Minor T, Klauke H, Isselhard W
Institute for Experimental Medicine, University of Cologne, Germany.
Eur Surg Res. 1997;29(4):246-53. doi: 10.1159/000129530.
In the present study a technique for isolated perfusion of rat intestines in vitro should be tested as an evaluative tool in the assessment of intestinal alterations related to ischemia and reoxygenation.
Segments of upper jejunum (15 cm) were isolated from Wistar rats with vascular pedicle (superior mesenteric artery, SMA and portal vein). The SMA was cannulated with polyethylene tubing and flushed with 10 ml of University of Wisconsin (UW) preservation solution. The intestinal lumen was rinsed with 10-15 ml of UW solution and the organ was stored immersed in UW solution at 4 degrees C for 4 or 18 h. After cold ischemic storage structural and functional integrity of the preparation was tested by biluminal perfusion with artificial buffer via SMA (5 ml/min modified Krebs-Henseleit buffer, 200 mg% glucose, 5% dextran 78, 0.06 mg% dexamethasone, 7 mg% atropine to counteract paralytic hypersecretion) and the intestinal lumen (0.5 ml/min NaCl 0.9% with 200 mg% of galactose). The in vitro model was validated by perfusion of control preparations harvested without ischemic alteration. It was seen that ischemic preservation of 4 h had only a minor impact on the recovery of cellular ATP content and enzyme release (LDH) upon reperfusion, whereas both parameters were significantly changed after 18 h of preservation. Functional parameters like transmucosal carbohydrate absorption and luminal water balance, however, were significantly impaired already after 4 h of ischemic storage of the gut, thus yielding sensitive criteria for the appreciation of the postischemic integrity of the gut.
It is concluded that the isolated gut preparation, being an inexpensive and technically feasible model, may be a useful tool in experimental research of intestinal ischemia/reperfusion.
在本研究中，一种大鼠肠体外单独灌注技术应作为一种评估工具，用于评估与缺血和再氧合相关的肠道改变。
从具有血管蒂（肠系膜上动脉，SMA和门静脉）的Wistar大鼠中分离出上段空肠（15厘米）段。用聚乙烯管将SMA插管并用10毫升威斯康星大学（UW）保存液冲洗。肠腔用10 - 15毫升UW溶液冲洗，器官在4℃下浸入UW溶液中保存4或18小时。冷缺血保存后，通过经SMA（5毫升/分钟改良的克雷布斯 - 亨泽莱特缓冲液，200毫克％葡萄糖，5％右旋糖酐78，0.06毫克％地塞米松，7毫克％阿托品以对抗麻痹性分泌过多）和肠腔（0.5毫升/分钟含200毫克％半乳糖的0.9％氯化钠）的双腔灌注来测试制剂的结构和功能完整性。通过灌注未发生缺血改变的对照制剂来验证体外模型。可以看出，4小时的缺血保存对再灌注时细胞ATP含量和酶释放（LDH）的恢复只有轻微影响，而保存18小时后这两个参数均发生了显著变化。然而，诸如跨粘膜碳水化合物吸收和肠腔水平衡等功能参数在肠道缺血保存4小时后就已受到显著损害，从而为评估缺血后肠道的完整性提供了敏感标准。
得出结论，分离的肠道制剂作为一种廉价且技术上可行的模型，可能是肠道缺血/再灌注实验研究中的一种有用工具。
