大鼠小肠移植排斥反应期移植肠基因表达、ICAM-1和IL-2R分子表达及上皮细胞凋亡的变化
李元新;李宁;李幼生;吴波;黎介寿
【期刊名称】《中华医学杂志（英文版）》 【年(卷),期】2001(114)010
【摘要】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rnIL-2R和ICAM-1表达，及移植肠上皮细胞凋亡的变化。rn方法选用近交系大鼠F344/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：rnWistar→Wistar；第3组：F344→Wistar；第4组：F344→Wistar+环孢霉素A(6 mg·kg-1·d-1，肌注)。术后第rn3、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、rnIFN-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rn移植肠上皮细胞凋亡的变化。rn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rn征象；②第1组各检测基因基本不表达。第3组IL-2 mRNA表达在术后第5天显著高于第2组(P＜rn0.05)；IFN-γ mRNA表达术后始终非常显著地高于第2、4组(P＜0.01)；穿孔素、粒细胞酶B mRNA表达在rn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rn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rn它三个对照组。rn结论 IL-2、IFN-γ、穿孔素和粒细胞酶B的转录，移植肠IL-2R和ICAM-1表达以及移植肠上皮细
胞凋亡在rn小肠移植排斥中发挥了重要作用；IFN-γ mRNA表达、IL-2R及上皮细胞凋亡可能成为临床上实用的早期、rn特异、敏感的诊断方法。通过改变这些分子的治疗策略可能促进目前抗排斥治疗策略的改进。%Objective To investigate the kinetics and the magnitude of intragraft gene expression of interleukin-2rn(IL-2), interferon-gamma (IFN-y), perforin and granzyme B, and intragraft expression of interieukin-2rnreceptor (IL-2R) and intercellular adhesion molecule-1 ( ICAM-1 ) during acute rejection episodes, and to rnanalyze the changes in apoptosis in small intestinal allograft
rejection.rnMethods
Heterotopic
small
intestine
transplantation was performed with inbred rats F344/N (RT11) and rnWistar/A (RT1-Ak, RT1-Ed). All recipients were divided into four groups: group 1 : Wistar, native control;rngroup 2: Wistar→Wistar; group 3: F344→Wistar and group 4: F344→Wistar + cyclosporine A rn(6 mg·kg-1 ·d-1 I.M. ). The grafts were harvested on postoperative days (PODs) 3, 5 and 7. All samples rnwere examined pathologically. Intragraft mRNA expression of IL-2, IFN-γ, perforin and granzyme B were rndetected with reverse transcriptase polymerase chain reaction (RT-PCR) and intragraft expression of IL-2R rnand ICAM-1 were stained using immunohistochemistry. We also analyzed the change in apoptosis rejection rnwith terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL).rnResults Mild acute rejection occurred on POD 3 in the ailograft group, moderate acute rejection on rnPOD 5, and
severe acute rejection on POD 7, while none of the isografts had histological evidence of acute rnrejection. Cyclosporine A could effectively control rejection. Gene expression was virtually negative in the rnnative control. Only on POD 5 was IL-2 mRNA expression of ailografts significantly higher than that of rnisografts ( P ＜ 0.05). IFN-γ mRNA expression was significantly higher than that of the control groups ( P ＜rn0.01 ) on PODs 3, 5 and 7, and the level of perforin and granzyme B mRNA expression reached rnsignificantly higher levels than in the other two control groups on POD 5 and POD 7. Intragraft IL-2Rrnexpression of the allograft was significantly higher than that of the other three control groups. Only on rnPOD 3 was intragraft ICAM-1 expression of allografts significantly higher than isografts. The number of rnapoptotic cells per crypt of allografts was significantly higher than that of the other three control groups on rnPOD 3 and POD 5 ( P ＜ 0.01 ).rnConclusion Transcription of IL-2, IFN-γ, perforin and granzyme B, and expression of IL-2R and ICAM-1rnas well as apoptosis of epithelial cells of the grafts play an important role in small intestine allograft rejection.rnIntragraft gene expression of IFN-γ and intragraft expression of IL-2R as well as apoptotic epithelial cells rnmay become a specific and sensitive diagnostic method of clinical value. Furthermore, therapeutic rnstrategies to alter these molecules in small intestine transplantation may improve the outcome of current rnantirejection
大鼠小肠移植排斥反应期移植肠基因表达、ICAM-1和IL-2R分子表达及上
