高迁移率族蛋白B1对血管内皮细胞ECV-304活化作用的研究  

Study on high mobility group box-1 protein inducing endothelial cell activation in vitro

作　　者：栾正刚[1] 张成[2] 葛春林[2] 马晓春[1] 郭仁宣[2] 

机构地区：[1]中国医科大学附属第一医院重症医学科,辽宁沈阳110001 [2]中国医科大学附属第一医院普通外科,辽宁沈阳110001

出　　处：《中国现代医学杂志》2008年第19期2798-2801,共4页China Journal of Modern Medicine

摘　　要：目的探讨高迁移率族蛋白B1(HMGB1)诱导血管内皮细胞活化的信号转导机制。方法体外培养人脐静脉内皮细胞(HUVECs)株ECV304,并将其分为对照组、HMGB1刺激组、丙酮酸乙酯(EP)处理组,应用酶联免疫吸附法(ELISA)检测细胞培养上清TNF-α水平变化、免疫荧光显微镜观察血管内皮细胞NF-κB活性的变化、凝胶迁移率实验(EMSA)检测细胞内NF-κB DNA结合活性的变化。结果HMGB1可诱导人脐静脉内皮细胞NF-κB在短时间内活化,促进TNF-α表达;EP处理后可显著降低NF-κB活性,抑制TNF-α表达。结论HMGB1可能通过活化血管内皮细胞NF-κB进而诱导TNF-α分泌,参与脓毒症的病理生理过程。EP可通过拮抗HMGB1刺激血管内皮细胞NF-κB活化从而发挥保护作用。[Objectives] To investigate the potential signal transduction mechanism in high mobihty group box-1 protein （HMGB1） activating vascular endothelial cell. [Methods] The cultured human umbilical vascular endothelial ceils （HUVECs） strain ECV-304 in vitro were divided into 3 groups： control group, HMGB1 group, HMGB1 plus ethyl pyruvate group. The quantities of TNF-α secreted in ECV-304 cells were measured by sandwich ELISA, the activation of nuclear faetor-KB （NF-KB） was observed by fluoreseenee microscope and the DNA binding activity of NF-KB in endothelial ceils was measured with eletrophoretic mobility shift assays （EMSA）. [Results] Incubation with HMGB1 was found to result in rapid increment of NF-kB in cultured HUVECs. HMGB1 markedly induced the expression of TNF-α from HUVECs. Ethyl pyruvate treatment could decrease the activity of NF-KB and the expression of TNF-α. [Conclusion] HMGB1 could enhance transient mobilization of NF-KB and markedly induce the release of TNF-α in endothelial cells, which suggested HMGB1 may play an important role in the occurrence and development of sepsis. Ethyl pyruvate is a potent inhibitor of NF-kB activation induced by HMGB1 in endothelial cells, which might explain its beneficial effect in endothelial cells.

关 键 词：血管内皮细胞 高迁移率族蛋白B1 丙酮酸乙酯 

分 类 号：R446.6[医药卫生—诊断学]