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

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18～28 d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allo-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.

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

Objective To explore the expression of protease-aetivated receptor (PAR)-1, 2 in a routine model of acute graft-versus-host disease (aGVHD). Methods A routine model of aGVHD after aUogeneic 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 immunohistoehemistry 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 eeUs and vascular endothelial cells of target organs of aGVHD. Conclusion Inereased 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.