核转录因子在细胞因子介导的川崎病血管内皮损伤中的作用机理  

Effect of NF-κB on endothelial damage mediated by inflammatory cytokines in Kawasaki disease

作　　者：付劲蓉[1] 李成荣[1] 周玉峰[1] 周雅德[2] 

机构地区：[1]深圳市儿童医院,518026 [2]重庆医科大学儿童医院

出　　处：《中华儿科杂志》2002年第2期72-74,共3页Chinese Journal of Pediatrics

基　　金：广东省医学科学技术研究基金资助 (A2 0 0 16 2 2 );深圳市卫生系统科研基金资助 ( 199940 0 6 )

摘　　要：目的　探讨核转录因子 (NF κB)在诱导川崎病炎性细胞因子及血管内皮细胞损伤中的作用及机理。方法　建立人脐静脉内皮细胞培养模型 ,酶联免疫吸附实验 (ELISA)双抗体夹心法检测川崎病患儿外周血单个核细胞 (PBMC)活化后培养上清液白细胞介素 6 (IL 6 )、白细胞介素 1β(IL 1β)、肿瘤坏死因子α(TNF α)的分泌量 ;采用AnnexinⅤ /PI双染色法 ,流式细胞仪检测川崎病患儿PBMC培养上清液所诱导的内皮细胞凋亡率 ;凝胶电泳迁移率转换实验 (EMSA)检测所活化PBMC核因子NF κB的活性。结果　川崎病组患儿PBMC体外经刺激后IL 6、IL 1β、TNF α等炎性细胞因子的分泌均明显高于正常对照组 (P <0 0 1) ,其培养上清液所诱导的内皮细胞凋亡率 [(38 4± 7 8) %]则较正常对照组 [(2 8± 0 8) %]明显升高。分别加入足量白细胞介素 6单克隆抗体 (IL 6McAb)、肿瘤坏死因子α单克隆抗体 (TNF αMcAb)、白细胞介素 1β受体a(IL 1ra)或联合加入上述阻断剂于川崎病患儿PBMC培养上清液中 ,可不同程度地逆转川崎病患儿PBMC培养上清液所诱导内皮细胞的凋亡。川崎病患儿PBMC刺激活化后NF κB活性显著增高。NF κB抑制剂SN50 明显抑制川崎病患儿PBMC上述细胞因子的分泌 。Objective Kawasaki disease (KD) is the leading cause of acquired heart diseases in children. Its etiology is unknown. In recent years, data have shown that the dysfunction of T cell and monocytes/macrophages plays a central role in the development of vasculitis. Nuclear factor-κB(NF-κB) is positioned to integrate information from immune signaling pathway that can regulate the function of T cell and monocytes/macrophages. But the role of NF-κB in endothelial damage of KD is unknown. Therefore, the objective of the study was to further explore the effects of NF-κB on the endothelial damage in KD. Methods Human umbilical vein endothelial cells were cultured in vitro and the production of tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1β) and interleukin-6 (IL-6) were measured by enzyme-linked immunosorbent assay ( ELISA). The proportion of apoptotic cells in endothelial cells induced by the supernatants of peripheral blood mononuclear cells (PBMCs) was detected by AnnexinⅤ/PI double-staining. Electrophoretic mobility shift assay (EMSA) was used to detect the activity of NF-κB. Results The levels of TNF-α, IL-1β and IL-6 were markedly increased in patients and the proportion of apoptotic cells in endothelial cells induced by the cultured supernatants of PBMCs was markedly elevated (38.45±7.80)% compared to (2.87±0.76)% in the control subjects. The apoptosis induced by the supernatants of cultured PBMCs could be reversed, to some degree, through alone anti- TNF-α, IL-1β or IL-6 monoclonal antibody (McAb), or the combination together. The activity of NF-κB in the activated PBMCs in patients with KD was distinctly increased and SN 50 , the blockade of NF-κB, could significantly inhibit the production of TNF-α, IL-1β, IL-6 and the apoptosis of endothelial cells induced by the supernatants of cultured PBMC. Conclusion NF-κB, which can trigger the transcription of inflammatory cytokines such, as TNF-α, IL-1β, IL-6, plays an important role in endothelial damage of KD.

关 键 词：川崎病 NF-κB 细胞活素类 血管内皮 发病机制 

分 类 号：R725.4[医药卫生—儿科]