这项工作得到了中国国家自然科学基金委员会（第81922076号、81973951号、81873373号）、上海市合作创新建设项目（第81922076号）的支持。ZYJKFW201811010，ZYJKFW201701005）。

所有动物实验均经上海中医药大学动物实验伦理委员会审核通过（伦理许可号PZSHUTCM190308020）。
1. 高平台的制造
<ol><li>将支撑装置的长度和宽度设置为 86 厘米 x 17 厘米（图 1）。</li>
	<li>垂直修复连接设备以连接到支持设备。连接装置应长约30厘米，以防止大鼠跑到高平台的顶部表面，宽度为14厘米。</li>
	<li>将水平踏板（5 厘米 x 5 厘米）固定到连接设备上。水平踏板底部的悬浮高度应大于或等于 50 厘米。 注:根据实验鼠的数量，可以使用多个水平踏板，每个踏板上放置一只大鼠。</li>
</ol>2. 建立哮喘的老鼠模型
<ol><li>用 OVA 使大鼠敏感<ol>
<li>在第 0 天准备感化解决方案。用1毫克OVA和10毫克氢氧化铝准备每1毫升的感化溶液，以普通盐水作为溶剂。</li>
		<li>在第0天给每只大鼠注射1mL的敏感溶液。使用合适的注射器绘制感性解决方案，供以后使用。</li>
		<li>抓住尾巴，抬起老鼠，放在粗糙的表面上，轻轻地把尾巴拉回来。用左拇指和食指抓住耳朵和脖子的皮肤，同时用剩下的三根手指和手掌紧紧握住背部皮肤（小心不要施加会窒息老鼠的过度力）。</li>
		<li>用右手，用1mL的感官溶液准备注射器，在大鼠中进行腹膜内注射。</li>
	</ol>
</li>
	<li>挑战大鼠与OVA<ol>
<li>选择带盖子的盒子（长度、宽度和高度为 39 厘米 x 28 厘米 x 23 厘米），并在侧面钻一个小孔。用橡胶管连接密封的盒子、雾化杯和压缩雾化器（补充图1）。 注:压缩雾化器的星云粒子大小为1-5微米，雾化率大于0.2 mL/min，最终压力为205 kPa。</li>
		<li>在敏感后的第14天，打开自制密封盒的盖子，将老鼠放在盒子里（盒子可以容纳5只老鼠），然后把密封的盒子放在一个生物安全柜里。</li>
		<li>准备挑战解决方案。将300毫克OVA与30毫升普通盐水彻底混合。</li>
		<li>打开超声波雾化器，将 30 mL 的挑战解决方案添加到雾化杯中。根据自制密封盒的体积调整挑战溶液的剂量和大鼠数量。</li>
		<li>重复数次（每天一次）或根据哮喘症状的严重程度调整程序。</li>
	</ol>
</li>
</ol>3. 针灸治疗
<ol><li>根据治疗哮喘的中医理论，选择包括GV14、双边BL12和双边BL13在内的穴位（补充图2）。在第2天进行手动针灸，每隔一天进行一次，共7次（图2）。
	<ol>
<li>针灸GV14位于第七颈椎和第一胸椎（肩骨的空心）之间，在背部中线（图3）。</li>
		<li>针灸 BL13 位于第三个胸椎以下，位于腹腔间空间的两侧（肩骨的下边缘）。</li>
		<li>针灸 BL12 位于第二个胸椎以下，位于间歇空间的两侧（在肩骨下方，根据 BL13 确定的位置）。</li>
	</ol>
</li>
	<li>抓住老鼠，把它们放在高高的平台上。</li>
	<li>用右手拇指和食指握住一次性不锈钢针（0.3毫米 x 13.0 毫米， 辅助图 3）手柄，用右手手掌按鼠背。 注:一次性针头必须仅一次性使用。因为反复使用会使针头钝化，从而增加大鼠的疼痛。</li>
	<li>触摸上边缘 （GV14） 或肩骨下边缘 （BL12 和 BL13） 中间的凹陷，然后用针尖轻轻刺破大鼠的皮肤以定位穴位。</li>
	<li>确定穴位的具体位置后，快速插入针头，并迅速移除右手。</li>
	<li>均匀地扭动针头约360°，速度为60次/分钟，10-20秒，每5分钟操纵一次，然后在20-30分钟后取出（补充图4）。</li>
</ol>4. 笔测量
<ol><li>准备甲基苯甲基苯甲酰:将100毫克甲基胆碱放入5mL离心管中，并将PBS添加到2mL（50毫克/mL甲基苯甲酰胆碱）的体积中。然后，将 50 毫克/mL 甲基胆碱溶液稀释到 25 毫克/mL、12.5 毫克/mL、6.25 毫克/mL 和 3.125 毫克/mL。</li>
	<li>仪器检查<ol>
<li>打开信号放大器（气溶胶分布系统和啮齿动物偏置流供应系统全部关闭，星云连接器关闭）。</li>
		<li>检查通道:单击 校准，将粗调整基线返回到 0，然后切换到直流 （DC） 位置。将基线微调到 0，用注射器注入 5 mL 气体。然后，切换到交替电流 （AC） 位置。</li>
		<li>检查通道后，确保误差值小于 0.5%。</li>
	</ol>
</li>
	<li>设置参数<ol>
<li>打开气溶胶分布系统和啮齿动物偏置流供应系统。根据汞球上的液体水平将流速调整到 4.5 mL。</li>
		<li>点击 创建研究选项 ，并选择 剂量响应研究，选择 老鼠 的物种，并点击 下一步。</li>
		<li>选择 简单的剂量列表，将 剂量数 设置为 6， 然后单击 下一个。</li>
		<li>选择 Penh 和 区域 参数:将动物适应时间设置为 5 分钟，雾化时间设置为 2 分钟，响应时间设置为 3 分钟，恢复时间设置为 2 分钟。</li>
		<li>依次向雾化器添加以下解决方案（每只大鼠 200 μL）:甲基胆碱（0 毫克/mL、3.125 毫克/mL、6.25 毫克/mL、12.5 毫克/mL、25 毫克/mL 和 50 毫克/mL）。可以观察到 Penh 的实时变化 （图 4）。</li>
		<li>在响应所有 6 种不同浓度的解决方案后，单击 "文件 "和 "结束会话 "以完成实验。</li>
	</ol>
</li>
</ol>

<ol>
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J., Li, J., Zeng, B. Y., Liang, F. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=History+of+acupuncture+research.">History of acupuncture research.</a> International Review of Neurobiology. 111, 1-23 (2013).</li><li>Yang, Y. Q., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Considerations+for+use+of+acupuncture+as+supplemental+therapy+for+patients+with+allergic+asthma.">Considerations for use of acupuncture as supplemental therapy for patients with allergic asthma.</a> Clinical Reviews in Allergy &amp; Immunology. 44 (3), 254-261 (2013).</li><li>Nurwati, I., 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=Reduction+of+interleukin-17+level+by+acupuncture+at+Feishu+(BL13)+is+strengthened+by+acupuncture+at+Zusanli+(ST36)+in+a+mouse+model+of+chronic+asthma:+An+experimental+study.">Reduction of interleukin-17 level by acupuncture at Feishu (BL13) is strengthened by acupuncture at Zusanli (ST36) in a mouse model of chronic asthma: An experimental study.</a> Medical Acupuncture. 27 (4), 278-282 (2015).</li><li>Nurwati, I., 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=Improvement+in+inflammation+and+airway+remodelling+after+acupuncture+at+BL13+and+ST36+in+a+mouse+model+of+chronic+asthma.">Improvement in inflammation and airway remodelling after acupuncture at BL13 and ST36 in a mouse model of chronic asthma.</a> Acupuncture in Medicine. 37 (4), 228-236 (2019).</li><li>Reddy, A. 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作者没有什么可透露的。

本研究中使用的GV14、双边BL12和双边BL13穴位位于背面，因此高平台适合固定大鼠。在其他研究中，应根据穴位15的位置选择适当的固定方法。对于腹部的穴位，大鼠可以向下放置在一个黑色的圆锥体（类似于面包师使用的糕点袋）17。链条连接到约束锥固定，大鼠的后腿也固定。这样，动物身体的前部被牢牢地固定在圆锥体中，而腹部和背部则暴露在外。对于腿部的穴位，大鼠可以垂直放置在一个特殊的透明桶中，将尾巴和后腿自然地放在水桶18之外。透明的桶盖住老鼠的身体，但方便在腿上进行针灸。
执行这些方法时应考虑几个关键步骤。在针灸治疗之前，大鼠应提前几天被放置在高平台上进行训练，直到大鼠在高平台上保持安静和平稳呼吸。对于针灸操作，右手的手掌越难按回大鼠，插入针头就越容易。在呼吸功能测量方面， WBP 系统测量了的变化，这些变化以非侵入性的方式反映大鼠的肺抵抗力变化，并最大限度地减少心理压力的影响。应尽量减少外部环境对大鼠的影响。大鼠应放在测试室，然后用塑料袋盖住测试室（在后部侧有1厘米×1厘米的孔进行通风），以减少外部环境造成的刺激。
这个实验的局限性是，如果大鼠在针灸过程中摆动，针头可能会脱落。因此，必须每 5 分钟操纵一次需要调整深度。此外，高平台适合大鼠，但不适合小动物，如老鼠。
综上所述，我们详细描述了在针灸过程中无限制地制造一个固定大鼠的高平台的方法。高平台可以完全暴露老鼠背部的穴位。使用高平台和非侵入性呼吸功能测量可促进针灸作为哮喘治疗的分析。研究针灸的治疗机制，可以为哮喘、慢性阻塞性肺病等呼吸道疾病带来新的治疗方法。

哮喘是一种慢性疾病，其特征是支气管收缩和气道反应过度。全世界约有3.394亿人被诊断患有哮喘2。全球哮喘患病率正在上升，预计未来十年将新增1亿例。吸入皮质类固醇和长效β 2-agonists不能有效地控制所有患者的症状4。因此，使用针灸等替代疗法治疗哮喘已受到越来越多的关注。治疗哮喘的有效方法是针GV14（大泽），双边BL12（丰门）和双边BL13（飞舒）6。针灸显著降低过敏性哮喘患者唾液和鼻分泌物中分泌免疫球蛋白A（SIgA）的水平，减少周围血液中嗜血球菌的计数，总有效率为85%6。慢性哮喘小鼠的BL13和ST36（Zusanli）针灸可降低IL-17水平40%，与椭圆形布明（OVA）组相比，平滑肌肉厚度降低33%，从而缓解炎症症状7、8。然而，哮喘针灸治疗的机制并不完全清楚。
动物模型是哮喘研究的重要工具9，因为动物模型可用于持续测量呼吸功能，以评估哮喘治疗10的有效性。同时，哮喘病理生理学的研究需要测试样本，包括气管、肺和支气管厕所的样本，以验证关键因素11水平的变化。哮喘的老鼠模型通常用于评估哮喘病理生理学，因为它们产生持久的气道反应，并显示即时和后期气道反应12。然而，由于反复的实验刺激，大鼠往往变得易怒。因此，需要一种合适的方法来修复老鼠。常用的老鼠固定方法包括麻醉和结合13。虽然麻醉方法提供了更好的暴露穴位，它可能会影响神经传导，并最终影响实验结果14。结合方法不会诱导与麻醉相同的生理效果，但受约束的老鼠会经历骨骼肌肉紧张。此外，针头不能轻易到达指定位置15。在这里，我们引入了一个高平台，可用于修复大鼠，而无需限制它们，并充分暴露背面的穴位。在针灸管理期间，大鼠可以放在高平台上。高平台只能容纳老鼠，因为它被放置在离地面一定距离的地方，老鼠不会移动，因为害怕高度16。
本文详细描述了高平台的制造、哮喘鼠模型的建立、高平台上需要大鼠的操作以及使用全身全息摄影（WBP）系统测量呼吸功能。

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			<td height="42" style="height:42px;width:267px;">0.3 mm x 13.0 mm acupuncture needle</td>
			<td style="width:192px;">China Suzhou Medical Device Factory</td>
			<td style="width:114px;"></td>
			<td style="width:154px;"></td>
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			<td height="25" style="height:24px;width:267px;">Al(OH)3</td>
			<td style="width:192px;">American Thermo</td>
			<td style="width:114px;">77161</td>
			<td></td>
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			<td height="42" style="height:42px;">Compression nebulizer</td>
			<td style="width:192px;">Jiangsu Lude Medical Electronics Co., Ltd.</td>
			<td style="width:114px;">NB-212C</td>
			<td></td>
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			<td height="21" style="height:21px;width:267px;">Fine Pointe V2.0</td>
			<td style="width:192px;">American Buxco</td>
			<td style="width:114px;"></td>
			<td></td>
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			<td height="21" style="height:21px;width:267px;">Laboratory gas drying unit</td>
			<td style="width:192px;">American Drierite</td>
			<td style="width:114px;">26800</td>
			<td></td>
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			<td height="21" style="height:21px;width:267px;">Methacholine</td>
			<td style="width:192px;">American Sigma</td>
			<td style="width:114px;">MKCF6054</td>
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			<td height="21" style="height:21px;width:267px;">Ovalbumin</td>
			<td style="width:192px;">American Sigma</td>
			<td style="width:114px;">A5503-25G</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:267px;">Phosphate Buffer solution (1x)</td>
			<td style="width:192px;">Gelifesciences</td>
			<td style="width:114px;">SH30256.01</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:267px;">Seal box</td>
			<td style="width:192px;">IRIS Corporation of Japan</td>
			<td style="width:114px;"></td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:267px;">Whole-body plethysmography respiratory function measurement system</td>
			<td style="width:192px;">American Buxco</td>
			<td style="width:114px;"></td>
			<td></td>
		</tr>
	</tbody>
</table>

acupuncture in a rat model of asthma

高平台可以不受限制地修复大鼠，并在针灸操作期间完全暴露背部的穴位。本文描述了制造高平台的方法，建立了哮喘的老鼠模型，并使用非侵入性和实时全身全息摄影（WBP）系统测量呼吸功能的变化。

在GV14、双边BL12和双边BL13点进行针灸时，大鼠保持一个容易的位置。老鼠只能把头扭到外面，不能扭到里面，因为一个方向被木板挡住了（图5）。
随着甲基苯甲酰浓度的增加，Penh 呈现出上升趋势，增长率逐渐增加。浓度为3.125毫克/千升、6.25毫克/千升、12.5毫克/mL、25毫克/mL和50毫克/千升， Penh 增长 18.40% ± 10.70%， 27.55% ± 26.30%， 88.10% ± 45.63 %，427.60%±172.92%和882.27%，±121.97%，显示出增加剂量的效果（图6）。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/61762/61762fig1.jpg"> 图1。高平台的制造。 高平台由支撑设备、三个连接设备和三个踏板组成。支撑装置的长度和宽度为86厘米×17厘米，连接装置的长度和宽度为30厘米×14厘米，踏板的长度和宽度为5厘米×5厘米。每个连接设备相隔 20 厘米， <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/61762/61762fig1large.jpg" target="_blank">请单击此处查看此图的较大版本。</a>
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/61762/61762fig2.jpg"> 图2。建立哮喘大鼠模型的议定书。 敏感在第 0 天进行，针灸每隔一天在高平台上进行一次，从第 2 天到第 14 天。挑战之后，将测量 Penh 的变化。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/61762/61762fig2large.jpg" target="_blank">请单击此处查看此图的更大版本。</a>
<img alt="Figure 3" class="xfigimg" src="/files/ftp_upload/61762/61762fig3.jpg"> 图3. GV14（大泽）、BL12（丰门）和BL13（飞舒）对大鼠的解剖位置。（A） 对显示肩骨、第一肋骨、第二肋骨、第三根肋骨和第二胸椎 （T2） 的老鼠背部进行解剖解剖。（B） 肋骨和胸椎的下垂平面。（C） 肩骨的横截面。（D） GV14:位于肩骨的空心处，后部中线:BL13:位于肩骨的下边缘;BL12:位于肩骨下方，根据BL13定位。<a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/61762/61762fig3large.jpg" target="_blank">请单击此处查看此图的更大版本。</a>
<img alt="Figure 4" class="xfigimg" src="/files/ftp_upload/61762/61762fig4.jpg"> 图4。呼吸功能的实时变化。 蓝色曲线显示 Penh 的实时变化。绿色曲线显示一定时期内潮汐体积 （TVb） 的总变化。紫色曲线显示 Penh 在一定时期内的总体变化。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/61762/61762fig4large.jpg" target="_blank">请单击此处查看此图的更大版本。</a>
<img alt="Figure 5" class="xfigimg" src="/files/ftp_upload/61762/61762fig5.jpg"> 图5。 GV14（大泽）、BL12（丰门）和BL13（飞树）在大鼠的高平台上针灸。（B） 在高平台上对针灸大鼠的横向视图。大鼠被放置在平台上，并在针灸期间保持在易发生位置。<a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/61762/61762fig5large.jpg" target="_blank">请单击此处查看此图的更大版本。</a>
<img alt="Figure 6" class="xfigimg" src="/files/ftp_upload/61762/61762fig6.jpg"> 图6。彭的测量。 WBP 用于测量正常大鼠的 Penh。随着甲基苯甲酰的浓度增加，边也逐渐增加。数据以平均± SEM（n = 3）的形式呈现。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/61762/61762fig6large.jpg" target="_blank">请单击此处查看此图的更大版本。</a>
补充图1。雾化器规格。压缩雾化器的星云颗粒大小为1 -5微米，雾化率大于0.2mL/min，最终压力为205 kPa。<a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/61762/61762supfig1large.jpg" target="_blank">请单击此处下载此文件。</a>
补充图2。GV14（大泽）、BL12（丰门）和BL13（飞舒）在大鼠中的示意图位置，<a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/61762/61762supfig2large.jpg" target="_blank">请点击这里下载此文件。</a>
补充 图3. 针头规格。 针的长度是 13 毫米， 直径是 0.3 毫米。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/61762/61762supfig3large.jpg" target="_blank">请点击这里下载这个文件。</a>
补充图4.针灸操作的示意图。插入针头后，均匀地扭曲针头约360°在不同的方向。<a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/61762/61762supfig4large.jpg" target="_blank">请单击此处下载此文件。</a>

Watch this Scientific Journal Video about Acupuncture in a Rat Model of Asthma at JoVE.com

Acupuncture in a Rat Model of Asthma

哮喘鼠模型中的针灸

A high platform can fix rats without restriction and completely expose the acupoints on the back during acupuncture manipulation. This article describes ...

Nanyang Technological University

Research

JoVE Journal

Medicine

5.9K 次观看.  Shanghai University of Traditional Chinese Medicine. 高平台可以不受限制地修复大鼠，并在针灸操作期间完全暴露背部的穴位。本文描述了制造高平台的方法，建立了哮喘的老鼠模型，并使用非侵入性和实时全身全息摄影（WBP）系统测量呼吸功能的变化。

视频: Acupuncture in a Rat Model of Asthma

Bronchial Thermoplasty:  A Novel Therapeutic Approach to Severe Asthma

Bronchial thermoplasty is a non-drug procedure for severe persistent asthma that delivers thermal energy to the airway wall in a precisely controlled manner to reduce excessive airway smooth muscle. Reducing airway smooth muscle decreases the ability of the airways to constrict, thereby reducing the frequency of asthma attacks. Bronchial thermoplasty is delivered by the Alair System and is performed in three outpatient procedure visits, each scheduled approximately three weeks apart. The first procedure treats the airways of the right lower lobe, the second treats the airways of the left lower lobe and the third and final procedure treats the airways in both upper lobes. After all three procedures are performed the bronchial thermoplasty treatment is complete.
Bronchial thermoplasty is performed during bronchoscopy with the patient under moderate sedation. All accessible airways distal to the mainstem bronchi between 3 and 10 mm in diameter, with the exception of the right middle lobe, are treated under bronchoscopic visualization. Contiguous and non-overlapping activations of the device are used, moving from distal to proximal along the length of the airway, and systematically from airway to airway as described previously. Although conceptually straightforward, the actual execution of bronchial thermoplasty is quite intricate and procedural duration for the treatment of a single lobe is often substantially longer than encountered during routine bronchoscopy. As such, bronchial thermoplasty should be considered a complex interventional bronchoscopy and is intended for the experienced bronchoscopist. Optimal patient management is critical in any such complex and longer duration bronchoscopic procedure. This article discusses the importance of careful patient selection, patient preparation, patient management, procedure duration, postoperative care and follow-up to ensure that bronchial thermoplasty is performed safely.
Bronchial thermoplasty is expected to complement asthma maintenance medications by providing long-lasting asthma control and improving asthma-related quality of life of patients with severe asthma. In addition, bronchial thermoplasty has been demonstrated to reduce severe exacerbations (asthma attacks) emergency rooms visits for respiratory symptoms, and time lost from work, school and other daily activities due to asthma.

Bronchial thermoplasty is a non-drug procedure for severe persistent asthma that delivers thermal energy to the airway wall in a precisely controlled manner to reduce excessive airway smooth muscle. Reducing airway smooth muscle decreases the ability of the airways to constrict, thereby reducing the frequency of asthma attacks.

支气管Thermoplasty:严重哮喘的一种新的治疗方法

Bronchial thermoplasty is a non-drug procedure for severe persistent asthma that delivers thermal energy to the airway wall in a precisely controlled ...

科研

医学

Angiogenesis in the Ischemic Rat Lung

The adult lung is perfused by both the systemic bronchial artery and the entire venous return flowing through the pulmonary arteries. In most lung pathologies, it is the smaller systemic vasculature that responds to a need for enhanced lung perfusion and shows robust neovascularization. Pulmonary vascular ischemia induced by pulmonary artery obstruction has been shown to result in rapid systemic arterial angiogenesis in man as well as in several animal models. Although the histologic assessment of the time course of bronchial artery proliferation in rats was carefully described by Weibel 1, mechanisms responsible for this organized growth of new vessels are not clear. We provide surgical details of inducing left pulmonary artery ischemia in the rat that leads to bronchial neovascularization. Quantification of the extent of angiogenesis presents an additional challenge due to the presence of the two vascular beds within the lung. Methods to determine functional angiogenesis based on labeled microsphere injections are provided.

The lung is perfused by both the systemic bronchial artery and pulmonary arteries. In most lung pathologies, it is the smaller systemic vasculature that shows robust neovascularization. Cessation of pulmonary blood flow promotes brisk bronchial angiogenesis. We provide surgical details of inducing left pulmonary artery ischemia that promotes bronchial neovascularization.

在缺血大鼠肺的血管生成

The  adult lung is perfused by both the systemic bronchial artery and the entire  venous return flowing through the pulmonary arteries. In most lung ...

A Model of Chronic Nutrient Infusion in the Rat

Chronic exposure to excessive levels of nutrients is postulated to affect the function of several organs and tissues and to contribute to the development of the many complications associated with obesity and the metabolic syndrome, including type 2 diabetes. To study the mechanisms by which excessive levels of glucose and fatty acids affect the pancreatic beta-cell and the secretion of insulin, we have established a chronic nutrient infusion model in the rat. The procedure consists of catheterizing the right jugular vein and left carotid artery under general anesthesia; allowing a 7-day recuperation period; connecting the catheters to the pumps using a swivel and counterweight system that enables the animal to move freely in the cage; and infusing glucose and/or Intralipid (a soybean oil emulsion which generates a mixture of approximately 80% unsaturated/20% saturated fatty acids when infused with heparin) for 72 hr. This model offers several advantages, including the possibility to finely modulate the target levels of circulating glucose and fatty acids; the option to co-infuse pharmacological compounds; and the relatively short time frame as opposed to dietary models. It can be used to examine the mechanisms of nutrient-induced dysfunction in a variety of organs and to test the effectiveness of drugs in this context.

A protocol for chronic infusions of glucose and Intralipid in rats is described. This model can be used to study the impact of nutrient excess on organ function and physiological parameters.

在大鼠模型慢性营养输液

Chronic exposure to excessive levels of nutrients is postulated to  affect the function of several organs and tissues and to contribute to the  ...

Intrarenal Injection of Escherichia coli in a Rat Model of Pyelonephritis

Pyelonephritis is a bacterial infection of the kidney and is most commonly caused by Escherichia coli. Recurrent infections can cause significant renal inflammation and fibrosis ultimately resulting in declining kidney function. Before improved clinical management and prevention of pyelonephritis can be instituted, a reliable animal model must be established in order to study the mechanisms of progression, recurrence, and therapeutic efficacy. The transurethral infection model closely mimics human pyelonephritis but exhibits considerable variation due to its reliance on urethral reflux to transport the bacteria to the kidney. Herein, a detailed surgical protocol for performing bacterial injections into the rat renal pelvis is provided and confirmed by non-invasive Magnetic Resonance Imaging (MRI). Using this protocol, animals receive direct exposure to a desired concentration of E. coli bacteria and can fully recover from the surgical procedure with adequate post-operative care. This facilitates subsequent longitudinal MRI assessments of the experimental animal models for comparison with saline (sham) controls. Using this direct delivery approach, the severity of infection is controllable and applicable for mechanistic studies of progression as well as development of novel treatment strategies.

Intrarenal Injection of Escherichia coli in a Rat Model of Pyelonephritis

This manuscript describes a rat surgical model of pyelonephritis using direct intra-renal infection by Escherichia coli into the renal pelvis. The experimental procedure can be utilized to study the pathogenesis of pyelonephritis as well as the associated inflammation and fibrosis.

肾内注射<em&gt;大肠杆菌</em&gt;在肾性肾炎大鼠模型中

Pyelonephritis is a bacterial infection of the kidney and is most commonly caused by Escherichia coli. Recurrent infections can cause significant renal ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and drugs. It is widely used in many studies on the physiological functions of vascular smooth muscle, the pathogenesis of vascular diseases such as hypertension, and the development of smooth muscle relaxant drugs. The mouse is a widely used model animal with a large pool of disease models and genetically modified strains. We introduced this method to measure the isometric contraction of mouse mesenteric artery in detail. A 1.4-mm segment of mouse mesenteric resistance artery was isolated and mounted on a myograph chamber by passing two steel wires through its lumen. After equilibration and normalization steps, the vessel segment was potentiated by a high-K+ solution twice prior to the contraction assay. As an example of the application of this method in drug development, we measured the relaxant effect of a novel natural substance, neoliensinine, isolated from a Chinese herb, embryos of the lotus seed (Nelumbo nucifera Gaertn.) on mouse mesenteric arteries. The vessel segments mounted on the myograph chamber were stimulated with a high-K+ solution. When the force tension reached a stable sustained phase, cumulative doses of neoliensinine were added to the chamber. We found that neoliensinine had a dose-dependent relaxant effect on smooth muscle contraction, thus suggesting that it bears potential activity against hypertension. In addition, as the vessel segment can survive at least 4 hours after mounting and maintain contractility induced by the high-K+ solution for many times, we suggest that the wire myograph system may be used for the time-consuming process of drug screening.

The wire myograph technique is used to investigate vascular smooth muscle functions and screen new drugs. We report a detailed protocol for measuring the isometric contractility of the mouse mesenteric artery and for screening new relaxants of vascular smooth muscle.

用线 Myography 测定小鼠肠系膜动脉的等轴收缩力

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...

Retinal Pathophysiological Evaluation in a Rat Model

A posterior segment eye disease like diabetic retinopathy alters the physiology of the retina. Diabetic retinopathy is characterized by a retinal detachment, breakdown of the blood-retinal barrier (BRB), and retinal angiogenesis. An in vivo rat model is a valuable experimental tool to examine the changes in the structure and function of the retina. We propose three different experimental techniques in the rat model to identify morphological changes of retinal cells, retinal vasculature, and compromised BRB. Retinal histology is used to study the morphology of various retinal cells. Also, quantitative measurement is performed by retinal cell count and thickness measurement of different retinal layers. A BRB breakdown assay is used to determine the leakage of extraocular proteins from the plasma to vitreous tissue due to the breakdown of BRB. Fluorescence angiography is used to study angiogenesis and leakage of blood vessels by visualizing retinal vasculature using FITC-dextran dye.

Diabetic retinopathy is one of the leading causes of blindness. Histology, blood-retinal barrier breakdown assay, and fluorescence angiography are valuable techniques to understand the pathophysiology of the retina, which could further enhance the efficient drug screening against diabetic retinopathy.

大鼠模型中的视网膜病理生理学评估

A posterior segment eye disease like diabetic retinopathy alters the physiology of the retina. Diabetic retinopathy is characterized by a retinal ...

A Model of Long-Term Ventricular Fibrillation in Isolated Rat Hearts

Ventricular fibrillation (VF) is a fatal arrhythmia with a high incidence in cardiac patients, but VF arrest under perfusion is a neglected method of intraoperative arrest in the field of cardiac surgery. With recent advances in cardiac surgery, the demand for prolonged VF studies under perfusion has increased. However, the field lacks simple, reliable, and reproducible animal models of chronic ventricular fibrillation. This protocol induces long-term VF through alternating current (AC) electrical stimulation of the epicardium. Different conditions were used to induce VF, including continuous stimulation with a low or high voltage to induce long-term VF and stimulation for 5 min with a low or high voltage to induce spontaneous long-term VF. The success rates of the different conditions, as well as the rates of myocardial injury and recovery of cardiac function, were compared. The results showed that continuous low-voltage stimulation induced long-term VF and that 5 min of low-voltage stimulation induced spontaneous long-term VF with mild myocardial injury and a high rate of recovery of cardiac function. However, the low-voltage, continuously stimulated long-term VF model had a higher success rate. High-voltage stimulation provided a higher rate of VF induction but showed a low defibrillation success rate, poor recovery of cardiac function, and severe myocardial injury. On the basis of these results, continuous low-voltage epicardial AC stimulation is recommended for its high success rate, stability, reliability, reproducibility, low impact on cardiac function, and mild myocardial injury.

This protocol presents a model of long-term ventricular fibrillation in rat hearts induced by continuous stimulation with low-voltage alternating current. This model has a high success rate, is stable, reliable, and reproducible, has a low impact on cardiac function, and causes only mild myocardial injury.

孤立大鼠心脏长期心室颤动模型

Ventricular fibrillation (VF) is a fatal arrhythmia with a high incidence in cardiac patients, but VF arrest under perfusion is a neglected method of ...

改进大鼠模型中的 FSN 治疗:最大限度地减少不适并最大限度地提高效率

Efficacy of Fu's Subcutaneous Needling on Sciatic Nerve Pain: Behavioral and Electrophysiological Changes in a Chronic Constriction Injury Rat Model

Fu&#39;s subcutaneous needling (FSN), an invented acupuncture technique from traditional Chinese medicine, is used worldwide for pain relief. However, the mechanisms of action are still not fully understood. During FSN treatment, the FSN needle is inserted and retained in the subcutaneous tissues for a long duration with a swaying movement. However, challenges arise from maintaining a posture while manipulating FSN in animal models (e.g., rats) for researchers. Uncomfortable treatment can lead to fear and resistance to FSN needles, increasing the risk of injury and may even affect research data. Anesthesia may also affect the study results too. Hence, there is a need for strategies in FSN therapy on animals that minimize injury during the intervention. This study employs a chronic constriction injury model in Sprague-Dawley rats to induce neuropathic pain. This model replicates the pain induced by nerve injury observed in humans through surgical constriction of a peripheral nerve, mimicking the compression or entrapment seen in conditions such as nerve compression syndromes and peripheral neuropathies. We introduce an appropriate manipulation for easily inserting an FSN needle into the subcutaneous layer of the animal&#39;s body, including needle insertion and direction, needle retention, and swaying movement. Minimizing the rat&#39;s discomfort prevents the rat from being tense, which causes the muscle to contract and hinder the entry of the needle and improves the study efficiency.

作者聚焦:揭示 FSN 治疗的治疗效果 – 桥接神经性疼痛的研究和临床应用

We present a protocol for using Fu's subcutaneous needling in a chronic constriction injury model to induce sciatic nerve pain in rats.

傅氏皮下针刺治疗坐骨神经疼痛的疗效:慢性缩窄损伤大鼠模型中的行为和电生理变化

Fu&#39;s subcutaneous needling (FSN), an invented acupuncture technique from traditional Chinese medicine, is used worldwide for pain relief. However, the ...