Melatonin protects against oxidized low-density lipoprotein-induced inhibition of nitric oxide production in human umbilical artery.

Abstract

We evaluated the antioxidative effect of melatonin on the oxidized low-density lipoprotein (LDL)-induced impairment of nitric oxide (NO) production in human umbilical artery, which may be the prime cause of endothelial dysfunction in pre-eclampsia. Umbilical artery sections with intact endothelium were obtained from healthy pregnant women who were delivered between 37 and 40 wk of gestation. The production of NO in the umbilical arteries was stimulated by adding L-arginine followed by incubation for 60 min. NO concentrations were estimated by measuring nitrite ions (NO(2) using high-performance liquid chromatography. LDL was oxidized by incubation with 5 microM CuSO(4) at 37 degrees C for 4 hr, followed by dialysis at 4 degrees C for 24 hr. Prior to the addition of L-arginine, the segments were treated with native or oxidized LDL (0, 50, 100, 200, 400 microg/mL), or were pre-treated with either mannitol (50 mM) or melatonin (20, 100, 500 microM) before adding oxidized LDL. Changes in L-arginine-induced NO(2)(-) production were expressed as a percentage of NO(2)(-) production at the end of pre-incubation. Treatment with oxidized LDL significantly reduced L-arginine-induced NO(2)(-) production (P<0.05), while NO(2)(-) production did not change by incubation with native LDL. Pre-treatment with melatonin significantly increased NO(2)(-) production that had been decreased by oxidized LDL (P<0.05). Similarly, pre-treatment with mannitol reversed the oxidized LDL-induced reduction in NO(2)(-) production (P<0.05). These results indicate that melatonin protects against oxidized LDL-induced inhibition of NO production in the endothelium of human umbilical arteries, most likely through its ability to scavenge hydroxyl radicals.