title: Hydrogen sulfide reduces RAGE toxicity through inhibition of its dimer formation.
abstract: RAGE is important in the development of neurodegenerative diseases. The present study was designed to investigate the effect of hydrogen sulfide (H2S, an endogenous gaseous mediator) on the cytotoxicity caused by RAGE activation during the chronic oxidative stress. Aβ1-42decreased cell viability and induced cell senescence in SH-SY5Y cells. Treatment with advanced glycation end products (AGEs) induced cell injury in HEK293 cells stably expressing RAGE (HEK293-RAGE) and stimulated inflammatory responses in SH-SY5Y cells. Pretreatment of SH-SY5Y cells with an H2S donor, NaHS, significantly attenuated the above harmful effects caused by Aβ1-42or AGEs. Western blotting analysis shows that oxidative stress enhanced RAGE protein expression which was attenuated by either NaHS or over-expression of cystathionine β-synthase (CBS), a critical enzyme for producing H2S in brain cells. Both Western blots and split GFP complementation analysis demonstrate that NaHS reduced H2O2-enhanced RAGE dimerization. Immunofluorescence analysis shows that H2O2up-regulated the membrane expression of wild-type RAGE. However, H2O2-enhanced expression of the RAGE harboring C259S/C310S double mutation (DM-RAGE) was observed in the endoplasmic reticulum. Treatment with NaHS attenuated the effects of H2O2on the protein expression of WT-RAGE, but not that of DM-RAGE. Cycloheximide chase and ubiquitination assays show that NaHS reduced the half-life of WT-RAGE to a similar level of DM-RAGE. S-sulfhydration assay with the tag-switch technique demonstrate that H2S may directly S-sulfhydrate the C259/C301 residues. Our data suggest that H2S reduces RAGE dimer formation and impairs its membrane stability. The lowered plasma membrane abundance of RAGE therefore helps to protect cells against various RAGE mediated pathological effects.
authors.affiliation: Department of Pharmacology, Yong Loo Lin School of Medicine, National University of Singapore, 117600 Singapore, Singapore.
authors.forename: Hong
authors.initials: H
authors.lastname: Zhou
authors.affiliation: Department of Pharmacology, Yong Loo Lin School of Medicine, National University of Singapore, 117600 Singapore, Singapore.
authors.forename: Lei
authors.initials: L
authors.lastname: Ding
authors.affiliation: Department of Pharmacology, Yong Loo Lin School of Medicine, National University of Singapore, 117600 Singapore, Singapore; Life Science Institute, National University of Singapore, Singapore.
authors.forename: Zhiyuan
authors.initials: Z
authors.lastname: Wu
authors.affiliation: Department of Pharmacology, Yong Loo Lin School of Medicine, National University of Singapore, 117600 Singapore, Singapore.
authors.forename: Xu
authors.initials: X
authors.lastname: Cao
authors.affiliation: Department of Pharmacology, Yong Loo Lin School of Medicine, National University of Singapore, 117600 Singapore, Singapore.
authors.forename: Qichun
authors.initials: Q
authors.lastname: Zhang
authors.affiliation: Laboratory of Cardiovascular Sciences, National Institute on Aging, Baltimore, MD 21224, USA.
authors.forename: Li
authors.initials: L
authors.lastname: Lin
authors.affiliation: Department of Pharmacology, Yong Loo Lin School of Medicine, National University of Singapore, 117600 Singapore, Singapore; Life Science Institute, National University of Singapore, Singapore. Electronic address: phcbjs@nus.edu.sg.
authors.forename: Jin-Song
authors.initials: JS
authors.lastname: Bian
mesh_terms: D000375:Aging / Q000378:metabolism / Q000473:pathology; D000951:Antigens, Neoplasm / Q000235:genetics* / Q000378:metabolism; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D016922:Cellular Senescence / Q000187:drug effects; D019281:Dimerization; D005609:Free Radicals / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D017127:Glycation End Products, Advanced / Q000008:administration & dosage / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D006862:Hydrogen Sulfide / Q000378:metabolism* / Q000494:pharmacology; D007249:Inflammation / Q000188:drug therapy / Q000378:metabolism* / Q000473:pathology; D020928:Mitogen-Activated Protein Kinases / Q000235:genetics* / Q000378:metabolism; D009410:Nerve Degeneration / Q000378:metabolism* / Q000473:pathology; D018384:Oxidative Stress / Q000187:drug effects / Q000235:genetics*; D015398:Signal Transduction / Q000187:drug effects
chemical_list: D000951:Antigens, Neoplasm; D005609:Free Radicals; D017127:Glycation End Products, Advanced; D006861:Hydrogen Peroxide; C498978:MOK protein, human; D020928:Mitogen-Activated Protein Kinases; D006862:Hydrogen Sulfide
keywords: Aging; Free radicals; Hydrogen sulfide; Neurodegeneration; Pharmacology