Amino Acid Mixture Acts as a Potent VEGF Lowering Agent in CHO-K1 Cells Exposed to High Glucose

Abstract

Background: Though the role of amino acids in Diabetes Mellitus is controversial, the beneficial effect of amino acids in Diabetes Mellitus has been reported based on its anti-glycating property and insulin potentiating effects. In the current study, we evaluated the ROS generation and VEGF expression in CHO-K1 cells induced by high glucose concentration. The effect of amino acids treatment was studied under this condition to evaluate the VEGF lowering effect.

Method: CHO-K1 cells were treated various concentration of glucose (7 mmol, 17 mmol and 27 mmol) with and without free amino acids (5 mmol) or the amino acids mixture (AAM). Intracellular reactive oxygen species (ROS) was estimated by fluorescein dye (DCFDA), nitric oxide (NO) by Griess reaction, hydrogen peroxide (H₂O₂) by fluorimetry using Amplex red dye, super oxide dismutase (SOD) by spectrophotometry and VEGF by immunoblotting.

Results: High glucose condition significantly induced the expression of VEGF and this was reduced significantly by AAM treatment (p = 0.004). AAM also significantly decreased the cellular levels of ROS, NO, H₂O₂ as well as the SOD activity in CHO-K1 cells exposed to high glucose condition (p <0.05).

Conclusion: The present study identified AAM as a potential VEGF lowering agent that intervenes at the level of oxidative stress in high glucose conditions as evaluated in CHO-K1 cells.

Keywords: Amino acids; ROS; SOD; VEGF.