RESUMO
Objective To investigate the effect of extracts from rabbit skins inflamed by Viccinia virus vaccine (analgecine) on the proliferation of human cancer cells and on the cytokine secretion in mouse spleen lymphocytes in vitro. Methods Five human tumor cell lines, HepG2, LM3, H460, A549 and HeLa were used and the effect of analgecine (1.63, 0.815, 0.326, 0.163 and0.0815 U/ml) on the cell proliferation was evaluated by the CCK-8 assay. The mouse spleen was isolated aseptically, and the spleen lymphocyte suspension was prepared and cultured with PRMI-1640 medium containing 10％ fetal bovine serum (FBS). For detection of the cytokine IL-2, IFN-γ, IL-4 and IL-12 level, the stimulant concanavalin A (ConA) or lipoplysaccharide (LPS) was added into the lymphocyte suspension, and the lymphocytes were cultured under the presence of analgecine at the final concentration of 0.815, 0.163 and 0.0815 U/ml for 24 hours. Then, the level of the cytokines in the supernatant was detected by the ELISA kit. On the other hand, the effect of supernatant of the spleen lymphocyte cultures under the presence of analgecine at 0.815 U/ml on the proliferation HepG2 cells was also evaluated by the CCK-8 assay. The CCK-8 assay was performed after cultivation of the HepG2 cells in the whole supernatant or in its dilution with fresh medium for 24 hours. Results Analgecine showed a dose-dependent inhibitory effect on the five tested cancer cell lines, with the inhibition rate of 58.95％, 55.08％, 57.28％, 45.80％ and 48.18％ at the 1.63 U/ml on the HepG2, LM3, H460, A549 and HeLa cells, respectively. Compared with the control group, the secretion of IL-2, IFN-γ and IL-4 was significantly increased in the 0.163 and 0.815 U/ml analgecine groups (P＜0.01), while the secretion level of IL-12 was increased in the 0.0815, 0.163 and 0.815 U/ml analgecine groups (P＜0.01). The supernatant of the mouse spleen lymphocyte cultures under the presence of0.163 U/ml analgecine could inhibit the HepG2 cell proliferation in a dose-dependent manner, and the inhibitory effect of the whole supernatant was significantly stronger than the effect of the same concentration analgecine 0.163 U/ml (P＜0.01). Conclusion Analgecine could inhibit the cell proliferation of the tested five human cancer cell lines, increased the secretion of IL-2, IFN-γ, IL-4 and IL-12 cytokines in mouse spleen lymphocytes, all in vitro, and its effect on the cytokine secretion may be related to the inhibitory effect on the human cancer cell proliferation.
RESUMO
Objective To investigate the antiplatelet effects of candidate drug W1 when combined with omeprazole,respec?tively.Methods The experimental rats were randomly divided into 5 groups:control group,clopidogrel(10 mg/kg)group,clopido?grel(10 mg/kg)+omeprazole(80 mg/kg)group,W1(3 mg/kg)group,and W1(3 mg/kg)+omeprazole(80 mg/kg)group.Four hours after oral administration of the drugs,the rats were measured for their platelet aggregation rate;Western blot was used to deter?mine the protein expressions of P2Y12 receptor,P-Akt and P-Erk.Results For the platelet aggregation rate,compared with the con?trol group,the 4 groups decreased significantly(P<0.01);the platelet aggregation rate in the clopidogrel + omeprazole group in?creased significantly than that in the clopidogrel group(42% and 20.4%,respectively,P<0.01);the platelet aggregoction rate (30.9%)in W1+omeprazole group was significantly higher(P<0.01)than that in the W1 group(20.5%),which was lower than that in the clopidogrel+omeprazole group(P<0.01).For the protein expression detected by the western blotting,there were no signif?icant changes in the expression of P2Y12 receptor on the platelet surface,in the clopidogrel or W1 group in comparison with the clopi?dogrel+omeprazole or W1+omeprazole group,respectively,while the phosphorylation level of Akt and Erk was up-regulated in the clopidogrel+omeprazole or W1+omeprazole group compared with the clopidogrel or W1 group,and the up-ragulatory effect of omepra?zole was weaker in the W1+omeprazole group than that in the clopidogrel+omeprazole group. Conclusion Combined use of omepra?zole could inhibit the antiplatelet activities of clopidogrel or W1,with the inhibitory effect weaker in W1 group than in clopidogrel group,suggesting that the risk for the combination of omeprazole with W1 is likely less than that for the combination with clopidogrel.