Korean J Urol.  2001 Nov;42(11):1204-1210.

Effects of Advanced Glycation End Products on Rat Bladder Fibroblast

• Chang IH
• Seong YW
• Myung SC
• Kim YS

Abstract

PURPOSE: The products of the irreversible non-enzymatic glycation is one of the important toxic components and it is called advanced glycation end products (AGEs). Because AGEs is related to development of neural toxicities, this study is directed to investigate the direct effects of AGEs on the rat bladder fibroblast cells in vitro.
MATERIALS AND METHODS
Fibroblast cells were cultured, using Sprague Dawley rat bladder. AGEs-bovine serum albumin (AGEs-BSA) was prepared, characterized by spectrophotometry, SDS-PAGE and ELISA. The viability of the fibroblast cells after AGEs-BSA treatment was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) assay. Receptor of advanced glycation end products (RAGE), procollagen type I, procollagen type III, and TGF-beta1, 2, 3 mRNA expression levels were measured by reverse transcriptase polymerase chain reaction (RT-PCR) and RNA PCR-Southern blotting.
RESULTS
Fibroblast cells were proliferated by high dose AGE (2,200nmol, p<0.05, statistical analysis by ANOVA). RAGE mRNA expression was confirmed in bladder fibroblast in vitro. Procollagen type I mRNA expression levels were increased by high dose AGEs (1,100, 2,200nmol FFI) treatment at 24 hours and 48 hours. TGF-beta2 mRNA expression levels were increased dose dependently at 72 hours. TGF-beta3 mRNA expression levels were suppressed in high dose AGEs treated cells from 24 hours.
CONCLUSIONS
AGEs increase procollagen type I mRNA, TGF-beta2, TGF-beta3 mRNA expression and proliferative activity of the bladder fibroblast cells in vitro. These results suggest that AGEs can direct toxic effects to bladder mesenchymal cell proper.

Keyword

Glycation; Diabetes mellitus; Urinary bladder; Rat; Fibroblast

MeSH Terms

Animals
Collagen Type I
Collagen Type III
Diabetes Mellitus
Electrophoresis, Polyacrylamide Gel
Enzyme-Linked Immunosorbent Assay
Fibroblasts*
Glycosylation End Products, Advanced*
Rage
Rats*
Reverse Transcriptase Polymerase Chain Reaction
RNA
RNA, Messenger
Serum Albumin
Spectrophotometry
Transforming Growth Factor beta1
Transforming Growth Factor beta2
Transforming Growth Factor beta3
Urinary Bladder*
Collagen Type I
Collagen Type III
Glycosylation End Products, Advanced
RNA
RNA, Messenger
Serum Albumin
Transforming Growth Factor beta1
Transforming Growth Factor beta2
Transforming Growth Factor beta3