白细胞介素-10在川崎病炎性细胞因子介导的血管损伤中的作用初探  

Effect of interleukin-10 on the 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]深圳市儿童医院分子免疫研究室,518026 [2]重庆医科大学附属儿童医院儿科研究室

出　　处：《中华风湿病学杂志》2001年第6期363-366,T002,共5页Chinese Journal of Rheumatology

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

摘　　要：目的　探讨白细胞介素 10 (IL 10 )在炎性细胞因子介导的川崎病血管内皮损伤中的作用及机制。方法　建立人脐静脉内皮细胞培养模型 ,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IL 6、IL 1β、肿瘤坏死因子 α(TNF α)、IL 10的分泌量 ;AnnexinV/PI双染色法 ,流式细胞仪检测川崎病患儿PBMC培养上清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活性。结果　川崎病组患者PBMC体外刺激活化后IL 6、IL 1β、TNF α等炎性细胞因子及IL 10的分泌量均明显高于正常对照组 (P <0 0 1) ,其培养上清所诱导内皮细胞凋亡率 (38 4± 7 8) %较正常组 (2 8± 0 8) %明显升高。抗IL 6、TNF αMcAb、IL 1ra加入川崎病患儿PBMC培养上清 ,可不同程度抑制川崎病患儿PBMC培养上清所诱导内皮细胞的凋亡 ,而抗IL 10McAb加入川崎病患儿PBMC培养上清所诱导内皮细胞的凋亡细胞反而增多 ,凋亡细胞的百分率高达 (5 8 6± 14 6 ) %。川崎病患儿PBMC刺激活化后核因子NF κB活性显著增高。于PBMC培养上清加入IL 10 ,可见伴随着IL 6、IL 1β、TNF α等炎性细胞因子在PBMC中的表达降低 ,NF κB活性也降低 ,同时内皮细胞凋亡明显减少。结论　IL 10抑制川崎病患儿免疫?Objective To explore the effects of Interleukin 10 (IL 10) on the endothelial damage mediated by inflammatory cytokines in Kawasaki disease (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),interleukin 6 (Il 6) and IL 10 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 Annexin V/PI double staining though flow cytometry.Electrophoretic mobility shift assay (EMSA) was used to detect the activity of nuclear factor κB (NF κB).Results The level of TNF α,IL 1β,IL 6 and IL 10 were markedly increased in KD patients and the proportion of apoptotic cells in endothelial cells induced by the cultured supernatants of PBMCs was markedly elevated (38 4±7 8)% compared to (2 8±0 8)% in the control subjects.The apoptosis of endothelial cells induced by the cultured supernatants of PBMCs could be reversed to some degree by anti TNF α,IL 1β and IL 6 monoclonal antibody (McAb),otherwise could be enhanced (58 6±14 6)% by anti IL 10 McAb.The activity of NF κB in the activated PBMCs in KD patients was distinctly increased and IL 10 could significantly inhibit the activity of NF κB,reduce the production of TNF α,IL 1β and IL 6 and the apoptosis of endothelial cells induced by the cultured supenatants of PBMCs in KD patients.Conclusion IL 10,which can inhibit the activity of NF κB and the transcription of inflammatory cytokines such as TNF α,IL 1β and IL 6,acts as an important protective factor in endothelial damage of KD.

关 键 词：川崎病 炎性细胞因子 黏膜皮肤淋巴结综合征 白细胞介素10 NF-κB 人脐静脉内皮细胞 血管内皮损伤 

分 类 号：R725.4[医药卫生—儿科]