FGF2(bFGF) glycation induced by TGF-beta1 in human lens epithelial cells.

Park, Hyun Kyung; Lee, Kwang Won; Wee, Won Ryang; Lee, Jin Hak; Joo, Choun Ki

Original Article

Journal of the Korean Ophthalmological Society 2001;42(10):1483-1489.
Published online October 1, 2001.

FGF2(bFGF) glycation induced by TGF-beta1 in human lens epithelial cells.

Hyun Kyung Park, Kwang Won Lee, Won Ryang Wee, Jin Hak Lee, Choun Ki Joo

¹Department of Ophthalmology and Visual science, College of Medicine, The Catholic university of Korea.
²Department of Ophthalmology and Visual science, , Seoul National University Hospital, Seoul.

수정체 상피세포에서 TGF-β1 에 의한 bFGF ( FGF2 ) 의 glycation

박현경(Hyun Kyung Park),이광원(Kwang Won Lee),위원량(Won Ryang Wee),주천기(Choun Ki Joo),이진학(Jin Hak Lee)

Abstract

PURPOSE
TGF-beta1 is a multifunctional cytokine that transforms epithelial cell to fibroblast like cell and its mechanism is diverse. In this experiment, we investigated whether TGF-beta1 produces ROS and induces intracellular proteins glycation, expecially bFGF glycation. MATERIALS AND METHODS: We cultured lens epithelial cell in MEM supplemented with 20% fetal bovine serum(FBS) at 37 degreesC. We examined the extracellular ROS after treatment with 1 ng/ml of TGF-beta1 and the intracellular ROS by confocal microscopy. Additionally, we investigated the glycation of total protein and bFGF by immunoblot analysis and proliferation assay by MTT. RESULTS: We found that both intracellualr and extracellular ROS were increased 1.2~1.5 fold after treatment with 1 ng/ml of TGF-beta1, and peaked at 6~8 hours. Intracellular ROS was continuously increased in 30minutes through 45 minutes and decreased after 60 minutes under confocal microscopic examination. Total advanced glycation end product (AGE) was increased by 1.5 fold. Changes of bFGF in lens epithelial cell by TGF-beta1were examined by Immunoprecipitation and western blot. Glycation of bFGF induced by 1ng/ml TGF-beta1 for 1 week was increased 4 fold higher than non-treated cell. Also B3 cells treated with glycated-bFGF showed about 50% decrease in proliferation effect. CONCLUSIONS: In this study, we found that TGF-beta1 produced both intracellular and extracellular ROSs inducing glycation of bFGF in lens epithelial cell B3. In addition, glycated intracellular bFGF resulted in loss of its original effect on proliferation.

Key Words: bFGF;Glycation;Lens epithelial cell;TGF-beta1