Study On Pharmacological Effects Of Polysaccharides From Artemisia Annual

Polysaccharide from Artenisia annua L.(HQG), the principal active component of southwood, which is belong to the genus Artemisia (Compositae), has rarely been studied neither at home or abroad the moment. The aim of this study was to investigate the pharmacological function of HQG on self-extracting HQG in vitro and in vivo, so as to provide theoretical basis for the clinical application and the development of new functions.Method details and results are showed as below:(1) HQG was extracted with papain and water, combining alcohol sedimenting techinique. The content of polysaccharide was determined by anthrone-sulfuric acid colorimetry, and the physicochemical property was analyzed by infrared spectrometric analysis instrument (ISAI).(2) The immunosuppression model induced by cyclophosphamide (CTX) in mice were established by intraperioneal injection at four different doses of25mg/kg,50mg/kg,100mg/kg or150mg/kg respectively. Counts of white blood cells (WBC) and the LZM levels in serum of the mice were determined and the thymus gland index (TI) and spleen index(SI)were calculated. The their morphological changes were observed. Five different doses of HQG (25mg/kg,50mg/kg,100mg/kg,150mg/kg or300mg/kg) were given to the normal mice by intraperioneal injection.The levels of IL-1β, IL-2, IL-6, IFN-y and LZM in the murine serum were determined.(3) HQG toxigenicity was observed by acute toxicity test (ATT) in mice, and LD50of HQG was determined by intraperitoneal injection according to improved Karber’s method.(4) The mouse ear acute inflammatory model was established by xylene, HQG at doses of25mg/kg,50mg/kg, or100mg/kg was given to the animal before. The ears of each animal were weighted, followed by the calculations of ear swelling rates and their repression rates.(5) In vitro induction of RAW264.7macrophages and porcine spleen cells were performed by Porcine circovirus type2(PCV2), and the values of NO, ROS, GSH, GSSG, MDA and T-SOD activity were determined to screen the optimum concentration of PCV2for establishment of the oxidative stress models. The RAW264.7macrophages and porcine spleen cells were treated at different concentration of HQG (25μg/mL,50μg/mL,100μg/mL or200μg/mL), then the values of NO, ROS, GSH, GSSG, MDA and T-SOD activity were determined.Results:(1) The content of saccharinity of HQG was24.74%, and HQG contained neither starch or amino acid. The spectroscopy proved that the structure of carbohydrate chain was galactopyranose type with an acetyl group.(2) significant efficiency of HQG in both anti-muscle atrophy function and lysozyme reduction induced by CTX had been observed in all4concentrations (P<0.05). Splenic nodule enlargement, splenocyte counts increase and cell proliferations were observed after treatments in murine thymus. The values of IL-6significantly increased at25mg/kg of HQG treatment, the increases both IL-1β level and IFN-y level has been observed at50mg/kg treatment.The IL-2level was increased by100mg/kg HQG while LZM levels were increased by4concentrations (P<0.05).(3) The LD50value of HQG was301.51mg/kg, and the95%confidence interval were ranged from270.80mg/kg to332.21mg/kg.(4)The mouse ear swelling rates were all significantly reduced with HQG treatment at dose of50mg/kg or100mg/kg (P<0.05); Compared to dexamethasone, the suppression rates were all significantly enhanced with HQG treatment at100mg/kg (P<0.05).(5) The optimum concentration of PCV2for RAW264.7oxidation stress model was10-2, while10-1was fit for pig splenocyte oxidation stress model. HQG at four concentrations in our study significantly reduced values of ROS during the induction of RAW264.7by PCV2infection in vitro.HQG treatment at50μg/mL or200μg/mL significantly reduced GSSG levsls, HQG at dose of25μg/mL50μg/mL or200μg/mL significantly reduced MDA level, while HQG at200μg/mL increased activities of T-SOD activity (P<0.05). In the other hand, HQG at50μg/mL or100μg/mL significantly reduced production of NO and ROS levels during the induction of porcine splenocytes by PCV2in vitro. HQG at50μg/mL 100μg/mL or200μg/mL of significantly reduced GSSG level, while25μg/mL of HQG increased the level of GSH (P<0.05).In conclusion:Polysaccharide of artenisia annua L. is a kind of low toxic substance and had immunoregulatory, anti-inflammatory and antioxidant functions.