The Transplant Institute, Sahlgrenska University Hospital, Gothenburg, Sweden.
Institute of Clinical Sciences, Laboratory for Transplantation and Regenerative Medicine, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden.
Am J Transplant. 2021 Jun;21(6):2220-2230. doi: 10.1111/ajt.16418. Epub 2021 Jan 18.
The organ damage incurred during the cold storage (CS) of intestinal grafts has short and long-term consequences. Animal studies suggest that additional luminal preservation (LP) with polyethylene glycol (PEG) may alleviate this damage. This study aims to validate these findings using human intestines. Ileal segments, perfused intravascularly with IGL-1 solution, were procured from 32 multiorgan donors and divided into two parts: one containing a PEG 3350-based solution introduced luminally (LP group) and another one without luminal treatment (control). Sampling was performed after 4 h, 8 h, 14 h, and 24 h of CS. Histology was assessed using the Chiu/Park score. Tight junctions (TJ), several inflammatory markers, and transcription factors were examined by immunofluorescence, ddPCR, and western blot. Tissue water content (edema) was also measured. Apoptotic activity was assessed with caspase -2, -3, and -9 assays. LP significantly lowered mucosal injury at all time points. Redistribution of TJ proteins occurred earlier and more severely in the control group. After 24 h of CS, LP intestines showed an emerging unfolding protein response. Increased caspase-3 and -9 activity was found in the control group. The current results indicate that luminal PEG is safe and effective in reducing damage to the intestinal epithelium during CS.
冷存储(CS)期间的器官损伤具有短期和长期的后果。动物研究表明,用聚乙二醇(PEG)进行额外的腔内保存(LP)可能会减轻这种损伤。本研究旨在使用人类肠道验证这些发现。从 32 名多器官供体中采集灌注血管内 IGL-1 溶液的回肠段,并将其分为两部分:一部分含有腔内引入的基于 PEG 3350 的溶液(LP 组),另一部分不含腔内处理(对照组)。在 CS 后 4 h、8 h、14 h 和 24 h 进行采样。使用 Chiu/Park 评分评估组织学。通过免疫荧光、ddPCR 和 Western blot 检查紧密连接(TJ)、几种炎症标志物和转录因子。还测量了组织含水量(水肿)。用半胱氨酸天冬氨酸蛋白酶 -2、-3 和 -9 测定评估凋亡活性。LP 在所有时间点均显著降低了黏膜损伤。TJ 蛋白的再分布在对照组中更早且更严重。CS 24 h 后,LP 肠道出现展开蛋白反应。在对照组中发现 caspase-3 和 caspase-9 活性增加。目前的结果表明,腔内 PEG 在 CS 期间减少肠道上皮损伤是安全有效的。