title: Morphological and proteomic analyses reveal that unsaturated guluronate oligosaccharide modulates multiple functional pathways in murine macrophage RAW264.7 cells.
abstract: Alginate is a natural polysaccharide extracted from various species of marine brown algae. Alginate-derived guluronate oligosaccharide (GOS) obtained by enzymatic depolymerization has various pharmacological functions. Previous studies have demonstrated that GOS can trigger the production of inducible nitric oxide synthase (iNOS)/nitric oxide (NO), reactive oxygen species (ROS) and tumor necrosis factor (TNF)-α by macrophages and that it is involved in the nuclear factor (NF)-κB and mitogen-activated protein (MAP) kinase signaling pathways. To expand upon the current knowledge regarding the molecular mechanisms associated with the GOS-induced immune response in macrophages, comparative proteomic analysis was employed together with two-dimensional electrophoresis (2-DE), matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/TOF MS) and Western blot verification. Proteins showing significant differences in expression in GOS-treated cells were categorized into multiple functional pathways, including the NF-κB signaling pathway and pathways involved in inflammation, antioxidant activity, glycolysis, cytoskeletal processes and translational elongation. Moreover, GOS-stimulated changes in the morphologies and actin cytoskeleton organization of RAW264.7 cells were also investigated as possible adaptations to GOS. This study is the first to reveal GOS as a promising agent that can modulate the proper balance between the pro- and anti-inflammatory immune responses, and it provides new insights into pharmaceutical applications of polysaccharides.
authors.affiliation: College of Life Science, Shenzhen Key Laboratory of Marine Bioresources and Ecology, Shenzhen University, Shenzhen 518060, China. xuxu@szu.edu.cn.
authors.forename: Xu
authors.initials: X
authors.lastname: Xu
authors.affiliation: College of Life Science, Shenzhen Key Laboratory of Marine Bioresources and Ecology, Shenzhen University, Shenzhen 518060, China. 215017325@qq.com.
authors.forename: De-Cheng
authors.initials: DC
authors.lastname: Bi
authors.affiliation: College of Life Science, Shenzhen Key Laboratory of Marine Bioresources and Ecology, Shenzhen University, Shenzhen 518060, China. 611279860@qq.com.
authors.forename: Chao
authors.initials: C
authors.lastname: Li
authors.affiliation: College of Life Science, Shenzhen Key Laboratory of Marine Bioresources and Ecology, Shenzhen University, Shenzhen 518060, China. 1210417703@qq.com.
authors.forename: Wei-Shan
authors.initials: WS
authors.lastname: Fang
authors.affiliation: College of Life Science, Shenzhen Key Laboratory of Marine Bioresources and Ecology, Shenzhen University, Shenzhen 518060, China. zhouruiswg@gmail.com.
authors.forename: Rui
authors.initials: R
authors.lastname: Zhou
authors.affiliation: College of Life Science, Shenzhen Key Laboratory of Microbial Genetic Engineering, Shenzhen University, Shenzhen 518060, China. msbiotools@gmail.com.
authors.forename: Shui-Ming
authors.initials: SM
authors.lastname: Li
authors.affiliation: National Glycoengineering Research Center, Shandong University, Jinan 250100, China. lianlichi@gmail.com.
authors.forename: Lian-Li
authors.initials: LL
authors.lastname: Chi
authors.affiliation: Division of Physiological Chemistry 2, Department of Medical Biochemistry and Biophysics, Karolinska Institute, Stockholm 17177, Sweden. min.wan@ki.se.
authors.forename: Min
authors.initials: M
authors.lastname: Wan
authors.affiliation: College of Life Science, Shenzhen Key Laboratory of Marine Bioresources and Ecology, Shenzhen University, Shenzhen 518060, China. slm@szu.edu.cn.
authors.forename: Li-Ming
authors.initials: LM
authors.lastname: Shen
mesh_terms: D000464:Alginates / Q000378:metabolism; D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D000975:Antioxidants / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D002240:Carbohydrate Sequence; D002467:Cell Nucleus / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D053145:Cell Nucleus Size / Q000187:drug effects; D048429:Cell Size / Q000187:drug effects; D015195:Drug Design; D005786:Gene Expression Regulation / Q000187:drug effects*; D020723:Glucuronic Acid / Q000378:metabolism; D006603:Hexuronic Acids / Q000378:metabolism; D006868:Hydrolysis; D020935:MAP Kinase Signaling System / Q000187:drug effects; D008262:Macrophage Activation / Q000187:drug effects; D008264:Macrophages / Q000166:cytology / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D051379:Mice; D008970:Molecular Weight; D009844:Oligosaccharides / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D010449:Peptide Mapping; D011133:Polysaccharide-Lyases / Q000378:metabolism; D011135:Polysaccharides, Bacterial / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D040901:Proteomics / Q000379:methods; D000067996:RAW 264.7 Cells
chemical_list: D000464:Alginates; D000894:Anti-Inflammatory Agents, Non-Steroidal; D000975:Antioxidants; D006603:Hexuronic Acids; D009844:Oligosaccharides; D011135:Polysaccharides, Bacterial; C111091:polyguluronic acid; D020723:Glucuronic Acid; D011133:Polysaccharide-Lyases; C016158:poly(beta-D-mannuronate) lyase
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