The effects and mechanism of high-salt medium on eNOS expression in human umbilical vein endothelial cell line

Objective: To investigate the effects of high-salt medium on the expression of endothelial nitric oxide synthase (eNOS) and related mechanisms in EA. Hy926 cell line. Methods: EA. hy926 cells were treated 48 h with different concentrations of high-salt medium; 137 mmol/L was selected as the target concentration intervention in further experiment. The cells were divided into four groups which received intervention for 48 h: DMEM high-glucose medium group (sodium chloride concentration of 109 mmol/L), high-salt medium (sodium chloride concentration of 137 mmol/L), high-salt medium (sodium chloride concentration of 137 mmol/L)+RNA interference, and hypertonic control group (mannitol concentration of 56 mmol/L). High-performance liquid chromatography was used to assay ADMA concentration in cell supernatant; real-time quantitative PCR and Western blot were used to assay the expressions and activities of PRMT-1, RhoA, ROCK and eNOS. Results: ADMA concentration, PRMT-1 expression, the mRNA expression of RhoA and ROCK activity increased significantly whereas the protein expression of eNOS significantly reduced in the high-salt group compared with those in the two control groups. High-salt medium + RNA interference restored the above biochemical indicators and upregulated the protein expression of eNOS significantly. Conclusion: High-salt medium induces ADMA-mediated RhoA/Rock-1 pathway activation and suppresses eNOS generation.