Upregulation of mesangial growth factor and extracellular matrix synthesis by advanced glycation end products via a receptor-mediated mechanism.
Enhanced advanced glycosylation end product (AGE) formation has been shown to participate in the pathogenesis of diabetes-induced glomerular injury by mediating the increased extracellular matrix (ECM) deposition and altered cell growth and turnover leading to mesangial expansion. These effects could be exerted via an AGE-receptor-mediated upregulation of growth factors, such as the IGFs and transforming growth factor-beta (TGF-beta). We tested this hypothesis in human and rat mesangial cells grown on nonglycated or native bovine serum albumin (BSA), glycated BSA with AGE formation (BSA-AGE), or glycated BSA in which AGE formation was prevented by the use of aminoguanidine (BSA-AM), in the presence or absence of an antibody, alpha-p60, directed against the p60/OST protein named AGE-receptor 1 (AGE-R1), or normal control (pre-immune) serum. The mRNA and/or protein levels of IGF-I, IGF-II, IGF receptors, IGF binding proteins (IGFBPs), TGF-beta1 and the ECM components fibronectin, laminin, and collagen IV were measured, together with cell proliferation. Both human and rat mesangial cells grown on BSA-AGE showed increased IGF-I and total and bioactive TGF-beta medium levels and enhanced IGF-I, IGF-II, and TGF-beta1 gene expression, compared with cells grown on BSA, whereas total IGFBP and IGFBP-3 medium content, IGF receptor density and affinity, and IGF-I receptor transcripts were unchanged. Moreover, cells grown on BSA-AGE showed increased ECM protein and mRNA levels versus cells cultured on BSA, whereas cell proliferation was unchanged in human mesangial cells and slightly reduced in rat mesangial cells. Growing cells on BSA-AM did not affect any of the measured parameters. Co-incubation of BSA-AGE with anti-AGE-R1, but not with pre-immune serum, prevented AGE-induced increases in IGF-I, TGF-beta1, and ECM production or gene expression; anti-AGE-R1 also reduced growth factor and matrix synthesis in cells grown on BSA. These results demonstrate that mesangial IGF and TGF-beta1 synthesis is upregulated by AGE-modified proteins through an AGE-receptor-mediated mechanism. The parallelism with increased ECM production raises the speculation that the enhanced synthesis of these growth factors resulting from advanced nonenzymatic glycation participates in the pathogenesis of hyperglycemia-induced mesangial expansion.
Pugliese
G
G
Department of Experimental Medicine, 2nd Institute of Internal Medicine, La Sapienza University, Rome, Italy.
Pricci
F
F
Romeo
G
G
Pugliese
F
F
Mené
P
P
Giannini
S
S
Cresci
B
B
Galli
G
G
Rotella
C M
CM
Vlassara
H
H
Di Mario
U
U
D000818:Animals; D002417:Cattle; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D003094:Collagen / Q000096:biosynthesis; D005109:Extracellular Matrix / Q000187:drug effects / Q000502:physiology*; D016326:Extracellular Matrix Proteins / Q000096:biosynthesis*; D005353:Fibronectins / Q000096:biosynthesis; D005786:Gene Expression Regulation / Q000187:drug effects; D005920:Glomerular Mesangium / Q000378:metabolism*; D017127:Glycation End Products, Advanced / Q000494:pharmacology*; D006146:Guanidines / Q000494:pharmacology; D006801:Humans; D018969:Insulin-Like Growth Factor Binding Proteins / Q000096:biosynthesis; D007334:Insulin-Like Growth Factor I / Q000096:biosynthesis* / Q000378:metabolism; D007335:Insulin-Like Growth Factor II / Q000096:biosynthesis* / Q000378:metabolism; D007700:Kinetics; D007797:Laminin / Q000096:biosynthesis; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D000067759:Receptor for Advanced Glycation End Products; D017526:Receptor, IGF Type 1 / Q000096:biosynthesis; D017527:Receptor, IGF Type 2 / Q000096:biosynthesis; D011971:Receptors, Immunologic / Q000187:drug effects / Q000502:physiology*; D012709:Serum Albumin / Q000494:pharmacology; D012710:Serum Albumin, Bovine / Q000494:pharmacology*; D014158:Transcription, Genetic / Q000187:drug effects; D016212:Transforming Growth Factor beta / Q000096:biosynthesis*; D015854:Up-Regulation
D016326:Extracellular Matrix Proteins; D005353:Fibronectins; D017127:Glycation End Products, Advanced; D006146:Guanidines; D018969:Insulin-Like Growth Factor Binding Proteins; D007797:Laminin; D012333:RNA, Messenger; D000067759:Receptor for Advanced Glycation End Products; D017527:Receptor, IGF Type 2; D011971:Receptors, Immunologic; D012709:Serum Albumin; D016212:Transforming Growth Factor beta; C081628:advanced glycation end products-bovine serum albumin; D012710:Serum Albumin, Bovine; D007334:Insulin-Like Growth Factor I; D007335:Insulin-Like Growth Factor II; D003094:Collagen; D017526:Receptor, IGF Type 1; C004479:pimagedine