61A-31 Nitric oxide production by polysaccharide isolated from ginseng marc in peritoneal macrophage

J. K. HWANG¹, K. Na², E. M. Choi, J. M. Yun, and T. S. Lim. (1) Bioproducts Research Center, Yonsei University, B138A Yonsei Engineering Complex, 134 Shinchon-dong Sudaemun-ku, Seoul, 120-749, South Korea, (2) Dept. of Biomaterials Science and Engineering, Yonsei University, B118CA Yonsei Engineering Complex, 134 Shinchon-dong Sudaemun-ku, Seoul, 120-749, South Korea

Many plant polysaccharides have been demonstrated to stimulate macrophage activity and hence the immune system. Nitric oxide (NO) production in macrophage activities was determined as biomarkers for immunostimulation. Activated macrophage-derived NO was known anti-microbial infection, tumor cell killing and protection against lethal endotoxemia. We hypothesize that the macrophage-activating polysaccharides may remain in by-products (ginseng marc; GM) after ethanol extract process of Panax ginseng C. A. Mayer. We identified NO productions of the polysaccharides isolated from GM in peritoneal macrophages under in vitro and in vivo conditions. Our objective was to demonstrate immunostimulating activity of peritoneal macrophage by ginseng marc polysaccharide (GMP) exposure in mouse. For isolation of GMP, hydolysates by commercial polysaccharidase were treated with 80% isopropanol (v/v) and insoluble fraction was recovered. Sugar composition of GMP was determined by high-performance anion exchange chromatography (HPAEC) and average molecular weight by gel permeation chromatography (GPC). Female Balb/c mice were used at the age of 7-8 weeks and received 50 งท/งธ body weight/day through oral administration in vivo. Cell number of macrophage was adjusted at 1ขฅ10⁶cell/งข. NO production was calculated by the calibration curve for NO₂^- determination using Griess reaction. The level of NO₂^- reflects NO synthesis. GMP isolated from isopropanol-insoluble fraction of ginseng marc hydrolysate possessed 29.1% arabinose, 22.7% galactose, 14.3% rhamnose and 33.9% uronic acid and its average molecular weight was 320000 D. In vitro the effect of NO production was observed dose-related increase in 50 and 100 งถ/งข and in vivo GMP alone could not induce the NO production, but it had synergy effect on 30% NO production when the macrophage were triggered with 20 งถ/งข lipopolysaccharide (LPS). These results suggest that GMP effectively increases the macrophage function in mice, and thus show high potent in the utilization of by-product.