小肠部分梗阻的小鼠模型
Summary
肠道阻塞是肠道的部分或全部堵塞, 可引起严重的腹痛、恶心、呕吐和防止大便的通过。该方法在小鼠肠道部分 obsructions 的建立是可靠的, 研究的机制, 病理细胞生长和死亡的肠道。
Abstract
肠道阻塞, 阻碍或阻断蠕动运动, 可能是由腹部粘连和大多数胃肠道 (胃肠) 疾病, 包括肿瘤生长引起的。然而, 由于肠道障碍而引起的细胞重塑机制被理解得很差。一些动物模型的肠道障碍已经开发, 但鼠标模型是最成本/时间的有效。老鼠模型使用的外科植入肠道部分梗阻 (PO), 如果它没有正确的执行率高死亡率。此外, 如果不使用适当的封锁或不适当放置, 接受 PO 手术的小鼠就无法发育肥大。在这里, 我们描述了一个详细的协议, 为 PO 手术, 产生可靠和可重复的肠道障碍, 死亡率非常低。该协议利用手术放置的硅胶环包围回肠, 部分阻断消化运动在小肠。部分堵塞使肠道因消化运动停止而扩张。肠道扩张会导致环的口腔侧平滑肌肥大, 逐渐发育超过2周, 直至死亡。手术 PO 鼠标模型提供了一个体内模型, 用于研究肠道细胞的病理变化, 包括平滑肌细胞 (SMC), Cajal (ICC), PDGFRα+和神经元细胞在肠道梗阻的发育过程中。
Introduction
肠道阻塞是小肠或大肠的部分或全部堵塞, 可防止消化的食物、液体和气体通过肠道移动1。由于梗阻, 堵塞导致肠道壁变厚, 缩小流明2。由于腹部或盆腔手术导致腹部粘连组织形成或胃肠道疾病 (如克罗恩病)、憩、疝、扭转、狭窄、肠套叠, 便秘, 粪便嵌塞, 伪梗阻, 癌症和肿瘤3,4,5。肠道阻塞在这些情况下经常导致肠道肌的肥大6。
PO 的流明诱发肠道扩张, 并增加平滑肌层厚度周围的梗阻, 以响应需要继续功能蠕动7,8,9,10, 11,12,13。开发了肠道动物模型, 研究小鼠的平滑肌肥厚7, 大鼠10, 豚鼠11, 狗12, 猫13 , 持续发展类似的肥厚在肠道肌肉层。
小肠 PO 的老鼠模型是产生和研究肠道障碍的最经济有效的方法在体内。小鼠小肠梗阻是通过使用硅胶环手术放置在回肠周围。在 po 手术后, po 小鼠的细胞数量 (增生) 和肌肉层厚度 (肥厚) 的增加较早,8,15。SMC 是主要的塑料细胞, 生长在平滑的肌肉层, 以响应肥厚条件14, 但其他细胞, 如 ICC 和 PDGFRα+细胞与 SMC 密切关联, 也填充。我们以前报告说, PO 小鼠在小肠发育肥大, 其中 SMC 被去分化肉瘤成PDGFRα + 细胞, 高度增生的7, 15, 16.相反, 在肠道 obsruction 的发育过程中, ICC 在肥厚平滑肌层中退化并丢失了7。PO 模型的另一个主要好处是它有能力诱发肠道神经系统的改变和传播神经源性运动模式。小鼠小肠的主要传播神经源性运动模式是迁移马达复合体 (MMC), 这是神经源性的, 不需要 ICC 或电慢波17。PO 模型可以清楚地了解复合材料和肠神经是如何通过局部梗阻重塑的。
在这里, 我们提出了一个小鼠的肠道 PO 手术的协议使用硅胶环。接受 PO 手术的小鼠在小肠肌中可靠地产生肥大。在肥厚的肌肉内, SMC, ICC, PDGFRα+, 神经元细胞被显著重塑。
Protocol
Representative Results
Discussion
我们表明, 小鼠接受肠道 PO 手术的持续和重现性发展肠平滑肌肥大, 这是模仿人类肠梗阻。为不同的动物开发了肠梗阻手术, 包括老鼠7, 老鼠10, 豚鼠11, 狗12和猫13。小鼠肠梗阻模型具有时间、成本、大小和表型优势, 优于其他大型动物模型。小鼠或大鼠肥大的发展只需要10-14 天10, 与豚鼠、狗…
Declarações
The authors have nothing to disclose.
Acknowledgements
作者要感谢本杰明 J Weigler, D.V.M., 博士和沃尔特曼德维尔, D.V.M. (动物资源 & 校园参加兽医, 内华达大学, 里诺) 为他们提供了优良的动物服务为老鼠并且他们的忠告在手术程序。
Materials
| Surgical drape | Medical and veterinary supplies | SMS40 | 40”X100 yards |
| Underpad, econ, pro plus | Medical and veterinary supplies | MSC281224 | 17×24” |
| Iris scissors | Braintree scientific, Inc | SC-i-130 | |
| Iris scissors | Vantage | V95-304 | |
| Dumont electronic & jeweler tweezers | Dumont | 98-180-3 | |
| Braided absorbable suture | Covidien polysorb | SL-5687G | 5-0, polyglactin |
| Nylon non-absorbable mono filament | AD surgical | S-N618R13 | 6-0, nylon |
| Surgical blade | Dynarex | No.15 | |
| Needle holder | Jacobson microvascular | 36-1342TC | 8.5 inch |
| Scalpel handle | Flinn scientific | AB1049 | |
| Microsurgical scissor | WPI | 503305 | |
| Petrolatum ophthalmic ointment | Puralube VET | 3.5 g | |
| Fluriso (isoflurane) | Vetone | V1 502017 | 250 ml |
| Steri-strip reinforced skin closure | 3M | R1547 | |
| Surgical gloves | Medline | MSG2270 | |
| Ear loop face mask | The safety zone | RS700 | |
| Avant gauze non-woven sponges | Caring | PRM25444 | |
| Surgical cup | Admiral craft OYC-2 | 725-A42 | 2.5 oz |
| Swabstick | ChloraPrep | 260103 | 2% w/v Chlorhexidine Gluconate (CHG) and 70% v/v Isopropyl Alcohol (IPA) |
| Cotton tipped applicator | Puritan | 806-WC | |
| Buprenorphine | Zoo pharm | BZ8069317 | 1 mg/ml |
| Gentamycin sulfate | Vetone | G-6336-04 | 100 mg/ml |
| Fast acting gel cream remover | Veet | 8111002 | |
| Syringe | AHS | AH01T2516 | 1 ml with needle |
| Silicon ring | VWR | 60985-720 | 6 mm in length, 4 mm exterior diameter, 3.5 mm interior diameter |
| C57BL/6 mice | The Jackson Laboratory | 4-6 weeks old |
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