Name: murine distal colostomy, a novel model of diversion colitis in c57bl/6 mice
Uploaded: 2018-07-12T13:30:00.000+00:00
Duration: 8 min 20 s
Description: Watch this Scientific Journal Video about Murine Distal Colostomy, A Novel Model of Diversion Colitis in C57BL/6 Mice at JoVE.com

这里描述的所有方法都已得到兽医政府当局 (Landesamt Landwirtschaft, Lebensmittelsicherheit 和 Fischerei, Vorpommern, LALLF M V) 的批准。
1. 动物的术前护理和准备
<ol><li>到达动物设施后, 将动物 (C57Bl/6) 分成相似大小的组, 将每个组保持在一起, 并在整个实验中保持组恒定。 注意: 使用相同性别的动物。用雄性小鼠得到了实验结果。 注意: 如果使用雄性动物, 笼群在7周的年龄开始, 这样就可以建立一个层次结构, 从而最大限度地减少实验期间攻击性行为的风险。</li>
	<li>手术前至少一周, 切换到高能 (> 14 兆焦耳/千克) 和高蛋白 (> 20%) 饲料含有所有必需的微量元素和维生素 (详情, 见材料清单)。 注: 确保所有小鼠在执行手术时重量至少25克。</li>
	<li>通过腹腔注射氯胺酮 (87 毫克/千克 ip) 和盐酸甲苯噻嗪 (13 毫克/千克 ip) 诱导麻醉和镇痛。等到老鼠容忍机械刺激, 如脚趾间距, 没有马达反应。</li>
	<li>将麻醉鼠标固定在位于操作台上的热垫上的仰卧位上, 保证在操作过程中稳定的定位, 避免全身热量的巨大损失。 注: 热衬底应具有36°c 到40°c 的表面温度;手术室温度应为21摄氏度。</li>
</ol>2. 远端结肠造口术
<ol><li>剃掉腹部的头发。在开始手术前, 使用酒精70% 和碘伏消毒手术现场三次。将操作现场悬垂, 保证无菌条件。</li>
	<li>执行15毫米中位剖腹手术通过切割腹部肌肉和腹膜沿 linea, 从而减少失血。</li>
	<li>使用两个迈克迪贝克非创伤性钳, 小心拉盲肠, 末端回肠, 和上升和横结肠从腹膜腔。 注意: 小心严格限制肠道的机械操作, 以防止对肠系膜结构的损伤。</li>
	<li>识别盲肠极, 上升结肠和小肠 (图1a 和 1b)。 注意: 正确识别上升结肠是正确放置结肠造口的基础。在回盲区解剖不明确的情况下, Peyer 斑块的存在识别小肠, 而形成的凳子的出现则是结肠的特点。</li>
	<li>使用标尺确定未来结肠造口的位置。它应该被放置在20毫米远端的回盲阀的远端结肠造口。</li>
	<li>在右上象限的腹壁上再做3毫米切口。通过这个切口提取先前识别的结肠段, 形成一个循环, 小心不要扭曲循环。</li>
	<li>小心通过乙状结肠22口径灵活的静脉注射套管。注意不要损伤肠系膜血管结构。</li>
	<li>将肠道恢复到腹膜腔。</li>
	<li>用简单的缝线和可吸收缝合 (例如, polyglactin 910 或 polyfil 4-0 1/2 c) 将软管的两端固定在皮肤上。</li>
	<li>在关闭剖腹手术前, 用腹腔注射0.5 毫升0.9% 生理盐水进行液体复苏。</li>
	<li>用可吸收缝合 (如polyglactin 910 或 polyfil 4-0 1/2 c) 闭合腹膜和肌肉层, 并进行连续缝合。用可吸收缝合 (如 polyglactin 910 或 polyfil 4-0 1/2 c), 用连续缝合法关闭皮肤。</li>
	<li>通过使用细剪刀执行小计横断来打开形象化冒号循环。避免所有的肠系膜损伤。不要完全横过结肠。</li>
	<li>使用 monofil, 可吸收缝线 (如polydioxanone 或 monofil 6-0 3/8) 修复每个结肠造口, 使用三张全厚针缝合腹膜和皮肤。传入回路, 这是一个功能性的末端结肠造口, 和传出循环, 这是一个粘液瘘, 在这一点上明显分离 (图 1c)。 注: 操作时间应少于20分钟, 以限制流体和热损失。</li>
	<li>完成手术后, 根据制造商的指示, 在超声波浴中使用无醛消毒液消毒仪器。</li>
</ol>3. 假操作 (Colotomy)
<ol><li>执行步骤1.1。到2.4。</li>
	<li>使用标尺确定未来的 colotomy 位置。colotomy 应定位与回盲阀的距离为实验组的结肠造口。</li>
	<li>用细剪刀打开结肠至少2/3 的周长。</li>
	<li>使用 monofil, 可吸收缝线 (例如, polydioxanone, monofil 6-0 3/8 s), 用单层、全厚度中断缝线闭合 colotomy。 注: 实验外科医生的手术时间应少于20分钟, 限制液体和热损失。</li>
	<li>执行步骤2.10。, 2.11。和2.14。</li>
</ol>4. 术后护理
<ol><li>把动物送回笼子里去。提供一个良好的气氛, 37 摄氏度 (如红外线灯), 直到老鼠完全清醒。然后, 保持老鼠在温度和湿度调节的环境 (21 °c; 30% 10% 相对湿度)。允许免费获得食物和饮用水。为促进流体吸收, 提供额外的水浸泡动物饲料。</li>
	<li>当动物表现出对机械刺激的反应时, 注射0.1 毫克/千克体重丁丙诺啡, 开始术后镇痛。小心避免呼吸压抑。</li>
	<li>用1毫克/毫升曲马多补充饮用水, 在术后第一周继续镇痛。</li>
	<li>为了补偿由于流动性降低而减少的液体摄入量, 在第一个术后的一周内在笼子里供应一个固体饮料垫。</li>
	<li>在第一周, 每第二天在第一个月的剩余时间, 每隔三天, 在第二个月, 使用佳中描述的疾病严重性评分, 衡量动物的体重, 并评分动物行为。6。</li>
</ol>

<ol>
	<li>Glotzer, D. J., Glick, M. E., Goldman, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Proctitis+and+colitis+following+diversion+of+the+fecal+stream.">Proctitis and colitis following diversion of the fecal stream.</a> Gastroenterology. 80, 438-441 (1981).</li><li>Morson, B., Dawson, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Gastrointestinal Pathology. , (1972).</li><li>Whelan, R. L., Abramson, D., Kim, D. S., Hashmi, H. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diversion+colitis.+A+prospective+study.">Diversion colitis. A prospective study.</a> Surg Endosc. 8, 19-24 (1994).</li><li>Haas, P. A., Fox, T. A., Szilagy, E. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endoscopic+examination+of+the+colon+and+rectum+distal+to+a+colostomy.">Endoscopic examination of the colon and rectum distal to a colostomy.</a> Am J Gastroenterol. 85, 850-854 (1990).</li><li>Valatas, V., Bamias, G., Kolios, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+colitis+models:+Insights+into+the+pathogenesis+of+inflammatory+bowel+disease+and+translational+issues.">Experimental colitis models: Insights into the pathogenesis of inflammatory bowel disease and translational issues.</a> Eur J Pharmacol. 759, 253-264 (2015).</li><li>Kleinwort, A., D&#246;ring, P., Hackbarth, C., Heidecke, C. D., Schulze, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Deviation+of+the+Fecal+Stream+in+Colonic+Bowel+Segments+Results+in+Increased+Numbers+of+Isolated+Lymphoid+Follicles+in+the+Submucosal+Compartment+in+a+Novel+Murine+Model+of+Diversion+Colitis.">Deviation of the Fecal Stream in Colonic Bowel Segments Results in Increased Numbers of Isolated Lymphoid Follicles in the Submucosal Compartment in a Novel Murine Model of Diversion Colitis.</a> Biomed Res Int. 2017, 13 (2017).</li><li>Tangjarukij, C., Navasumrit, P., Zelikoff, J. T., Ruchirawat, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effects+of+pyridoxine+deficiency+and+supplementation+on+hematological+profiles,+lymphocyte+function,+and+hepatic+cytochrome+P450+in+B6C3F1+mice.">The effects of pyridoxine deficiency and supplementation on hematological profiles, lymphocyte function, and hepatic cytochrome P450 in B6C3F1 mice.</a> J Immunotoxicol. 6, 147-160 (2009).</li><li>Soave, O., Brand, C. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Coprophagy+in+animals:+a+review.">Coprophagy in animals: a review.</a> Cornell Vet. 81, 357-364 (1991).</li><li>Ebino, K. Y., Yoshinaga, K., Suwa, T., Kuwabara, Y., Takahashi, K. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+prevention+of+coprophagy+on+pregnant+mice--is+coprophagy+beneficial+on+a+balanced+diet.">Effects of prevention of coprophagy on pregnant mice--is coprophagy beneficial on a balanced diet.</a> Jikken Dobutsu. 38, 245-252 (1989).</li><li>Babickova, J., Tothova, L., Lengyelova, E., Bartonova, A., Hodosy, J., Gardlik, R., Celec, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sex+Differences+in+Experimentally+Induced+Colitis+in+Mice:+a+Role+for+Estrogens.">Sex Differences in Experimentally Induced Colitis in Mice: a Role for Estrogens.</a> Inflammation. 38, 1996-2006 (2015).</li><li>Lee, S. M., Kim, N., Son, H. J., Park, J. H., Nam, R. H., Ham, M. H., Choi, D., Sohn, S. H., Shin, E., Hwang, Y. J., 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=The+Effect+of+Sex+on+the+Azoxymethane/Dextran+Sulfate+Sodium-treated+Mice+Model+of+Colon+Cancer.">The Effect of Sex on the Azoxymethane/Dextran Sulfate Sodium-treated Mice Model of Colon Cancer.</a> J Cancer Prev. 21, 271-278 (2016).</li><li>Angele, M. K., Pratschke, S., Hubbard, W. J., Chaudry, I. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gender+differences+in+sepsis:+cardiovascular+and+immunological+aspects.">Gender differences in sepsis: cardiovascular and immunological aspects.</a> Virulence. 5, 12-19 (2013).</li><li>Brown, R. Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Social+Behaviour,+reproduction,+and+population+changes+in+the+house+mouse+(Mus+musculus+L).">Social Behaviour, reproduction, and population changes in the house mouse (Mus musculus L).</a> Eccological Monographs. 23, 788-795 (1953).</li><li>Poole, T. B., Morgan, H. D. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differences+in+aggressive+behaviour+betweeen+male+mice+(Mus+musculus+L.)+in+colonies+of+different+sizes.">Differences in aggressive behaviour betweeen male mice (Mus musculus L.) in colonies of different sizes.</a> Animal Behaviour. 21, 788-795 (1973).</li><li>Pasquarelli, N., Voehringer, P., Henke, J., Ferger, B. <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+a+change+in+housing+conditions+on+body+weight,+behavior+and+brain+neurotransmitters+in+male+C57BL/6J+mice.">Effect of a change in housing conditions on body weight, behavior and brain neurotransmitters in male C57BL/6J mice.</a> Behav Brain Res. 333, 35-42 (2017).</li><li>Langgartner, D., Foertsch, S., Fuchsl, A. M., Reber, S. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Light+and+water+are+not+simple+conditions:+fine+tuning+of+animal+housing+in+male+C57BL/6+mice.">Light and water are not simple conditions: fine tuning of animal housing in male C57BL/6 mice.</a> Stress. 20, 10-18 (2017).</li><li>Nicholson, A., Malcolm, R. D., Russ, P. L., Cough, K., Touma, C., Palme, R., Wiles, M. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+response+of+C57BL/6J+and+BALB/cJ+mice+to+increased+housing+density.">The response of C57BL/6J and BALB/cJ mice to increased housing density.</a> J Am Assoc Lab Anim Sci. 48, 740-753 (2009).</li><li>Buchler, G., Wos-Oxley, M. L., Smoczek, A., Zschemisch, N. H., Neumann, D., Pieper, D. H., Hedrich, H. J., Bleich, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Strain-specific+colitis+susceptibility+in+IL10-deficient+mice+depends+on+complex+gut+microbiota-host+interactions.">Strain-specific colitis susceptibility in IL10-deficient mice depends on complex gut microbiota-host interactions.</a> Inflamm Bowel Dis. 18, 943-954 (2012).</li><li>Nakanishi, M., Tazawa, H., Tsuchiya, N., Sugimura, T., Tanaka, T., Nakagama, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+strain+differences+in+inflammatory+responses+of+colonic+mucosa+induced+by+dextran+sulfate+sodium+cause+differential+susceptibility+to+PhIP-induced+large+bowel+carcinogenesis.">Mouse strain differences in inflammatory responses of colonic mucosa induced by dextran sulfate sodium cause differential susceptibility to PhIP-induced large bowel carcinogenesis.</a> Cancer Sci. 98, 1157-1163 (2007).</li><li>Mahler, M., Bristol, I. J., Leiter, E. H., Workman, A. E., Birkenmeier, E. H., Elson, C. O., Sundberg, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differential+susceptibility+of+inbred+mouse+strains+to+dextran+sulfate+sodium-induced+colitis.">Differential susceptibility of inbred mouse strains to dextran sulfate sodium-induced colitis.</a> Am J Physiol. 274, 544-551 (1998).</li><li>Mekada, K., Abe, K., Murakami, A., Nakamura, S., Nakata, H., Moriwaki, K., Obata, Y., Yoshiki, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetic+differences+among+C57BL/6+substrains.">Genetic differences among C57BL/6 substrains.</a> Exp Anim. 58, 141-149 (2009).</li><li>Sellers, R. S., Clifford, C. B., Treuting, P. M., Brayton, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Immunological+variation+between+inbred+laboratory+mouse+strains:+points+to+consider+in+phenotyping+genetically+immunomodified+mice.">Immunological variation between inbred laboratory mouse strains: points to consider in phenotyping genetically immunomodified mice.</a> Vet Pathol. 49, 32-43 (2012).</li><li>Mills, C. D., Kincaid, K., Alt, J. M., Heilman, M. J., Hill, A. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=M-1/M-2+macrophages+and+the+Th1/Th2+paradigm.">M-1/M-2 macrophages and the Th1/Th2 paradigm.</a> J Immunol. 164, 6166-6173 (2000).</li></ol>

所有作者都宣称他们没有相互竞争的兴趣。

本协议所提出的小鼠模型可靠地再现了人 dc 的组织病理学特征 ( 例如, 在炎症性肠段粘膜下层淋巴滤泡的发育, 隐窝缩短, 以及减少杯状细胞数)。除了这个优势, 这个模型是由一个非常相似的触发因素引起的, 并呈现出一个中等到轻度的严重性的临床过程, 正如在大多数受影响的人的情况。
为获得重现性结果和可接受的围手术期致死率, 需要一些显微外科技术的实践。然而, 结肠造口和假手术 (colotomy) 可以执行非常低的围手术期死亡率 (< 10%)。一些操作步骤是非常重要的: 1) 积极识别结肠和末端回肠。在回盲区解剖不清楚的情况下, 结肠可以通过存在 Peyer 的补丁和回肠末端来识别。2) exteriorizing 结肠造口回路时, 保证肠系膜不扭曲, 避免肠系膜缺血和机械肠梗阻。3) 用软管穿刺肠系膜时, 确保选择无血管区, 从而预防出血和肠系膜缺血。4) 当在腹壁上创建小切口时, 检查是否有足够的止血。如有必要, 可使用热凝固停止肌肉血管出血。5) 确保结肠环至少在其周长的2/3 处开放, 后壁充分形象化, 从而保证大便通道完全堵塞到远端肠段。6) 确保结肠造口术的两端都能很好地固定在皮肤上, 以避免由于粪便流不能排出而导致感染风险增高或机械性肠梗阻的开放性通道。正常情况下, 所有手术程序都可以不用放大镜或手术显微镜进行。一般而言, 这些设备的使用需要进一步的培训。
即使手术过程的复杂性是适度的, 也有许多必要的手术和围手术期的问题, 以成功终止实验的可重现的结果。
它是必不可少的长期生存与永久性结肠造口, 老鼠有一个完整的盲肠。与回盲阀相邻的近端 colostomies 的实验组显示, 术后一周的死亡率为 100%, 原因是不可逆转的体重损失 (未显示数据)。因此, 最重要的是选择结肠造口部位至少20毫米远端从 Bauhin 的阀门。这种观察可能是由于啮齿动物消化的生理。从近端结肠到盲肠的逆行转运发生在这些物种中。盲肠是由细菌菌群 occurs8 合成养分的地方。正常的 MCV 和妇幼保健的价值在长期随访表明, 动物与远端 colostomies 不遭受严重的维生素缺乏。此外, coprophagy 是啮齿动物在自然环境中营养行为的重要组成部分。在建立模型时, 我们高度关注的是, 由于存在永久性结肠造口, 扰动 coprophagy 可能导致营养不足 (如维生素 B)。然而, 虽然这是一个问题, 在获得营养的条件是有限的, 以前已经表明, coprophagy 失去它的营养价值在实验条件下, 自由过剩的水和营养均衡的食物9.在实验条件下, 优化术后营养摄入的条件至关重要。我们使用了蛋白质含量高、蛋白质品质优化、能量密度提高的动物饲料。动物在手术前至少两周适应这种饮食习惯。术后, 给有利于食物和液体摄入提供了libidum的水浸泡饲料。
在实验性和假组中利用同种性别的小鼠是至关重要的。性已经表明, 不仅对全身炎症的过程, 而且对肠道炎症的影响, 在各种化学诱发的结肠炎模型10,11,12。一般情况下, 雄性动物会产生更明显的疾病, 炎症更大, 更隐窝损伤, 再生少, 促炎性细胞因子水平增高, 体重减轻10。使用雄性动物的另一个好处是更好的年龄匹配的术前体重, 从而更好地补偿最初的术后减肥期。然而, 如果要解决特定的问题 (例如性激素对 DC 表现的影响), 雌性小鼠可以在《议定书》中使用。使用雄性老鼠需要更多的努力来维持稳定的住房条件。雄性小鼠更倾向于积极的相互作用导致伤害和死亡。雄性小鼠与一个显性男性生活在既定的层次上。一旦团队中的层次结构建立起来, 持续的战斗就不那么常见了13,14。由于这个原因, 我们在动物设施的老鼠到达前至少两周就形成了实验组, 直到实验结束, 这些小组才得以维持。一般来说, 实验组的5到7只动物被保存在一个笼子里。维持稳定的社会和住房条件对于尽量减少实验动物的攻击行为至关重要。住房条件已经显示影响体重增加以及免疫参数15,16,17。
对化学诱导和基因改良的结肠炎模型的易感性在小鼠菌株18,19,20之间有很大差异。本议定书是建立在 C57Bl/6 小鼠, 一个小鼠菌株与明确的遗传背景和良好的免疫系统21,22,23显示中间的强炎症反应在葡聚糖硫酸钠诱导的结肠炎模型20和被认为是一个典型的 Th1 菌株23。当需要更严重的疾病活动或 Th2 环境时, 可以考虑使用其他老鼠菌株。
总之, 我们的议定书是评估各种免疫细胞类型、细胞因子、趋化因子和其他信号分子在 DC 发病中的作用的宝贵工具。大量的基因修饰小鼠模型可与 C57BL/6 背景结合, 并可与我们的模型 (例如, 小鼠缺乏 IL-17, IL-10, 趋化因子 CXCL13, 因子受体 CXCR5 和 CCR7, sphingosine-1-phosphate受体 4)。在 C57BL/6 背景下, 同类系小鼠菌株的可用性在很大程度上促进了转移实验, 从而确定了不同细胞类型在 DC 病因学中的作用。最后, 该模型提供了机会, 以评估当地干预 (如修改当地微生物和局部抗炎治疗) 对黏膜免疫的影响和非受影响的肠段和系统性免疫平衡。

近年来, 大量的非传染性结肠炎实体不同于典型的炎症性肠病 (IBDs;i., 克罗恩病或结肠炎溃疡性皮肤) 已临床和病理的特点在人类。导致这些结肠炎形式的发展的病理生理学机制部份地不完全地被了解, 因为适当的动物模型是稀缺的。分流性结肠炎是最近被描述的实体之一。虽然这个词在1980年由 Glotzer1创造了, 相似的表型的最初的描述在1972年被给了 Morson2。该病在50% 至91% 的转移肠患者中发展, 其临床强度变化3,4。鉴于美国每年约有12万口结肠造口患者的发病率, 这一疾病实体构成了一个重要的健康问题。
制定这项议定书的总目标是提供一个小鼠 dc 模型, 它依赖于类似于人类 DC 中所看到的结肠炎触发, 再现人类疾病的主要病理学特征。与其他小鼠结肠炎模型相比, 我们模型中的结肠炎诱导不需要转基因动物 (例如, IL-7 转基因小鼠, n-钙粘蛋白显性阴性小鼠, 或 TGFβ鼠), 应用化学刺激性物质 (例如, 葡聚糖硫酸盐钠 (DSS) 诱发结肠炎, 或硝基苯磺酸 (TNBS) 诱发结肠炎), 或在免疫缺陷小鼠 (如 CD45RB高转移的特定细胞群的转移结肠炎模型) (综述, 见5)。与其他模型相比, 我们的 dc 模型的完整免疫系统允许评估在 dc 发展中所涉及的免疫机制。粘膜炎症对被排除的肠段的限制, 允许评估其对胃肠道其他部位粘膜免疫的影响, 对肠道其他免疫室的免疫平衡 (例如,, Peyer 的补丁和 mesenteriale 淋巴结), 以及整个生物体的免疫平衡。最后, 我们的模型构成了一个适当的工具来研究控制局部炎症刺激的机制, 这是由于当地微生物和消化抗原的正常当地环境的变化。

<table><tbody><tr><td>&lt;em&gt;Operation material&lt;/em&gt;</td><td></td><td></td><td></td></tr><tr><td>heat underlay, 6 watt</td><td>ThermoLux, Witte + Sutor GmbH</td><td>461265</td><td></td></tr><tr><td>Ethanol 70% (with methylethylketon)</td><td>Pharmacie of the University Hospital Greifswald</td><td></td><td></td></tr><tr><td>Wooden applicators, small cotton head onesided, wooden stick, 150 mm length</td><td>Centramed GmbH, Germany</td><td>8308370</td><td>for disinfection of operation field</td></tr><tr><td>Scissor</td><td>Aesculap (BRAUN)</td><td>BC064R</td><td></td></tr><tr><td>Atraumatic DeBakey forceps</td><td>Aesculap (BRAUN)</td><td>OC021R</td><td></td></tr><tr><td>Needle holder</td><td>Aesculap (BRAUN)</td><td>BM012R</td><td></td></tr><tr><td>Vasofix Safety &lt;em&gt;i.v.&lt;/em&gt; cannula 22G</td><td>Braun Melsungen AG, Germany</td><td>4268091S-01</td><td></td></tr><tr><td>VICRYL Plus 4-0 violet braided; V-5 17 m 1/2c; 70 cm</td><td>ETHICON, Inc.</td><td>VCP994H</td><td></td></tr><tr><td>PDS II 6-0 monofil; V-18 13 mm 3/8c; 70 cm</td><td>ETHICON, Inc.</td><td>Z991H</td><td></td></tr><tr><td>BD Plastipak 1 mL (Syringes with needle with sterile interior)</td><td>BD Medical</td><td>305501</td><td></td></tr><tr><td>Triacid-N (N-Dodecylpropan-1,3-diamin)</td><td>ANTISEPTICA, Germany</td><td>18824-01</td><td>disinfection of surgical instruments in ultrasonic bath</td></tr><tr><td>&lt;em&gt;Medications&lt;/em&gt;</td><td></td><td></td><td></td></tr><tr><td>ketamine 10%, 100 mg/mL (ketamine hydrochloride)</td><td>selectavet, Dr. Otto Fischer GmbH</td><td>9089.01.00</td><td>87 mg/kg &lt;em&gt;i.p.&lt;/em&gt;</td></tr><tr><td>Xylasel, 20 mg/mL (xylazine hydrochloride)</td><td>selectavet, Dr. Otto Fischer GmbH</td><td>400300.00.00</td><td>13 mg/kg &lt;em&gt;i.p.&lt;/em&gt;</td></tr><tr><td>NaCl 0.9%</td><td>Braun Melsungen AG, Germany</td><td>6697366.00.00</td><td></td></tr><tr><td>Buprenovet 0.3 mg/mL (buprenorphine)</td><td>Bayer, Germany</td><td>PZN: 01498870</td><td>0.1 mg/kg &lt;em&gt;s.c.&lt;/em&gt;</td></tr><tr><td>Tramal Drops, 100 mg/mL (tramadol hydrochloride)</td><td>Gr&amp;uuml;nenthal GmbH, Germany</td><td>10116838</td><td>1 mg/mL drinking water</td></tr><tr><td>Ceftriaxon-saar 2 g (ceftriaxone)&amp;nbsp;</td><td>Cephasaar GmbH, Germany</td><td>PZN: 08844252</td><td>25 mg/kg body weight &lt;em&gt;i.p.&lt;/em&gt;</td></tr><tr><td>metronidazole 5 mg/mL</td><td>Braun Melsungen AG, Germany</td><td>PZN: 05543515</td><td>12.5 mg/kg body weight &lt;em&gt;i.p.&lt;/em&gt;</td></tr><tr><td>&lt;em&gt;Food&lt;/em&gt;</td><td></td><td></td><td></td></tr><tr><td>ssniff M-Z Ereich</td><td>ssniff, Germany</td><td>&amp;nbsp;V1184-3</td><td></td></tr><tr><td>Solid Drink Dehyprev Vit BIO, pouches</td><td>TripleATrading, the Netherlands</td><td>SDSHPV-75</td><td></td></tr><tr><td>Equipment</td><td></td><td></td><td></td></tr><tr><td>Nikon Eclipse Ci-L</td><td>Nikon Instruments Europe BV, Germany</td><td></td><td>light microscopy</td></tr><tr><td>VetScan HM 5&amp;nbsp;</td><td>Abaxis, USA</td><td>770-9000</td><td>Veterinary hematology analyzer of 50 &amp;micro;l venous EDTA-blood</td></tr><tr><td>Bandelin SONOREX (Ultrasonic bath)</td><td>Bandelin electronics, Germany</td><td>RK 100 H</td><td>disinfection of surgical instruments</td></tr><tr><td>&lt;em&gt;Software&lt;/em&gt;</td><td></td><td></td><td></td></tr><tr><td>NIS-Element BR4 software</td><td>Nikon Instruments Europe BV, Germany</td><td></td><td></td></tr><tr><td>GraphPad Prism Version 6</td><td>GraphPad Software, Inc.</td><td></td><td></td></tr></tbody></table>

murine distal colostomy, a novel model of diversion colitis in c57bl/6 mice

分流性结肠炎 (DC) 是一种常见的临床情况发生在肠段的病人排除在粪便流由于分流肠。这种疾病的病因仍然不明确, 但似乎有别于传统的炎症性肠病, 如克罗恩病和溃疡结肠炎。研究旨在破译导致这种疾病发展的病理生理学机制, 由于缺乏合适的小鼠模型而受到严重阻碍。该协议生成了小鼠 dc 模型, 有利于研究免疫系统在 dc 发育过程中的作用及其与微生物的相互作用。在这个模型使用 C57BL/6 动物, 结肠远端部分被排除在粪便流通过创造一个远端结肠造口, 触发轻度到中度炎症在排除的肠段, 并再现人类的标志性病变的发展DC 具有适度的全身炎症反应。与大鼠模型相比, 大量的基因修饰的小鼠模型在 C57BL/6 的背景下可用。这些动物与我们的模型相结合, 可以使个体细胞因子、趋化因子或生物活性分子受体 (如白细胞介素 (IL)-17) 发挥潜在作用;IL-10, 趋化因子 CXCL13, 趋性因子受体 CXCR5 和 CCR7, sphingosine-1-phosphate 受体 4) 被评估在 DC 的发病机制。在 C57BL/6 背景下, 同类系小鼠菌株的可用性在很大程度上促进了转移实验, 从而确定了不同细胞类型在 DC 病因学中的作用。最后, 该模型提供了机会, 以评估当地干预 (如修改当地微生物或局部抗炎治疗) 对黏膜免疫的影响和非受影响的肠段和对系统性免疫平衡。

手术在实验性 (结肠造口) 和假 (colotomy) 组均耐受良好。手术和围手术期处理正确执行时, 围手术期死亡率不应超过10%。在术后第一周, 实验性和假组均有明显的体重下降。假组的动物通常在第四术后的一天达到其体重最低点, 但在实验组的一天后发生。体重减轻在实验组更明显, 达到21.7% 的初始体重。相比之下, 假动物失去了10.8% 的初始体重 (图 2)6。术后第一周体重降至最低点后, 两个实验组的体重均持续上升, 但实验组的坡度较慢。严重肠道炎症的体征和症状 (即血流量或液体肠运动均不发生在实验性或假组6)。术后头60天总死亡率在结肠造口组约 40%, colotomy 组约10%。大多数死亡发生在术后第一周 (实验组 61%, 66% 人在假组)。死亡原因如图 3所示。
小鼠远端结肠造口术的形成主要是淋巴细胞性结肠炎的发生, 再现了人 DC 的特点。这些改变随着肠道排斥时间的延长而增加。隐窝长度在被排除的肠段中明显缩短。这一缩短达到统计学意义后14天的粪便分流 (图 3a)。在被排除的肠段30天后, 杯状细胞的隐窝长度减少 (图 3b)。14天后, 在被排除的肠段内的墓穴中, 绝对杯状细胞数也明显减少 (图 3c)。DC 的标志性病变, 粘膜淋巴滤泡的发育, 需要较长的大便偏差时间。虽然淋巴滤泡的数量在两周内可以观察到, 但两个月后的差异变得显著 (图 5 a c)。前面描述的所有病理学变化比被排除的肠段的近端区域更明显。典型的中性渗透作为急性炎症的征兆通常没有被观察 (图 5 d e)。
作为局部肠道炎症的系统性反应的标志, 在手术后的14天内, 结肠造口动物的中性粒细胞计数明显增加。这种差异维持到观察期结束 (60 天)。60天后结肠转移动物的血小板计数略有增加 (图 6)。结肠造口组术后14天血红蛋白水平和红细胞压积减少, 与假组比较。
结果表明, 啮齿类动物的维生素缺乏导致平均红细胞量减少 (MCH) 和平均红细胞血红蛋白 (MCV) 值7。在我们的模型中, 我们看到两个参数在14和30天后的初始增加。在更长的随访之后, MCV 和 MCH 都返回正常值 (图 7)。这表明远端结肠造口术在长期随访中不会导致临床上严重的维生素缺乏。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/57616/57616fig1.jpg"> 图 1: 盲肠区域的解剖和外科手术。(a) 术后解剖学的图形化表现。(b) 盲肠的地形和解剖地标。(c) 结肠造口的图形化表现。图 1a已从佳等6修改。<a href="/files/ftp_upload/57616/57616fig1large.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/57616/57616fig2.jpg"> 图 2: 体重发展。体重以术前体重的百分比显示。结肠造口和假 (colotomy) 动物体重曲线显示, 结肠造口组的术后早期体重损失明显高于假组 (p < 0.001)。值是指平均每组21-26 只动物的标准误差 (21 只动物接受 colostomies; 26 只动物在假组中)。这个数字已从佳等6修改。<a href="/files/ftp_upload/57616/57616fig2large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 3" class="xfigimg" src="/files/ftp_upload/57616/57616fig3.jpg"> 图 3: 死亡原因。并发症表现为全死因死亡率的百分比, 并在饼图 ((a) 结肠造口组 (b) 假组) 中加以说明。术后并发症由尸检决定。浪费综合征被定义为连续体重损失大于33% 的初始体重, 没有其他发现在尸检。尸检诊断为肠梗阻和吻合口瘘。气孔并发症被定义为 peristomal 脓肿和黏膜分离。其他并发症包括剖腹手术切口裂开, 盲肠缺血, 尸检不清楚的病例。两组患者术后并发症分布有显著性差异 (p = 0.021, 用费舍尔精确试验分析了多达 6 x 6 个应急表)。结肠造口组有39只动物, 假组29例。这个数字已从佳等6修改。<a href="/files/ftp_upload/57616/57616fig3large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 4" class="xfigimg" src="/files/ftp_upload/57616/57616fig4.jpg"> 图 4: 隐窝长度和杯状细胞数量。用直肠石蜡切片在周期性酸性席夫碱染色后, 确定了窖长和杯状细胞数。(a) 在转移性结肠炎 (DC) 的动物直肠中, 隐窝长度明显减少。(b) 土窖的杯状细胞承载区域的长度被测量并设置为与整个墓穴长度的比值。术后三十和六十天, DC 组内隐窝长度的杯状细胞的百分比降低。(c) 与假组相比, DC 动物的杯状细胞数明显减少。图 (a) 通过 (c) 显示每组5至9只动物的手段和标准误差 (结肠造口14和30天, 假14天: n = 8; 结肠造口60天: n = 5; 假30和60天: n = 9);∗p < 0.05;∗∗p < 0.01。这个数字已从佳等6修改。<a href="/files/ftp_upload/57616/57616fig4large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 5" class="xfigimg" src="/files/ftp_upload/57616/57616fig5.jpg"> 图 5: 淋巴滤泡和炎症浸润。(a) 被转移的冒号和假动物的淋巴滤泡被计数在直肠的石蜡切片染色苏木精和伊红。该图显示了每组5至9只动物的方法和标准误差 (结肠造口14和30天, 假14天: n = 8; 结肠造口60天: n = 5; 控制30和60天: n = 9), ∗p < 0.05;∗∗p < 0.01。(b) 和 (c) 结肠造口 (b) 和假 (c) 动物的直肠部分的典型例子, 术后60天染有苏木精和伊红。一个突出的淋巴卵泡 (∗) 存在于结肠造口小鼠的黏膜中。刻度条代表100µm. (d) 和 (e) 在石蜡切片上进行了氯乙酸酯酶反应染色, 以检测中性粒细胞颗粒。在转运直肠的横断面上, 在结肠造口组 (d) 和假动物 (e) 术后60天内均观察到急性炎症中性浸润。刻度条代表100µm。这个数字已从佳等6修改。<a href="/files/ftp_upload/57616/57616fig5large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 6" class="xfigimg" src="/files/ftp_upload/57616/57616fig6.jpg"> 图 6: 中性粒细胞和血小板计数。用兽医血液分析仪测定静脉血标本的血计数。(a) 结肠造口组与假动物术后60天相比, 中性粒细胞计数明显增加。(b) 术后60天内, 结肠造口组的血小板计数略有升高。该图显示了每组5至9只动物的方法和标准误差 (结肠造口14和30天, 假14天: n = 8; 结肠造口60天: n = 5; 假30和60天: n = 9), ∗p < 0.05;∗∗p < 0.01。<a href="/files/ftp_upload/57616/57616fig6large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 7" class="xfigimg" src="/files/ftp_upload/57616/57616fig7.jpg"> 图 7: 红细胞参数。用兽医血液分析仪测定静脉血标本的血计数。(a) 结肠造口组与假组相比, 血红蛋白和 (b) 红细胞压积显著降低。(c) 结肠造口组与假组相比, 术后平均红细胞容积 (MCH) 显著升高14和30天, 但在60天后恢复正常。(d) 平均红细胞血红蛋白 (MCV) 值在30天后升高, 但在结肠造口组术后60天后不再与假动物相比。该图显示了每组5至9只动物的方法和标准误差 (结肠造口14和30天, 假14天: n = 8; 结肠造口60天: n = 5; 假30和60天: n = 9), ∗p < 0.05;∗∗p < 0.01。<a href="/files/ftp_upload/57616/57616fig7large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>

Watch this Scientific Journal Video about Murine Distal Colostomy, A Novel Model of Diversion Colitis in C57BL/6 Mice at JoVE.com

Murine Distal Colostomy, A Novel Model of Diversion Colitis in C57BL/6 Mice

小鼠远端结肠造口术, 一种新的 C57BL/6 老鼠转移性结肠炎模型

小鼠远端结肠造口为人分流型结肠炎提供了鼠模型, 一种主要的淋巴细胞结肠炎在结肠段被排除从粪流。

Diversion colitis (DC) is a frequent clinical condition occurring in patients with bowel segments excluded from the fecal stream as a result of a ...

murine-distal-colostomy-novel-model-diversion-colitis-c57bl6

Universidad San Sebastian

Research

JoVE Journal

Medicine

12.8K 次观看.  Universitätsmedizin Greifswald. 小鼠远端结肠造口为人分流型结肠炎提供了鼠模型, 一种主要的淋巴细胞结肠炎在结肠段被排除从粪流。

视频: Murine Distal Colostomy, A Novel Model of Diversion Colitis in C57BL/6 Mice

A Genetically Engineered Mouse Model of Sporadic Colorectal Cancer

Despite the advantages of easy applicability and cost-effectiveness, colorectal cancer mouse models based on tumor cell injection have severe limitations and do not accurately simulate tumor biology and tumor cell dissemination. Genetically engineered mouse models have been introduced to overcome these limitations; however, such models are technically demanding, especially in large organs such as the colon in which only a single tumor is desired.
As a result, an immunocompetent, genetically engineered mouse model of colorectal cancer was developed which develops highly uniform tumors and can be used for tumor biology studies as well as therapeutic trials. Tumor development is initiated by surgical, segmental infection of the distal colon with adeno-cre virus in compound conditionally mutant mice. The tumors can be easily detected and monitored via colonoscopy. We here describe the surgical technique of segmental adeno-cre infection of the colon, the surveillance of the tumor via high-resolution colonoscopy and present the resulting colorectal tumors.

A protocol for the establishment of a genetically engineered mouse model of colorectal cancer by segmental adeno-cre infection and its surveillance via high-resolution colonoscopy is presented.

遗传工程小鼠模型的散发性结肠直肠癌

Despite the advantages of easy applicability and cost-effectiveness, colorectal cancer mouse models based on tumor cell injection have severe limitations ...

科研

癌症研究

Modeling Colitis-Associated Cancer with Azoxymethane (AOM) and Dextran Sulfate Sodium (DSS)

Individuals with inflammatory bowel disease (IBD), such as Crohn's disease (CD) or ulcerative colitis (UC) are at increased risk of developing colorectal cancer (CRC) over healthy individuals. This risk is proportional to the duration and extent of disease, with a cumulative incidence as high as 30% in individuals with longstanding UC with widespread colonic involvement.1 Colonic dysplasia in IBD and colitis associated cancer (CAC) are believed to develop as a result of repeated cycles of epithelial cell injury and repair while these cells are bathed in a chronic inflammatory cytokine milieu.2 While spontaneous and colitis-associated cancers share the quality of being adenocarcinomas, the sequence of underlying molecular events is believed to be different.3 This distinction argues the need for specific animal models of CAC.
Several mouse models currently exist for the study of CAC. Dextran sulfate sodium (DSS), an agent with direct toxic effects on the colonic epithelium, can be administered in drinking water to mice in multiple cycles to create a chronic inflammatory state. With sufficient duration, some of these mice will develop tumors.4 Tumor development is hastened in this model if administered in a pro-carcinogenic setting. These include mice with genetic mutations in tumorigenesis pathways (APC, p53, Msh2), as well as mice pre-treated with genotoxic agents (azoxymethane [AOM], 1,2-dimethylhydrazine [DMH]).5 
The combination of DSS with AOM as a model for colitis associated cancer has gained popularity for its reproducibility, potency, low price, and ease of use. Though they have a shared mechanism, AOM has been found to be more potent and stable in solution than DMH. While tumor development in other models generally requires several months, mice injected with AOM and subsequently treated with DSS develop adequate tumors in as little as 7-10 weeks.6, 7 Finally, AOM and DSS can be administered to mice of any genetic background (knock out, transgenic, etc.) without cross-breeding to a specific tumorigenic strain. Here, we demonstrate a protocol for inflammation-driven colonic tumorigenesis in mice utilizing a single injection of AOM followed by three seven-day cycles of DSS over a 10 week period. This model induces tumors with histological and molecular changes closely resembling those occurring in human CAC and provides a highly valuable model for the study of oncogenesis and chemoprevention in this disease.8

Modeling Colitis-Associated Cancer with Azoxymethane AOM and Dextran Sulfate Sodium DSS

We demonstrate a protocol in which administration of the genotoxic agent azoxymethane (AOM) followed by three cycles of the pro-inflammatory agent dextran sulfate sodium (DSS) rapidly and consistently generates colon tumors in mice with morphologic and molecular similarities to those seen in human colitis-associated cancer.

建模结肠炎相关的癌症与​​氧化偶氮甲烷（AOM）和葡聚糖硫酸钠（DSS）

Individuals with inflammatory bowel disease (IBD), such as Crohn's disease (CD) or ulcerative colitis (UC) are at increased risk of developing colorectal ...

医学

The Citrobacter rodentium Mouse Model: Studying Pathogen and Host Contributions to Infectious Colitis

This protocol outlines the steps required to produce a robust model of infectious disease and colitis, as well as the methods used to characterize Citrobacter rodentium infection in mice. C. rodentium is a gram negative, murine specific bacterial pathogen that is closely related to the clinically important human pathogens enteropathogenic E. coli and enterohemorrhagic E. coli. Upon infection with C. rodentium, immunocompetent mice suffer from modest and transient weight loss and diarrhea. Histologically, intestinal crypt elongation, immune cell infiltration, and goblet cell depletion are observed. Clearance of infection is achieved after 3 to 4 weeks. Measurement of intestinal epithelial barrier integrity, bacterial load, and histological damage at different time points after infection, allow the characterization of mouse strains susceptible to infection.
The virulence mechanisms by which bacterial pathogens colonize the intestinal tract of their hosts, as well as specific host responses that defend against such infections are poorly understood. Therefore the C. rodentium model of enteric bacterial infection serves as a valuable tool to aid in our understanding of these processes. Enteric bacteria have also been linked to Inflammatory Bowel Diseases (IBDs). It has been hypothesized that the maladaptive chronic inflammatory responses seen in IBD patients develop in genetically susceptible individuals following abnormal exposure of the intestinal mucosal immune system to enteric bacteria. Therefore, the study of models of infectious colitis offers significant potential for defining potentially pathogenic host responses to enteric bacteria. C. rodentium induced colitis is one such rare model that allows for the analysis of host responses to enteric bacteria, furthering our understanding of potential mechanisms of IBD pathogenesis; essential in the development of novel preventative and therapeutic treatments.

The Citrobacter rodentium Mouse Model: Studying Pathogen and Host Contributions to Infectious Colitis

Citrobacter rodentium infection provides a valuable model to study enteric bacterial infections as well as host immune responses and colitis in mice. This protocol outlines the measurement of barrier integrity, pathogen load and histological damage allowing for the thorough characterization of pathogen and host contributions to murine infectious colitis.

“<em&gt;枸橼酸rodentium</em&gt;鼠标产品型号：研究感染性结肠炎的病原体和宿主的贡献

This protocol  outlines the steps required to produce a robust model of infectious disease and  colitis, as well as the methods used to characterize ...

免疫与感染

Flexible Colonoscopy in Mice to Evaluate the Severity of Colitis and Colorectal Tumors Using a Validated Endoscopic Scoring System

The use of modern endoscopy for research purposes has greatly facilitated our understanding of gastrointestinal pathologies. In particular, experimental endoscopy has been highly useful for studies that require repeated assessments in a single laboratory animal, such as those evaluating mechanisms of chronic inflammatory bowel disease and the progression of colorectal cancer. However, the methods used across studies are highly variable. At least three endoscopic scoring systems have been published for murine colitis and published protocols for the assessment of colorectal tumors fail to address the presence of concomitant colonic inflammation. This study develops and validates a reproducible endoscopic scoring system that integrates evaluation of both inflammation and tumors simultaneously. This novel scoring system has three major components: 1) assessment of the extent and severity of colorectal inflammation (based on perianal findings, transparency of the wall, mucosal bleeding, and focal lesions), 2) quantitative recording of tumor lesions (grid map and bar graph), and 3) numerical sorting of clinical cases by their pathological and research relevance based on decimal units with assigned categories of observed lesions and endoscopic complications (decimal identifiers). The video and manuscript presented herein were prepared, following IACUC-approved protocols, to allow investigators to score their own experimental mice using a well-validated and highly reproducible endoscopic methodology, with the system option to differentiate distal from proximal endoscopic colitis (D-PECS).

Over the past few years, new generation endoscopes have emerged as important diagnostic research aids for evaluating murine colitis and colorectal tumors. We present herein a detailed protocol for endoscopic assessment of inflammation and colorectal tumors in mice, as well as a novel scoring system that uses decimal identifiers to document the endoscopic severity of colitis and colorectal tumors.

灵活的结肠镜检查在小鼠使用经过验证的内镜评分系统来评估结肠炎和结肠直肠肿瘤的严重程度

The use of modern endoscopy for research purposes has greatly facilitated our understanding of gastrointestinal pathologies. In particular, experimental ...

Murine Colitis Modeling using Dextran Sulfate Sodium (DSS)

Colitis can occur from viral or bacterial infections, ischemic insult, or autoimmune disorders; most notably Ulcerative Colitis and the colonic variant of Crohn&rsquo;s Disease - Crohn&rsquo;s Colitis. Acute colitis may present with abdominal pain and distention, malabsorption, diarrhea, hematochezia and mucus in the stool. We are beginning to understand the complex interactions between the environment, genetics, and epithelial barrier dysfunction in Inflammatory Bowel Disease and animal models of colitis have been essential in advancing our understanding of this disease. One popular model involves supplementing the drinking water of mice with low-molecular weight Dextran Sodium Sulfate (DSS), resulting in epithelial damage and a robust inflammatory response in the colon lasting several days 1.Variations of this approach can be used to model acute injury, acute injury followed by repair, and repeated cycles of DSS interspersed with recovery modeling chronic inflammatory diseases 2. After a single four-day treatment of 3% DSS in drinking water, mice show signs of acute colitis including weight loss, bloody stools, and diarrhea. Mice are euthanized at the conclusion of the treatment course and at necropsy dissected colons are processed and can be 'Swiss rolled" 3 to allow microscopic analysis of the entire colon or infused with formalin as "sausages" to allow macroscopic analysis. Tissue is then embedded in paraffin, sectioned, and stained for histologic review.

Murine Colitis Modeling using Dextran Sulfate Sodium DSS

Dextran sulfate sodium (DSS) administered in the drinking water is an established murine inflammatory injury model of acute colitis. This protocol outlines the method for DSS treatment and the preparation of tissues.

小鼠结肠炎模型使用葡聚糖硫酸钠（DSS）

Colitis can occur from viral or bacterial infections, ischemic insult, or autoimmune disorders; most notably Ulcerative Colitis and the colonic variant of ...

生物学

Creation of Colonic Anastomosis in Mice

Intestinal anastomoses are commonly performed in both elective and emergent operations. Even so, anastomotic leaks are a highly feared complications of colonic surgeries and can occur in up to 26% of surgical anastomoses, with mortality being up to 39% for patients with such a leak. Currently, there remains a paucity of data detailing the cellular mechanisms of anastomotic healing. Devising preventative strategies and treatment modalities for anastomotic leak could be greatly potentiated by a better understanding of appropriate anastomotic healing.
A murine model is ideal as previous studies have shown that the murine anastomosis is the most clinically similar to the human case as compared with other animal models. We offer an easily reproducible murine model of colonic anastomosis in mice that will allow for further illustration of anastomotic healing.

Anastomotic leak or breakdown after surgery is a major cause of postoperative morbidity and mortality. Our surgery for creating a colonic anastomosis is a reliable and reproducible method for studying the healing mechanisms of anastomoses.

小鼠结肠吻合术的建立

Intestinal anastomoses are commonly performed in both elective and emergent operations. Even so, anastomotic leaks are a highly feared complications of ...

Murine Appendectomy Model of Chronic Colitis Associated Colorectal Cancer by Precise Localization of Caecal Patch

The human appendix has been recently implicated to play important biological roles in the pathogenesis of various complex diseases, such as colorectal cancer, inflammatory bowel disease, and Parkinson&#8217;s disease. To study the function of the appendix, a gut disease-associated murine appendectomy model has been established and its step-by-step protocol is described here. This report introduces a facile protocol for caecal patch removal in mice followed by the chemical induction of chronic colitis-associated colorectal cancer using a combination of dextran sulfate sodium (DSS) and azoxymethane (AOM). IgA specific cells and IgA concentration were significantly reduced upon removal of the caecal patch in male C57BL/6 mice&#160;compared to those in the sham group. Simultaneously administering 2% DSS and AOM resulted in nearly 80% mice survival in both sham and appendectomy groups without significant body weight loss. Histological results confirmed colonic inflammation and different degrees of adenocarcinoma. This model can be used for the study of the functional role of the appendix in maintaining gut microbiota homeostasis and pathogenesis of gut colitis and malignancies, as well as for the potential development of drug targeting therapies.

The presented protocol describes a facile surgical removal of the appendix (caecal patch) in a mouse followed by the induction of inflammatory bowel disease-associated colorectal cancer. This murine appendectomy model enables investigation of the biological role of the appendix in the pathogenesis of human gastrointestinal disease.

慢性结肠炎相关结肠直肠癌的肿瘤切除术模型，通过卡卡贴片的精确定位

The human appendix has been recently implicated to play important biological roles in the pathogenesis of various complex diseases, such as colorectal ...

Single-Anastomosis Duodeno-Ileal Bypass with Sleeve Gastrectomy Model in Mice

Obesity is a major health issue worldwide. As a response, bariatric surgeries have emerged to treat obesity and its related comorbidities (e.g., diabetes mellitus, dyslipidemia, non-alcoholic steatohepatitis, cardiovascular events, and cancers) through restrictive and malabsorptive mechanisms. Understanding the mechanisms by which these procedures allow such improvements often require their transposition into animals, especially in mice, because of the ease of generating genetically modified animals. Recently, the single-anastomosis duodeno-ileal bypass with sleeve gastrectomy (SADI-S) has emerged as a procedure that uses both restrictive and malabsorptive effects, which is being used as an alternative to gastric bypass in case of major obesity. Thus far, this procedure has been associated with strong metabolic improvements, which has led to a marked increase in its use in daily clinical practice. However, the mechanisms underlying these metabolic effects have been poorly studied as a result of a lack of animal models. In this article, we present a reliable and reproducible model of SADI-S in mice, with a special focus on perioperative management. The description and use of this new rodent model will be helpful for the scientific community to better understand the molecular, metabolic, and structural changes induced by the SADI-S and to better define the surgical indications for clinical practice.

Single-anastomosis duodeno-ileal bypass (SADI-S) is an emerging bariatric procedure with important metabolic effects. In this article, we present a reliable and reproducible model of SADI-S in mice.

小鼠单吻合十二指肠回肠旁路与袖状胃切除术模型

Obesity is a major health issue worldwide. As a response, bariatric surgeries have emerged to treat obesity and its related comorbidities (e.g., diabetes ...

A Mouse Model of Intestinal Partial Obstruction

Intestinal obstructions, that impede or block peristaltic movement, can be caused by abdominal adhesions and most gastrointestinal (GI) diseases including tumorous growths. However, the cellular remodeling mechanisms involved in, and caused by, intestinal obstructions are poorly understood. Several animal models of intestinal obstructions have been developed, but the mouse model is the most cost/time effective. The mouse model uses the surgical implantation of an intestinal partial obstruction (PO) that has a high mortality rate if it is not performed correctly. In addition, mice receiving PO surgery fail to develop hypertrophy if an appropriate blockade is not used or not properly placed. Here, we describe a detailed protocol for PO surgery which produces reliable and reproducible intestinal obstructions with a very low mortality rate. This protocol utilizes a surgically placed silicone ring that surrounds the ileum which partially blocks digestive movement in the small intestine. The partial blockage makes the intestine become dilated due to the halt of digestive movement. The dilation of the intestine induces smooth muscle hypertrophy on the oral side of the ring that progressively develops over 2 weeks until it causes death. The surgical PO mouse model offers an in vivo model of hypertrophic intestinal tissue useful for studying pathological changes of intestinal cells including smooth muscle cells (SMC), interstitial cells of Cajal (ICC), PDGFR&#945;+, and neuronal cells during the development of intestinal obstruction.

Intestinal obstructions are a partial or complete blockage of the intestine that can cause severe abdominal pain, nausea, vomiting, and preventing the passage of stool. This procedure for creating intestinal partial obsructions in mice is reliable in studying the mechanisms underlying pathological cell growth and death in the intestine.

小肠部分梗阻的小鼠模型

Intestinal obstructions, that impede or block peristaltic movement, can be caused by abdominal adhesions and most gastrointestinal (GI) diseases including ...

A Technique to Generate a Murine Model of Dextran Sulfate Sodium-Induced Colitis

Source: Du, Y., et al. <a href="https://app-jove-com.bdigitaluss.remotexs.co/v/58668">Effects of Taste Signaling Protein Abolishment on Gut Inflammation in an Inflammatory Bowel Disease Mouse Model.</a> J. Vis. Exp. (2018)
This video demonstrates a technique to generate a chemical-induced murine model of colitis &#8212; an inflammatory bowel disease. The mouse is allowed to consume a dextran sodium sulfate (DSS) solution for an adequate duration. Upon consumption, DSS damages the mucus and epithelial layer barrier in the colon, allowing gut microbes to enter the underlying tissue &#8212; where they are sensed by the resident immune cells to mediate an exaggerated immune response, resulting in inflammation.

一种生成硫酸葡聚糖钠诱导的结肠炎小鼠模型的技术

Source: Du, Y., et al. Effects of Taste Signaling Protein Abolishment on Gut Inflammation in an Inflammatory Bowel Disease Mouse Model. J. Vis. Exp. ...

小鼠远端结肠造口术, 一种新的 C57BL/6 老鼠转移性结肠炎模型

摘要

探索更多视频

此视频中的章节

遗传工程小鼠模型的散发性结肠直肠癌

建模结肠炎相关的癌症与氧化偶氮甲烷（AOM）和葡聚糖硫酸钠（DSS）

“

灵活的结肠镜检查在小鼠使用经过验证的内镜评分系统来评估结肠炎和结肠直肠肿瘤的严重程度

小鼠结肠炎模型使用葡聚糖硫酸钠（DSS）

小鼠结肠吻合术的建立

慢性结肠炎相关结肠直肠癌的肿瘤切除术模型，通过卡卡贴片的精确定位

小鼠单吻合十二指肠回肠旁路与袖状胃切除术模型

小肠部分梗阻的小鼠模型

一种生成硫酸葡聚糖钠诱导的结肠炎小鼠模型的技术

小鼠远端结肠造口术, 一种新的 C57BL/6 老鼠转移性结肠炎模型

摘要

探索更多视频

此视频中的章节

遗传工程小鼠模型的散发性结肠直肠癌

建模结肠炎相关的癌症与​​氧化偶氮甲烷（AOM）和葡聚糖硫酸钠（DSS）

“

灵活的结肠镜检查在小鼠使用经过验证的内镜评分系统来评估结肠炎和结肠直肠肿瘤的严重程度

小鼠结肠炎模型使用葡聚糖硫酸钠（DSS）

小鼠结肠吻合术的建立

慢性结肠炎相关结肠直肠癌的肿瘤切除术模型，通过卡卡贴片的精确定位

小鼠单吻合十二指肠回肠旁路与袖状胃切除术模型

小肠部分梗阻的小鼠模型

一种生成硫酸葡聚糖钠诱导的结肠炎小鼠模型的技术

建模结肠炎相关的癌症与氧化偶氮甲烷（AOM）和葡聚糖硫酸钠（DSS）