Investigative ophthalmology & visual science · 2002
Effect of high glucose on fibronectin expression and cell proliferation in trabecular meshwork cells.
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Sato Tsuyoshi, Roy Sayon
Аннотация
PURPOSE: Increased fibronectin accumulation in the trabecular meshwork of glaucomatous eyes may contribute to the resistance of aqueous outflow and the development of primary open-angle glaucoma (POAG). Because the glucose level is increased in the aqueous humor of patients with diabetes, this study was conducted to determine whether a high-glucose condition alters fibronectin expression and contributes to cell loss in trabecular meshwork.
METHODS: The fibronectin mRNA level was determined using RT-PCR in bovine trabecular meshwork cells grown in normal (5 mM) or high (30 mM)-glucose medium for 7 days, and cell counts were measured during this period. Distribution and the relative amount of fibronectin protein were determined in these cells by immunofluorescence microscopy and Western blot analysis.
RESULTS: Fibronectin mRNA level in cells grown in high-glucose medium was significantly upregulated two- to threefold compared with cells grown in normal medium (P < 0.05). In cells grown in high-glucose medium, fibronectin immunofluorescence was more intense, and the relative amount of fibronectin protein was significantly increased (131% +/- 15% of control, P < 0.05) compared with the amount in cells grown in normal medium. A moderate decrease in cell number was observed in cells grown in high-glucose medium (78% +/- 7% of control, P < 0.05)
CONCLUSIONS: These findings indicate that a high glucose level in aqueous humor of patients with diabetes may increase fibronectin syntheses and accumulation in trabecular meshwork and accelerate the depletion of trabecular meshwork cells, a characteristic feature of the outflow system in POAG. The striking similarity between high glucose-induced alterations in trabecular meshwork cells and those of vascular endothelial cells may represent a common biochemical link in the pathogenesis of POAG and diabetic microangiopathy.
METHODS: The fibronectin mRNA level was determined using RT-PCR in bovine trabecular meshwork cells grown in normal (5 mM) or high (30 mM)-glucose medium for 7 days, and cell counts were measured during this period. Distribution and the relative amount of fibronectin protein were determined in these cells by immunofluorescence microscopy and Western blot analysis.
RESULTS: Fibronectin mRNA level in cells grown in high-glucose medium was significantly upregulated two- to threefold compared with cells grown in normal medium (P < 0.05). In cells grown in high-glucose medium, fibronectin immunofluorescence was more intense, and the relative amount of fibronectin protein was significantly increased (131% +/- 15% of control, P < 0.05) compared with the amount in cells grown in normal medium. A moderate decrease in cell number was observed in cells grown in high-glucose medium (78% +/- 7% of control, P < 0.05)
CONCLUSIONS: These findings indicate that a high glucose level in aqueous humor of patients with diabetes may increase fibronectin syntheses and accumulation in trabecular meshwork and accelerate the depletion of trabecular meshwork cells, a characteristic feature of the outflow system in POAG. The striking similarity between high glucose-induced alterations in trabecular meshwork cells and those of vascular endothelial cells may represent a common biochemical link in the pathogenesis of POAG and diabetic microangiopathy.