Establishment of new cold preservation method for intestinal graft

本研究探讨诱导表达热休克蛋白70是否能减轻离体小肠保存损伤。将肠上皮细胞（IEC）-18分为两个实验组。第1组为热应激组，热休克蛋白-70诱导组。第2组：37℃培养，不加热应激。在43℃烘箱中进行热休克30分钟。然后将细胞在37℃下放置2小时，然后在威斯康星州大学（UW）溶液中于4℃下保存24小时。热应激2小时后，第1组细胞被抗热休克蛋白（hsp）-70抗体染色，而第2组染色较弱。第1组的冷保存损伤程度较第2组轻。冷保存后，第1组的存活细胞数高于第2组。冷保存2 h后，第2组培养液中LDH含量高于第1组。结果表明，经hsp-70诱导后，IEC细胞对冷保存损伤具有一定的抵抗力，细胞凋亡减少。

In this study, we investigate whether the induction of hsp-70 could reduce small intestinal preservation injury in vitro. Intestinal epiethelial cell (IEC)-18 cells were divided two experimental groups. In group 1, the cells were thermally stressed and hsp-70 was induced. In group 2, the cells were cultured at 37℃ without thermal stress. Heat shock was performed in an oven at 43℃ for 30minutes. Cells were then placed in at 37℃ for 2 hours and then preserved at 4℃ in University of Wisconsin (UW) solution for 24 hours. In group 1, the cells were stained with anti-heat shock protein (hsp)-70 antibody 2 hours after thermal stress, but staining was weak in group 2. Magnitude of cold preservation injury was weak in group 1 than in group 2. Survival cell number after cold preservation was higher in group 1 than in group 2. LDH in culture medium 2 hours after cold preservation was higher in group 2 than in group 1. Apoptosis 24 hours after cold preservation was reduced in group 1 when compared with group 2.These results suggest that after inducion of hsp-70 IEC cells become resistant to cold preservation injury, reducing apoptosis.