| Objective: The effects of different concentrations of tea polysaccharide mice respectively B16F10 cell tumor treatment and prevention effects, discuss tea polysaccharides inhibit B16F10 melanoma growth and its mechanism.Methods: 1. The experiment conducted in two parts, namely therapeutic and preventive experimental test. Therapeutic experimental groups in B16F10 cells replicate mouse tumor model based on the screening and 48 were randomized to the control group NS, TPS high dose group(800mg / kg.d), TPS dose group(400mg/ kg.d) and TPS low dose group(200mg / kg.d), n = 12; and preventive approach to experimental groups: from healthy C57 BL /6 female mice were 48, were random Ly divided into four groups, the group name with preventive test.2. Treatment of experimental tumor model mouse cell: Right footpath inoculated subcutaneously concentration to 1×106 B16F10 tumor cells 0.2m L establish a mouse model, and the morphological and HE staining methods such proof B16F10 cell tumor modeling success.3.Administration:treatment experiment against different tumor-bearing mice fed tea polysaccharide solvent 28 days, while the control group received NS normal saline; preventive test is normal mice fed tea polysaccharide solvent first 28 days, the first 29 days of each mouse, respectively, of 2×105 B16F10 tumor cells attack and continue treatment until the 56 th day, to observe and record the prophylactic effect in mice tumor cells in the body.4. Detection indicators: mouse spleen weight was weighed to calculate spleen index; mouse tumor mass weighing weight inhibition rate was calculated; the use of anti-NK cell activity assay; determination of NO by nitrate reductase content; ELISA was used to detect serum levels IL-4, IFN-γ content; real-time PCR to detect apoptotic gene bcl-2m RNA, baxm RNA RT level; Western blot assay withered death protein bcl-2, bax expression levels.Results: 1. The results of treatment1.1 B16F10 cell tumor replication model:The tumor model mice will be varying degrees of weight loss, slow, loss of appetite, unresponsive, and even some alopecia.After HE staining cells of mice will find disordered different size and shape, with atypia; and normal tissue cell size, morphology basically the same, indicating that tumor model successfully established.1.2 TPS quality of treatment and control group differences were statistically significant(P <0.05) before and after the experimental group tumor-bearing mice tumor drugs, in both dose groups were the most significant(P <0.01).1.3 NK cell activity was significantly different in TPS treatment group from that of the controls when the optimal effector-target ratio was set at 20:1(P<0.05), and the activity reached 39.06% in mice administered with medium dosage(P<0.01).1.4 The level of IFN-γ in serum of TPS group were higher than the control group(P <0.05), and the middle dose group of TPS was highest(P <0.01).while IL-4 expression in serum level showed no obvious change in the each groups.1.5 The bcl-2m RNA expression of tumor cells was down-regulation,and baxm RNA expression was up-regulation in TPS group,and had a significant difference compared with the control group(P <0.05), and the middle dose group of TPS had a most significant difference(P <0.01).Apoptotic protein bcl-2, bax and NS of the group had a significant difference compared with the control group(P <0.05), bcl-2 synthesis was inhibited, while bax showed increased,the middle dose had a most significant difference(P <0.01).2.Prevention of experimental results2.1 TPS preventive experimental groups appear to slow the proliferation of B16F10 cell tumors, tumor mass is reduced by the NS control group, the difference was statistically significant(P <0.05), of which the most obvious dose group(P <0.01).2.2 TPS serum NO prevention in the experimental group were higher than NS control group(P <0.05), while TPS dose group was significantly higher than the control group(P <0.01).2.3 Prevention of experimental polysaccharide in experimental group effect target ratio of 100:1, 20:1 and 10:1,the killing activity of NK cells was more than NS control group.and when in the effect target ratio of 20:1,the killing activity of NK cells had a significant difference(P <0.05), the middle dose had a most significant difference(P <0.01).2.4 The level of IFN-γ in serum of prevention of experimental polysaccharide in experimental group were higher than the NS control group(P <0.05), and the middle dose group was highest(P <0.01). IL-4 expression in serum level had a significant difference between prevention of experimental polysaccharide in experimental group and NS control group(P <0.05).2.5 The bcl-2m RNA and baxm RNA expression had a significant difference between prevention of experimental polysaccharide in experimental group and NS control group(P <0.05),and the bcl-2m RNA expression of tumor cells was down-regulation,and baxm RNA expression was up-regulation.The middle dose group had a most significant difference(P <0.01).Prevention of experimental polysaccharide in experimental group compare with NS control group,tumor tissue extracts of bcl-2 protein bands smaller average gray value, and bax protein bands average gray value is larger, of which the most obvious effect dose group(P <0.01).Conclusion:1. Tea Polysaccharides on B16F10 mice inhibit the proliferation of tumor cells, showing a better therapeutic and prophylactic effect, in which the TPS dose group(400mg /kg.d) effect is more obvious;2. Tea Polysaccharides on B16F10 cell tumor of mice has a significant preventive effect, which may be related to enhanced immunity in mice Function and promote tumor cell apoptosis related. |
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