The ability of neonatal and maternal erythrocytes to produce reactive oxygen species in response to oxidative stress

The ability of neonatal and maternal erythrocytes to produce reactive oxygen species (ROS) in response to oxidative stress was investigated using the chemiluminescence probe Cypridina luciferin analogue. The chemiluminescence probe, based on 2-methyl-6-[p-methoxyphenyl]-3,7-dihydro-imidazo[1,2-alpha]pyrazin-3-one (MCLA), is highly specific and sensitive to superoxide anion (O(2)(-)) and singlet oxygen (1O(2)). Blood from 11 mothers who experienced no complications and their healthy full-term newborns was collected and heparinized. MCLA was put into the washed erythrocytes suspension, and phenylhydrazine (PH) was added to cause oxidative stress. Chemiluminescence was measured using an Argus 50 image processing system. It was found that erythrocytes in neonatal blood had a 2.0-fold greater maximum chemiluminescence than did those in maternal blood. There was no change in the emission after the addition of NaN(3), but there was complete suppression with superoxide dismutase (SOD), demonstrating that these were O(2)(-).The present results demonstrated that neonatal erythrocytes produce about twice as much O(2)(-) as do adult erythrocytes in response to oxidative stress. With this method, it is very simple to measure the amount of O(2)(-) produced in erythrocytes and the rate at which they are produced. This method may therefore be very useful for determining the effects of antioxidants.