Effects of Advanced Glycation End Products on Rat Bladder Fibroblast.
- Author:
In Ho CHANG
1
;
Yong Wan SEONG
;
Sun Chul MYUNG
;
Young Sun KIM
Author Information
1. Department of Urology, Chung-Ang University College of Medicine, Seoul, Korea. caucih@dreamwiz.com
- Publication Type:In Vitro ; Original Article
- Keywords:
Glycation;
Diabetes mellitus;
Urinary bladder;
Rat;
Fibroblast
- MeSH:
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*
- From:Korean Journal of Urology
2001;42(11):1204-1210
- CountryRepublic of Korea
- Language:Korean
-
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.
