大鼠移植小肠冷保存/再灌注后基因表达谱的变化

Gene expression profile in rat small intestinal allografts after cold preservation/reperfusion

目的 研究冷保存 /再灌注后大鼠移植小肠基因表达谱的变化 ,以探讨移植物损伤的机制。方法 建立改良式大鼠异位节段小肠移植和正常对照模型。抽提对照组正常小肠和移植肠冷保存 /再灌注 1h后总RNA并纯化mRNA将两组等量抽提纯化的小肠mRNA逆转录合成cDNA荧光探针 ,与cDNA芯片杂交。严格洗片后扫描芯片荧光信号图像 ,分析比较两组中差异表达基因。结果 在 40 96条基因中 ,两组间存在差异表达的基因共 82条 ,新发现基因 18条 ,表达序列标识 (EST ) 3 3条 ,已报道基因 3 1条。该 3 1条差异表达基因与移植物冷保存 /再灌注损伤存在相关性。结论 移植小肠冷保存 /再灌注损伤与中性多形核白细胞和微血管内皮细胞异常黏附、移植物细胞能量和葡萄糖。

Objective To determine the changes of gene expression profile in small intestinal allograft in rats after cold preservation/reperfusion and investigate the mechanism of allograft injury. Methods Heterotopic segmental small bowel transplantation in rats was performed with a modified technique. Total RNAs was extracted from the allografts (experimental group) and normal intestines (control group) 1h after cold preservation /reperfusion, and then purified to mRNA. Equal mRNAs of the two groups were as then reversely transcribed to cDNA and to prepare hybridization probes. The mixed probes were hybridized to the cDNA microarray . After high-stringent washing, the fluorescent signals on cDNA microarray were scanned and analyzed . Results Among the 4096 target genes, 82 differentially expressed genes were identified between the two groups. There were 18 novel genes and 33 expression sequence tags. 31 genes reported and differentially expressed could be related to the preservation/reperfusion injury of grafts. Conclusions Abnormal adhesion between polymorphonuclear cells and microvascular endothelial cells, and failure in energy, glucose and protein metabolism of grafts may contribute to cold preservation/reperfusion injury of small intestinal allografts.