| AIDS caused by the human immunodeficiency virus (HIV), has lead to huge disaster to human health. The possibility of highly active anti-retroviral therapy (HAATR) was confirmed in AIDS treatment, however it could not eliminate the virus, and causes a variety of side effects. In recent years, the incidence of HSV also increased significantly, which shows great threat to human being. ACV exhibits ideal performance in treating HSV infection, but trigger resistance drug strains easily. At present, all commonly used antiviral agents show side effect and cause drug resistance. These defeats can be avoided by using natural products as agents which exhibit multi-target in anti-virus and show lower toxicity. Therefore, it is amazing field to search the new anti-viral preparation around the abundant plant resources.STG seeds were crushed and dried in the sun. Then petroleum ether was used for degreasing to get the STG powder. STG agglutinin was separated, which can agglutinate red blood cell. Subsequent procedure was taken to evaluate the total activity and specific activity. Pure STG agglutinin was obtained through continuous manipulation:1)extracting with saline at 4℃,2)precipitating with the ammonium sulfate,3)dialyzing with distilled water,4)Sephadex G-25 gel filtration,5) HITrap Q Sepharose XL ionic exchange chromatographic. The purity of STG agglutinin was analyzed with the SDS-PAGE, which showed a single band with relative molecular mass of 63 400.STG agglutinin could agglutinate rabbit erythrocytes and the lowest agglutination concentration is 38μg/ml. It also agglutinates chicken, pig, human blood group A, B, AB and O. The agglutination of rabbit erythrocytes could be inhibited by Glucose, D-fructose, D-mannose, D-arabinose in different level. The influence of temperature and pH to agglutination activity was analyzed. STG agglutinin show strong tolerance to temperature and pH because the agglutination activity was not affected when the temperature is below 60℃and pH is between 5 and 8. In addition, the effect of metal ion, organic solvent and denaturant on agglutinating activity were investigated. The results indicate that Ca2+, Mg2+ affect the agglutinating activity; Phenol and isopropyl alcohol inhibit agglutinating activity; the inhibition of urea is reversible, but SDS is irreversible.The anti-HIV-1ⅢB and anti-HSV-2 effect of the STG agglutinin was analyzed in this study. According to the drug screening method in vitro, the C8166 and Vero were choosed as the target cell respectively to examine the anti-HIV and anti-HSV activity and anti-viral mechanism of STG agglutinin. The STG agglutinin has anti-HIV-1ⅢB activity, IC50 is 49.8μg/mL, SI is 36. Base on the result of time-addition assay, fusion assay, STG agglutinin may affect the prophase of HIV life cycle. On the other hand, STG agglutinin shows the activity of anti-HSV-2, IC50 is 26.28μg/mL, SI is 45. The anti-HSV mechanism experiment indicated that the STG agglutinin inhibits the HSV-2 by affecting different HSV-2 life-cycles. |
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