STUDY OF IMMUNOMODULATORY EFFECT OF WATER EXTRACTS OF FIVE-FLOWER REMEDY AND ITS COMPOSITIONS ON MACROPHAGES

Abstract

Five-flower remedy is a traditional Thai remedy used for promoting health and reducing fever. It consists of the flowers of Jasminum sambac, Nelumbo nucifera, Mesua ferrea, Mimusops elengi and Mammea siamensis, in equal weight. This study aimed to investigate effects of the water extracts of the remedy and its components on macrophage J774A.1 cells. Effects of these extracts on production of nitric oxide (NO) were determine by Griess reaction. The extracts of the remedy, J. sambac and N. nucifera, all at 6.25-100 µg/ml, potently increased NO production. The extracts of M. ferrea, M. elengi and M. siamensis had less stimulating activities than the extracts mentioned before. Effects of all extracts on mRNA expression of some markers of M1 and M2 activated macrophages were also evaluated by RT- PCR. The extracts of the remedy and J. sambac at 6.25 – 25 µg/ml, the extract of N.nucifera at 25 – 100 µg/ml, and the extracts of M. ferrea, M. elengi and M. siamensis at 50 – 200 µg/ml were used. These extracts up-regulated mRNA expression of TNF-α, IL-1, and inducible nitric oxide synthase (iNOS) which are markers of M1 macrophages and IL-10 which is a marker of M2 macrophages. The extracts of the remedy, J. sambac, N. nucifera, and M. ferrea stimulated the expression of a M1 macrophage marker IL-6 and Arg-1, a marker of M2 macrophages. All six extracts activated macrophage J774A.1 cells in similar manner to LPS which is known to activate M1 macrophages. The extracts of remedy and J. sambac had the highest activating activity, followed by the extracts N. nucifera and M. ferrea, respectively. The extracts of M. elengi and M. siamensis demonstrated the lowest stimulating effects. These results suggest that the water extracts of five-flower remedy and its four components may stimulate macrophage J774A.1 cells to M1 activated phenotype with different potencies. It is predicted that all components synergistically activate macrophages when they are combined as the remedy. Activation of macrophages may be a part of pharmacological properties of this five-flower remedy for health promotion.