| AIM: Resercheres are more and more interested in polysaccharide sulphates for its anti-virus effects, especially for its anti-HIV activity. Angelica sinensis is a tonic traditional Chinese medicine. In our previous studies, Angelica polysaccharide sulphates (APS) were synthesized and were demonstrated having immunomodulative effects. This study was to investigate the therapeutic effect of APS on a murine AIDS model and its cooperation with anti-AIDS drug Combivir (Azidothymidine+Lamivudine), and further to discuss the possibility of APS and its combination with anti-virus drug on AIDS therapy. The initial mechanism of APS was also investigated.L6565 murine leukemia virus (L6565 MuLV) to set up a murine AIDS model. 4 weeks later, some of them were inspected to verify the model. Then APS(ip) and/or Combivir(ig) were given for 30 days. The anti-virus effects of APS and/or Combivir were examined as follows: RT-PCR method was used to measure viral load in plasma and the electrophoresed bands of PCR products were quantitated by image analyzing; the percentage of CD4+ and CD8+ lymphacytes in blood cells was estimated by flowcytometric method and the ratio of CD4+/CD8+ was calculated; haematological examination was also carried out as well as marrow karyote and index of immune organs were weighed and calculated. The 14 days acute toxicity of Combivir was also detected when used independently or combined with APS. APS was given intraperitoneally for 5 days followed by a LD50 of Combivir intragastricly, then the mouse auto-activity was measured, 14 days survival was recorded, and haematological examination was carried out.The Hela cells were cultured conventionally, then APS was added and cultured together in different concentration for 24h followed by a oxidation injury: treatment with 0.1 mmol·L-1 H2O2 for 1h, or irradiation with UV in 25 cm distance for 4 min. The anti-oxidation injury effects of APS were detected as follows: cell viability was measured by MTT assay; colorimetric analysis was used to determine SOD activity, GSH and MDA level in cytoplasm.in mouse plasma was not detected in control. While in the model, the viral load was at high level (3365±181), the percentage of CD4+ lymphocyte (22.3±4.1 vs 32.8 + 5.7 in control), the number of RBC and marrow karyote, and thymus index were significantly decreased, while the number of WBC and spleen index were remarkably increased (P<0.05 or P<0.01 vs control).After administration of APS for 30 days, the level of viral load in plasma in model was even higher (4165 + 198). The percentage of CD4+ lymphacyte (18.2±4.9) , the ratio of CD4+ /CD8+ (1.4±0.3), the number of RBC and marrow karyote, and thymus index were significantly decreased in the model mice, while the number of WBC and spleen index were remarkable increased compared with control(P<0.05 or P<0.01). Compared with model, at the dose of 10 and 30 mg-kg-1, the viral load in plasma (3328±103 and 3394± 171), WBC and spleen index were significantly decreased by APS, while the percentage of CD4+ lymphacyte (52.6 ± 5.8 and 57.0 ± 5.5) , the ratio of CD4+ /CD8+ (3.6 + 0.6 and 3.4+0.5) , the number of RBC and marrow karyote were remarkably increased by APS (P<0.05 or P<0.01) . Some of the items even recovered to normal level.Compared with model, the plasma viral load of APS+Combivir groups were significantly decreased at the dose of 10+90 mg·kg-1 and 30+90 mg·kg-1 respectively (2035+243 and 1763±103) (P<0.01) , even though that of APS+Combivir at 30+90 mg·kg-1 group was further decreased. As far as the percentage of CD4+T lymphocyte, it was remarkably increased by Combivir at 90 mg·kg-1 (27.0 + 2.1)compared with model, and increased more by APS+Combivir at the dose of 10+90 mg·kg-1 and 30+90 mg·kg-1 respectively (38.1 ±9.7 and 44.8±6.5) (P<0.01 vs Combivir group) . Although combination of APS and Combivir improved pathological changes of the model, such as decreases of RBC, marrow karyote, thymus index and increases of WBC and spleen index, only RBC and marrow karyote improvements were significant compared with Combivir group. Furthermore, APS not only increased the survival when mice were given LD50 of Combivir but also improved RBC, HB, PLT reduction, which were the side effects of Combivir.Treatment with H2O2 or UV radiation significantly decreased cell viability, GSH and SOD activity in cytoplasm, while increased MDA in cytoplasm. At the range of 0.3-100 mg·ml-1, APS markedly increased cell viability, GSH and SOD activity, while decreased MDA in a dose dependent manner (P<0.05 or P<0.01).CONCLUSION Some symptoms like that of AIDS patients were shown in mice infected with L6565 murine leukemia virus, which suggested that this murine AIDS model could be used to evaluate the effect of anti-AIDS drugs. The results of treatment of model mice with APS and/or Combivir suggest that APS has not only anti-virus effects, but also immunomodulative effect, and APS has cooperation effect with Combivir. Since APS can decrease virus load and increase CD4+T lymphacytes, it may be a potential anti-AIDS drug candidate. APS has anti-oxidation injury effect, which may be one of its anti-AIDS... |
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