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

分子生物学方法对活体小肠移植适宜移植部位的探讨

基本信息

批准号：
09470388
负责人：
TAGUCHI Tomoaki
金额：
$ 3.14万
依托单位：
KYUSHU UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 2000
项目状态：
已结题

项目摘要

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.

从分子生物学方面研究了合适的部位作为生活相关的移植物以及尸体小肠移植。合成性节段性肠道移植是使用刘易斯对刘易斯大鼠组合进行的。肠道组织是从0分钟，30分钟，4小时，24小时和72小时后进行的。使用Western印迹法检查了C-Fos和C-Jun即时伯爵基因的表达作为适应的参数。将空肠移植物的结果与回肠移植物的结果进行了比较。移植后4小时可见C-FOS表达，而回肠移植物的表达比空肠植物表现更强，更早。这种表达持续到24小时，并在72小时消失。在移植后仅观察到C-Jun表达，然后在24小时消失。回肠移植物的表达也比空肠移植物更强。这些结果表明，在小肠移植中，回肠的适应能力比jejunum更大。肠道肠道移植中，C-FOS和C-JUN mRNA的表达是在缺血性渗透性 - 渗透 - 渗透促进后测量后的（ I/r）在大鼠模型中，使用RT-PCR。 C-FOS表达瞬时激活，在再灌注后30分钟时达到峰值，并立即下降。 C-Jun的激活较慢，在再灌注后90分钟时达到峰值，其表达延长了比C-FOS更长。 PCNA阳性细胞的数量在60至360分钟时增加。再灌注后。在再灌注后120至360分钟时，与PCNA阳性细胞相比，TUNEL阳性细胞的增加。在I/R应力之后，在小肠上皮中观察到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有关，例如增殖和凋亡。