Fatima Javairiah, Houghton Scott G, Sarr Michael G
Department of Surgery, Gastroenterology Research Unit (AL 2-435), Mayo Clinic, 200 First Street SW, Rochester, MN 55905, USA.
J Gastrointest Surg. 2007 Aug;11(8):1052-6. doi: 10.1007/s11605-007-0185-0.
Small bowel transplantation (SBT) is associated with poorly understood enteric dysfunction. The study of SBT in mice is hindered by the technical difficulty of orthotopic SBT in the mouse. Our aim was to develop an easy preparation of extrinsic denervation of the entire jejunoileum in mice as a model of orthotopic SBT. All neurolymphatic tissues accompanying the superior mesenteric artery (SMA) and vein (SMV) were ligated just distal to the middle colic vessels. The SMA and SMV were then stripped of investing adventitia, and the mesentery to jejunum and colon were transected radially. Jejunum and colon were not transected and reanastomosed. To confirm extrinsic denervation 1, 3, and 6 months later, segments of small bowel were stained for protein gene product 9.5 (PGP9.5) and tyrosine hydroxylase (TH). Tyrosine hydroxylase immunoreactive intensity was then quantified using a semiquantitative analysis. Immunohistochemical fluorescence showed persistence of PGP9.5 immunoreactivity confirming enteric nerves in jejunoileum; however, there was no TH immunoreactivity in jejunoileum in denervated mice despite the expected preservation of TH immunoreactivity in the still-innervated duodenum at 1 month. At 3 months, sparse immunoreactivity for TH was present, and by 6 months, reinnervation of TH-containing nerves appeared similar to controls. Quantification of intensity at each time-point further confirmed this trend. This technique in the mouse accomplishes a complete extrinsic denervation of jejunoileum early postoperatively (1 and 3 months); reinnervation occurs by 6 months. This is an easily learned murine model of orthotopic SBT.
小肠移植（SBT）与尚未完全了解的肠道功能障碍有关。小鼠原位SBT的技术难度阻碍了对其的研究。我们的目的是开发一种简便的小鼠全空肠回肠去交感神经制备方法，作为原位SBT的模型。在中结肠血管远侧结扎所有伴随肠系膜上动脉（SMA）和肠系膜上静脉（SMV）的神经淋巴组织。然后剥去SMA和SMV的外膜，并沿径向切断空肠和结肠的系膜。空肠和结肠未横断和重新吻合。为了在1、3和6个月后确认去交感神经，对小肠段进行蛋白质基因产物9.5（PGP9.5）和酪氨酸羟化酶（TH）染色。然后使用半定量分析对酪氨酸羟化酶免疫反应强度进行量化。免疫组织化学荧光显示PGP9.5免疫反应性持续存在，证实空肠回肠中有肠神经；然而，去神经小鼠的空肠回肠中没有TH免疫反应性，尽管在1个月时仍有神经支配的十二指肠中TH免疫反应性如预期保留。3个月时，出现稀疏的TH免疫反应性，到6个月时，含TH神经的再支配情况与对照组相似。对每个时间点的强度进行量化进一步证实了这一趋势。该小鼠技术在术后早期（1个月和3个月）实现了空肠回肠完全去交感神经；6个月时出现再支配。这是一种易于掌握的原位SBT小鼠模型。
