THE INVESTIGATION CONCERNING THE SUITABLE SITE FOR GRAFT IN LIVING-RELATED SMALL INTESTINAL TRANSPLANTATION USING MOLECULAR BIOLOGICAL METHOD

从分子生物学角度探讨活体及尸体小肠移植的适宜移植部位，采用刘易斯-刘易斯大鼠组合进行同系节段性小肠移植。在移植后0分钟、30分钟、4小时、24小时和72小时后取肠组织。用Western印迹法检测了直接免疫调节基因c-Fos和c-Jun的表达，作为适应的参数。移植后4h即可见c-Fos表达，回肠移植组表达早于空肠移植组，且表达强于空肠移植组。这种表达持续到24小时，并在72小时消失。c-Jun在移植后4h开始表达，24h后表达消失。结果表明，回肠移植后的肠组织中c-fos和c-jun mRNA的表达明显高于空肠移植后的肠组织，表明回肠移植后的肠组织中c-fos和c-jun mRNA的表达明显高于空肠移植后的肠组织，表明回肠移植后的肠组织中c-fos和c-jun mRNA的表达明显高于空肠移植后的肠组织。c-fos表达短暂激活，再灌注30分钟达高峰，随后迅速下降。c-jun的激活较慢，在再灌注后90分钟达到高峰，其表达持续时间长于c-fos。再灌注60~360min，PCNA阳性细胞数增加。TUNEL阳性细胞在再灌注后120~360分钟明显高于PCNA阳性细胞。I/R应激后，小肠上皮中观察到c-fos和c-jun的转录激活。这些基因的顺序表达模式被认为与细胞反应如增殖和凋亡有关。

The suitable site as a graft of living-related, as well as, cadaveric small intestinal transplantation was investigated from the aspects of molecular biology.The syngeneic segmental intestinal transplantation was performed using Lewis to Lewis rat combination. Intestinal tissues were taken from after 0 minute, 30 minutes, 4 hours, 24 hours, and 72 hours after transplantation. The expression of c-Fos and c-Jun, immediate earl genes, was examined as parameters of adaptation, using western blotting method. The results of jejunal grafts were compared with those of ileal grafts.The c-Fos expression was seen 4 hours after transplantation, and the ileal grafts showed stronger and earlier expression than the jejunal grafts. This expression continued until 24 hours and disappeared at 72 hours. The c-Jun expression was seen only 4 hours after transplantation, and then disappeared at 24 hours. The ileal grafts also showed stronger expression than the jejunal grafts.These results suggested that the ileum has bigger adaptation capacity than the jejunum in small intestinal transplantation.Furthermore, the expression of c-fos and c-jun mRNA was measured after ischemia-reperfusion (I/R) in rat model, using RT-PCR. The c-fos expression was activated transiently, came to a peak at 30 minutes after reperfusion, and immediately declined. Activation of c-jun was slower, peaked at 90 minutes after reperfusion, and its expression prolonged longer than c-fos. The number of PCNA positive cells increased at 60 to 360 min. after reperfusion. And TUNEL positive cells increased later than PCNA positive cells, at 120 to 360 minutes after reperfusion. After I/R stress, transcriptional activation of c-fos and c-jun was observed in the small intestinal epithelium. Sequential expression patterns of those genes are thought to be related to the cellular response such as proliferation and apoptosis.