Cibicek Norbert, Ehrmann Jiri, Proskova Jitka, Vecera Rostislav
Department of Medical Chemistry and Biochemistry, Faculty of Medicine and Dentistry, Palacky University, Olomouc, Czech Republic.
Department of Histology and Embryology, Faculty of Medicine and Dentistry, Palacky University, Olomouc, Czech Republic.
PLoS One. 2018 Jan 11;13(1):e0191041. doi: 10.1371/journal.pone.0191041. eCollection 2018.
Sensors able to record large bowel physiology and biochemistry in situ in awake rodents are lacking. Microdialysis is a mini-invasive technique that may be utilized to continuously deliver or recover low-molecular substances from various tissues. In this experiment we evaluated the feasibility of in vivo microdialysis to monitor extracellular fluid chemistry in the descending colon submucosa of conscious, freely moving rodents. Following surgical implantation of a microdialysis probe, male Wistar rats were housed in metabolic cages where they were analgized and clinically followed for four days with free access to standard diet and water. To assess local microcirculation and probe function, glucose, lactate, glucose-to-lactate ratio and urea clearance were determined in the dialysates from the three postoperative days with focus on the final 24-h period. In an attempt to mitigate the expected tissue inflammatory response, one group of animals had the catheters perfused with 5-aminosalicylic acid-enriched medium with final concentration 1 μmol/L. For verification of probe position and the assessment of the surrounding foreign body reaction, standard histological and immunohistochemical methods were employed. Microdialysis of rat gut is associated with considerable technical challenges that may lead to the loss of probe function and high drop-out rate. In this setting, limited data did not allow to draw any firm conclusion regarding local anti-inflammatory effectiveness of 5-aminosalicylic acid perfusion. Although intestinal microdialysis may be suitable for larger anesthetized animals, low reproducibility of the presented method compromises its routine experimental use in awake and freely moving small-sized rodents.
目前尚缺乏能够在清醒啮齿动物体内原位记录大肠生理和生化指标的传感器。微透析是一种微创技术，可用于持续从各种组织中输送或回收低分子物质。在本实验中，我们评估了在清醒、自由活动的啮齿动物降结肠黏膜下层进行体内微透析以监测细胞外液化学成分的可行性。在手术植入微透析探针后，将雄性Wistar大鼠饲养在代谢笼中，使其处于无疼痛状态并进行临床观察4天，大鼠可自由获取标准饮食和水。为评估局部微循环和探针功能，在术后三天的透析液中测定葡萄糖、乳酸、葡萄糖与乳酸的比值以及尿素清除率，重点关注最后24小时。为减轻预期的组织炎症反应，一组动物的导管用终浓度为1 μmol/L的富含5-氨基水杨酸的培养基灌注。为验证探针位置并评估周围异物反应，采用了标准的组织学和免疫组化方法。大鼠肠道的微透析存在相当大的技术挑战，可能导致探针功能丧失和高脱落率。在此情况下，有限的数据无法就5-氨基水杨酸灌注的局部抗炎效果得出任何确凿结论。尽管肠道微透析可能适用于较大的麻醉动物，但所提出方法的低重现性限制了其在清醒、自由活动的小型啮齿动物中的常规实验应用。
