de Souza Pinto Raphael, Castilho Gabriela, Paim Bruno Alves, Machado-Lima Adriana, Inada Natalia M, Nakandakare Edna Regina, Vercesi Aníbal Eugênio, Passarelli Marisa
Lipids Laboratory-LIM 10, Faculty of Medical Sciences, University of São Paulo, Av. Dr Arnaldo 455-Room 3305, São Paulo 01246-000, Brazil.
Lipids. 2012 May;47(5):443-50. doi: 10.1007/s11745-011-3647-9. Epub 2012 Jan 21.
We investigated the role of aminoguanidine and benfotiamine on the inhibition of reactive oxygen species (ROS) generation in macrophages induced by advanced glycated albumin (AGE-albumin) and its relationship with cell cholesterol homeostasis, emphasizing the expression of the ATP binding cassette transporter A-1 (ABCA-1). AGE-albumin was made by incubating fatty acid-free albumin with 10 mM glycolaldehyde. ROS production and ABCA-1 protein level were determined by flow cytometry in J774 macrophages treated along time with control (C) or AGE-albumin alone or in the presence of aminoguanidine or benfotiamine. Mitochondrial function was evaluated by oxygraphy. Compared to C-albumin, AGE-albumin increased ROS production in macrophages, which was ascribed to the activities of NADPH oxidase and of the mitochondrial system. Mitochondrial respiratory chain activity was reduced in cells incubated with AGE-albumin. ROS generation along time was associated with the reduction in macrophage ABCA-1 protein level. Aminoguanidine prevented ROS elevation and restored the ABCA-1 content in macrophages; on the other hand, benfotiamine that promoted a lesser reduction in ROS generation was not able to restore ABCA-1 levels. Inhibition of oxidative stress induced by AGE-albumin prevents disturbances in reverse cholesterol transport by curbing the reduction of ABCA-1 elicited by advanced glycation in macrophages and therefore may contribute to the prevention of atherosclerosis in diabetes mellitus.
我们研究了氨基胍和苯磷硫胺对晚期糖基化白蛋白（AGE-白蛋白）诱导的巨噬细胞活性氧（ROS）生成的抑制作用及其与细胞胆固醇稳态的关系，重点关注ATP结合盒转运蛋白A-1（ABCA-1）的表达。通过将无脂肪酸白蛋白与10 mM乙醇醛孵育制备AGE-白蛋白。采用流式细胞术测定J774巨噬细胞在单独使用对照（C）或AGE-白蛋白或在氨基胍或苯磷硫胺存在下长时间处理后的ROS生成和ABCA-1蛋白水平。通过氧电极法评估线粒体功能。与C-白蛋白相比，AGE-白蛋白增加了巨噬细胞中的ROS生成，这归因于NADPH氧化酶和线粒体系统的活性。用AGE-白蛋白孵育的细胞中线粒体呼吸链活性降低。长时间的ROS生成与巨噬细胞ABCA-1蛋白水平的降低有关。氨基胍可防止ROS升高并恢复巨噬细胞中的ABCA-1含量；另一方面，促进ROS生成减少程度较小的苯磷硫胺无法恢复ABCA-1水平。抑制AGE-白蛋白诱导的氧化应激可通过抑制巨噬细胞中晚期糖基化引起的ABCA-1减少来防止逆向胆固醇转运紊乱，因此可能有助于预防糖尿病中的动脉粥样硬化。
