The Study On Immuno-Enhancing Activity And Mechanism Of Nine Chinese Herbal Medicine Ingredients

In order to screen the novel immunostimulant from Chinese Herbal Medicine ingredients (CHMIs), we extracted nine bio-active ingredients from five kinds of CHMs, and investigated on their mechanism with the technology of immunity. Effects of the nine ingredients on the specific and non-specific factors in immunology, immunocytes and immuno-molecular in vivo and in vitro were studied, and their immno-enhancing activity in clinic were compared. Then several ingredients of these CHMIs were selected as the novel immunostimulant.Section 1. Nine bio-active ingredients were extracted from five kinds of CHMs, including astragaluspolysaccharide (APS), propolis polysaccharide (PPS), epimedium polysaccharide (EPS), Chinese angelica polysaccharide (CAPS), isatis root polysaccharide(IRPS), astragalus flavone(AF), propolis flavone(PF), epimedium flavone(EF), astragaloside ( AS). And net content (%) of every ingredients are 77. 28, 82.48, 82.94, 66.38, 51.41, 46.92, 38.37, 1.39, 72.88 respectively.Section 2. To detect the immune activities and the target cells of the CHMIs, the stimulating activities of these CHMIs on peripheral blood and splenic lymphocytes in mice were compared. All CHMIs could be arranged as EPS, APS, PF, GS, CAPS, PPS, IRPS, EF, AS, according to which they stimulated lymphocytes to proliferate in vivo. In this study, we also selected the best stimulating dose in vivo of every CHMI by the difference of their stimulating activities, and there are 20mg/Kg .d in polysaccharides, 5 mg/Kg.d in f lavones, Ginsenoside and Astrogaloside.Section 3. The stimulating activities of nine CHMIs on peripheral blood and spleenic lymphocytes cultured in vitro were compared. All CHMIs could be made a turn about EPS, APS, GS, PF, EF, CAPS, AS, according to which they stimulated lymphocytes to proliferate in vitro, and which is similar to that of in vivo. In this study, we also selected the best stimulating dose in vitro of every CHMIs by the difference of their stimulating activity, and there are 20ug/ml in polysaccharides, 5 ug/ml in flavones, Ginsenoside and Astrogaloside.Section 4. In this study, the IL-2mRNA levels in normal mice T cells stimulated by nine CHMIs was determined using a reverse transcription and semi-quantification polymerase chain reaction. The results showed that all CHMIs would be arranged in turn by the level of IL-2mRNA they stimulated like this: APS>EPS>PF>CAPS>AS>PPS>IRPS>GS>BF.Section 5. The acticity of natural killer(NK) cell cytotoxic and murine peritoneal macrophages(PM) in mice were determined after the mice were injected the nine CHMIs, respectively. The results showed that APS, EPS, CAPS, PF, EF and GS enhanced PM activity significantly, furthermore, the six and PPS improved NK cells cytotoxic activity markedly.Section 6. 50 mice were divided into 10 groups and immunized APS, EPS, PPS, CAPS, IRPS, PF, EF, AS and GS, respectively, then the production of spleen ASC were observed for all groups 4 days after immunization by SRBC, in the way of quantitative haemolysis of SRBC. It was found that all CHMIs could be made in a turn of GS>PF>APS>EF>EPS>AS.Section 7. Nine CHMIs in the proper dose and (or) LPS added into spleen lymphocytes cultured in vitro, and the IgG levels were measured 5 days after with sandwich ELISA. In this study, all CHMIs would be arranged in turn by the level of IgG they stimulated like this: GS>APS, PF>EPS>CAPS>EF.Section 8. Healthy young rabbits were injected the nine CHMIs three times in 3 days before they were immunized with the vaccine of RVHD, respectively, and HI employed to determine the level of antibody immunity. It was found that they would be arranged like GS, EPS, APS, PF, CAPS and EF according to their enhancing activity.All of these results suggest that these CHMIs have different stimulating activities and different target cells in mice. APS, GS, EPS, PF were screened clinically for the animal immune enhancers, and CAPS, EF, AS second.