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目的 明确大鼠小肠移植排斥反应期移植肠粘膜IL 2、IFN γ、穿孔素、粒细胞酶B的mRNA表达 ,移植肠IL 2R和ICAM 1表达 ,及移植肠上皮细胞凋亡的变化。方法 选用近交系大鼠F34 4 /N和Wistar/A进行全小肠异位移植。实验分为 4组 ,第 1组 :Wistar;第 2组 :Wistar→Wistar ;第 3组 :F34 4→Wistar ;第 4组 :F34 4→Wistar +环孢霉素A (6mg·kg 1·d 1,肌注 )。术后第 3、5、7天取各组动物移植肠标本进行病理学检查 ,应用逆转录多聚酶链反应 (RT PCR)检测移植肠IL 2、IFN γ、穿孔素、粒细胞酶B的mRNA表达 ,免疫组化检测移植肠IL 2R和ICAM 1表达 ,应用TUNEL法检测移植肠上皮细胞凋亡的变化。结果 ①病理学检查显示第 3组大鼠在术后第 3、5、7天分别符合轻、中、重度排斥 ,第 2、4组无明显排斥征象 ;②第 1组各检测基因基本不表达。第 3组IL 2mRNA表达在术后第 5天显著高于第 2组 (P <0 0 5 ) ;IFN γmRNA表达术后始终非常显著地高于第 2、4组 (P <0 0 1) ;穿孔素、粒细胞酶BmRNA表达在术后第 5、7天均高于第 2、4组 ,且具有统计学意义 ;③第 3组移植肠IL 2R表达术后始终显著高于三个对照组 ,仅在第 3天ICAM 1表达高于对照组 ;④术后第 3、5天第 3组移植肠上皮细胞凋亡非常显著高于其它三个对照组。结论
OBJECTIVE: To determine the expression of IL-2, IFN-γ, perforin, granzyme B mRNA, the expressions of IL-2R and ICAM-1 in intestine, and the changes of apoptosis in transplanted intestinal epithelial cells after small intestinal transplantation in rats. Methods Inbred rats F34 4 / N and Wistar / A were selected for total intestinal allotransplantation. The experiment was divided into 4 groups, group 1: Wistar; group 2: Wistar → Wistar; group 3: F34 4 → Wistar; group 4: F34 4 → Wistar + cyclosporine A 1, intramuscular injection). The animals were sacrificed on the 3rd, 5th, 7th day after operation for histological examination. RT-PCR was used to detect the mRNA expressions of IL 2, IFNγ, perforin and granzyme B The expressions of IL 2R and ICAM 1 were detected by immunohistochemistry. The apoptosis of transplanted intestinal epithelial cells was detected by TUNEL. Results ① The pathological examination showed that the rats in group 3 were mild, moderate and severe rejection on the 3rd, 5th and 7th day after operation, respectively, and no signs of rejection were found in groups 2 and 4; ② The detected genes in group 1 were basically not expressed . The expression of IL-2 mRNA in group 3 was significantly higher than that in group 2 on the 5th postoperative day (P <0 05). The expression of IFN γ mRNA was always significantly higher than those in groups 2 and 4 after operation (P <0.01); Perforin and granzyme B mRNA expression were significantly higher than those of the second and fourth groups on the 5th and 7th day postoperatively. (3) The expression of IL-2R in the third group was significantly higher than that in the three control groups , And the expression of ICAM 1 was higher than that of the control group only on the third day. ④ The apoptosis of intestinal epithelial cells in the third group was significantly higher than the other three control groups on the 3rd and 5th day after operation. in conclusion