| Two different active components have been isolated from Kudzuvine Root(Radix puerariae), one was the ethanol extraction portion- total flavone (TFP) and another was the water extraction portion-total polysaccharide(TPP). For exploring the biological efficacy and the mechanism of Bi-direction modulation effects of different Pueraria lobata active components, a series studies have been carried out by using chemiluminescence> Biochip> Cell culture^ flow cytometry technology and animal experiments.The results showed as following:1. Extraction and indentifcation of total flavone(TFP) and total polysaccharide of Puerariae(TPP): ethanol soluble portion and water soluble portion were saperated simultaneously from Puerariae by using a improved chemical technology. They were proved to be TFP and TPP by using qualitative as well as quantitative analysis according to thestandards of Chinese herbal medicine identification.2. Different concentration of two active componentshad been studied by using PMA stimulated immunol cells initiated chemiluminescent system, to evaluate the inhibitory or stimulation effects of TFP and TPP on respiratory burst emitted ROS. The results indicated that TPP could enhance chemiluminescence in concentration dependently whereas TFP could inhibit the chemiluminescence significantly.3. H202 initiated PCi2 cell cytotoxicity had been usedas experimental model to investigate the biological effects of TFP and TPP on the cultured PCi2 cells. The activity of cultured PCi2 cell was monitored by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assays, the antioxidant activity of the culture media was monitored by hypoxanthine/xanthine oxidase initiated chemiluminescent system and the nitritewas measured by Griess reagents. The results showed thatTFP could protect the PC12 cells from the oxidative damages of H2O2 at 1-100 ng/ml while TPP could enhance the oxidative damages on the cells at 1-1000 ng/ml.4. To investigate the preventive effect of TFP and TPP on asthmatic model rats, 64 rats were randomly divided into eight groups. Control group(Group A): the rats was raised in normal condition; asthmatic instant stimulated group(Group B); TFP control group(Group C): 300mg/kg TFP only was administrated 28 for days TFP protection group(Group D): As Group B, 300mg/kg TFP was administrated before the rats were inhaled with 1% OA aerosol. TPP control group(Group E): 900mg/kg TPP was administrated for 28 days. TPP intervention group(Group F): As Group B, 900mg/kg TPP was administrated before the rats were inhaled with 1% OA aerosol. L-arginine control group(Group G): 100mg/kg TPP wasadministrated for 28 days. L-arginine intervention group(Group H): As Group B, 100mg/kg L-arginine was administrated before the rats were inhaled with 1% OA aerosol. T cells subsets in spleen were examined and the changes of lung tissues in rat asthmatic model were observed using histopathologic methods. Control rats did not have eosinophils in their submucosal of airways, whereas eosinophils as well as lymphocytes were apparently increased in the airways of Group B, and the percentages of CD4+ as well as the ratio of CD4+/CD8+ were significantly higher in group B than Control group; The application of TFP in Group D not only inhibited the increase of EOS, IL-4> IL-5> CD4\ CD4*VCD8+ , but also depress the airway inflammation, the differences were statistically significant. Results showed that TFP can prevent allergic asthma by inhibit the increasing of EOSx IL-4, IL-5^ CD4\ CD4+/CD8+; On the contrary,TPP can enhance the immunity of normal rats, increasing CD4+> CD4+/CD8+, but have no effects on hyperactive immune rats;5. Effects of TFP and TPP on blood gas of asthmaticrats: PaO2 . pH of asthmatic rats decreased sharply , while PaCO2 increased (p<0.01), which showed that the asthmatic rats was in serious anoxic situation, In group D, PaO2> pH and PaCO2 have no significant difference compared to Control group; in group F, the value of PaO2 ^ pH and PaCO2 were si... |
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