Effects of Deer Antler Extract on Serum IGF-I, Bone Growth and Splenocyte Proliferation in Growing Rats

Although it has traditionally known that deer antler and medicinal herbs extract contain some functional components for health promotion, the nutritional significance remains to be elucidated. This study examined the efficacy of deer antler extract (DA) , medicinal herbs extract (MH) and their mixture (DAMH) on serum IGF-I, bone growth with growing rats in vivo and splenocyte proliferation with spleen cells in vitro. Three week-old young female rats (Sprague-Dawley) were divided into 4 groups and then fed basal diet (AIN-93G) or experimental diets containing DA, MH, DAMH, respectively, for 7 weeks. We collected blood, liver, kidney, spleen, femur and tibia from rats. There was no significant difference in weight gain, but food intake increased in DA- and MH-fed groups. There were no signs of liver and kidney damage in the DA, MH and DAMH-fed groups compared to basal diet group. In femur and tibia, wet weights, breaking forces and bone minerals (Ca, Mg and Zn) were significantly higher in the DA-fed group than in the other groups. Serum alkaline phosphatase (ALP) , bone-specific alkaline phosphatase (BALP) activities were significantly lower in the DA, MH, DAMH-fed groups than in basal diet group. Also, serum insulin-like growth factor-I (IGF-I) concentrations were significantly increased in DA-fed group compared to the other groups. Therefore DA was shown to have an activity of bone growth promotion by increasing the IGF-I, a major bone growth factor. The deer antler extract showed an enhanced immune action on the primary cultured-cells from spleen of rats, representing that splenocytes were proliferated by lipopolysaccharide (LPS) , but not by concanavalin A (Con A) . These results indicate that deer antler extract has beneficial effects on bone growth via IGF-I and on splenocyte activation.

Production of IL-15 and lts Functional Study in Mouse Splenocyte Activation / 대한면역학회지

After the synthesis of IL-15 cDNA from the total RNA of mouse spleen, it was inserted into the prokaryotic expression vector, pRseta, and eukaryotic expression vector, pcDNA3.0, respectively. Subsequently, the insertion of gene and open reading frame were confirmed by sequencing of each plasmid, respectively. Using pRseta- IL-15 plasmid, the recombinant IL-15 protein was induced by IPTG under BL21 (DE 3) host cells and recombinant IL-15 was expressed at 14.5 KDa with time. Then, IL- 15 was separated by His-tag affinity chromatography and analyzed by SDS-PAGE to yield soluble IL-15 at 14.5 KDa as monomer and 29.0 KDa as dimer. In order to inspect the function and contribution of IL-15, the in vitro experiment was established using mononuclear cells separated from the mouse spleen. After 48h exposure of PHA to mouse splenocyte and 24h treatment with recombinant IL-15, the effects of cytokine inductions inspected against IL-2, IL-6, IL-10, IL-12, IFN-r, and GM-CSF. The results showed that comparing with the control, IL-6 increased, IL-2, IL-12 and IFN-r increased and similar, and GM-CSF decreased. In addition, the direct injection of pcDNA3.0-IL-15 plasmid into mice gave the similar results to in vitro studies. Namely, IL-6 and IL-12 increased, and IL-2, IFN-r and GM-CSF were similar or decreased. IL-10 was not induced in in vitro and in vivo experiments. These results suggested that the IL-15 induce the splenocyte activation and can be an important factor in proliferation and fuction recovery of weakened T-cell.