JNK、P38-MAPK、IκB-α在异基因CD4^+T细胞致人脐静脉内皮细胞损伤中的表达及与TF的关系  

Expressions of JNK,P38-MAPK and IκB-α in Allogeneic CD4^+T Lymphocyte-induced HUVECs Injury and Their Correlation with Tissue Factor

作　　者：韩红[1,2] 黎纬明[1] 邹萍[1] 樊晓武[3] 黄畦[3] 

机构地区：[1]华中科技大学同济医学院附属协和医院血液病研究所,武汉430022 [2]华中科技大学同济医学院附属梨园医院血液科,武汉430077 [3]华中科技大学同济医学院附属梨园医院心血管外科,武汉430077

出　　处：《华中科技大学学报（医学版）》2013年第5期515-519,共5页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：国家自然科学基金青年基金资助项目(No.30600570)

摘　　要：目的检测磷酸化的c-Jun氨基末端激酶(c-Jun N-terminal kinase,JNK)、P38-丝裂原活化的蛋白激酶(P38-mitogen activated protein kinase,P38-MAPK)、核转录因子抑制因子(IκB-α)及组织因子(tissue factor,TF),在异基因及同基因CD4+T细胞与人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)混合培养时在HUVECs中的表达,分析JNK、P38-MAPK、IκB-α与TF的相关性,探讨TF在异基因造血干细胞移植急性移植物抗宿主病(aGVHD)血管内皮损伤中可能的作用机制。方法体外原代分离培养HUVECs,用人T细胞亚群阴性分离柱试剂盒,从健康志愿者周围血中分离出CD4+T细胞作为异基因实验组,以同基因CD4+T细胞为对照组,用γ干扰素预处理HUVECs后与CD4+T细胞混合培养,分别在培养前及培养后不同时间点用流式细胞仪检测HUVECs中TF抗原的表达,实时定量PCR检测TF-mRNA基因的转录状况,Western blot检测磷酸化JNK、P38-MAPK、IκB-α的蛋白含量。结果异基因组TF及磷酸化JNK、P38-MAPK、IκB-α的表达明显升高,与对照组在不同时间点比较差异均有统计学意义(均P<0.05)。相关性分析显示,TF与磷酸化JNK、P38-MAPK表达呈正相关,而与磷酸化IκB-α表达无相关性。结论 TF可能通过JNK、P38-MAPK信号通路参与异基因造血干细胞移植时aGVHD血管内皮细胞的活化与损伤。Objective To detect the expressions of c-Jun N-terminal kinase （JNK）,P38-mitogen activated protein kinase （P38-MAPK）,IκB-α and tissue factor （TF） in human umbilical vein endothelial cells （HUVECs） co-cultured with allogeneic and autologous CD4＋ T lymphocytes and explore the correlation of JNK,P38-MAPK and IκB-α with TF and the role of TF in vascular endothelial cell injury in acute graft versus host disease （aGVHD）. Methods HUVECs were isolated and cultured in vitro. CD4＋ T lymphocytes were isolated from the peripheral blood of healthy volunteers by using human T cell negative isolation reagent. HUVECs pretreated with interferon （IFN）-γ were co-cultured with allogeneic CD4＋ T lymphocytes （experimental group） or autologous CD4＋T lymphocytes （control group）. Before and at different time points after co-culture, the expression of TF on the membrane of HUVECs was determined by flow cytometry, the mRNA expression of TF by real-time quantitative polymerase chain reaction （RQ-PCR） and the protein expression of phosphorylated JNK, P38-MAPK and IκB-α by Western blot. Results The expression of TF, phosphorylated JNK, P38-MAPK and IκB-α was significantly increased in experimental group relative to control group with significant difference found at each time point （P〈0.05 for all）. TF expression was positively associated with the expression of phosphorylated JNK and P38-MAPK. No positive correlation was found between the ex pression of TF and IκB-α. Conclusion TF may participate in the endothelial cell injury in aGVHD during allogeneic hematopoietic stem cell transplantation （HSCT） via JNK and P38-MAPK signal pathways.

关 键 词：C-JUN氨基末端激酶 P38-丝裂原活化的蛋白激酶 核转录因子抑制因子 组织因子 磷酸化 混合淋巴细胞反应 急性移植物抗宿主病 

分 类 号：R349.6[医药卫生—基础医学]