中文摘要：

目的探讨组织因子（TF）在移植物抗宿主病（GVHD）血管内皮损伤中的作用及其机制。方法用实时定量PCR和Western blot方法检测TF在异基因造血干细胞移植（allo—HSCT）和自体造血干细胞移植（auto-HSCT）小鼠各组织中基因和蛋白水平的表达。用流式细胞术、实时定量PCR和Western blot方法分别检测异基因CD4^＋或CD8^＋ T淋巴细胞对脐静脉内皮细胞（HUVEC）中TF、VCAM-1、TNF-α、IFN-γ、IL-6以及MAPK表达的影响，观察抗TF抗体、p38MAPK和JNK阻滞剂对异基因T淋巴细胞诱导的HUVEC中细胞因子表达的影响。结果①在allo—HSCT后发生GVHD小鼠的皮肤、胃、小肠和肝组织中，TF在基因水平的表达为对照组的（15．1±2．1），（5．5±1．4），（9．7±2．3），（14．3±2．9）倍，TF蛋白的表达水平分别为对照组的（13．5±2．7），（6．2±0．9），（7．9±1．6），（15．3±3．2）倍；②异基因T淋巴细胞可显著诱导HUVEC中TF、VCAM-1、TNF—α、IFN-γ和IL-6的表达；异基因T淋巴细胞作用后，HUVEC中p38MAPK和JNK磷酸化增强，而磷酸化ERK无变化。p38MAPK和JNK阻滞剂可显著降低异基因CD4^＋和CD8^＋T淋巴细胞诱导HUVEC的TF表达。抗TF抗体和p38MAPK、JNK阻滞剂均可下调异基因T淋巴细胞诱导的HUVEC中VCAM-1、TNF-α、IFN-γ和IL-6的表达。结论TF通过细胞内信号通路p38MAPK和JNK参与了GVHD导致的血管内皮细胞损伤和活化。

英文摘要：

Objective To study the pathogenetic role of tissue factor （TF） in endothelial-injury in GVHD. Methods Gene and protein expressions of TF in the organs of allogenic hematopoietic stem cell transplantation（allo-HSCT） and autologous HSCT （auto-HSCT） mice were determined by real-time PCR and Western blot. The effect of allogeneic T lymphocytes on the expression of TF and other cytokines and activation of MAPKs in human umbilical vein endothelial cells （HUVECs） was detected by flow cytometry, real- time PCR or Western blot. The influence of TF antibodies （SB203580 and SP600125 ） on allogeneic T lymphoeytes-induced cytokines expression was also tested. Results ①TF gene and protein expression in the liver, skin, small intestine and stomach of allo-HSCT mice was significantly elevated about 15.1 ± 2.1, 5.5±1.4, 9.7 ± 2.3, 14.2± 2.9 folds and 13.5±2.7, 6.2 ± 0.9, 7.9±1.6, 15.3 ± 3.2 folds respectively compared with that of auto-HSCT mice. ②Allogeneic CD4^＋ CD8 ^＋ T lympbocytes significantly enhanced TF, VCAM-1, TNF-α, IFN-γ and IL-6 expression in TNF-α prestimulated HUVECs. ③Aflogeneic T lymphocytes enhanced p38MAPK and JNK phosphorylation in HUVECs, but did not affect ERK phosphorylation, p38 MAPK JNK inhibitors SB203580 and SP600125 reduced allogeneic T lymphocytes-induced TF expression in HUVECs. ④ SB203580 and SP600125 down-regulated allogeneie T lymphocytes-induced VCAM-1, TNF-α, IFN-γ, IL-6 expression in HUVECs. Conclusion TF mediates vascular endothelial-injury and activation in GVHD via phosphorylation of p38MAPK and JNK.