谷氨酰胺在小肠保存中作用的实验研究

Role of glutamine on preservation of small intestine

目的 :研究谷氨酰胺 (Gln)在小肠保存中的作用。 方法 :根据保存液的不同将SD大鼠随机分为三组 ,即UW液组、WMO液组和添加Gln的WMO G液组。根据保存时间的不同 ,每组又分为保存 8h和保存 1 2h的两个实验亚组。从大鼠腹主动脉置管灌入灌注液 ,直至小肠组织变白 ,然后用 4℃的 5 %甲硝唑溶液冲洗肠腔 ,取材。测定组织ATP的含量 ,观察细胞凋亡 (TUNEL法 )的情况。另外 ,取一部分小肠组织进行体外培养 ,观察小肠粘膜上皮细胞的增生活力 (增殖细胞核抗原 ,PCNA) ,并观察组织学病理变化。 结果 :WMO G组的ATP含量高于UW组和WMO组 ,尤其是在保存 1 2h后 ,与其他两组的差别明显。WMO G组的细胞凋亡数密度低于其他两组 ,而UW组和WMO组之间差异不明显。但是从切片上可以看出 ,UW组的病理变化轻于WMO组。组织培养后可见小肠隐窝处的细胞增生 ,WMO G组的PCNA阳性细胞多于其他两组。 结论 :在保存液中加入Gln可为小肠粘膜上皮细胞提供能量。在组织保存过程中 ,可减轻小肠组织的损伤 ,并可观察到Gln可促进小肠粘膜上皮细胞的增生。

Objectives: To investigate the effect of the addition of glutamine to WMO solution on the small bowel preservation. Methods:According to preservation solutions, the rats were divided randomly into three groups: University of Wisconsin solution(UW group), WMO solution (WMO group) and WMO solution with the addition of glutamine(WMO G group). And each group was redivided into two subgroups in terms of preservation time of 8 h or 12 h. The intestine was perfused by intubation via abdomial aorta, then the gut was flushed with metronidazole solution(4℃,5%). The proliferation of small intestine was observed through tissue culture. The histology, immunohistochemistry (TUNEL, PCNA) of intestinal mucosa and determination of ATP were used to evaluate the results. Results: Compared with UW group and WMO group, ATP contents of WMO G group were significantly higher, particularly in 12 h subgroup. Apoptosis in WMO G group was slighter than those in UW group and in WMO group. The difference in the two latter was not obviously, although pathological change in UW group was slighter than that in WMO group. The number of positive PCNA cells in WMO G group was more than that in other groups. Conclusions: The addition of glutamine to WMO solution could decrease injury of small bowel induced by cold ischemia, provide energy for the small bowel,and promote the proliferation of small bowel mocosal cell.