Name: ileectomy-induced bile overaccumulation in mouse intestine
Uploaded: 2017-08-21T13:00:00
Description: Watch this Scientific Journal Video about Ileectomy-induced Bile Overaccumulation in Mouse Intestine at JoVE.com

这项研究支持由汤姆皮特森基金会和美国国立卫生研究院授予 R01 HL119780 （耆那教、 MK）。

动物议定书 》 进行了审查和批准机构动物护理和使用委员会在凯斯西储大学医学院和按照国家卫生研究院 (NIH) 指南进行维护和使用的实验室动物 （2011 年第 8 版）。小鼠处死的方法符合美国兽医医疗协会 (AVMA) 准则为动物安乐死 （2013年版）。在本议定书中使用 C57BL/6J，男，8-16 周老小鼠。老鼠被安置在 12 h 黑暗/光照周期环境。
 1.手术前准备 <ol><li>转移小鼠到干净的笼子?...

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为了执行成功的回肠，必须预先结扎肠系膜上动脉来阻止对切除部分的血液供应。缺血性回肠段结扎后会变成暗紫色。然后必须完全切除回肠段。保留的两端必须保证正常的血液供应。这是必须避免出血和不当的去除，这可以很容易地导致手术失败由于缺血性坏死后缝合。期间吻合，它是重要的回肠末端一起加入他们自然的解剖位置。肠系膜被认为是为初始的缝合线，以避免可能发生的扭曲两端?...

在现代生物医学研究中，基因操纵的动物模型被广泛的应用，以收集洞察人类疾病。尤其是，组织或细胞特异性基因的增益损失功能已被用于研究分子调控，以及诱发的生物学效应。尽管操纵目标基因在体内的进步，有挥之不去的限制。第一，许多细胞或组织的特定缺失会影响多个器官。例如上, 皮基因缺失将消除多种组织中上皮细胞的表达。此外，即使删除仅限于为特定的组织，空间的控制是很少可行的。例如，在像肠组织，不同的部分进行十分具体的职能不能操纵与精度的体内。在这些情况下，基因包含组织切除术被认为是更有效的学习方法，以确定组织传播的机械和功能意义。
回肠是克罗恩病的主要是用于的患者和涉及回肠1，，23的炎症性疾病。回肠通常会产生几个能量存储激素样成纤维细胞生长因子 15/19 (FGF15/19)、 肽 (PYY) 和胰高血糖素样肽-1/2 (GLP1/2);这些激素许多生物学功能4，，56中发挥了重要的地方和内分泌作用。这些激素间 FGF15 已被确定为肝脏的胆汁酸合成鲁棒内分泌抑制剂。一旦进入回肠肠吸收，胆汁酸激活核受体法尼酯 X 受体 (FXR) 来刺激Fgf15的表达，随后导致的肝胆汁酸合成7反馈抑制。在最近的一项研究中，我们介绍了小鼠回肠模型为研究回肠 kruppel 样因子 (KLF15) 15-Fgf15信号调节昼夜节律的胆汁酸生产中肝8的轴。最重要的是，我们新的家庭，kruppel 样因子，特别是 KLF15，引入胆汁酸生物学。我们决定通过一种间接的非肝机制，KLF15 上调胆汁酸合成包括回肠手术的功能研究的基础。最后，回肠 KLF15 也被称为Fgf15第一内源性负性调节。
从近端到远端区域降序的肠段是负责吸收不同的营养。回肠是负责胆汁酸和维生素 B12 (VB12) 吸收9的主要部分。早先的研究中采用小鼠模型的近端肠切除术研究短肠综合征;提出了各种切除长度、 饮食和缝合类型，以保持最佳的手术后生存率10。此外，最近的审查表明，回肠切除通常会导致更严重的疾病比其他胃肠道 (GI) 段切除术由于剩余道11的自适应能力下降。本主题获得了集约化利益的基础和临床研究群体，而复苏和有效的治疗方法的了解仍然很有限。
胆汁酸腹泻结果从胆汁酸肝肠循环12，13稳态失衡。它可以是回肠切除术、 胃肠道疾病，或结果特发性胆汁酸吸收不良的后果。已发现了超过 80%的患者，切除回肠切除14后提出与腹泻。回肠有潜力成为重要手术模型为胆汁酸腹泻的调查。在此研究中，回肠切除一系列提供梯度评估 FGF15 不足以及肠胆盐吸收不良、 积滞和毒性损害。

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			<td height="21" style="width:248px;height:21px;">Dissection microscope</td>
			<td style="width:195px;">Olympus</td>
			<td style="width:97px;">SZ61</td>
			<td style="width:183px;">For surgery&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:248px;height:21px;">Animal temperature controller</td>
			<td style="width:195px;">Physitemp Instruments, Inc.</td>
			<td style="width:97px;">TCAT-2LV</td>
			<td style="width:183px;">For body temperature control</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:248px;height:21px;">Isoflurane anesthetic vaporizer</td>
			<td style="width:195px;">VetEquip</td>
			<td style="width:97px;">&#160;911104</td>
			<td style="width:183px;">For anesthesia</td>
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			<td height="21" style="width:248px;height:21px;">Dissection forceps</td>
			<td style="width:195px;">&#160;Fine Science Tools, Inc.</td>
			<td style="width:97px;">11274-20</td>
			<td style="width:183px;">For surgery&#160;</td>
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			<td height="21" style="width:248px;height:21px;">Scissors</td>
			<td style="width:195px;">&#160;Fine Science Tools, Inc.</td>
			<td style="width:97px;">14084-08</td>
			<td style="width:183px;">For surgery&#160;</td>
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		<tr height="21">
			<td height="21" style="width:248px;height:21px;">Needle holder</td>
			<td style="width:195px;">&#160;Roboz Surgical Instrument Co.</td>
			<td style="width:97px;">RS-7882</td>
			<td style="width:183px;">For surgery&#160;</td>
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		<tr height="21">
			<td height="21" style="width:248px;height:21px;">Micro knives-needle blade</td>
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			<td style="width:97px;">10318-14</td>
			<td style="width:183px;">For surgery&#160;</td>
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		<tr height="21">
			<td height="21" style="width:248px;height:21px;">6-0 monofilament suture</td>
			<td style="width:195px;">Ethicon</td>
			<td style="width:97px;">1698G</td>
			<td style="width:183px;">For abdominal skin closure</td>
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		<tr height="21">
			<td height="21" style="width:248px;height:21px;">7-0 silk suture</td>
			<td style="width:195px;">Ethicon</td>
			<td style="width:97px;">766G</td>
			<td style="width:183px;">For ligation</td>
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		<tr height="21">
			<td height="21" style="width:248px;height:21px;">8-0 monofilament suture</td>
			<td style="width:195px;">Ethicon</td>
			<td style="width:97px;">1714G</td>
			<td style="width:183px;">For anastomosis</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:248px;height:21px;">Surgical sponges</td>
			<td style="width:195px;">Dynarex Corp.</td>
			<td style="width:97px;">3333</td>
			<td style="width:183px;">For surgery&#160;</td>
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			<td height="21" style="width:248px;height:21px;">Small cotton-tipped applicators&#160;</td>
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			<td style="width:97px;">23-400-118</td>
			<td style="width:183px;">For surgery&#160;</td>
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			<td height="21" style="width:248px;height:21px;">Isoflurane</td>
			<td style="width:195px;">Piramal Healthcare Limited</td>
			<td style="width:97px;">66794-013-25</td>
			<td style="width:183px;">For anasthesia</td>
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			<td height="21" style="width:248px;height:21px;">Buprenorphine hydrochloride</td>
			<td style="width:195px;">Reckitt-Benckiser Pharmaceuticals</td>
			<td style="width:97px;">12496-0757-1</td>
			<td style="width:183px;">For analgesia</td>
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			<td height="21" style="width:248px;height:21px;">0.9% sodium chloride Injection</td>
			<td style="width:195px;">B. Braun Medical Inc.</td>
			<td style="width:97px;">0264-7800-10</td>
			<td style="width:183px;">For washing/injection</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:248px;height:21px;">Povidone iodine prep solution</td>
			<td style="width:195px;">Dynarex Corp.</td>
			<td style="width:97px;">1413</td>
			<td style="width:183px;">For skin preparation</td>
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		<tr height="21">
			<td height="21" style="width:248px;height:21px;">Puralube vet ointment</td>
			<td style="width:195px;">Dechra Veterinary Products</td>
			<td style="width:97px;">17033-211-38</td>
			<td style="width:183px;">For eye pretection</td>
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			<td height="21" style="width:248px;height:21px;">Hair remover lotion</td>
			<td style="width:195px;">Church &#38; Dwight Co., Inc.</td>
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			<td style="width:183px;">For skin preparation</td>
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			<td height="21" style="width:248px;height:21px;">Intensive care unit</td>
			<td style="width:195px;">ThermoCare</td>
			<td style="width:97px;">FW-1</td>
			<td style="width:183px;">For post-surgery recovery</td>
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			<td height="21" style="width:248px;height:21px;">DietGel recovery</td>
			<td style="width:195px;">ClearH2O</td>
			<td style="width:97px;">72-06-5022</td>
			<td style="width:183px;">For post-surgery recovery</td>
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			<td height="21" style="width:248px;height:21px;">Aurum total RNA fatty and fibrous tissue kit</td>
			<td style="width:195px;">Bio-Rad</td>
			<td style="width:97px;">7326830</td>
			<td style="width:183px;">For RNA isolation</td>
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		<tr height="31">
			<td height="31" style="width:248px;height:31px;">iScript reverse transcription supermix for RT-qPCR</td>
			<td style="width:195px;">Bio-Rad</td>
			<td style="width:97px;">1708841</td>
			<td style="width:183px;">For reverse transcription assay</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:248px;height:21px;">TaqMan fast advanced master mix</td>
			<td style="width:195px;">Applied Biosystems/Life Technologies</td>
			<td style="width:97px;">4444965</td>
			<td style="width:183px;">For QPCR analysis</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:248px;height:21px;">Total bile acid assay kit</td>
			<td style="width:195px;">Genzyme Diagnostic</td>
			<td style="width:97px;">DZ042A-K01</td>
			<td style="width:183px;">For bile acid assay</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:248px;height:21px;">C57BL/6J&#160;</td>
			<td style="width:195px;">The Jackson Laboratory</td>
			<td style="width:97px;"></td>
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ileectomy-induced bile overaccumulation in mouse intestine

肠段切除术是常见的治疗方法，为人类的疾病，如肥胖、 炎症性肠病、 克罗恩病和结肠癌，往往导致严重的短肠综合征样不良反应包括胆汁酸腹泻脱水、 电解质紊乱、 营养吸收不良。在这里我们介绍一种小鼠回肠切除模型，称为回肠，评价组织通信和系统稳态的维持。回肠切除术后血液循环是永久没有回肠特定内分泌激素成纤维细胞生长因子 15 (FGF15)，释放出肝脏的胆汁酸合成及其内分泌抑制。与增加的生产和胆汁酸后删除回肠的废除的吸收相结合，接受手术的老鼠患胆盐积滞在肠和相关性的腹泻、 发病率和死亡率。小说使用的这项研究中引入的手术模型可能洞察机械和功能回肠控制的系统性代谢调控的生理和疾病。

回肠的程序如图 1所示。第一步包括制备小鼠腹部皮肤，使腹部的切口，并利用牵开器充分暴露肠 (图 1A C)。接下来，鼠标盲肠是位于 (图 1D);给出了它的大小和形状使其很容易辨认的一个里程碑。盲肠、 回肠与远端空肠的一部分被曝光并移除从腹腔内 (图 1<s...

Watch this Scientific Journal Video about Ileectomy-induced Bile Overaccumulation in Mouse Intestine at JoVE.com

Ileectomy-induced Bile Overaccumulation in Mouse Intestine

小肠依赖胆汁酸的重吸收和反馈抑制肝胆汁酸合成系统稳态和健康至关重要。在此研究中，我们描述为回肠切除回肠致胆汁吸收不良，积滞，和毒性小鼠肠道中的小鼠模型。

回肠致胆汁积滞在小鼠肠道中

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Hyperinsulinemic-euglycemic Clamps in Conscious, Unrestrained Mice

在自觉的，奔放的小鼠正常血糖高胰岛素 - 夹钳

Type 2 diabetes is characterized by a defect in insulin action. The hyperinsulinemic-euglycemic clamp, or insulin clamp, is widely considered the "gold ...

科研

医学

3-Dimensional Resin Casting and Imaging of Mouse Portal Vein or Intrahepatic Bile Duct System

树脂浇铸和3维成像的小鼠门静脉或肝内胆管系统

In  organs, the correct architecture of vascular and ductal structures is  indispensable for proper physiological function, and the formation and  ...

A Simple Method of Mouse Lung Intubation

一个简单的方法小鼠肺插管

A  simple procedure to intubate mice for pulmonary function measurements would  have several advantages in longitudinal studies with limited numbers or  ...

Technical Aspects of the Mouse Aortocaval Fistula

在的鼠标Aortocaval瘘的技术方面

Technical aspects of creating an arteriovenous fistula in the mouse are discussed. Under general anesthesia, an abdominal incision is made, and the aorta ...

Isolating and Using Sections of Bovine Mesenteric Artery and Vein as a Bioassay to Test for Vasoactivity in the Small Intestine

隔离和使用的牛肠系膜动脉和静脉部分，作为生物测定在小肠测试的Vasoactivity

Mammalian gastrointestinal systems are constantly exposed to compounds (desirable and undesirable) that can have an effect on blood flow to and from that ...

Bile Duct Ligation in Mice: Induction of Inflammatory Liver Injury and Fibrosis by Obstructive Cholestasis

胆管结扎小鼠：感应炎性肝损伤和纤维化的阻塞性胆汁淤积

In most vertebrates, the liver produces bile that is necessary to emulsify absorbed fats and enable the digestion of lipids in the small intestine as well ...

Intrauterine Telemetry to Measure Mouse Contractile Pressure In Vivo

Intrauterine Telemetry to Measure Mouse Contractile Pressure In Vivo

宫内遥测来衡量鼠标收缩压力<em&gt;在体内</em

A complex integration of molecular and electrical signals is needed to transform a quiescent uterus into a contractile organ at the end of pregnancy. ...

A Simple Device to Rapidly Prepare Whole Mounts of the Mouse Intestine

一个简单的设备，迅速准备小鼠肠的全坐骑

Preparing whole mounts of the mouse small intestine and colon for subsequent analysis or quantification can be time consuming and difficult. We describe ...

Isolation and Profiling of MicroRNA-containing Exosomes from Human Bile

分离和剖析人力胆汁含外来体的MicroRNA-

Exosome research in the last three years has greatly extended the scope towards identification and characterization of biomarkers and their therapeutic ...