练之后，图4C显示了肌肉结构中肌源纤维的紧密结构和局部肌肉节段的对称排列。这表明静态力量训练可以调节 T2DM 小鼠的骨骼肌形态。另一方面，在对照小鼠的腓肠肌中，线粒体局部分布，膜完整，线粒体局部分裂或融合（用红色箭头表示）。同样，在静态力量训练后的小鼠骨骼肌中，线粒体的数量是正常的;其中一些显然是融合或分裂的（红色箭头）。相比之下，在小鼠的T2DM模型中，线粒体的数量较少，并且活性较低。（见图4）这表明静态力量训练可能会影响骨骼肌细胞中的线粒体功能和活动。
<img alt="Figure 4" class="xfigimg" src="/files/ftp_upload/65556/65556fig04.jpg"> 图4：静态训练对T2DM小鼠骨骼肌的影响。 （A）对照组;（B） T2DM模型组;（三）静态训练组。M代表线粒体。白色刻度条代表50μm，红色刻度条代表10μm。底部图像是顶部图像的部分放大。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/65556/65556fig04large.jpg" target="_blank">请点击这里查看此图的较大版本.</a>

Watch this Scientific Journal Video about Author Spotlight: Advancing Diabetes Research with Static Exercise Training in Mice at JoVE.com

作者聚焦：通过小鼠静态运动训练推进糖尿病研究

该协议提供了一种为小鼠制作静态训练设备的简单方法。该装置维持小鼠四肢肌肉的等长收缩，以验证传统运动对2型糖尿病（T2DM）的干预效果，为T2DM的临床治疗提供新的运动疗法。

2型糖尿病小鼠静态力量训练方法

The treatment of type 2 diabetes mellitus (T2DM) is a major difficulty in improving patient health. Exercise is one of the main interventions for T2DM. ...

运动疗法在世界范围内被公认为治疗糖尿病的重要手段。在之前的临床试验中，我们证实，非传统的中式运动可以有效缓解患者的二型糖尿病症状。因此，我们设计了这项研究，试图弄清楚传统运动是如何运作的。为了研究运动对糖尿病的影响并获得更多的见解，我们使用代谢混合和转录混合等技术进行研究。影响二型糖尿病的分子机制非常复杂，包括几种遗传、代谢和炎症途径，但挑战在于确定和选择某种途径以最佳方式治疗疾病而又不产生副作用。

static-strength-training-method-for-type-2-diabetic-mice

Nanyang Technological University

Research

JoVE Journal

Medicine

Static Strength Training Method for Type 2 Diabetic Mice

403 次观看.  Nanjing University of Chinese Medicine. 运动疗法在世界范围内被公认为治疗糖尿病的重要手段。在之前的临床试验中，我们证实，非传统的中式运动可以有效缓解患者的二型糖尿病症状。因此，我们设计了这项研究，试图弄清楚传统运动是如何运作的。为了研究运动对糖尿病的影响并获得更多的见解，我们使用代谢混合和转录混合等技术进行研究。影响二型糖尿病的分子机制非常复杂，包括几种遗传、代谢?...

视频: 作者聚焦：通过小鼠静态运动训练推进糖尿病研究

Measuring the Strength of Mice

Kondziela7 devised the inverted screen test and published it in 1964. It is a test of muscle strength using all four limbs. Most normal mice easily score maximum on this task; it is a quick but insensitive gross screen, and the weights test described in this article will provide a finer measure of muscular strength.
There are also several strain gauge-based pieces of apparatus available commercially that will provide more graded data than the inverted screen test, but their cost may put them beyond the reach of many laboratories which do not specialize in strength testing. Hence in 2000 a cheap and simple apparatus was devised by the author. It consists of a series of chain links of increasing length, attached to a "fur collector" a ball of fine wire mesh sold for preventing limescale build up in hard water areas. An accidental observation revealed that mice could grip these very tightly, so they proved ideal as a grip point for a weight-lifting apparatus. A common fault with commercial strength meters is that the bar or other grip feature is not thin enough for mice to exert a maximum grip. As a general rule, the thinner the wire or bar, the better a mouse can grip with its small claws.
This is a pure test of strength, although as for any test motivational factors could potentially play a role. The use of scale collectors, however, seems to minimize motivational problems as the motivation appears to be very high for most normal young adult mice.

Deficits in muscular strength occur in many clinical conditions such as motor neuron disease. The inverted screen and weight lifting tests described here measure strength in mice almost exclusively, with minimal influence of factors such as coordination.

强度测量小鼠

Kondziela7 devised the inverted screen test and published it in 1964. It is a test of muscle strength using all four limbs. Most normal mice easily score ...

科研

医学

Assessment of Murine Exercise Endurance Without the Use of a Shock Grid: An Alternative to Forced Exercise

Using laboratory mouse models, the molecular pathways responsible for the metabolic benefits of endurance exercise are beginning to be defined. The most common method for assessing exercise endurance in mice utilizes forced running on a motorized treadmill equipped with a shock grid. Animals who quit running are pushed by the moving treadmill belt onto a grid that delivers an electric foot shock; to escape the negative stimulus, the mice return to running on the belt. However, avoidance behavior and psychological stress due to use of a shock apparatus can interfere with quantitation of running endurance, as well as confound measurements of postexercise serum hormone and cytokine levels. Here, we demonstrate and validate a refined method to measure running endurance in na&#239;ve C57BL/6 laboratory mice on a motorized treadmill without utilizing a shock grid. When mice are preacclimated to the treadmill, they run voluntarily with gait speeds specific to each mouse. Use of the shock grid is replaced by gentle encouragement by a human operator using a tongue depressor, coupled with sensitivity to the voluntary willingness to run on the part of the mouse. Clear endpoints for quantifying running time-to-exhaustion for each mouse are defined and reflected in behavioral signs of exhaustion such as splayed posture and labored breathing. This method is a humane refinement which also decreases the confounding effects of stress on experimental parameters.

A method to assess exercise endurance in laboratory mice without the use of a shock grid is demonstrated. This method is a humane refinement that can decrease the confounding effects of stress on experimental parameters.

小鼠运动耐力的评估没有一个冲击电网的使用:另一种强迫运动

Using laboratory mouse models, the molecular pathways responsible for the metabolic benefits of endurance exercise are beginning to be defined. The most ...

行为学

A Murine Model of Muscle Training by Neuromuscular Electrical Stimulation

Neuromuscular electrical stimulation (NMES) is a common clinical modality that is widely used to restore1, maintain2 or enhance3-5 muscle functional capacity. Transcutaneous surface stimulation of skeletal muscle involves a current flow between a cathode and an anode, thereby inducing excitement of the motor unit and the surrounding muscle fibers. 
NMES is an attractive modality to evaluate skeletal muscle adaptive responses for several reasons. First, it provides a reproducible experimental model in which physiological adaptations, such as myofiber hypertophy and muscle strengthening6, angiogenesis7-9, growth factor secretion9-11, and muscle precursor cell activation12 are well documented. Such physiological responses may be carefully titrated using different parameters of stimulation (for Cochrane review, see 13). In addition, NMES recruits motor units non-selectively, and in a spatially fixed and temporally synchronous manner14, offering the advantage of exerting a treatment effect on all fibers, regardless of fiber type. Although there are specified contraindications to NMES in clinical populations, including peripheral venous disorders or malignancy, for example, NMES is safe and feasible, even for those who are ill and/or bedridden and for populations in which rigorous exercise may be challenging. 
Here, we demonstrate the protocol for adapting commercially available electrodes and performing a NMES protocol using a murine model. This animal model has the advantage of utilizing a clinically available device and providing instant feedback regarding positioning of the electrode to elicit the desired muscle contractile effect. For the purpose of this manuscript, we will describe the protocol for muscle stimulation of the anterior compartment muscles of a mouse hindlimb.

A murine model of neuromuscular electrical stimulation (NMES), a safe and inexpensive clinical modality, to the anterior compartment muscles is described. This model has the advantage of modifying a readily available clinical device for the purpose of eliciting targeted and specific muscle contractions in mice.

一个小鼠模型，通过神经肌肉电刺激肌肉训练

Neuromuscular electrical stimulation (NMES) is a common clinical modality that is widely used to restore1, maintain2 or enhance3-5 muscle functional ...

Alleviation of Diabetic Tendon Injury via Activation of Tendon Fibroblasts Autophagy under Berberine Treatment

Berberine (BBR) is an isoquinoline alkaloid isolated from Coptis chinensis and possesses valuable pharmacological activities, including anti-inflammatory, anti-tumor, and alleviating several complications of type 2 diabetes mellitus (T2DM). However, the role of BBR in regulating diabetic tendon injury remains poorly understood. In this study, a rat model of T2DM was constructed, and cell apoptosis and autophagy were assessed in tendon tissues after BBR treatment through TdT-Mediated dUTP nick-end labeling (TUNEL) assay and immunohistochemical analysis. Tendon fibroblasts were obtained from the rat Achilles tendon, and the role of BBR in regulating cell apoptosis, the production of inflammatory cytokines, and autophagy activation were assessed using flow cytometry, quantitative real-time PCR (qRT-PCR), and western blot analysis. We demonstrated that BBR treatment significantly increased autophagy activation and decreased cell apoptosis in tendon tissues of T2DM rats. In tendon fibroblasts, BBR repressed High glucose (HG)-induced cell apoptosis and production of proinflammatory cytokines. HG treatment resulted in a decrease of autophagy activation in tendon fibroblasts, whereas BBR restored autophagy activation. More important, pharmacological inhibition of autophagy by 3-MA weakened the protective effects of BBR against HG-induced tendon fibroblasts injury. Taken together, the current results demonstrate that BBR helps relieve diabetic tendon injury by activating autophagy of tendon fibroblasts.

Berberine (BBR) is an isoquinoline alkaloid isolated from Coptis chinensis and possesses valuable pharmacological activities, including anti-inflammatory, ...

生物化学

Improving Strength, Power, Muscle Aerobic Capacity, and Glucose Tolerance through Short-term Progressive Strength Training Among Elderly People

This protocol describes the simultaneous use of a broad span of methods to examine muscle aerobic capacity, glucose tolerance, strength, and power in elderly people performing short-term resistance training (RET). Supervised progressive resistance training for 1 h three times a week over 8 weeks was performed by RET participants (71&#177;1 years, range 65-80). Compared to a control group without training, the RET showed improvements on the measures used to indicate strength, power, glucose tolerance, and several parameters of muscle aerobic capacity. Strength training was performed in a gym with only robust fitness equipment. An isokinetic dynamometer for knee extensor strength permitted the measurement of concentric, eccentric, and static strength, which increased for the RET group (8-12% post- versus pre-test). The power (rate of force development, RFD) at the initial 0-30 ms also showed an increase for the RET group (52%). A glucose tolerance test with frequent blood glucose measurements showed improvements only for the RET group in terms of blood glucose values after 2 h (14%) and the area under the curve (21%). The blood lipid profile also improved (8%). From muscle biopsy samples prepared using histochemistry, the amount of fiber type IIa increased, and a trend towards a decrease in IIx in the RET group reflected a change to a more oxidative profile in terms of fiber composition. Western blot (to determine the protein content related to the signaling for muscle protein synthesis) showed a rise of 69% in both Akt and mTOR in the RET group; this also showed an increase in mitochondrial proteins for OXPHOS complex II and citrate synthase (both ~30%) and for complex IV (90%), in only the RET group. We demonstrate that this type of progressive resistance training offers various improvements (e.g., strength, power, aerobic capacity, glucose tolerance, and plasma lipid profile).

The effect of short-term resistance training on elderly people was investigated through the simultaneous use of several methods. Compared to a control group, many improvements were seen, including on muscle aerobic capacity, glucose tolerance, strength, power, and muscle quality (i.e., protein involved in cell signaling and muscle fiber type composition).

通过长期进步力量训练提高力量，力量，肌肉有氧能力和葡萄糖耐量

This protocol describes the simultaneous use of a broad span of methods to examine muscle aerobic capacity, glucose tolerance, strength, and power in ...

Dosage-Adjusted Resistance Training in Mice with a Reduced Risk of Muscle Damage

Progressive resistance training (PRT), which involves performing muscle contractions against progressively greater external loads, can increase muscle mass and strength in healthy individuals and in patient populations. There is a need for precision rehabilitation tools to test the safety and effectiveness of PRT to maintain and/or restore muscle mass and strength in preclinical studies on small and large animal models. The PRT methodology and device described in this article can be used to perform dosage-adjusted resistance training (DART). The DART device can be used as a standalone dynamometer to objectively assess the concentric contractile torque generated by the ankle dorsiflexors in mice or can be added to a pre-existing isokinetic dynamometry system. The DART device can be fabricated with a standard 3D printer based on the instructions and open-source 3D print files provided in this work. The article also describes the workflow for a study to compare contraction-induced muscle damage caused by a single bout of DART to muscle damage caused by a comparable bout of isometric contractions (ISOM) in a mouse model of limb-girdle muscular dystrophy type 2B/R2 (BLAJ mice). The data from eight BLAJ mice (four animals for each condition) suggest that less than 10% of the tibialis anterior (TA) muscle was damaged from a single bout of DART or ISOM, with DART being less damaging than ISOM.

The present protocol describes a unique technique called dosage-adjusted resistance training (DART), which can be incorporated into precision rehabilitation studies performed in small animals, such as mice.

降低肌肉损伤风险的小鼠剂量调整阻力训练

Progressive resistance training (PRT), which involves performing muscle contractions against progressively greater external loads, can increase muscle ...

神经科学

In vivo Measurement of Knee Extensor Muscle Function in Mice

Skeletal muscle plasticity in response to countless conditions and stimuli mediates concurrent functional adaptation, both negative and positive. In the clinic and the research laboratory, maximal muscular strength is widely measured longitudinally in humans, with knee extensor musculature the most reported functional outcome. Pathology of the knee extensor muscle complex is well documented in aging, orthopedic injury, disease, and disuse; knee extensor strength is closely related to functional capacity and injury risk, underscoring the importance of reliable measurement of knee extensor strength. Repeatable, in vivo assessment of knee extensor strength in pre-clinical rodent studies offers valuable functional endpoints for studies exploring osteoarthritis or knee injury. We report an in vivo and non-invasive protocol to repeatedly measure isometric peak tetanic torque of the knee extensors in mice across time. We demonstrate consistency using this novel method to measure knee extensor strength with repeated assessment in multiple mice producing similar results.

Quantification of knee extensor maximal strength is imperative to understand functional adaptations to aging, disease, injury, and rehabilitation. We present a novel method to repeatedly measure in vivo knee extension isometric peak tetanic torque.

小鼠膝盖外展肌肉功能的体内测量

Skeletal muscle plasticity in response to countless conditions and stimuli mediates concurrent functional adaptation, both negative and positive. In the ...

生物学

Versatility of Protocols for Resistance Training and Assessment Using Static and Dynamic Ladders in Animal Models

Resistance training is a physical exercise model with profound benefits for health throughout life. The use of resistance exercise animal models is a way to gain insight into the underlying molecular mechanisms that orchestrate these adaptations. The aim of this article is to describe exercise models and training protocols designed for strength training and evaluation of resistance in animal models and provide examples. In this article, strength training and resistance evaluation are based on ladder climbing activity, using static and dynamic ladders. These devices allow a variety of training models as well as provide precise control of the main variables which determine resistance exercise: volume, load, velocity, and frequency. Furthermore, unlike resistance exercise in humans, this is a forced exercise. Thus, aversive stimuli must be avoided in this intervention to preserve animal welfare. Prior to implementation, a detailed design is necessary, along with an acclimatization and learning period. Acclimatization to training devices, such as ladders, weights, and clinical tape, as well as to the manipulations required, is necessary to avoid exercise rejection and to minimize stress. At the same time, the animals are taught to climb up the ladder, not down, to the resting area on the top of the ladder. Resistance evaluation can characterize physical strength and permit adjusting and quantifying the training load and the response to training. Furthermore, different types of strength can be evaluated. Regarding training programs, with appropriate design and device use, they can be sufficiently versatile to modulate different types of strength. Furthermore, they should be flexible enough to be modified depending on the adaptive and behavioral response of the animals or the presence of injuries. In conclusion, resistance training and assessment using ladders and weights are versatile methods in animal research.

The present protocol describes resistance training and testing using static and dynamic ladders in animal models.

在动物模型中使用静态和动态梯子进行阻力训练和评估方案的多功能性

Resistance training is a physical exercise model with profound benefits for health throughout life. The use of resistance exercise animal models is a way ...

A Simple and Inexpensive Running Wheel Model for Progressive Resistance Training in Mice

Previously developed rodent resistance-based exercise models, including synergistic ablation, electrical stimulation, weighted-ladder climbing, and most recently, weighted-sled pulling, are highly effective at providing a hypertrophic stimulus to induce skeletal muscle adaptations. While these models have proven invaluable for skeletal muscle research, they are either invasive or involuntary and labor-intensive. Fortunately, many rodent strains voluntarily run long distances when given access to a running wheel. Loaded wheel running (LWR) models in rodents are capable of inducing adaptations commonly observed with resistance training in humans, such as increased muscle mass and fiber hypertrophy, as well as stimulation of muscle protein synthesis. However, the addition of moderate wheel load either fails to deter mice from running great distances, which is more reflective of an endurance/resistance training model, or the mice discontinue running nearly entirely due to the method of load application. Therefore, a novel high-load wheel running model (HLWR) has been developed for mice where external resistance is applied and progressively increased, enabling mice to continue running with much higher loads than previously utilized. Preliminary results from this novel HLWR model suggest it provides sufficient stimulus to induce hypertrophic adaptations over the 9 week training protocol. Herein, the specific procedures to execute this simple yet inexpensive progressive resistance-based exercise training model in mice are described.

This procedure describes a translatable progressive loaded running wheel resistance training model in mice. The primary advantage of this resistance training model is that it is entirely voluntary, thus reducing stress for the animals and the burden on the researcher.

一种简单且廉价的跑轮模型，用于小鼠的渐进式阻力训练

Previously developed rodent resistance-based exercise models, including synergistic ablation, electrical stimulation, weighted-ladder climbing, and most ...

<meta charset="utf8"></head><body>高脂饮食和 STZ 注射治疗糖尿病心肌病的小鼠模型

Modeling and Evaluation of Murine Diabetic Cardiomyopathy Model

The underlying pathophysiological mechanisms of diabetic cardiomyopathy (DbCM), a leading cause of mortality among patients with type 2 diabetes mellitus (T2DM), remain poorly understood. The myocardial toxicity associated with T2DM is attributed to factors such as lipotoxicity, glucotoxicity, oxidative stress, reduced cardiac efficiency, and lipoapoptosis. Compared to rats, mice offer greater accessibility, cost-effectiveness, and broader applicability for animal experiments. Insulin resistance and impaired insulin secretion are crucial factors in the pathophysiology of T2DM. We introduce a novel nongenetic murine model that replicates the progression of human DbCM induced by a combination of high-fat diet (HFD) feeding and streptozotocin (STZ) injection. In this study, we used wild-type C57BL/6J mice, administering an HFD regimen for 12 weeks, followed by intraperitoneal injections of STZ for an additional 12 weeks to induce characteristic manifestations of T2DM. We conducted oral glucose tolerance tests and measured serum insulin concentrations to confirm the development of insulin resistance and insufficient insulin secretion. Cardiac structure and function were rigorously assessed through noninvasive transthoracic echocardiography. Pathological characteristics were evaluated through Masson's trichrome staining and wheat germ agglutinin (WGA) staining, revealing pathological features related to DbCM. Therefore, we provide a robust and versatile method for establishing a nongenetic murine model of DbCM.

<meta charset="utf8"></head><body>小鼠糖尿病心肌病模型的建模与评价

This protocol outlines a method for inducing diabetic cardiomyopathy through a combination of high-fat diet feeding and streptozotocin injection. This approach aims to provide a reliable framework for scientific investigation into diabetic cardiomyopathy and to explore potential avenues for clinical treatment applications.

小鼠糖尿病心肌病模型的建模与评价

The underlying pathophysiological mechanisms of diabetic cardiomyopathy (DbCM), a leading cause of mortality among patients with type 2 diabetes mellitus ...