ACTIVITY OF LONGEVITY REMEDY WATER EXTRACT ON MURINE MACROPHAGES

Abstract

Longevity remedy is a Thai traditional remedy which has been used to promote and restore health for longevity. This remedy consists of six herbs including Cyperus rotundus root, Piper nigrum fruit, Streblus aspera seed, Tinospora crispa stem, Diospyros rhodocalyx bark, and Albizia procera bark, in the same proportion. This study aimed to investigate the stimulatory activities of the water extracts of longevity remedy and its components on macrophage J774A.1 cells. Effects of the water extracts on nitric oxide (NO) production were measured by Griess reaction assay after 24 h of treatment. The water extracts of longevity remedy, C. rotundus and P. nigrum at 1.56–25 μg/ml significantly increased NO production in a concentration-dependent manner. The extracts of S. aspera, T. crispa and D. rhodocalyx at 12.5–100 μg/ml demonstrated lower stimulatory effects on NO production than the extracts above. The extract of A. procera did not stimulate NO production. All water extracts at all concentrations used in this study had no effects on J774A.1 cell viability. Effects of all water extracts on mRNA expression of several markers of macrophage activation were also evaluated by RT-PCR after 4 and 24 h of treatment. The water extracts of longevity remedy, C. rotundus and P. nigrum at 3.13, 6.25 and 12.5 μg/ml as well as the extracts of S. aspera, T. crispa and D. rhodocalyx at 25, 50 and 100 μg/ml were used. These extracts increased mRNA expression of IL-1β, IL-6, TNF-α, and iNOS which are often used as the markers of M1 activated macrophages. The extracts also up-regulated mRNA expression of IL-10 and argenase-1 which are the markers of M2 activated macrophages. The extract of A. procera had no effect on mRNA expression of all markers. The results from this study demonstrated that the water extracts of the longevity remedy and almost all of its components can activate macrophage J774A.1 cells with different potencies. The extracts of the remedy and C. rotundus had the highest potency, followed by the extract of P. nigrum. The other three extracts including S. aspera, T. crispa and D. rhodocalyx had lower potencies on macrophage activation than the extracts of the remedy, C. rotundus, and P. nigrum. This immunostimulatory activity may be a part of beneficial effects of the use of this longevity remedy.