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JoVE Encyclopedia of Experiments
Biology
小鼠回肠袢模型:研究小鼠肠道通透性的手术模型
小鼠回肠袢模型:研究小鼠肠道通透性的手术模型
Encyclopedia of Experiments
Biology
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Encyclopedia of Experiments Biology
Mouse Ileal Loop Model: A Surgical Model to Study Intestinal Permeability in the Mouse

小鼠回肠袢模型:研究小鼠肠道通透性的手术模型

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3,421 Views
04:58 min
July 8, 2025
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Transcript

在

病理条件下,上皮和内皮屏障的崩解增加了肠道通透性,导致肠内容物渗漏到周围的脉管系统和腹腔中。可以使用回肠袢模型模拟肠道通透性。

首先,准备一只仰卧位的麻醉小鼠。在腹部做一个中线切口。将蘸有适当缓冲液的无菌棉纱布放在手术区域,以在手术过程中保持器官水分。

将

回肠(肠道的远端部分)外化,并将其铺在纱布上以露出肠系膜。确定回肠中的两个结扎部位,它们在肠系膜中间隔适当且没有任何关键血管。然后,刺穿肠系膜,在确定的部位添加结扎线。

接下来,解剖回肠以生成一个孤立的回肠袢,具有完整的血液供应和肠系膜。此后,将合适的缓冲液注入环中以冲洗废物内容物。

现在,结扎回肠袢的开口端。使用注射器将趋化因子(一种趋化性蛋白质悬浮液)注射到环中,使其膨胀。将器官返回腹部。

缝合切口。让鼠标恢复。最终,趋化因子分子穿过回肠壁进入相邻的血管,引起炎症和白细胞流入。

达到手术麻醉平面后,首先用酒精棉签或浸有 70% 乙醇的纱布海绵擦洗腹部中线的皮毛。不要用酒精弄湿大面积的皮毛,以防止体温过低。使用剪刀进行中线剖腹手术。在腹部中间做一个水平切口,露出腹膜,注意不要伤害腹腔内器官。

将

预先剪好的湿棉纱布放在暴露的腹腔上。用湿棉签将盲肠调动和外部化,并小心地将其放在湿棉纱布上。然后,动员并轻轻地将回肠外移。在湿棉纱布上放置至少 6 厘米的回肠末端,而不会破坏肠系膜血管和血液供应。

用温热的 HBSS 始终保持暴露的组织湿润。确定靠近盲肠的肠系膜中供应回肠的主要动脉。然后,在肠系膜中找到两个没有关键血管的结扎部位。用钝组织镊子牢牢抓住回肠末端,用细尖镊子开窗肠系膜,避开血管。

将丝线缝合穿过穿孔并打一个手术结以形成第一个结扎。用尺子测量距离第一根连字 4 厘米的距离,然后创建第二条连字。用细剪刀小心地剪开每个结扎旁边,以隔离 4 厘米的回肠袢,保持血液供应和肠系膜完好无损。

使用连接到 10 毫升注射器的柔性黄色饲管,用温热的 HBSS 轻轻冲洗回肠袢段的内容物。确保将管腔内容物冲出腹腔,以保持手术部位清洁。用丝线结扎冲洗回肠袢的两端。

使用带有 30 号针头的 1 毫升注射器将 250 微升试剂(例如 FITC-葡聚糖或趋化因子)缓慢注入肠腔。回肠袢会膨胀,导致粘膜适度扩张。

使用持针器、解剖镊子和带有反向切割针的 3.0 不可吸收丝缝合线闭合腹壁。将动物干燥以防止体温过低后,将其放入温度调节的麻醉室中进行潜伏期。

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