The protective effect of Arginase inhibitor on retinal microvascular endothelial cells in high glucose cultures

Abstract

Objective To investigate the effect of arginase (Arg) inhibitor N-ω-Hydroxy-L nor-Arginine (nor-NOHA) on high glucose cultured rhesus macaque retinal vascular endothelial cell line (RF/6A) in vitro. Methods The RF/6A cells were divided into the following 4 groups: normal control group (5.0 mmol/L of glucose, group A), high glucose group (25.0 mmol/L, group B), high glucose with 125 mg/L nor-NOHA group (group C), and high glucose with 1% DMSO group (group D). The proliferation, migration ability and angiogenic ability of RF/6A cells were measured by Methyl thiazolyl tetrazolium (MTT), transwell chamber and tube assay respectively. The express of Arg I, eNOS, iNOS mRNA of RF/6A cells were measured by real-time polymerase chain reaction (RT-PCR), Enzyme-linked immuno sorbent assay (ELISA) was used to detect the expression of NO and interleukine (IL)-1b of RF/6A cells. Results The proliferation, migration, and tube formation ability of group A (t=2.367, 5.633, 7.045; P<0.05) and group C (t=5.260, 6.952, 8.875; P<0.05) were significantly higher than group B. RT-PCR results showed the Arg I and iNOS expression in group B was higher than that in group A (t=6.836, 3.342; P<0.05) and group C (t=4.904, 7.192; P<0.05). The eNOS expression in group B was lower than that in group A and group C (t=4.165, 6.594; P<0.05). ELISA results showed NO expression in group B was lower than that in group A and group C (t=4.925, 5.368; P<0.05). IL-1b expression in group B was higher than that in group A and group C (t=5.032, 7.792; P<0.05). Conclusions Nor-NOHA has a protective effect on cultured RF/6A cells in vitro and can enhance its proliferation, migration and tube formation. The mechanism may be inhibiting the oxidative stress by balancing the expression of Arg/NOS. Key words: Arginase/antagonists & inhibitors; Retinal Vessels/cytology; Endothelial cells/physiology; Cells, cultured