Houghton Scott G, Nicholson Virginia D, Sarr Michael G
Department of Surgery, Mayo Clinic College of Medicine, Rochester, MN 55905, USA.
J Surg Res. 2006 Mar;131(1):53-7. doi: 10.1016/j.jss.2005.07.029. Epub 2005 Nov 14.
Our aim was to develop and validate a technically easy, reliable, and reproducible method of complete jejunoileal denervation in the rat to allow study of the physiologic effects of intestinal transplantation devoid of immunologic phenomena and ischemia/reperfusion injury.
Six adult Sprague-Dawley rats underwent transection and reanastomosis of the proximal jejunum and proximal colon, transection of all neurolymphatic tissues at the base of the mesentery, stripping adventitia off the superior mesenteric artery and vein, and radial transection of the intervening mesenteries, thereby denervating the jejunoileum in situ without disrupting blood flow. Three rats each were sacrificed 1 and 6 months later. Intestinal smooth muscle from the still-innervated duodenum and the denervated jejunum, mid-small bowel, and ileum was compared to corresponding tissues from a normal rat for tyrosine hydroxylase immunohistochemistry, a marker of extrinsic innervation.
One and six months after denervation, all duodenal samples demonstrated normal tyrosine hydroxylase immunostaining. In contrast, tyrosine hydroxylase immunoreactivity was undetectable in jejunum, mid-small bowel, or ileum of rats at 1 month and 2 of the 3 rats at 6 months; 1 rat at 6 months had low levels of tyrosine hydroxylase immunoreactivity at the mesenteric border of jejunum and mid small bowel.
This simple technique of in situ neural isolation effectively and reproducibly achieves complete extrinsic denervation of the entire rat jejunoileum. Low levels of neural regeneration may be present 6 months after denervation.
我们的目的是开发并验证一种技术上简便、可靠且可重复的大鼠全空肠回肠去神经支配方法,以便在无免疫现象和缺血/再灌注损伤的情况下研究肠道移植的生理效应。
六只成年Sprague-Dawley大鼠接受了近端空肠和近端结肠的横断与再吻合、肠系膜根部所有神经淋巴组织的横断、肠系膜上动脉和静脉外膜的剥离以及中间肠系膜的放射状横断,从而在不中断血流的情况下原位去空肠回肠的神经。分别在1个月和6个月后处死三只大鼠。将仍有神经支配的十二指肠以及去神经支配的空肠、中小肠和回肠的肠道平滑肌与正常大鼠的相应组织进行酪氨酸羟化酶免疫组化比较,酪氨酸羟化酶是外在神经支配的标志物。
去神经支配1个月和6个月后,所有十二指肠样本均显示酪氨酸羟化酶免疫染色正常。相比之下,去神经支配1个月时大鼠的空肠、中小肠或回肠以及6个月时三只大鼠中的两只均未检测到酪氨酸羟化酶免疫反应性;6个月时一只大鼠在空肠和中小肠的肠系膜边界处酪氨酸羟化酶免疫反应性水平较低。
这种简单的原位神经隔离技术能有效且可重复地实现大鼠整个空肠回肠的完全外在去神经支配。去神经支配6个月后可能存在低水平的神经再生。