摘要：目的观察二羰基化合物对人血管内皮细胞(VEC)核因子(NF)κB的激活作用，以探讨该类物质病理生物学作用的机制。方法从人脐静脉分离VEC并与不同浓度的二羰基化合物共同培养。以氧化敏感的荧光染料2，7-二氢二氯荧光素(DCFH)染色，用流式细胞仪测定细胞内氧化水平；用间接免疫荧光法观察NF-κDp65在细胞内的分布，判断NF-κB的活化。结果二羰基化合物能够导致细胞内氧化水平升高，并激活人VECNF-κB；抗氧化剂(N-乙酰半胱氨酸和丙丁酚)以及羰基化合物清除剂(氨基胍)能够抑制二羰基化合物导致的细胞氧化应激，同时抑制NF-κB的活化。结论二羰基化合物通过诱导细胞氧化应激，激活人VECNF-κB，这一作用可能参与了慢性肾功能衰竭和糖尿病时血管病变的发生。

Activation of nuclear transcription factor-κB by dicarbonyl compounds in cultured human vascular endothelial cells

LIANG Min ,HOU Fanfan ,ZHANG Xun

(Division of Nephrology,Nanfang Hospital,the First Minitary Medical Unirersity,Guangzhou 510515,China)

Abstract：Objective To elucidate the effect of dicarbonyl compounds on activation of nuclear transcription factor-κB(NF-κB ) in human vascular endothelial cells (VEC).Methods VECs were isolated from human umbilical vein and incubated with two kinds of dicarbonyl compounds,3-deoxyglucosone (3-DG) or methylglyoxal(MGO).Intracellular oxidative levels were measured by flow cytometric assay using an oxidant sensitive dye-2,7-dichlorefluoresin (DCFH).Immunofluorescent staining was performed using polyclonal antibody against human NF-κB p65 to detect the translocation of NF-κB in VECs.Results When VECs were incubated with 3-DG or MGO,the levels of intracellular oxidation increased immediately (P＜0.05).Simultaneously,the amounts of the cells in which NF-κB p65 translocated into nucleuses increased significantly as compared with the unstimulated group (P＜0.01).Both activation of NF-κB and oxidative stress of VECs induced by 3-DG and MGO were inhibited when a carbonyl scavanger aminoguanidine(AG) was included into the cultures,or pre-incubated VECs with oxidative inhibitors,N-acetylcysteine (NAC) or probucol.Conclusions NF-κB in VECs can be activated by dicarbonyl compounds in company with induction of cellular oxidative stress,which can be partially blocked by both antioxidants and earbonyl scavanger.Increased level of plasma dicarbonyl compounds may be involved in the pathogenesis of vascular diseases seen in diabetes and chronic renal failure.

Keywords：Vascular endothelium;Nuclear factor-κB;Oxidative stress;Dicarbonyl compounds

基金项目：国家自然科学基金资助项目(39970341)；广东省自然科学基金团队项目

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收稿日期：2000-08-18