Inhibitory effect of Rumex Japonicus Houtt on the porphyrin photooxidative reaction.

Rumex Japonicus Houtt (RJH), a plant indigenous to Okinawa, Japan, has been used traditionally by the local people for treatment of acute and chronic cutaneous diseases; however, its pharmacological effect is not clearly understood. To investigate its active function, we examined the antioxidative effect of RJH on the hematoporphyrin-induced photooxidative reaction. We measured intracellular reactive oxygen species (ROS) in cultured transformed human vascular endothelial cells (ECV-304) by flow cytometry, lipid peroxide (LPO) in erythrocyte ghosts by spectrofluorometry, and hemolysis by spectrophotometry. Results showed the generation of intracellular ROS in ECV-304, LPO production in erythrocyte ghosts, and hemolysis after visible light irradiation in the presence of hematoporphyrin. In the RJH root extract treated group, generation of intracellular ROS, LPO production, and hemolysis were inhibited significantly. It has been reported that the RJH root contains emodin and chrysophanic acid. In our study, three fractions were separated from the RJH root extract by thin layer chromatography (TLC). We found that fraction 1 (near the origin) showed a strong inhibitory effect on LPO levels; however, fraction 2 (equal to emodin) and fraction 3 (equal to chrysophanic acid) showed no inhibitory effect on LPO levels. Our experiments verified that RJH has an antioxidative effect against photohemolysis and supports our hypothesis that RJH may have potential as a protective agent against oxidative stress associated with some diseases such as porphyrias.