[Diphenylene iodonium and apocynin reduce the translocation and level of p47phox in PBMCs of premature infants to inhibit reactive oxygen species production].

To observe the effects of NADPH oxidase inhibitor diphenylene iodonium (DPI) and apocynin on the generation of reactive oxygen species (ROS) induced by p47phox and the mechanism of p47phox-induced ROS production under hyperoxic conditions. Peripheral blood mononuclear cells (PBMCs) were isolated from the peripheral blood (2 mL) of premature infants of less than 32 weeks without oxygen uptake. The isolated cells were divided into four groups, control group, hyperoxia group, hyperoxia and DPI group, hyperoxia and apocynin group. The control group was cultured in incubator with 50 mL/L CO(2) at 37°, and the other groups were cultured in 950 mL/L O(2) and 50 mL/L CO(2) mixed gas. After 48 hours, ROS was detected by Mitosox Red staining under a confocal laser scanning microscope; malondialdehyde (MDA) was measured by thiobarbituric acid colorimetry; the location and translocation rate of p47phox was observed by immunofluorescence staining; the level of p47phox protein was tested by Western blotting. Compared with the hyperoxia group, the remaining three groups showed significantly decreased ROS and MDA levels and reduced translocation rate and level of p47phox. Compared with the control group, both the hyperoxia and DPI group and the hyperoxia and apocynin group were not significantly different in the above indexes. DPI and apocynin can reduce hyperoxia-induced ROS production by decreasing the translocation and level of p47phox.