作者感谢克里斯蒂安Pahrmann的技术援助。 SS收到的资金从德意志研究联合会（DFG）（SCHR992/3-2和SCHR992/4-2）。

所有的动物工作应按照有关动物护理指引进行。获得批准的机构动物工作开始前协议。 1，肌内膜增生模型（一） 对于血管内膜增生模型，购买C57BL/6J（股票编号000664，C57BL/6J）小鼠在8周体重大约25克的年龄。房子老鼠常规条件下这些实验;喂小鼠标准鼠标饲料，并提供水随意 。 <ol><li> 鼠标的准备：介绍鼠标采用感应腔麻醉状态，异氟醚（2％）。 <ol><li>用头发剪取出腹毛，奠定了鼠标在它的后面，覆盖它的鼻子和嘴的面罩以保证麻醉。 </li><li>与普罗沃碘，消毒腹部区域普遍，其次是80％的乙醇，在两个周期。 </li><li>确保不存在反射捏住后肢监测麻醉深度足够的。 </li><li>分离皮肤和肌肉沿两个步骤，露出腹部器官的白线。 </li><li>躺在肠子在盐水滋润手套。环绕肠子的手套，以保持湿润。 </li><li>认真清除脂肪组织压片腹主动脉。暴露下腹主动脉至其分叉处，小心不要伤到任何分支血管。 </li><li>首先，放置一个显微外科钳上的近端，下腹主动脉关闭血流量。现在，适用旁边主动脉分叉第二远端钳。 </li><li>当血液流动被中断，使一个小的切口进入主动脉靠近近侧钳。 </li><li>插入球囊导管尖，这已被加湿用0.9％生理盐水中，并前进朝向主动脉的远端夹具。 </li><li>对于内皮损伤，通过虚报了巴鲁剥夺主动脉Ñ​​小心缓慢地拉出导管进入近端夹具的方向。 </li><li>重复剥蚀机动两次。 </li><li>取出导管和使用10-0普理灵缝线运行关闭主动脉切口。 </li><li>小心打开远端钳​​。若在织脉切开术部位出血，再次关闭夹，找到出血，并把额外的中断针，是必要的。 </li><li>如果没有出血可以观察，慢慢打开近端钳。 </li><li>主动脉脉冲应该​​从切口向远侧可见。现在肠子可以安排回原位 。 </li><li>冲洗，利用预热的无菌生理盐水腹腔。 </li><li>关闭腹壁开始与肌肉层用6-0 Prolene线连续缝合。 </li><li>接下来适应皮肤使用5-0普理灵进行连续缝合下。 </li><li>在当时的鼠标还在麻醉，适用4-5毫克/公斤卡洛芬通过皮下麟蹄CTION。 </li><li>加入安乃近的饮用水（每100毫升50毫克安乃近）进行疼痛控制和持续3天手术后。监测动物在整个恢复。 注：观察期为这个模型是28天。 </li></ol></li></ol> 2，动脉粥样硬化模型（B） 对于动脉粥样硬化模型，购买载脂蛋白E缺陷（ApoE基因- / - ）小鼠（股份编号002052，B6.129P2-Apoetm1Unc / J）在4周龄时，再喂4-6个月。房子老鼠常规条件下这些实验;小鼠喂高脂饮食西部抵达时（哈伦实验室TD.88137）和4至6个月期间提供自由饮水 。 <ol><li> 鼠标的准备： <ol><li>喂的ApoE - / - 小鼠与西方的饮食开始，在4周龄。饮食控制应直接抵达时开始。 </li><li>维持西方饮食在整个实验持续时间。&lt;/ LI&gt; </ol></li></ol> 3。分析<ol><li> 肌内膜增生模型（一） 在此模型中感兴趣的区域是“病变区域”。分析选项包括管腔闭塞，I / M比值（内膜/中膜比），最大myointima厚度，肌内膜面积，三色染色纤维化，和H＆E染色的单个核细胞（如如图1A-1C）。根据科学问题众多其他染色像天狼猩红胶原蛋白，免疫组织化学和共焦的方法可以被应用。 </li><li> 动脉粥样硬化模型（B） 在此模型中感兴趣的区域是主动脉瓣，主动脉弓以及降主动脉。分析选项：苏丹红染色血脂和主动脉瓣，主动脉弓和降主动脉（ 如图1D）的三色染色。 这种模式也产生了各种像其他染色选项例如油红O为中性脂肪，天狼猩红，免疫组织化学和激光共聚焦方法。 <ol><li>苏丹红染色：对刚收获的主动脉组织执行苏丹红染色。 <ol><li>收获前去除饮食6小时。 </li><li>介绍鼠标利用感应腔麻醉状态，用异氟烷（2％）。按捏后肢监测麻醉深度足以确保不存在反射消失。 </li><li>打开胸腔;切断心脏的顶点和通过左心室灌注，先用生理盐水（3毫升）中，随后加入4％PFA（3ml）中通过主动脉根部修复主动脉。 </li><li>从根释放到主动脉分叉处。尽量去除脂肪组织，尽可能多地。 </li><li>切断心脏和主动脉根部，并把它们在4％PFA存储的组织学。 </li><li>用于组织学的主动脉根部有仔细放置在石蜡块在垂直位置，以允许AOR的切割抽动根生成幻灯片，显示所有3瓣膜小叶。 </li><li>切开主动脉树的一端固定到硅层用细销。纵向打开主动脉和​​修复主动脉组织。 </li><li>冲洗的组织用50％的乙醇。浸入所述组织与约15分钟苏丹III染色溶液。 </li><li>用50％乙醇冲洗轻轻除去过量的苏丹III。 （注：50％的乙醇可洗染料的路程。）冲洗DH 2 O，并拍照作进一步的分析。 </li></ol></li></ol></li></ol>

<ol>
	<li>Manuel, D. G., Leung, M., Nguyen, K., Tanuseputro, P., Johansen, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Burden+of+cardiovascular+disease+in+Canada.">Burden of cardiovascular disease in Canada.</a> Can J Cardiol. 19, 997-1004 (2003).</li><li>ER, O. B., Ma, X., Simard, T., Pourdjabbar, A., Hibbert, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pathogenesis+of+neointima+formation+following+vascular+injury.">Pathogenesis of neointima formation following vascular injury.</a> Cardiovasc Hematol Disord Drug Targets. 11, 30-39 (2011).</li><li>Ross, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+pathogenesis+of+atherosclerosis:+a+perspective+for+the+1990s.">The pathogenesis of atherosclerosis: a perspective for the 1990s.</a> Nature. 362, 801-809 (1993).</li><li>Meng, X., Zhang, F., Valji, K., 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=Ultrasound+guidance+for+the+creation+of+a+small+animal+model+of+aortic+injury.">Ultrasound guidance for the creation of a small animal model of aortic injury.</a> J Vasc Interv Radiol. 22, 1193-1197 (2011).</li><li>Deuse, T., Koyanagi, T., Erben, R. G., 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=Sustained+inhibition+of+epsilon+protein+kinase+C+inhibits+vascular+restenosis+after+balloon+injury+and+stenting.">Sustained inhibition of epsilon protein kinase C inhibits vascular restenosis after balloon injury and stenting.</a> Circulation. 122, 170-178 (2010).</li><li>Schrepfer, S., Deuse, T., Sultan, K. R., 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=Inhibition+of+restenosis+development+after+mechanical+injury:+a+new+field+of+application+for+malononitrilamides.">Inhibition of restenosis development after mechanical injury: a new field of application for malononitrilamides.</a> Cardiology. 108, 128-137 (2007).</li><li>Tao, M., Mauro, C. R., Yu, P., 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=A+simplified+murine+intimal+hyperplasia+model+founded+on+a+focal+carotid+stenosis.">A simplified murine intimal hyperplasia model founded on a focal carotid stenosis.</a> Am J Pathol. 182, 277-287 (2013).</li><li>Kumar, A., Lindner, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Remodeling+with+neointima+formation+in+the+mouse+carotid+artery+after+cessation+of+blood+flow.">Remodeling with neointima formation in the mouse carotid artery after cessation of blood flow.</a> Arterioscler Thromb Vasc Biol. 17, 2238-2244 (1997).</li><li>Hua, X., Deuse, T., Michelakis, E. D., 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=Human+internal+mammary+artery+(IMA)+transplantation+and+stenting:+a+human+model+to+study+the+development+of+in-stent+restenosis.">Human internal mammary artery (IMA) transplantation and stenting: a human model to study the development of in-stent restenosis.</a> J Vis Exp. , 3663 (2012).</li><li>Daugherty, A., Whitman, S. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantification+of+atherosclerosis+in+mice.">Quantification of atherosclerosis in mice.</a> Methods Mol Biol. , (2003).</li><li>Nakashima, Y., Plump, A. S., Raines, E. W., Breslow, J. L., Ross, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=ApoE-deficient+mice+develop+lesions+of+all+phases+of+atherosclerosis+throughout+the+arterial+tree.">ApoE-deficient mice develop lesions of all phases of atherosclerosis throughout the arterial tree.</a> Arterioscler Thromb. 14, 133-1340 (1994).</li><li>Zhang, Y., Li, L., You, J., Cao, J., Fu, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+7-difluoromethyl-5,+4'-dimethoxygenistein+on+aorta+atherosclerosis+in+hyperlipidemia+ApoE(-/-)+mice+induced+by+a+cholesterol-rich+diet.">Effect of 7-difluoromethyl-5, 4'-dimethoxygenistein on aorta atherosclerosis in hyperlipidemia ApoE(-/-) mice induced by a cholesterol-rich diet.</a> Drug Des Devel Ther. 7, 233-242 (2013).</li><li>Rudolph, T. K., Rudolph, V., Edreira, M. 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=Nitro-fatty+acids+reduce+atherosclerosis+in+apolipoprotein+E-deficient+mice.">Nitro-fatty acids reduce atherosclerosis in apolipoprotein E-deficient mice.</a> Arterioscler Thromb Vasc Biol. 30, 938-9345 (2010).</li><li>Rondelet, B., Kerbaul, F., Motte, S., 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=Bosentan+for+the+prevention+of+overcirculation-induced+experimental+pulmonary+arterial+hypertension.">Bosentan for the prevention of overcirculation-induced experimental pulmonary arterial hypertension.</a> Circulation. 107, 1329-1335 (2003).</li><li>Zeibig, S., Li, Z., Wagner, S., 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=Effect+of+the+oxLDL+binding+protein+Fc-CD68+on+plaque+extension+and+vulnerability+in+atherosclerosis.">Effect of the oxLDL binding protein Fc-CD68 on plaque extension and vulnerability in atherosclerosis.</a> Circ Res. 108, 695-703 (2011).</li><li>Painter, T. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Myointimal+hyperplasia:+pathogenesis+and+implications.+2.+Animal+injury+models+and+mechanical+factors.">Myointimal hyperplasia: pathogenesis and implications. 2. Animal injury models and mechanical factors.</a> Artificial organs. 15, 103-118 (1991).</li><li>Nitschke, Y., Weissen-Plenz, G., Terkeltaub, R., Rutsch, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Npp1+promotes+atherosclerosis+in+ApoE+knockout+mice.">Npp1 promotes atherosclerosis in ApoE knockout mice.</a> Journal of cellular and molecular. 15, 2273-2283 (2011).</li></ol>

作者什么都没有透露。

虽然这两种疾病导致在患者类似的症状，斑块发展的基本机制，因此，治疗方法有很大的不同2。在不同形式的动脉狭窄的病人的临床表现以及斑块发展的时间表取决于底层机制。 取决于在患者中的临床发现，样品的分析方法的不同，必须执行5,15。有必要分析适应于这种疾病的特征的研究结果，是被模仿由所选择的模型14。反之亦然，这取决于疾病正?...

在工业化国家，缺血性心脏疾病的死亡率仍1的主要原因。起因于冠状动脉狭窄是多方面的，包括肌内膜增生和动脉粥样硬化，血管再通与剩余的最常见的治疗策略2。研究模型内膜增生和动脉粥样硬化的机制途径明确区分是必不可少的探讨动脉斑块发展的病理生物学和病理生理过程。在这段视频中用于研究肌内膜增生或动脉粥样硬化无论是发展的两个不同的小鼠模型进行了介绍。 肌内膜增生是假设，形成通过最初描述为动脉粥样硬化3“响应到伤害”的范式。内皮细胞层的机械破坏导致一个强烈的重塑通过旁分泌作用增强。有几种二fferent动物模型通常用于研究肌内膜增生。有些团体使用的金属器件在大鼠4的升主动脉。用气囊导管进行的主动脉剥脱模型也常用于实验室大鼠5,6。在实验室老鼠7执行的血管内膜增生模型实现了颈动脉结扎诱导肌内膜病变8。在我们的实验室中，腹主动脉剥脱模型-研究肌内膜增生的发展-以及一个人性化的支架内再狭窄模型9常用。这部影片强调，选择合适的实验动物模型是动脉狭窄的机械或病理生理的研究是至关重要的。 肌内膜增生模型必须从动脉粥样硬化模型的区别。对于后者，载脂蛋白E缺陷（ApoE基因 - / - ）小鼠与西方饮食组合常用于诱导atheroscl色情病变10,11,12,13。对于这两种类型的小鼠肌内膜疾病的诱导例子如下所示，随着不同的模型适应选择，分析血管狭窄14。

<table border="0" cellpadding="0" cellspacing="0" style="width:617px;" width="618">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="17">
			<td height="17" style="height:17px;width:156px;">Name</td>
			<td style="width:183px;">Company</td>
			<td style="width:139px;">Catalog number</td>
			<td style="width:140px;">Comments</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Thyoglycollate Broth 3%</td>
			<td>Fluka</td>
			<td align="right">70157</td>
			<td>powder</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">PFA 4%</td>
			<td>Electron Microscopy Sciences</td>
			<td>#157135S</td>
			<td align="right">20%</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Sudan III staining solution</td>
			<td>Sigma Aldrich</td>
			<td>S4131</td>
			<td>powder</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">mouse C57BL/6J</td>
			<td>Jackson Laboratories</td>
			<td>&#160;Stock # 0006664</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">mouse ApoE-/-</td>
			<td>Jackson Laboratories</td>
			<td>&#160;Stock #002052</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Western Diet</td>
			<td>Harlan Laboratories</td>
			<td>TD.88137</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">hair clipper</td>
			<td>WAHL</td>
			<td>8786-451A ARCO SE</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Forene</td>
			<td>Abbott</td>
			<td></td>
			<td>Isoflurane</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">microsurgical clamp</td>
			<td>Fine Science Tools</td>
			<td>18055-04</td>
			<td>Micro-Serrefine - 4mm</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">clamp applicator</td>
			<td>Fine Science Tools</td>
			<td>18056-14</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">catheter</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">10-0 prolene suture</td>
			<td>Ethicon</td>
			<td>788G</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">6-0 prolene suture</td>
			<td>Ethicon</td>
			<td>8709H</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">5-0 prolene suture</td>
			<td>Ethicon</td>
			<td>EH7229H</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Rimadyl</td>
			<td>Pfizer&#160;</td>
			<td></td>
			<td>Carprofen</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Metacam 1.5mg/ml</td>
			<td>Boehringer Ingelheim</td>
			<td></td>
			<td>Metamizol</td>
		</tr>
	</tbody>
</table>

inducing myointimal hyperplasia versus atherosclerosis in mice: an introduction of two valid models

已经建立了各种体内实验室啮齿动物模型中对动脉狭窄的诱导以模仿包括动脉斑块的形成和狭窄，如在缺血性心脏疾病观察例如疾病。两个高度可重复的小鼠模型 - 既导致动脉狭窄，但每个底层不同的途径发展 - 在这里介绍。该模型代表动脉狭窄的两种最常见的原因;即会显示一个鼠标型号为每个肌内膜增生，动脉粥样硬化。为了诱导肌内膜增生，进行腹主动脉球囊导管损伤。对于粥样硬化斑块的发展，ApoE基因 - / - 与西方脂肪的饮食组合的小鼠模型被使用。对结果的衡量和评估不同的模式适应选项将在本手稿命名和描述。这两款车型的引进和比较提供了信息离子为科学家选择在按照要求科学问题的相应动脉狭窄模型。

图1显示了不同的分析选项，并为这两个模型中引起的疾病状态。对于内膜增生模型，示出了在三色染色的代表横截面的分析。从这些横截面，如I / M比，最大斑块厚度，最大内膜厚度在管腔内，以百分比管腔闭塞，以及斑块面积的结果可以被测量和计算。 为了评估动脉粥样硬化模型中的结果，可以使用所有上面提到的方法。此外，定量斑块面积在腹主动脉，斑?...

Watch this Scientific Journal Video about Inducing Myointimal Hyperplasia Versus Atherosclerosis in Mice: An Introduction of Two Valid Models at JoVE.com

Inducing Myointimal Hyperplasia Versus Atherosclerosis in Mice: An Introduction of Two Valid Models

该视频显示两个模型内膜斑块的发展在小鼠动脉，并强调在肌内膜增生和动脉粥样硬化的差异。

诱导肌内膜增生对战动脉粥样硬化小鼠：两种有效的模型介绍

Various in vivo laboratory rodent models for the induction of artery stenosis have been established to mimic diseases that include arterial plaque ...

inducing-myointimal-hyperplasia-versus-atherosclerosis-mice-an

Research

JoVE Journal

Medicine

12.0K 次观看.  University Hospital Hamburg. 该视频显示两个模型内膜斑块的发展在小鼠动脉，并强调在肌内膜增生和动脉粥样硬化的差异。 

视频: Inducing Myointimal Hyperplasia Versus Atherosclerosis in Mice: An Introduction of Two Valid Models

Implantation of a Carotid Cuff for Triggering Shear-stress Induced Atherosclerosis in Mice

It is widely accepted that alterations in vascular shear stress trigger the expression of inflammatory genes in endothelial cells and thereby induce atherosclerosis (reviewed in 1 and 2). The role of shear stress has been extensively studied in vitro investigating the influence of flow dynamics on cultured endothelial cells 1,3,4 and in vivo in larger animals and humans 1,5,6,7,8. However, highly reproducible small animal models allowing systematic investigation of the influence of shear stress on plaque development are rare. Recently, Nam et al. 9 introduced a mouse model in which the ligation of branches of the carotid artery creates a region of low and oscillatory flow. Although this model causes endothelial dysfunction and rapid formation of atherosclerotic lesions in hyperlipidemic mice, it cannot be excluded that the observed inflammatory response is, at least in part, a consequence of endothelial and/or vessel damage due to ligation.
In order to avoid such limitations, a shear stress modifying cuff has been developed based upon calculated fluid dynamics, whose cone shaped inner lumen was selected to create defined regions of low, high and oscillatory shear stress within the common carotid artery 10. By applying this model in Apolipoprotein E (ApoE) knockout mice fed a high cholesterol western type diet, vascular lesions develop upstream and downstream from the cuff. Their phenotype is correlated with the regional flow dynamics 11 as confirmed by in vivo Magnetic Resonance Imaging (MRI) 12: Low and laminar shear stress upstream of the cuff causes the formation of extensive plaques of a more vulnerable phenotype, whereas oscillatory shear stress downstream of the cuff induces stable atherosclerotic lesions 11. In those regions of high shear stress and high laminar flow within the cuff, typically no atherosclerotic plaques are observed.
 
In conclusion, the shear stress-modifying cuff procedure is a reliable surgical approach to produce phenotypically different atherosclerotic lesions in ApoE-deficient mice.

The constricting cuff presented in this article is designed to induce atherosclerosis in the murine common carotid artery. Due to the conical shape of its inner lumen the implanted cuff generates well-defined regions of low, high and oscillatory shear stress triggering the development of atherosclerotic lesions of different inflammatory phenotypes.

植入小鼠触发剪应力致动脉粥样硬化的颈袖

It is widely accepted that alterations in vascular shear stress trigger the expression of inflammatory genes in endothelial cells and thereby induce ...

科研

医学

Quantitative Analysis and Characterization of Atherosclerotic Lesions in the Murine Aortic Sinus

Atherosclerosis is a disease of the large arteries and a major underlying cause of myocardial infarction and stroke. Several different mouse models have been developed to facilitate the study of the molecular and cellular pathophysiology of this disease. In this manuscript we describe specific techniques for the quantification and characterization of atherosclerotic lesions in the murine aortic sinus and ascending aorta. The advantage of this procedure is that it provides an accurate measurement of the cross-sectional area and total volume of the lesion, which can be used to compare atherosclerotic progression across different treatment groups. This is possible through the use of the valve leaflets as an anatomical landmark, together with careful adjustment of the sectioning angle. We also describe basic staining methods that can be used to begin to characterize atherosclerotic progression. These can be further modified to investigate antigens of specific interest to the researcher. The described techniques are generally applicable to a wide variety of existing and newly created dietary and genetically-induced models of atherogenesis.

We describe procedures to quantify and characterize atherosclerotic lesions in mouse models using precision sectioning of the aortic sinus and ascending aorta combined with histochemical and immunohistochemical analysis.

Atherosclerosis is a disease of the large arteries and a major underlying cause of myocardial infarction and stroke. Several different mouse models have ...

Acute Brain Trauma in Mice Followed By Longitudinal Two-photon Imaging

Although acute brain trauma often results from head damage in different accidents and affects a substantial fraction of the population, there is no effective treatment for it yet. Limitations of currently used animal models impede understanding of the pathology mechanism. Multiphoton microscopy allows studying cells and tissues within intact animal brains longitudinally under physiological and pathological conditions. Here, we describe two models of acute brain injury studied by means of two-photon imaging of brain cell behavior under posttraumatic conditions. A selected brain region is injured with a sharp needle to produce a trauma of a controlled width and depth in the brain parenchyma. Our method uses stereotaxic prick with a syringe needle, which can be combined with simultaneous drug application. We propose that this method can be used as an advanced tool to study cellular mechanisms of pathophysiological consequences of acute trauma in mammalian brain in vivo. In this video, we combine acute brain injury with two preparations: cranial window and skull thinning. We also discuss advantages and limitations of both preparations for multisession imaging of brain regeneration after trauma.

Acute brain trauma is a severe injury that has no adequate treatment to date. Multiphoton microscopy allows studying longitudinally the process of acute brain trauma development and probing therapeutical strategies in rodents. Two models of acute brain trauma studied with in vivo two-photon imaging of brain are demonstrated in this protocol.

急性脑外伤小鼠其次是纵向双光子成像

Although acute brain trauma often results from head damage in different accidents and affects a substantial fraction of the population, there is no ...

Minimal Invasive Surgical Procedure of Inducing Myocardial Infarction in Mice

Myocardial infarction still remains the main cause of death in western countries, despite considerable progress in the stent development area in the last decades. For clarification of the underlying mechanisms and the development of new therapeutic strategies, the availability of valid animal models are mandatory. Since we need new insights into pathomechanisms of cardiovascular diseases under in vivo conditions to combat myocardial infarction, the validity of the animal model is a crucial aspect. However, protection of animals are highly relevant in this context. Therefore, we establish a minimally invasive and simple model of myocardial infarction in mice, which assures a high reproducibility and survival rate of animals. Thus, this models fulfils the requirements of the 3R principle (Replacement, Refinement and Reduction) for animal experiments and assure the scientific information needed for further developing of therapeutical strategies for cardiovascular diseases.

A highly reproducible model for myocardial infarction in mice with minimal invasive manipulations is described. The model can be easily performed, resulting in a high reproducibility and survival rate. Thus, the described model will reduce the number of required animals as requested by the 3R principle (Replacement, Refinement and Reduction).

在诱导小鼠心肌梗死的微创手术过程

Myocardial infarction still remains the main cause of death in western countries, despite considerable progress in the stent development area in the last ...

Surgical Models of Gastroesophageal Reflux with Mice

Multiple surgical procedures have been reported to induce gastroesophageal reflux in animals. Herein, we report three surgical models with mice aiming to induce reflux of gastric contents, duodenal contents or mixed contents. Surgical procedures and general principles have been described in detail. A researcher with surgical experience should be able to grasp the technique after a short period of practice. After surgery, most mice can survive and develop reflux esophagitis similar to that in humans. However, it should be noted that histological differences between mouse and human esophagus are the inherent limitations of these surgical models. If used for research on Barrett&#8217;s esophagus and adenocarcinoma, these procedures may need to be combined with genetic modifications.

This article demonstrates surgical procedures of gastroesophageal reflux with mice. These models are useful tools for research on mechanisms and treatment of gastroesophageal reflux disease and potentially Barrett&#8217;s esophagus and esophageal adenocarcinoma.

胃食管反流与小鼠手术模型

Multiple surgical procedures have been reported to induce gastroesophageal reflux in animals. Herein, we report three surgical models with mice aiming to ...

An Improved Method for Rapid Intubation of the Trachea in Mice

Despite some anatomical and physiological differences, mouse models continue to be an essential tool for studying human lung disease. Bleomycin toxicity is a commonly used model to study both acute lung injury and fibrosis, and multiple methods have been developed for administering bleomycin (and other toxic agents) into the lungs. However, many of these approaches, such as transtracheal instillation, have inherent drawbacks, including the need for strong anesthetics and survival surgery. This paper reports a quick, reproducible method of intratracheal intubation that involves mild inhaled anesthesia, visualization of the trachea, and the use of a surrogate spirometer to confirm exposure. As a proof of concept, 8-12 week old C57BL/6 mice were administered either 2.0 U/kg of bleomycin or an equivalent volume of PBS, and both damage and fibrotic endpoints were measured post-exposure. This procedure allows researchers to treat a large cohort of mice in a relatively short period with little expense and minimal post-procedure care.

This article presents a rapid and simple method for administering bleomycin directly into the mouse trachea via intubation. Key advantages of this method are that it is highly reproducible, easy to master, and does not require specialized equipment or lengthy recovery times.

在小鼠气管插管快速的改进方法

Despite some anatomical and physiological differences, mouse models continue to be an essential tool for studying human lung disease. Bleomycin toxicity ...

Glaucoma-inducing Procedure in an In Vivo Rat Model and Whole-mount Retina Preparation

Glaucoma is a disease of the central nervous system affecting retinal ganglion cells (RGCs). RGC axons making up the optic nerve carry visual input to the brain for visual perception. Damage to RGCs and their axons leads to vision loss and/or blindness. Although the specific cause of glaucoma is unknown, the primary risk factor for the disease is an elevated intraocular pressure. Glaucoma-inducing procedures in animal models are a valuable tool to researchers studying the mechanism of RGC death. Such information can lead to the development of effective neuroprotective treatments that could aid in the prevention of vision loss. The protocol in this paper describes a method of inducing glaucoma - like conditions in an in vivo rat model where 50 &#181;l of 2 M hypertonic saline is injected into the episcleral venous plexus. Blanching of the vessels indicates successful injection. This procedure causes loss of RGCs to simulate glaucoma. One month following injection, animals are sacrificed and eyes are removed. Next, the cornea, lens, and vitreous are removed to make an eyecup. The retina is then peeled from the back of the eye and pinned onto sylgard dishes using cactus needles. At this point, neurons in the retina can be stained for analysis. Results from this lab show that approximately 25% of RGCs are lost within one month of the procedure when compared to internal controls. This procedure allows for quantitative analysis of retinal ganglion cell death in an in vivo rat glaucoma model.

Glaucoma-inducing Procedure in an In Vivo Rat Model and Whole-mount Retina Preparation

Glaucoma is characterized by damage to retinal ganglion cells. Inducing glaucoma in animal models can provide insight into the study of this disease. Here, we outline a procedure that induces loss of RGCs in an in vivo rat model and demonstrates the preparation of whole-mount retinas for analysis.

青光眼诱导在程序<em&gt;在体内</em&gt;大鼠模型和全安装视网膜准备

Glaucoma is a disease of the central nervous system affecting retinal ganglion cells (RGCs). RGC axons making up the optic nerve carry visual input to the ...

Induction of Accelerated Atherosclerosis in Mice: The "Wire-Injury" Model

Atherosclerosis is a proliferative fibro-inflammatory disease developing in the arterial wall, inducing a deficient blood flow or a lack of blood flow. Moreover, by rupture of the defective vascular wall, atherosclerosis induces occlusive thrombus formation, which represents the main cause of myocardial infarction or stroke and the most frequent cause of death. Despite the advances in the cardiovascular field, many questions remain unanswered, and additional basic research is essential to improve our understanding of the molecular mechanisms during atherosclerosis and its effects. Due to limited clinical studies, there is a need for representative animal models recreating atherosclerotic conditions such as neointima formation after stent implantation, balloon angioplasty, or endarterectomy. Since the mouse presents many advantages and is the most frequently used model for studying molecular processes, the current study proposes an invasive procedure of endothelial denudation, also known as the wire-injury model, which is representative of the human condition of neointima formation in arteries after revascularization procedures.

Induction of Accelerated Atherosclerosis in Mice: The &quot;Wire-Injury&quot; Model

This study describes an invasive procedure for the induction of accelerated atherosclerosis in mice. In comparison to other methods using electric- or cryo-induced injury, mechanical-induced injury mimics the human condition of restenosis after revascularization therapies and is ideal for the study of the molecular mechanisms involved.

小鼠加速动脉硬化的诱导："电线损伤"模型

Atherosclerosis is a proliferative fibro-inflammatory disease developing in the arterial wall, inducing a deficient blood flow or a lack of blood flow. ...

Balloon-based Injury to Induce Myointimal Hyperplasia in the Mouse Abdominal Aorta

The use of animal models is essential for a better understanding of MH, one major cause for arterial stenosis.In this article, we demonstrate a murine balloon denudation model, which is comparable with established vessel injury models in large animals. The aorta denudation model with balloon catheters mimics the clinical setting and leads to comparable pathobiological and physiological changes. Briefly, after performing a horizontal incision in the aorta abdominalis, a balloon catheter will be inserted into the vessel, inflated, and introduced retrogradely. Inflation of the balloon will lead to intima injury and overdistension of the vessel. After removing the catheter, the aortic incision will be closed with single stiches. The model shown in this article is reproducible, easy to perform, and can be established quickly and reliably. It is especially suitable for evaluating expensive experimental therapeutic agents, which can be applied in an economical fashion. By using different knockout-mouse strains, the impact of different genes on MH development can be assessed.

This article demonstrates a murine model to study the development of myointimal hyperplasia (MH) after aortic balloon injury.

球囊损伤诱发小鼠腹主动脉 Myointimal 增生的研究

The use of animal models is essential for a better understanding of MH, one major cause for arterial stenosis.In this article, we demonstrate a murine ...

Inducing Acute Lung Injury in Mice by Direct Intratracheal Lipopolysaccharide Instillation

Airway administration of lipopolysaccharide (LPS) is a common way to study pulmonary inflammation and acute lung injury (ALI) in small animal models. Various approaches have been described, such as the inhalation of aerosolized LPS as well as nasal or intratracheal instillation. The presented protocol describes a detailed step-by-step procedure to induce ALI in mice by direct intratracheal LPS instillation and perform FACS analysis of blood samples, bronchoalveolar lavage (BAL) fluid, and lung tissue. After intraperitoneal sedation, the trachea is exposed and LPS is administered via a 22 G venous catheter. A robust and reproducible inflammatory reaction with leukocyte invasion, upregulation of proinflammatory cytokines, and disruption of the alveolo-capillary barrier is induced within hours to days, depending on the LPS dosage used. Collection of blood samples, BAL fluid, and lung harvesting, as well as the processing for FACS analysis, are described in detail in the protocol. Although the use of the sterile LPS is not suitable to study pharmacologic interventions in infectious diseases, the described approach offers minimal invasiveness, simple handling, and good reproducibility to answer mechanistic immunological questions. Furthermore, dose titration as well as the use of alternative LPS preparations or mouse strains allow modulation of the clinical effects, which can exhibit different degrees of ALI severity or early vs. late onset of disease symptoms.

Presented here is a step-by-step procedure to induce acute lung injury in mice by direct intratracheal lipopolysaccharide instillation and to perform FACS analysis of blood samples, bronchoalveolar lavage fluid, and lung tissue. Minimal invasiveness, simple handling, good reproducibility, and titration of disease severity are advantages of this approach.

通过直接内切内脂多糖灌输诱导小鼠急性肺损伤

Airway administration of lipopolysaccharide (LPS) is a common way to study pulmonary inflammation and acute lung injury (ALI) in small animal models. ...