We thank Gilles Mithieux and Aude Barataud (INSERM U1213, Lyon, France) and Marie Liabeuf and Stephanie Lemarchand-Minde (Animal facility, l'Institut du Thorax, Nantes, France) for their help with the animal care protocol. This work was supported by grants from La r&#233;gion des Pays de la Loire, the Fondation d'Avenir, and the Casden Bank. We would like to thank Catherine Postic, Fadila Benhamed and Michelle Ca&#252;zac from l'institut Cochin for their hospitality and their help during the filming process.

动物和住房:获取8周龄C57BL / 6雄性小鼠。在10周龄，给水C57BL / 6小鼠免费接入和高脂肪的饮食（DIO饮食:脂肪35％千卡，25.8％酪蛋白，1.30％的矿物AIN，1.30％的维生素，1.70％，磷酸氢钙，0.7 ％碳酸钙，2.10％柠檬酸钾，0.026％胆碱酒石酸氢盐，8.9％蔗糖，0.384％胱氨酸，6.5％的纤维素，31.7％猪油，3.3％大豆油，和16.29％麦芽糖糊精）8（SG）或14（RYGB）之前的减肥手术周。促进体重增加，给专用于RYGB手术（假＆RYGB）含水20％果糖的小鼠（重量/体积）除了高脂肪的饮食。 伦理学声明:下述所有协议的步骤如下道德委员会对动物实验的指导方针下，自付批准文号01953.01卢瓦尔广场。 1.一般术前准备<ol><li>快小鼠在手术前6小时。凝胶迪等食品给予手术前和手术专用的前一天3天。 （0.4升/分钟双氧）中，用5％异氟醚（0.4升/分钟）和O 2的室诱导麻醉。执行脚趾捏测试，以确认该麻醉是有效的。管理通过皮下注射止痛剂（0.1毫克/千克丁丙诺啡），抗生素（10毫克/千克marbofloxacine），和亲动力学（1毫克/千克胃复安）。 </li><li>分配给外科手术（远离实验室的交通和换气扇），具体的工作区。请用专用的硬表面消毒手术区。广场上的工作空间区域加热垫手术过程中保持鼠标homeothermy。应用清洁，吸水性底垫在加热垫。 </li><li>从胸骨刮腹部用电动剃须刀到骨盆。包装在无菌保鲜膜（ 图1A和2A）鼠标的腹部，并放置在仰卧位将鼠标放在加热p广告。 </li><li>对眼睛应用兽医药膏，以防止干燥时的麻醉下。鼠标放置在鼻锥和保持用2％异氟醚（0.4升/分钟）和O 2（0.4升/分钟）麻醉。 </li><li>在开始手术前，戴上无菌检查袍，一次性磨砂帽，口罩和无菌手套。打开包含消毒手术器械一套，这是需要手术的所有步骤蒸压包。切灭菌保鲜膜一个窗口，并用消毒2次连续聚维酮碘溶液皮肤。 </li></ol> 2.胃切除术:手术过程<ol><li> 剖腹探查术中位数（图1B） <ol><li>下双目显微镜和用手术刀，执行从胸骨正中切口腹部打开腹腔（不损坏腹肌）的中间。保护带37℃的生理盐水溶液中浸泡过的无菌压缩皮肤。 </li></ol></li><li> 胃曝光（图1C） <ol><li>轻轻地调动使用浸湿，棉签胃。不受用浸湿的棉签或微型剪刀其横向紧密结缔组织附件的胃。 </li><li>轻轻地，完全外部化胃和放置止血胶原压缩胃的后方。缝合沿胃大弯侧幽门和食道艘8.0非吸收缝合线，以避免将来出血（ 图1D和E）。注意:使用止血剂可诱发组织损伤，确保只使用它的组织，旨在切除。 </li></ol></li><li> 胃的心脏区域的切除术（眼底） <ol><li>在目前的幽门区（主体）和胃（眼底）的心脏区域（ 图1F）之间的解剖行执行胃造瘘术。 </li><li>与对待每2浸湿的棉签去除残留的食物胃的一面。采用微型剪刀，切胃沿胃底和胃（ 图1G）的主体之间的边界。 </li><li>按止血胶原大出血的情况下压缩2分钟。关闭从胃oesophagal结用8.0不可吸收缝线的胃开口的切口（ 图1H）结束。 </li></ol></li><li> 胃幽门区切除术（阴茎） <ol><li>完成沿大弯胃切除，以便除去胃中（ 图1I）的大约80％。规范SG的宽度至3毫米。按止血胶原大出血的情况下压缩2分钟。 </li><li> 8.0非吸收缝合线（ 图1J）关闭胃幽门区的开放。轻轻摇动朝向胃的每侧2浸湿的棉签，以确保缝合是防漏。 </li><li>在泄漏的情况下，8.0非吸收缝合线完成。除去止血胶原压缩和胃返回到腹腔。发表止血胶原压缩对缝线。 </li></ol></li><li> 关腹 <ol><li>用5.0不可吸收缝合线（ 图1K）关闭腹壁的肌肉层。通过减少异氟烷浓度为1％减少麻醉。关闭使用5.0非吸收缝合线（ 图1L）皮肤。 </li></ol></li></ol> 3.空肠Roux-en-Y胃旁路术:手术过程<ol><li> 剖腹探查术中位数 <ol><li>下双目显微镜和用手术刀，执行从胸骨正中切口腹部打开腹腔（不损坏腹肌）的中间。保护带37℃的生理盐水溶液（ 图2B）浸过的无菌压缩皮肤。 </li></ol> &lt;/ LI&gt; <li> 胰肢体和消化道肢体 <ol><li>外部化肠道（ 图2C）。测量从幽门8厘米并用5.0不可吸收缝合线（图2D）进行肠两个连字。 </li><li>切两结扎线（ 图2E）之间的肠。放置两端的肢体近端在腹部的左上象限。注意:这将被用来作为消化道肢体（ 图2F）。 </li><li>放置面向消化道臂6厘米肢体近端（ 图2F）下面的两个端部的远端肢体。 注意:这将被用作胆肢。 </li></ol></li><li> 空肠-空肠吻合术 <ol><li>切两个肢体近端和肠道回路采用微型剪刀和执行相同长度的两个系膜切口（ 图2G和2H）。 </li><li>执行边侧吻合的两个8.0不可吸收的缝线。执行背侧吻合第一（ 图2I），其次是腹侧吻合（ 图2J）。 </li><li>轻轻摇动朝着吻合的每侧2浸湿的棉签，以确保缝合是防漏。在泄漏的情况下，8.0非吸收缝合线完成。 </li></ol></li><li> 胃空肠吻合术 <ol><li>用浸湿的棉签轻轻地调动了肚子里。不受用浸湿的棉签或微型剪刀其横向紧密结缔组织附件的胃。 </li><li>轻轻地，完全外部化的肚子，将重新吸收止血胶原压缩胃的后方。执行使用通过采用弧形微型镊子（ 图2K和2L）网膜通过了5.0非吸收缝合线幽门连字。 </li><li>切都胃的腹侧，从幽门1.5厘米，和远侧肢体采用微型剪刀，创建相同的长度（ 图2M）两种切口。 </li><li>具有两个8.0不可吸收的缝合线进行侧 - 侧吻合术。先从背部侧吻合（ 图2N），然后执行腹侧侧吻合（ 图20）。轻轻摇动2朝着吻合的每一侧润湿-棉签，以确保缝合是防漏。在泄漏的情况下，8.0非吸收缝合线完成。 </li></ol></li><li> 关腹 <ol><li>收使用5.0不可吸收缝合线（ 图2P）腹壁的肌肉层。通过减少异氟烷浓度为1％降低麻醉。通过皮下注射关闭.Administer 25毫升/公斤的温生理盐水溶液用5.0不可吸收缝合线（ 图2Q）皮肤。 </li></ol></li></ol> 4.一般术后护理<ol><li>停止异氟醚和0.8 L / min的氧气流量继续，直到鼠标完全清醒。直到它重新获得电机控制，由动物开始显示移动在笼子里，并能够站立并没有掉下来走，不要离开鼠标无人值守。 </li><li>将鼠标（仅每笼一只小鼠）在培养箱中30℃的温度条件下，进行5天。 </li><li>维持铁（0.5毫克/公斤/天;皮下注射）和维生素（800毫克/ 180毫升水）补剂在RYGB小鼠直到协议的结束。 </li><li>返回到凝胶减肥食品免费访问:手术后5天（高脂肪凝胶饮食10％猪油，10％液糖，57％的水）。重新引入了坚实的饮食手术后3天。 </li><li>皮下注射丁丙诺啡（0.1毫克/公斤，每日两次，从第0天至手术后第3天），美洛昔康（1毫克/千克，从第0天至手术后第3天），胃复安（1毫克/千克，从第0天到手术后第5天），和marbofloxacine（10毫克/千克，从第0天至第3天AFTE<html> - [R手术）。 </li></ol> 5.术后代谢参数评估<ol><li> 食物摄入量的测量 <ol><li>允许小鼠在手术后1周恢复。屋里的老鼠每笼之一。测量放置在笼子里的固体食物量。重量剩下的食物24小时后。只要需要重复此步骤。 </li></ol></li><li> 口服葡萄糖耐受性试验 <ol><li>口服丸前禁食6小时的老鼠。收获从尾部末端的血液滴，并将其应用在血糖仪插入确定时间0的血糖值的葡萄糖条。 </li><li>辖的20％D-葡萄糖溶液（2克/公斤）口服丸剂。测量血液中葡萄糖水平在15，30，60，和120分钟后管饲法。 </li></ol></li><li> 血红蛋白水平分析 <ol><li>使用血液系统，测量新鲜血液20微升品尝血红蛋白水平。 </li></ol></li></ol>"&gt; 6。安乐死<ol><li>通过腹膜内注射甲苯噻嗪/氯胺酮溶液（10/80毫克/千克）麻醉小鼠。执行颈椎脱位。通过确保心脏没有跳动和眼球眨眼反射消失验证完整的安乐死。 </li></ol>

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The authors declare no conflicts of interest.

为了克服肥胖的日益流行，第一减肥手术程序在20世纪60年代出现在美国。自那时以来，全世界每年进行的程序的数量仍增加，今天，这些技术代表了病态肥胖6的管理的最佳治疗选择。之间制定的程序，SG和RYGB是在临床实践4中使用的两种最常用的方法。动物模型，特别是啮齿动物，已被用于破译后面减肥手术后观察到的一般健康状况改善的机制。 破译维持减肥手术的有益的代谢效应的分子机制，我们开发了减肥手术的两种小鼠模型:SG，它通常被定义为一个限制性技术，并且RYGB，其被描述为一个限制性和吸收不良手术。这些小鼠减肥的实现程序需要显微外科经验。这些模型是使用专门的动物手术和人类内脏外科医生的技术人员开发的。从以往的经验，外科培训是必要的RYGB后达到75％的存活率。掌握SG不需要同样多的时间，和学习曲线较短，死亡率在95％左右。相比之下，RYGB是一个更复杂的技术，并且需要6至8周的培训，以达到可接受的术后存活率。 在文献中，有各种各样的啮齿动物减肥手术是先前已经描述7，8，9。在此SG技术中，我们取出胃的心脏区域（也称为眼底），不存在于人类。其目的是要去除胃的约80％。 RYGB模型大鼠的袋尺寸范围为&lt;5％到&gt;初始胃尺寸的20％。一些STudies使用鼠标RYGB模型，呈现胃10中 ，一个小胃囊5，或总胃改道5的任一个完整保存。在一个非常全面的研究，邓平贤和他的同事评估了RYGB的不同小鼠模型，并报告说，小鼠的胃没有被解剖适应于建立一个胃囊5。对于我们的RYGB手术中，我们选择不伐胃进行空肠Roux-en-Y。作为限制，我们的模型不诱导食物摄入，其中对比了它在RYGB人模型中观察到的减少。 RYGB手术后的存活率大鼠很少通过文献中，作者提到的，但它似乎平均11达到约30％至35％。在我们的手中，死亡主要是由于吻合口瘘或狭窄或术后晚期gastrojejunal吻合小号tenosis，导致慢性阻塞持续性过度减肥，这损害动物福利。 RYGB需要特定的围手术期管理，包括在前三术后天凝胶饮食和疼痛管理制度。手术后改善长期生存，我们添加维生素术后，特别是脂溶性维生素和铁，预防贫血。相反，老鼠并不需要脂溶性维生素补充剂SG之后，因为它只是一个限制性程序。在以前的文章中，作者并没有术后过程中描述手术动物的健康状况。因为它是一个大手术，我们不能假设在这些研究动物的系统福祉。 总之，无论是限制性的（SG）的小鼠模型或限制性-吸收不良（RYGB）减肥手术是可行的，但需要显微训练用的学习曲线，特别是为RYGB手术。柯EP身体健康，围手术期管理是RYGB程序中非常重要，尤其是在疼痛方面的动物，而且在补充维生素条款。通过实现更快的，无菌的手术，并通过使用显微训练，可以大大改善术后存活率。这些模型是非常有用和必要的工具来了解减肥手术的代谢作用。

肥胖和糖尿病的全球大流行病是在严重性毁灭性的。全球超过两十亿成年人（占总人口的30％）是超重（BMI&gt; 25公斤/米2）或肥胖（BMI&gt; 30公斤/米2）1。这可以与代谢的并发症，如2型糖尿病，高血压，和血脂异常一起走，导致增加的发病率和死亡率。肥胖会增加整体死亡率和癌症2的患病率。由于缺乏任何有效的无创治疗，减肥手术表示可以导致长期的，持续的体重减轻3,4是唯一的选择。许多不同的手术方法已被开发，但在套筒胃切除术（SG）和空肠Roux-en-Y胃旁路术（RYGB）是在临床实践中最常使用的两个过程。在SG过程，胃的初始体积的80％是拆除;因此，该技术是提高饱限制性手术之一。该RYGB是的制约，吸收不良的技术之一。 RYGB中，创建了一个小胃囊（总胃体积的1-2％）和肠被重新布置成Y形，其中延迟消化和营养物质的吸收。这两种技术导致显著体重减少和普遍改善在频繁相关的合并症（ 例如，高血压，2型糖尿病和血脂异常）3，与RYGB看到一个更高的效率。然而，后面的减肥手术的多效性有益效果的分子机制常常没有完全阐明。由于可用的众多基因改造的菌株，小鼠模型是最方便的动物模型来研究这些分子机制。 然而，减肥手术很难直接适应小动物模型和requirË高外科灵巧。而SG可以容易地在啮齿类动物中具有很好的存活率进行RYGB是在小鼠致死因严重肠梗阻5。不同改性RYGB技术已经被提出来抵消这个问题，特别是oesojejunostomy 5。在这里，我们提出了另一种选择:无胃切除胃空肠造口。本变形RYGB再现大多数的人类中观察到的有益效果（即一个显著体重减少和在葡萄糖和脂质体内稳态的改善）。 此稿件旨在总结和讨论小鼠SG与RYGB的技术和实验细节，并促进这些程序与视频的帮助。具体的亮点将有关的术前和术后保健的协议，使维生素和铁缺乏减少优化进行。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Drugs</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>High Fat diet</td>
			<td>DIO diet</td>
			<td>Safe</td>
			<td></td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>Forane</td>
			<td>Baxter</td>
			<td></td>
		</tr>
		<tr>
			<td>Buprenorphin</td>
			<td>Buprecare</td>
			<td>Animalcare</td>
			<td></td>
		</tr>
		<tr>
			<td>Marbofloxacine</td>
			<td>Marbocyl</td>
			<td>Vetoquinol</td>
			<td></td>
		</tr>
		<tr>
			<td>Ammonium iron citrate, vitamins PP-B12</td>
			<td>Fercobsang</td>
			<td>Vetoquinol</td>
			<td></td>
		</tr>
		<tr>
			<td>Vitamins A-D3-E-K-B</td>
			<td>Vita Rongeur</td>
			<td>Virbac</td>
			<td></td>
		</tr>
		<tr>
			<td>NaCl 0,9%</td>
			<td>NaCl 0,9%</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Povidone solution</td>
			<td>Betadine Scrub</td>
			<td>Betadine</td>
			<td></td>
		</tr>
		<tr>
			<td>Povidone solution</td>
			<td>Betadine Solution</td>
			<td>Betadine</td>
			<td></td>
		</tr>
		<tr>
			<td>Carboptol 980 NF</td>
			<td>Ocrygel</td>
			<td>TVM</td>
			<td></td>
		</tr>
		<tr>
			<td>Name</td>
			<td>References</td>
			<td>Company</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>Sutures</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Prolene&#174;</td>
			<td>8.0, 6,5 mm</td>
			<td>Ethicon</td>
			<td></td>
		</tr>
		<tr>
			<td>Prolene&#174;</td>
			<td>5.0, 13 mm</td>
			<td>Ethicon</td>
			<td></td>
		</tr>
		<tr>
			<td>Name</td>
			<td>References</td>
			<td>Company</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>Surgical equipments</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Scissors</td>
			<td></td>
			<td>FST</td>
			<td></td>
		</tr>
		<tr>
			<td>Needle holder</td>
			<td>Olsen-Hegar</td>
			<td>FST</td>
			<td></td>
		</tr>
		<tr>
			<td>Micro scissors</td>
			<td>Vannas</td>
			<td>FST</td>
			<td></td>
		</tr>
		<tr>
			<td>Micro forceps&#160;</td>
			<td>Graefe</td>
			<td>FST</td>
			<td></td>
		</tr>
		<tr>
			<td>Micro forceps curved</td>
			<td>Graefe</td>
			<td>FST</td>
			<td></td>
		</tr>
		<tr>
			<td>Curved micro needle holder</td>
			<td>Castroviejo</td>
			<td>FST</td>
			<td></td>
		</tr>
		<tr>
			<td>Hemostatic collagen compress</td>
			<td>Pangen</td>
			<td>Urgo</td>
			<td></td>
		</tr>
		<tr>
			<td>Absorbent underpads</td>
			<td></td>
			<td>VWR</td>
			<td></td>
		</tr>
		<tr>
			<td>Name</td>
			<td>References</td>
			<td>Company</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>Specific equipments</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Hematology system&#160;</td>
			<td>Hemavet 950FS</td>
			<td>Hemavet</td>
			<td></td>
		</tr>
		<tr>
			<td>Glucose strips and glucometer</td>
			<td>One touch Verio</td>
			<td>Life scan</td>
			<td></td>
		</tr>
		<tr>
			<td>Stereo microscope</td>
			<td>MZ6</td>
			<td>Leica&#160;</td>
			<td></td>
		</tr>
	</tbody>
</table>

techniques of sleeve gastrectomy and modified roux-en-y gastric bypass in mice

肥胖是一个重大的公共健康问题，以4〜28％的男性和6.2〜36.5％的妇女在欧洲（2003年至2008年）患病。病态肥胖通常与代谢并发症，如2型糖尿病，高血压和血脂异常相关，降低寿命和质量。在没有任何有效的非侵入性的治疗，减肥手术是治疗病态肥胖（体重指数（BMI）"40公斤/米2）一个有价值的治疗选择，从而导致长期的，持续的代谢并发症体重减轻和改善。然而，维持减肥手术的有益效果的底层的细胞和分子机制尚未完全清楚。由于可用众多遗传修饰的菌株，所述的小鼠模型是最方便的动物模型，探讨背后减肥手术的多效性有益效果的分子机制。在这里，我们详细介绍了优化healthc是在小鼠中的两个最广泛使用的减肥手术的方法和手术的协议:该套筒胃和改性空肠Roux-en-Y胃旁路。破译底层减肥手术的治疗作用的分子机制提供了新的识别目标的疗法的承诺。

大体情况 平均手术时间为SG程序是49.3±1.5分钟。我们除去62.8±5.0毫克胃的，它代表了胃的约80％的百分比。没有老鼠在手术过程中或七天死亡。一个鼠标（7.1％）在11 日术后第二天死了，因为引起的牛黄胃梗阻。 平均手术时间为RYGB手术为89.1±2.8分钟。死亡率RYGB后（P = 0.038 66.6％:25％ 与 RYGB假）是显著高。术后死亡的主要原因是吻合口漏（N = 8），吻合口狭窄（N = 5），麻醉并发症（4例），术后出血（N = 2）。我们还观察到两项长期死于胃肠吻合狭窄慢性阻塞和营养不良。正如图3所示，我们注意到了手术后的存活率的逐步改善整个协议，表示密集外科训练掌握RYGB手术技术要求。 体重，食物摄入，和葡萄糖稳态 老鼠在手术前也有类似的体重。相比于假手术对照组（; P &lt;0.01; 图4A -16.0％:-6.6％与SG假）SG证明手术后14天显著的减肥效果。第14天，手术后-7.9％，并在28日手术后与RYGB -5.1％:第14天，手术后-22.9％-25.5和在％RYGB与来自术后14天（假显著消瘦第28天手术后; 图4B）。在14天的外周的每日食物摄取量减少了35％<html> OD的SG（ 图4C）以下。相比之下，我们没有观察到假和RYGB小鼠（ 图4D）之间的这种食欲减退作用。我们还报告下列SG和RYGB手术（ 图4E，4F）葡萄糖耐受的改善。 补铁对RYGB诱发贫血有益作用 我们没有观察 （未示出数据）SG手术后14天血红蛋白血症的显著下降。与此相反，严重贫血在RYGB手术的小鼠进行了测定，但它从特定补铁（ 图5A）那样的好处。每日补充铁确实关键和RYGB手术（ 图5B）在28天后避免显著贫血。 /54905/54905fig1.jpg"/&gt; 图1: 袖长胃切除术过程。 （ 一 ）术前准备。 （B）中位数剖腹手术。 （C）胃曝光。 （D）幽门血管缝合。 （ 五 ）食管血管缝合。 （F）初始胃造瘘术。胃的心脏区域的（G）切除术。从胃食管交界处胃开口的切口的端部的（H）的封闭。胃幽门区（I）切除术。幽门区的开放的（J）闭幕。腹壁的肌肉层的（K）的封闭。 （L）皮肤闭幕。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54905/54905fig1large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> 图2:空肠Roux-en-Y旁路过程。 （ 一 ）术前准备。 （B）中位数剖腹手术。小肠（ 三 ）外部化。 （D），胆胰和消化道肢体连字。 （ 五 ）结扎线之间的小肠部分。 （F）肠肢体定位为空肠-空肠吻合。 （G - 高 ）肢体近端和肠道循环系膜切口。 （Ⅰ）背侧-侧吻合术。 （J）的腹侧边侧吻合。 （K - L）幽门结扎。胃腹侧的（M）切口。远端肢体和背侧侧吻合的（N）的切口。 （O）腹侧-侧吻合术。 （P </st荣&gt;）腹壁肌肉层闭幕。皮肤（Q）闭幕。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54905/54905fig2large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图3" src="/files/ftp_upload/54905/54905fig3.jpg" /> 图3:RYGB手术后7天小鼠存活率。数据显示在RYGB手术的小鼠的存活率外科训练的好处。的值显示为平均值±SEM表示。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54905/54905fig3large.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/54905/54905fig4.jpg" /> 图4:SG 的影响（A，C，D）和RYGB（B，D，F）第Bodyweight，每日进食量，及糖耐量。数据显示为平均值±SEM表示。 NS:不显著; * P &lt;0.05; ** P &lt;0.01（Mann-Whitney检验，ANOVA检验）。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54905/54905fig4large.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/54905/54905fig5.jpg" /> 图5:RYGB 后血浆血红蛋白水平的小鼠无（A）或（B）补铁。数据显示为平均值±SEM表示。 NS:不显著; * P &lt;0.05（Mann-Whitney检验）。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54905/54905fig5large.jpg" target="_blank">请点击此处查看该图的放大版本。</a>

Watch this Scientific Journal Video about Techniques of Sleeve Gastrectomy and Modified Roux-en-Y Gastric Bypass in Mice at JoVE.com

Techniques of Sleeve Gastrectomy and Modified Roux-en-Y Gastric Bypass in Mice

Bariatric surgery is the most efficient way to reduce body weight and the deadly metabolic complications (diabetes, obesity, and dyslipidemia) frequently associated with morbid obesity. Mouse models of bariatric surgery represent a unique asset for deciphering molecular mechanisms behind the beneficial effects of these surgeries on diabetes, hypertension, and dyslipidemia.

袖胃切除术的技术和改良的小鼠空肠Roux-en-Y胃旁路

Obesity is a major public health issue, with a prevalence of 4 to 28% for men and 6.2 to 36.5% for women in Europe (from 2003 to 2008). Morbid obesity is ...

techniques-sleeve-gastrectomy-modified-roux-en-y-gastric-bypass

Nanyang Technological University

Research

JoVE Journal

Medicine

11.9K 次观看.  INSERM, CNRS, UNIV Nantes. Bariatric surgery is the most efficient way to reduce body weight and the deadly metabolic complications (diabetes, obesity, and dyslipidemia) frequently associated with morbid obesity. Mouse models of bariatric surgery represent a unique asset for deciphering molecular mechanisms behind the beneficial effects of these surgeries on diabetes, hypertension, and dyslipidemia.

视频: Techniques of Sleeve Gastrectomy and Modified Roux-en-Y Gastric Bypass in Mice

Modified Technique for Coronary Artery Ligation in Mice

Myocardial infarction (MI) is one of the most important causes of mortality in humans1-3. In order to improve morbidity and mortality in patients with MI we need better knowledge about pathophysiology of myocardial ischemia. This knowledge may be valuable to define new therapeutic targets for innovative cardiovascular therapies4. Experimental MI model in mice is an increasingly popular small-animal model in preclinical research in which MI is induced by means of permanent or temporary ligation of left coronary artery (LCA)5. In this video, we describe the step-by-step method of how to induce experimental MI in mice.
The animal is first anesthetized with 2% isoflurane. The unconscious mouse is then intubated and connected to a ventilator for artificial ventilation. The left chest is shaved and 1.5 cm incision along mid-axillary line is made in the skin. The left pectoralis major muscle is bluntly dissociated until the ribs are exposed. The muscle layers are pulled aside and fixed with an eyelid-retractor. After these preparations, left thoracotomy is performed between the third and fourth ribs in order to visualize the anterior surface of the heart and left lung. The proximal segment of LCA artery is then ligated with a 7-0 ethilon suture which typically induces an infarct size ~40% of left ventricle. At the end, the chest is closed and the animals receive postoperative analgesia (Temgesic, 0.3 mg/50 ml, ip). The animals are kept in a warm cage until spontaneous recovery.

The surgical procedure used to induce experimental myocardial infarction in mice begins with left thoracotomy between the third and the fourth ribs in order to visualize the anterior surface of the heart and left lung. The left coronary artery is ligated, the chest is closed and the mouse is allowed to recover spontaneously.

改性技术在小鼠冠状动脉结扎

Myocardial infarction (MI) is one of the most important causes of mortality in humans1-3. In order to improve morbidity and mortality in patients with MI ...

科研

医学

Roux-en-Y Gastric Bypass Operation in Rats

Currently, the most effective therapy for the treatment of morbid obesity to induce significant and maintained body weight loss with a proven mortality benefit is bariatric surgery1,2. Consequently, there has been a steady rise in the number of bariatric operations done worldwide in recent years with the Roux-en-Y gastric bypass (gastric bypass) being the most commonly performed operation3. Against this background, it is important to understand the physiological mechanisms by which gastric bypass induces and maintains body weight loss. These mechanisms are yet not fully understood, but may include reduced hunger and increased satiation4,5, increased energy expenditure6,7, altered preference for food high in fat and sugar8,9, altered salt and water handling of the kidney10 as well as alterations in gut microbiota11. Such changes seen after gastric bypass may at least partly stem from how the surgery alters the hormonal milieu because gastric bypass increases the postprandial release of peptide-YY (PYY) and glucagon-like-peptide-1 (GLP-1), hormones that are released by the gut in the presence of nutrients and that reduce eating12.
During the last two decades numerous studies using rats have been carried out to further investigate physiological changes after gastric bypass. The gastric bypass rat model has proven to be a valuable experimental tool not least as it closely mimics the time profile and magnitude of human weight loss, but also allows researchers to control and manipulate critical anatomic and physiologic factors including the use of appropriate controls. Consequently, there is a wide array of rat gastric bypass models available in the literature reviewed elsewhere in more detail 13-15. The description of the exact surgical technique of these models varies widely and differs e.g. in terms of pouch size, limb lengths, and the preservation of the vagal nerve. If reported, mortality rates seem to range from 0 to 35%15. Furthermore, surgery has been carried out almost exclusively in male rats of different strains and ages. Pre- and postoperative diets also varied significantly.
Technical and experimental variations in published gastric bypass rat models complicate the comparison and identification of potential physiological mechanisms involved in gastric bypass. There is no clear evidence that any of these models is superior, but there is an emerging need for standardization of the procedure to achieve consistent and comparable data. This article therefore aims to summarize and discuss technical and experimental details of our previously validated and published gastric bypass rat model.

Numerous studies using gastric bypass rat models have been recently conducted to uncover the underlying physiological mechanisms of Roux-en-Y gastric bypass operations. This article aims to demonstrate and discuss the technical and experimental details of our published gastric bypass rat model to understand advantages and limitations of this experimental tool.

空肠Roux-en-Y胃搭桥手术大鼠

Currently, the most effective therapy for the treatment of morbid obesity to induce significant and maintained body weight loss with a proven mortality ...

Assessment of Gastric Emptying in Non-obese Diabetic Mice Using a [13C]-octanoic Acid Breath Test

Gastric emptying studies in mice have been limited by the inability to follow gastric emptying changes in the same animal since the most commonly used techniques require killing of the animals and postmortem recovery of the meal1,2. This approach prevents longitudinal studies to determine changes in gastric emptying with age and progression of disease. The commonly used [13C]-octanoic acid breath test for humans3 has been modified for use in mice4-6 and rats7 and we previously showed that this test is reliable and responsive to changes in gastric emptying in response to drugs and during diabetic disease progression8. In this video presentation the principle and practical implementation of this modified test is explained. As in the previous study, NOD LtJ mice are used, a model of type 1 diabetes9. A proportion of these mice develop the symptoms of gastroparesis, a complication of diabetes characterized by delayed gastric emptying without mechanical obstruction of the stomach10.
This paper demonstrates how to train the mice for testing, how to prepare the test meal and obtain 4 hr gastric emptying data and how to analyze the obtained data. The carbon isotope analyzer used in the present study is suitable for the automatic sampling of the air samples from up to 12 mice at the same time. This technique allows the longitudinal follow-up of gastric emptying from larger groups of mice with diabetes or other long-standing diseases.

Assessment of Gastric Emptying in Non-obese Diabetic Mice Using a [13C]-octanoic Acid Breath Test

Determination of gastric emptying with a non-invasive [13C]-octanoic acid breath test for tracking gastroparesis in female NOD LtJ mice.

使用的非肥胖型糖尿病小鼠胃排空的评估[<sup&gt; 13</sup&gt; C-辛酸呼气试验

Gastric emptying studies in mice have been limited by the inability to follow gastric emptying changes in the same animal since the most commonly used ...

Using the Sleeve Technique in a Mouse Model of Aortic Transplantation - An Instructional Video

Orthotopic aortic transplantation using the sleeve technique reduces injury to the aorta with failure rate of only 10-20%. The time to anastomose the aorta in mice using the sleeve method was short and easy averaging 20 min, permitting studies of iso/allo grafts. The following article describes the aortic transplantation procedure used in our laboratory. The mice were anesthetized with a mixture of 1.5% volume isoflurane and 100% oxygen through a face mask. At this point, the segment of the aorta between the renal arteries and its bifurcation was separated from the vena cava, freely prepared and clampedat the proximal and distal segments with a single silk suture. Prior to the removal of the aorta, a saline solution containing heparin was injected into the inferior vena cava. Then the aorta was cut between the clamps and a saline heparin solution was used to flush the lumen. The sleeve technique with monofilament sutures was used in order to transplant the abdominal aorta in the orthotopic position.

We present an orthotopic aortic transplantation model using the sleeve technique in mice. It is a very rapid anastomosis method, which can be employed in studies of vascular disease.

套筒技术在主动脉移植小鼠模型中的应用-教学视频

Orthotopic aortic transplantation using the sleeve technique reduces injury to the aorta with failure rate of only 10-20%. The time to anastomose the ...

Cardiopulmonary Bypass in a Mouse Model: A Novel Approach

As prolonged cardiopulmonary bypass becomes more essential during cardiac interventions, an increasing clinical demand arises for procedure optimization and for minimizing organ damage resulting from prolonged extracorporal circulation. The goal of this paper was to demonstrate a fully functional and clinically relevant model of cardiopulmonary bypass in a mouse. We report on the device design, perfusion circuit optimization, and microsurgical techniques. This model is an acute model, which is not compatible with survival due to the need for multiple blood drawings. Because of the range of tools available for mice (e.g., markers, knockouts, etc.), this model will facilitate investigation into the molecular mechanisms of organ damage and the effect of cardiopulmonary bypass in relation to other comorbidities.

This paper describes how to perform cardiopulmonary bypass in mice. This novel model will facilitate the investigation of the molecular mechanisms involved in organ damage.

小鼠体外循环模型: 一种新方法

As prolonged cardiopulmonary bypass becomes more essential during cardiac interventions, an increasing clinical demand arises for procedure optimization ...

One-anastomosis Gastric Bypass (OAGB) in Rats

The goal of this protocol is to set up a preclinical model of bariatric surgery and, more specifically, OAGB in obese rats. Based on this preclinical model, longitudinal studies can be carried out to provide an improved understanding of the mechanisms underlying the outcomes seen after bariatric surgery in humans. For this purpose, rats are operated on through a laparotomy under general anesthesia with isoflurane. First, the surgeon creates a long and tubular gastric pouch: after greater curve and hiatal dissection, the nonglandular stomach is stapled and removed. Then, the remaining stomach is also stapled in order to create a gastric tube and exclude the antrum of the stomach. After that, the surgeon performs a single end-to-side gastrojejunostomy 35 cm from the duodenojejunal angle. This limb length has been chosen in order to reproduce the same ratio between the biliopancreatic limb (BPL) and common limb (CL) length as in human bariatric surgery. The operation ends by aponeurotic and cutaneous closure. The early postoperative management consists of subcutaneous hydration, an intramuscular prophylactic antibiotic injection, a parietal injection of xylocaine, the administration of painkillers, and a progressive reintroduction of diet.

One-anastomosis Gastric Bypass OAGB in Rats

This protocol is for the performance of one-anastomosis gastric bypass (OAGB) on rat. The operator performs a long and tubular-stapled gastric pouch followed by hand-sewn anastomosis. For this model, the operator reproduces the same ratio between biliopancreatic and common limb in humans; therefore, the biliopancreatic limb measures 35 cm.

大鼠胃旁路 (oagb) 的一次吻合

The goal of this protocol is to set up a preclinical model of bariatric surgery and, more specifically, OAGB in obese rats. Based on this preclinical ...

Sleeve Gastrectomy in Mice using Surgical Clips

The number of people who are overweight and obese is continually increasing both in the adult and adolescent populations. This coincides with the increased universal phenomenon of type 2 diabetes (T2D) and other metabolic problems. Bariatric surgery, such as SG, is currently one of the most effective and commonly used long-term treatment for obesity and T2D, but the association between them is not completely explored yet. The mechanisms underlying the outcomes seen after bariatric surgery in humans can be investigated based on preclinical animal studies. The SG reduces body weight, glucose levels and many metabolic parameters, and is easy to perform with a low incidence of complications. The goal of this work is to provide a simple method and an uncomplicated preclinical model of bariatric surgery in animals for researchers.

The prevalence of diabetes and obesity is continuously increasing worldwide. The mechanisms between diabetes, obesity and their associated mortality and co-morbidities need to be further investigated. Here, we present a protocol for sleeve gastrectomy (SG) in animals as an uncomplicated preclinical model of bariatric surgery.

使用手术夹的小鼠的套管胃切除术

The number of people who are overweight and obese is continually increasing both in the adult and adolescent populations. This coincides with the ...

A Protocol for Roux-en-Y Gastric Bypass in Rats using Linear Staplers

Roux-en-Y gastric bypass (RYGB) is commonly performed for the treatment of severe obesity and type 2 diabetes. However, the mechanism of weight loss and metabolic changes are not well understood. Multiple factors are thought to play a role, including reduced caloric intake, decreased nutrient absorption, increased satiety, the release of satiety-promoting hormones, shifts in bile acid metabolism, and alterations in the gut microbiota.
The rat RYGB model presents an ideal framework to study these mechanisms. Prior work on mouse models have had high mortality rates, ranging from 17 to 52%, limiting their adoption. Rat models demonstrate more physiologic reserve to surgical stimulus and are technically easier to adopt as they allow for the use of surgical staplers. One challenge with surgical staplers, however, is that they often leave a large gastric pouch which is not representative of RYGB in humans. 
In this protocol, we present a RYGB protocol in rats that result in a small gastric pouch using surgical staplers. Utilizing two stapler fires which remove the forestomach of the rat, we obtain a smaller gastric pouch similar to that following a typical human RYGB. Surgical stapling also results in better hemostasis than sharp division. Additionally, the forestomach of the rat does not contain any glands and its removal should not alter the physiology of RYGB.
Weight loss and metabolic changes in the RYGB cohort were significant compared to the sham cohort, with significantly lower glucose tolerance at 14 weeks. Furthermore, this protocol has an excellent survival of 88.9% after RYGB. The skills described in this protocol can be acquired without previous microsurgical experience. Once mastered, this procedure will provide a reproducible tool for studying the mechanisms and effects of RYGB.

Roux-en-Y gastric bypass (RYGB) is performed to treat obesity and diabetes. However, the mechanisms underlying RYGB's efficacy are not fully understood, and studies are limited by technical difficulty leading to high mortality in animal models. This article provides instructions on how to perform RYGB in rats with high success rates.

使用线性压花器在老鼠身上进行胃旁路的规程

Roux-en-Y gastric bypass (RYGB) is commonly performed for the treatment of severe obesity and type 2 diabetes. However, the mechanism of weight loss and ...

Robotic Central Pancreatectomy with Roux-en-Y Pancreaticojejunostomy

Central pancreatectomy is a parenchyma-sparing alternative to distal pancreatectomy in patients with a benign or low-grade malignant tumor in the body of the pancreas. The aim of central pancreatectomy is to prevent postoperative life-long endocrine and exocrine insufficiency. The downside of central pancreatectomy is the high rate of postoperative pancreatic fistula, which is the main reason that many surgeons do not routinely use central pancreatectomy in eligible patients. Most studies report open or laparoscopic central pancreatectomy with a pancreatico-gastrostomy anastomosis in adults. This is the first description of a standardized approach to robotic central pancreatectomy with Roux-en-Y pancreaticojejunostomy reconstruction in an adolescent (16-year-old boy) with a pseudopapillary tumor in the body of the pancreas. The operation time was 248 min with 20 mL of blood loss. The postoperative course was uneventful except for the short-term medical treatment for a grade B pancreatic fistula. Robotic central pancreatectomy can be safely applied in selected patients in experienced centers.

Robotic central pancreatectomy may be used in selected patients in experienced centers. This protocol presents all steps and the feasibility of a robotic central pancreatectomy with Roux-en-Y pancreaticojejunostomy in a 16-year-old adolescent patient.

机器人中央胰腺切除术与鲁恩-Y胰腺切除术

Central pancreatectomy is a parenchyma-sparing alternative to distal pancreatectomy in patients with a benign or low-grade malignant tumor in the body of ...

A Modified Simple Method for Induction of Myocardial Infarction in Mice

Myocardial infarction (MI) represents one of the leading causes of death. MI models are widely used for investigating the pathomechanisms of post-MI remodeling and evaluation of novel therapeutics. Different methods (e.g., isoproterenol treatment, cryoinjury, coronary artery ligation, etc.) have been used to induce MI. Compared with isoproterenol treatment and cryoinjury, coronary artery ligation may better reflect the ischemic response and chronic remodeling after MI. However, traditional methods for coronary ligation in mice are technically challenging. The current study describes a simple and efficient process for induction of MI in mice with readily available materials. The mouse chest skin was cut open under stable anesthesia. The heart was immediately externalized through the intercostal space after blunt separation of the pectoralis major and pectoralis minor. The left anterior descending branch (LAD) was ligated with a 6-0 suture 3 mm from its origin. Following LAD ligation, staining with 2,3,5-Triphenyltetrazolium chloride (TTC) indicated successful induction of MI and temporal changes of post-MI scar size. Meanwhile, survival analysis results showed overt mortality within 7 days after MI, mainly due to cardiac rupture. Moreover, post-MI echocardiographic assessment demonstrated successful induction of contractile dysfunction and ventricular remodeling. Once mastered, an MI model can be established in mice within 2-3 min with readily available materials.

Under adequate anesthesia, the mouse heart was externalized through the intercostal space, and myocardial infarction was successfully induced by ligating the left anterior descending artery (LAD) using materials readily available in most laboratories.

一种改良的小鼠心肌梗死诱导简易方法

Myocardial infarction (MI) represents one of the leading causes of death. MI models are widely used for investigating the pathomechanisms of post-MI ...