核转录因子-κB通路在CD40-CD154诱导血管内皮细胞活化中的作用

目的 探讨血管内皮细胞(VEC)内核转录因子-κB(NF-κB)通路在CD40-CD154相互作用诱导VEC活化中的作用.方法 应用重组人肿瘤坏死因子α(rhTNF-α)和重组人γ干扰素(rhIFN-γ)刺激人主动脉VEC,通过流式细胞仪(FACS)检测CD40和粘附因子表达水平;通过表达CD154的细胞株D1.1和VEC共同培养后,观察VEC粘附因子的表达水平;通过抗CD154单克隆抗体阻断CD40-CD154间的反应;应用NF-κB阻断剂BAY11-7082观察阻断NF-κB通路后对CD40-CD154诱导VEC活化的影响.结果 未活化的VEC膜表达低水平的CD40、CD54和CD106,但不表达CD62E,经白细胞介素刺激后可上调这些蛋白分子的表达水平.D1.1细胞株和VEC共同培养后可诱导VEC活化并上调CD62E、CD54和CD106的表达水平;抗CD154抗体可阻断CD40-CD154间的反应所诱导的VEC活化;NF-κB阻断剂可阻断rhTNF-α诱导的VEC活化以及CD40-CD154间的反应所诱导的VEC活化.结论 VEC中CD40和T淋巴细胞中CD154之间的相互作用在诱导VEC活化过程中起重要作用;CD40-CD154相互作用并通过NF-κB通路诱导VEC活化;NF-κB通路阻断剂可抑制VEC活化.

Role of NF-κB pathway in CD40-CD154 interaction-induced human aortic endothelial cell activation

Objective To determine human endothelial cells(VEC) activation induced by CD40-CD154 interaction and the role of NF-κB pathway in CD40-CD154 interaction. Methods VEC were isolated from aorta,and subcultures were stimulated by recombinant human TNF-α(rhTNF-α)or IFN-γ(rhIFN-γ). The expression of CD40 and adhesion molecules was detected by using flow cytometry. D1.1 cells expressing CD154 were co-incubated with VEC with or without CD154 specific monoclonal antibody, and the effects of CD40-CD154 interaction in induceding VEC activation were determined. In an additional study,BAY 11-7082,a NF-κB pathway inhibitor,was used to study whether CD40-CD154 interaction-induced VEC activation was NF-κB pathway-dependent. Results Resting VEC demonstrated weak expression of CD40,CD54 and CD106 but undetectable CD62E. VEC stimulated with rhTNF-α or rhINF-γ up-regulated the expression of CD40,CD62E,CD54 and CD106 respectively. VEC demonstrated up-regulation of CD62E, CD54 and CD106 after co-incubation with D1.1 cells,and VEC activation induced bv CD40-CD154 interaction was inhibited by CD154 specific monoclonal antibody. NF-κB could block the VEC activation induced bv rhTNF-α stimulation or by CD40-CD154 interaction. Conclusions The interaction between VEC-derived CD40 and CD154 expressing T cells plays a critical role in VEC activation. Inhibition of NF-κB during CD40-CD154 interaction prevents VEC activation.