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[Effect of insulin on VEGF expression in bovine retinal microvascular endothelial cells exposed to normal or high glucose].

Haixiang Wu, Xin Xia, Kun Liu, Zhi Jie Zheng, Dongqing Zhu, Xun Xu, Qing Gu

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Abstract

OBJECTIVE: To investigate the effect of insulin or (and) high glucose on the vascular endothelial growth factor (VEGF) expression in bovine retinal microvascular endothelial cells (BREC). METHODS: It was a experimental study. BREC were cultured with selective culture media. Passaged cells were cultured in normal (5 mmol/L) or high glucose (30 mmol/L) medium for 3 days. Equimolar mannitol was supplemented for osmotic controls. Cells were serum-deprived overnight in Dulbecco's modified Eagle's medium containing 0.2% w/v bovine serum albumin and then incubated in the absence or presence of 100 nmol/L insulin for 24 hrs. Cells were harvested and the expression of VEGF mRNA was analyzed by Real-time PCR. VEGF protein level was determined by HUVEC proliferation assay, immunofluorescence and western-blot RESULTS: Insulin or (and) high glucose significantly increased VEGF mRNA (F = 5.67, 9.04; P <0.05) and protein (F = 5.50, 5.57; P <0.05) expression. However, the combined effect of insulin and high glucose is weak in contrast to insulin or high glucose alone. CONCLUSIONS: High glucose attenuated insulin-induced VEGF expression. Therefore, VEGF may be not the major factor that lead to transient worsening of diabetic retinopathy after acute insulin therapy in diabetes.

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What this paper is about

OBJECTIVE: To investigate the effect of insulin or (and) high glucose on the vascular endothelial growth factor (VEGF) expression in bovine retinal microvascular endothelial cells (BREC). METHODS: It was a experimental study. BREC were cultured with selective culture media. Passaged cells were cultured in normal (5 mmol/L) or high glucose (30 mmol/L) medium for 3 days. Equimolar mannitol was supplemented for osmotic controls. Cells were serum-deprived overnight in Dulbecco's modified Eagle's medium containing 0.2% w/v bovine serum albumin and then incubated in the absence or presence of 100 nmol/L insulin for 24 hrs. Cells were harvested and the expression of VEGF mRNA was analyzed by Real-time PCR. VEGF protein level was determined by HUVEC proliferation assay, immunofluorescence and western-blot RESULTS: Insulin or (and) high glucose significantly increased VEGF mRNA (F = 5.67, 9.04; P <0.05) and protein (F = 5.50, 5.57; P <0.05) expression. However, the combined effect of insulin and high glucose is weak in contrast to insulin or high glucose alone. CONCLUSIONS: High glucose attenuated insulin-induced VEGF expression. Therefore, VEGF may be not the major factor that lead to transient worsening of diabetic retinopathy after acute insulin therapy in diabetes.

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Available abstract

OBJECTIVE: To investigate the effect of insulin or (and) high glucose on the vascular endothelial growth factor (VEGF) expression in bovine retinal microvascular endothelial cells (BREC). METHODS: It was a experimental study. BREC were cultured with selective culture media. Passaged cells were cultured in normal (5 mmol/L) or high glucose (30 mmol/L) medium for 3 days. Equimolar mannitol was supplemented for osmotic controls. Cells were serum-deprived overnight in Dulbecco's modified Eagle's medium containing 0.2% w/v bovine serum albumin and then incubated in the absence or presence of 100 nmol/L insulin for 24 hrs. Cells were harvested and the expression of VEGF mRNA was analyzed by Real-time PCR. VEGF protein level was determined by HUVEC proliferation assay, immunofluorescence and western-blot RESULTS: Insulin or (and) high glucose significantly increased VEGF mRNA (F = 5.67, 9.04; P <0.05) and protein (F = 5.50, 5.57; P <0.05) expression. However, the combined effect of insulin and high glucose is weak in contrast to insulin or high glucose alone. CONCLUSIONS: High glucose attenuated insulin-induced VEGF expression. Therefore, VEGF may be not the major factor that lead to transient worsening of diabetic retinopathy after acute insulin therapy in diabetes.

Key concepts: Internal medicine, Endocrinology, Insulin, L-Glucose, Diabetic retinopathy, Vascular endothelial growth factor, Diabetes mellitus, Bovine serum albumin

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[Effect of insulin on VEGF expression in bovine retinal microvascular endothelial cells exposed to normal or high glucose]. — Research Paper | ScholarLens