Screening of sulfated polysaccharide and sugar-related compounds as the regulator of advanced glycation end-products uptake by macrophage

[Speaker] Takashi Nishinaka:1
[Co-author] Takuro Kobori:1, Yui Yamazaki:1, Shinichi Hamasaki:2, Atsuhiro Kitaura:2, Atsuko Niwa:1, Shuji Mori:3, Masahiro Nishibori:4, Hideo Takahashi:1
1:Department of Pharmacology, Faculty of Medicine, Kindai University, Japan, 2:Department of Anesthesiology, Faculty of Medicine, Kindai University, Japan, 3:School of Pharmacy, Shujitsu University, Japan, 4:Department of Pharmacology, Okayama University, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Japan

Advanced glycation end-products (AGEs) which result from the prolonged exposure of proteins to sugars accumulate in the blood and tissues of diabetic patients. Accumulated evidence suggests that the AGEs are associated with both microvascular and macrovascular complications including retinopathy, nephropathy, neuropathy and cardiovascular disease in diabetic mellitus. Especially among AGEs, toxic AGEs derived from glyceraldehyde (AGE-2) and glycolaldehyde (AGE-3) have potent activity compared to other AGEs. Macrophages are capable of ingesting extracellular particles by receptor-mediated endocytosis. AGEs can interact with two types of cell surface receptors on macrophages. Scavenger receptors including type I and type II macrophage scavenger receptors, such as class B scavenger receptor (CD36), scavenger receptors-1 class A (SR-A, CD204), and lectin-like oxidized low-density lipoprotein receptor 1 (LOX-1), are predominantly involved in AGE capture, removal, and degradation. These receptors contribute to the development of atherosclerosis. Recently, we have demonstrated that the SR-A is involved in the AGEs uptake by macrophage. Furthermore, we found that the fucoidan which is a complex sulfated polysaccharide and a SR-A antagonist suppresses the AGEs uptake. These results raise the possibility that the sulfated polysaccharide or sugar-related compounds are candidate for the treatment of atherosclerosis. In the present study, we screened the sulfated polysaccharide or sugar related compounds as the regulator of AGES uptake by macrophage.
AGEs were prepared from BSA incubated with D-glyceraldehyde or D-glycoaldehyde at 37°C and fluorescently labelled by using Alexa Fluor 488 C5 maleimide. The mouse macrophage cell line, RAW264.7 were grown in Dulbecco's Modified Eagle Medium (DMEM) containing 2 mM glutamine and 10% heat-inactivated fetal bovine serum (FBS) at 37°C and 5% CO2. The cells were concomitantly treated with fluorescently labelled-AGEs and each compound for 1 h, then the AGEs uptake was evaluated by the flow cytometric analysis.

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