Braun F, Hosseini M, Wieland E, Sattler B, Laabs S, Lorf T, Müller A R, Fändrich F, Kremer B, Ringe B
Klinik für Allgemeine Chirurgie und Thoraxchirurgie, Universität Schleswig-Holstein, Campus Kiel, Kiel, Germany.
Transplant Proc. 2004 Mar;36(2):265-6. doi: 10.1016/j.transproceed.2004.01.081.
Ischemia-reperfusion injury (IRI) can result in severe organ dys- or nonfunction. Interaction of leukocytes and endothelial cells mediated by E-selectin appears to be a key step for disturbed microcirculation. Therefore we studied gene and protein expression as well as localization of E-selectin during intestinal IRI.
Intestinal tissue samples were obtained from extracorporeal perfused intestines (cold ischemia time [CIT] 2 or 20 hours, each n = 5) and additionally in intestinal transplanted pigs (CIT 2 or 20 hours, each n = 1). Mucosal damage was graded according to the Chiu classification. E-selectin mRNA was determined by PCR and quantitative RT-PCR. Localization of E-selectin mRNA was performed by in situ hybridization and of the protein by immunohistochemistry.
Histologically, mucosal damage occurred during reperfusion and was earlier and more severe after 20 hours of CIT. E-selectin mRNA expression was detected by PCR already after laparotomy and was elevated after reperfusion. Interestingly, mRNA expression was already increased after 20 hours of CIT. E-selectin mRNA was localized to the luminal surface of muscular, submucosal, and mucosal endothelial cells and the protein was detected on submucosal arterial endothelium as early as 2 hours after reperfusion.
Prolongation of CIT results in more severe mucosal damage during reperfusion, which is associated with protein expression of E-selection that might be used as a marker for activated endothelial cells. Increased E-selectin mRNA at end of 20 hours of CIT might indicate a preactivated state of endothelial cells potentially triggered by bacterial translocation or products.
缺血再灌注损伤（IRI）可导致严重的器官功能障碍或功能丧失。由E-选择素介导的白细胞与内皮细胞的相互作用似乎是微循环紊乱的关键步骤。因此，我们研究了肠道IRI期间E-选择素的基因和蛋白表达以及定位。
从体外灌注肠（冷缺血时间[CIT]2或20小时，每组n = 5）以及另外从肠道移植猪（CIT 2或20小时，每组n = 1）获取肠道组织样本。根据Chiu分类法对黏膜损伤进行分级。通过PCR和定量RT-PCR测定E-选择素mRNA。通过原位杂交进行E-选择素mRNA的定位，通过免疫组织化学进行蛋白的定位。
组织学上，黏膜损伤在再灌注期间发生，并且在CIT 20小时后更早且更严重。剖腹手术后通过PCR已检测到E-选择素mRNA表达，再灌注后升高。有趣的是，在CIT 20小时后mRNA表达就已经增加。E-选择素mRNA定位于肌层、黏膜下层和黏膜内皮细胞的腔面，并且在再灌注后2小时最早在黏膜下动脉内皮上检测到该蛋白。
CIT延长导致再灌注期间更严重的黏膜损伤，这与E-选择素的蛋白表达相关，E-选择素可能用作活化内皮细胞的标志物。CIT 20小时结束时E-选择素mRNA增加可能表明内皮细胞的预激活状态，这可能由细菌易位或产物触发。
