Mantke R, Schubert D, Röcken C, Paege I, Halangk W, Peters B, Lippert H, Schulz H-U
Department of General, Gastroenterological and Vascular Surgery, Otto von Guericke University, Magdeburg, Germany.
Langenbecks Arch Surg. 2009 Mar;394(2):363-9. doi: 10.1007/s00423-008-0401-8. Epub 2008 Aug 9.
Early events in the pathogenesis of experimental acute pancreatitis are intensively studied using isolated cells or animal models. However, the results and their interpretations are dependent on the complexity of biological structures. Therefore, we proposed that studies on isolated perfused pancreas can give additional information about processes leading to acinar cell injury. This hypothesis was examined adapting the well-established caerulein hyperstimulation model and the taurocholate model of acute pancreatitis to the extracorporeal perfused isolated rat pancreas.
The pancreas was removed with the duodenum including the arterial supply. A continuous perfusion of the organ was performed with a modified Krebs-Ringer bicarbonate buffer. Intraarterial caerulein application or an intraductal taurocholate (3.5%) application were used to induce acinar cell injury which was determined as the release of amylase, lipase and lactate dehydrogenase into the portal outflow medium and into the transudation fluid and by examination of histological alterations. Trypsinogen release and activation was followed by analysis of trypsinogen activation peptide (TAP) in the transudation fluid and in pancreatic tissue.
Perfusion of isolated rat pancreas with supramaximal concentrations of caerulein or retrograde injection of taurocholate (3.5%) resulted in acinar cell injury indicated by elevated levels of amylase and lipase into the perfusate and into the transudation fluid. TAP levels in the transudation fluid significantly increased after perfusion with caerulein or retrograde injection of taurocholate (3.5%). The histological alterations after taurocholate application include oedema and necrosis and show significant differences to the control perfusion. Extensive pancreatic necroses were not observed after caerulein hyperstimulation.
The isolated perfused rat pancreas is a useful model to investigate pathophysiological mechanisms which are relevant for the early phase of acute pancreatitis. The caerulein and the taurocholate models are transferable to the isolated rat pancreas. Studies on isolated perfused rat pancreas enable pathophysiological investigations of the exocrine pancreas without influence of systemic components, but with preserved morphology.
实验性急性胰腺炎发病机制的早期事件已通过分离细胞或动物模型进行了深入研究。然而，结果及其解释取决于生物结构的复杂性。因此，我们提出对离体灌注胰腺的研究可以提供有关导致腺泡细胞损伤过程的额外信息。采用成熟的蛙皮素过度刺激模型和急性胰腺炎牛磺胆酸盐模型对体外灌注的离体大鼠胰腺进行研究，以验证这一假设。
将胰腺连同十二指肠及动脉供应一并切除。用改良的 Krebs-Ringer 碳酸氢盐缓冲液对器官进行持续灌注。通过动脉内注射蛙皮素或导管内注射牛磺胆酸盐（3.5%）诱导腺泡细胞损伤，损伤程度通过淀粉酶、脂肪酶和乳酸脱氢酶释放到门静脉流出液和渗出液中以及组织学改变来确定。通过分析渗出液和胰腺组织中的胰蛋白酶原激活肽（TAP）来追踪胰蛋白酶原的释放和激活情况。
用超最大浓度的蛙皮素灌注离体大鼠胰腺或逆行注射牛磺胆酸盐（3.5%）导致腺泡细胞损伤，表现为灌注液和渗出液中淀粉酶和脂肪酶水平升高。用蛙皮素灌注或逆行注射牛磺胆酸盐（3.5%）后，渗出液中的 TAP 水平显著升高。应用牛磺胆酸盐后的组织学改变包括水肿和坏死，与对照灌注有显著差异。蛙皮素过度刺激后未观察到广泛的胰腺坏死。
离体灌注大鼠胰腺是研究急性胰腺炎早期相关病理生理机制的有用模型。蛙皮素和牛磺胆酸盐模型可转移至离体大鼠胰腺。对离体灌注大鼠胰腺的研究能够对外分泌胰腺进行病理生理研究，不受全身因素影响，但保留了形态学特征。
