蛋白酶激活受体-1，2在小鼠急性移植物抗宿主病模型中的表达

Gene and protein expression of protease-activated receptor-1, 2 in a murine model of acute graft-versus- host disease

目的研究蛋白酶激活受体-1，2（PAR-1，2）在小鼠急性移植物抗宿主病（aGVHD）模型中的表达。方法建立小鼠异基因造血干细胞移植（allo-HSCT）后aGVHD模型，并用同基因HSCT小鼠作为对照。观察小鼠aGVHD表现；实时荧光定量聚合酶链反应、蛋白印迹和免疫组织化学法检测移植小鼠各种组织中PAR-1，2在基因和蛋白水平的表达。结果allo-HSCT小鼠在移植后18-28d出现了典型的aGVHD临床和病理表现。PAR-1基因在allo—HSCT组小鼠的皮肤、肝脏和小肠组织中的相对表达量较对照组显著增高（皮肤：0．039±0．013和0．008±0．002，P〈0．01；肝脏：0．165±0．084和0．017±0．006，P〈0．01；小肠：0．215±0．109和0．016±0．009，P〈0．01）；PAR-2基因在a11o-HSCT组小鼠的肝脏和小肠组织中表达较对照组增高（肝脏：0．010±0．003和0．003±0．002，P〈0．01；小肠：0．006±0．002和0．003±0．002，P〈0．05）；PAR-1，2基因在其余器官组织中的表达与对照组比较，差异无统计学意义。PAR-1，2在蛋白水平的表达与基因表达的检测结果一致。免疫组织化学染色显示PAR-1，2主要在aGVHD靶器官的上皮细胞和内皮细胞中表达增强。结论PAR-1，2表达增高很有可能参与异基因造血干细胞移植后aGVHD的病理生理过程。

Objective To explore the expression of protease-activated receptor （PAR）-1, 2 in a murine model of acute graft-versus-host disease （aGVHD）. Methods A murine model of aGVHD after allogeneic hematopoietic stem cell transplantation （allo-HSCT） was established, and the syngeneic HSCT mice were used as the controls. Quantitative real-time PCR, Western blot and immunohistochemistry were done to detect the gene and protein expression of PAR-1, 2 in multiple organs of allo-HSCT mice and the controls. Results Allo-HSCT mice showed classical symptoms and histological changes of aGVHD. PAR-1 mRNA expression was significantly increased in the skin, liver, small intestine of allo-HSCT mice （skin： 0. 039 ± 0. 013 vs. controls： 0. 008 ± 0. 002,P〈0. 01 ; liver： 0. 165± 0.084, vs. controls： 0.017± 0.006, P〈0.01; small intestine： 0.215 ± 0. 109 vs. controls： 0. 016 ±0. 009, P〈0. 01）, but not in the stomach, lung and kidney of allo-HSCT mice （P 〈0. 05 ）. PAR-2 mRNA expression in the liver and small intestine of allo-HSCT mice was significantly elevated （liver： 0. 010± 0. 003 vs. controls： 0. 003 ± 0. 002, P〈0. 01; small intestine： 0. 006 ± 0. 002 vs. controls： 0. 003 ± 0. 002,P〈0. 05）, but not in the other organs （P〈0. 05）. The protein expression of PAR-1, 2 was in accordance with the mRNA expression. Immunohistochemistry revealed the PAR-1, 2 expression was increased in the epithelial cells and vascular endothelial cells of target organs of aGVHD. Conclusion Increased expression of PAR-1, 2 in the target organs of aGVHD suggests PAR-1, 2 may contribute to the pathogenesis of aGVHD after allo-HSCT.