The Plasma Membrane is Involved in the Visible Light–Tissue Interaction

Abstract

Objective: The aim of the present study was to determine whether the plasma membrane is also involved in the light–tissue interaction because of the nicotinamide adenine dinucleotide phosphate (NADPH) oxidase electron chain, which can serve as a photosensitizer. Background data: It has been suggested that the mechanism of photobiostimulation involves light-induced low levels of reactive oxygen species (ROS) that serve as signal transduction messengers. Production of ROS following visible-light irradiation was verified by the electron paramagnetic resonance (EPR) spin-trapping technique, and the mitochondrial cytochromes were suggested as the main cellular target for visible-light absorption. Methods: Isolated sperm membranes were illuminated with visible light and the increase in oxygen radical production was measured using the EPR spin-trapping technique coupled with the probe 5,5-dimethyl-1-pyrroline-N-oxide (DMPO). A broadband visible light source (400–800 nm) at 40–130 mW/cm² with appropriate filters provided the illumination. In order to determine whether the light-induced ROS production is a result of a photo-accelerated electron transfer in the enzyme-catalyzed reaction with oxygen in the plasma membrane, or resulted from a photochemical reaction of the chromophores alone with oxygen, denatured membranes were irradiated as well. Results: Visible-light–induced oxyradicals were detected in isolated sperm membranes. Blue light was found to be more effective than red. Illuminated denatured membranes produced the same amount of ROS as non-denatured membranes. Conclusions: Visible-light illumination, especially in the blue region, increases ROS levels in isolated plasma membranes. The mechanism of ROS formation is probably a photochemical reaction of the membranal chromophhores, for example, cytochromes or flavins with oxygen, and not an enzyme-catalyzed photochemical reaction.