Katouzian Fourogh, Sblattero Daniele, Not Tarcisio, Tommasini Alberto, Giusto Elena, Meiacco Daniela, Stebel Marco, Marzari Roberto, Fasano Alessio, Ventura Alessandro
Department of Reproductive and Development Science and IRCCS, Instituto per I'Infanzia Burlo Garofolo Clinica Pediatrica University of Trieste, Via dell'Istria 65/1, IT-34100, Trieste, Italy.
Clin Chim Acta. 2005 Feb;352(1-2):191-7. doi: 10.1016/j.cccn.2004.09.023.
Intestinal permeability is determined by measuring nonmetabolized sugars. In animals, intestinal permeability is determined in urine, using cumbersome and expensive metabolic cages. We developed an HPLC method for determining concentrations of lactulose (L) and L-rhamnose (R) in blood-drop of rabbits and mice, and we compared these results with the procedure based on sugars excreted in urine. We measured the intestinal permeability induced by a fragment (DeltaG) of the zonula occludens toxin which opens the paracellular pathway.
The animals received sugar solution and later received the same solution+DeltaG. Five-hour urine collection and timed blood tests were performed after ingestion of sugars. Sugars were measured with HPLC, and the percentage of recovered sugars was expressed as L/R ratio.
At 60 min after administration of sugars, the mean L/R ratio for rabbits and mice was 0.026 and 0.052, respectively. At 60 min after administration of sugars+DeltaG, the mean L/R ratio for rabbits and mice was 0.22 and 0.53. The mean L/R ratio in the urine was 0.023 at basal condition and 0.25 after DeltaG ingestion.
Testing small serum samples for sugar permeability is effective for monitoring changes in permeability of the gut in animals. This cheap simple method allows us to measure in vivo the biological activity of other molecules which modulate the paracellular pathway.
肠道通透性通过测量非代谢性糖来确定。在动物中,肠道通透性是在尿液中测定的,使用的是笨重且昂贵的代谢笼。我们开发了一种高效液相色谱法(HPLC)来测定兔和小鼠血滴中乳果糖(L)和L-鼠李糖(R)的浓度,并将这些结果与基于尿液中排出糖的方法进行比较。我们测量了由紧密连接毒素片段(DeltaG)诱导的肠道通透性,该片段可打开细胞旁通道。
动物先接受糖溶液,随后接受相同溶液加DeltaG。摄入糖后进行5小时尿液收集和定时血液检测。用HPLC测量糖,并将回收糖的百分比表示为L/R比值。
给予糖后60分钟,兔和小鼠的平均L/R比值分别为0.026和0.052。给予糖加DeltaG后60分钟,兔和小鼠的平均L/R比值分别为0.22和0.53。基础状态下尿液中的平均L/R比值为0.023,摄入DeltaG后为0.25。
检测小血清样本中的糖通透性可有效监测动物肠道通透性的变化。这种廉价简单的方法使我们能够在体内测量其他调节细胞旁通道分子的生物活性。