title: Advanced glycation end products (AGEs) increase human mesangial foam cell formation by increasing Golgi SCAP glycosylation in vitro.
abstract: Advanced glycation end products (AGEs) is one of the causative factors of diabetic nephropathy, which is associated with lipid accumulation in glomeruli. This study was designed to investigate whether N(ε)-(carboxymethyl) lysine (CML; a member of the AGEs family) increases lipid accumulation by impairing the function of sterol-regulatory element binding protein (SREBP) cleavage-activating protein (SCAP) in human mesangial cells (HMCs). Intracellular cholesterol content was assessed by Oil Red O staining and quantitative assay. The expression of molecules controlling cholesterol homeostasis was examined using real-time quantitative RT-PCR and Western blotting. The activity of Golgi-processing enzymes was determined using enzyme-based methods, and the translocation of SCAP from the endoplasmic reticulum (ER) to the Golgi was detected by confocal microscopy. CML increased cholesterol accumulation in HMCs. Exposure to CML increased expression and abnormal translocation of SCAP from the ER to the Golgi even in the presence of a high concentration of LDL. The increased SCAP translocation carried more SREBP-2 to the Golgi for activation by proteolytic cleavages, enhancing transcription of 3-hydroxy-3-methylclutaryl-CoA reductase and the LDL receptor. CML increased Golgi mannosidase activity, which may enhance glycosylation of SCAP. This prolonged the half-life and enhanced recycling of SCAP between the ER and the Golgi. The effects of CML were blocked by inhibitors of Golgi mannosidases. AGEs (CML) increased lipid synthesis and uptake, thereby causing foam cell formation via increasing transcription and protein glycosylation of SCAP in HMCs. These data imply that inhibitors of Golgi-processing enzymes might have a potential renoprotective role in prevention of mesangial foam cell formation.
authors.affiliation: Dept. of Endocrinology, Zhongda Hospital, Southeast University, No. 87 Dingjiaqiao, Nanjing, Jiangsu Province 21009, P.R. China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Yuan
authors.forename: Lei
authors.initials: L
authors.lastname: Zhao
authors.forename: Yaxi
authors.initials: Y
authors.lastname: Chen
authors.forename: John F
authors.initials: JF
authors.lastname: Moorhead
authors.forename: Zac
authors.initials: Z
authors.lastname: Varghese
authors.forename: Stephen H
authors.initials: SH
authors.lastname: Powis
authors.forename: Shane
authors.initials: S
authors.lastname: Minogue
authors.forename: Zilin
authors.initials: Z
authors.lastname: Sun
authors.forename: Xiong Z
authors.initials: XZ
authors.lastname: Ruan
mesh_terms: D015153:Blotting, Western; D002478:Cells, Cultured; D002784:Cholesterol / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000378:metabolism; D005487:Foam Cells / Q000502:physiology*; D005920:Glomerular Mesangium / Q000166:cytology / Q000378:metabolism*; D017127:Glycation End Products, Advanced / Q000494:pharmacology*; D006031:Glycosylation; D006056:Golgi Apparatus / Q000502:physiology*; D006801:Humans; D006903:Hydroxymethylglutaryl CoA Reductases / Q000378:metabolism; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism*; D008239:Lysine / Q000031:analogs & derivatives / Q000494:pharmacology; D008565:Membrane Proteins / Q000378:metabolism*; D018613:Microscopy, Confocal; D008861:Microsomes / Q000378:metabolism; D010957:Plasmids; D012313:RNA / Q000096:biosynthesis / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D011973:Receptors, LDL / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D043323:alpha-Mannosidase / Q000378:metabolism
chemical_list: D017127:Glycation End Products, Advanced; D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins; D012333:RNA, Messenger; D011973:Receptors, LDL; C102196:SREBP cleavage-activating protein; D012313:RNA; C048496:N(6)-carboxymethyllysine; D002784:Cholesterol; D006903:Hydroxymethylglutaryl CoA Reductases; D043323:alpha-Mannosidase; D008239:Lysine