Experimental Study On Effects Of Antiangiogenesis With Taohongsiwu Decoction

Background: Taohongsiwu decoction(THSWD)has been identified as a potent prescription to treat all kinds of diseases with blood stasis syndrome in Traditional Chinese Medicine. Also,it was belived that a high correlation between angiogenesis diseases and blood stasis syndrome. So far, however,THSWD has not been studied on its effects of antiangiogenesis.Objectives: 1. To screen antiangiogenesis drugs in THSWD and its separated prescriptions decoction with Chick Chorioallantoic Membrane (CAM)assay in vivo. 2. To investigate the effects of THSWD on the proliferation of human umbilical vein vascular endothelial cells ECV304 in vitio. 3. To evaluate the antiangiogenesis efficiency of THSWD in B16 melanoma model and to investigate the potential mechanism of its effects. Methods: 1. CAM model was bulit to screen antiangiogenesis drugs. Fertilized white chicken eggs under constant humidity at 37℃. On day 7, the experimental groups were treated with THSWD or its separated prescription decoction. Protamine injection was used as the positive control. On day 10, photographed in ovo with camera . The amount of blood vessel (N)were counted with a stereomicroscope at 10×. The blood vessel's area (VA)and the ratio between the vessel area and the CAM area( AA)were analyzed by computer image analysis systerm. 2. MTT assay was used to investigate the effect of THSWD on proliferation of ECV304. The experimental groups were treated respectively with THSWD 10mg/ml,25mg/ml,50mg/ml,100mg/ml,200mg/ml . Protamine injection was used as the positive control. 3. C57BL/6J mice bearing B16 melanoma were used in this study. The experimental groups were treated respectively with THSWD 0.25g/kg/day, 0.5g/kg/day,1g/kg/day and THSWD 1g/kg/day + cyclophosphamide(CTX)25mg/kg. The positive control was treated with CTX 50mg/kg . Immunohistochemistry assay was used to observe the expression of KDR/flk-1 and MVD. Results: 1. The promoting blood flow prescriptions and herbs of THSWD were obviously inhibited the N,VA and AA of the CAM than saline control group (p<0.05) ,on the contrary, the invigorationg blood prescriptions and herbs of THSWD obviously stimulated the N of the CAM than saline control group (p<0.05). 2. ECV304 were treated directly with THSWD 100mg/ml,200mg/ml, the absorbance (A ) was significantly lower than that in saline control group (P<0.05) . 3,THSWD 0.5g/kg/d,1.0g/kg/d and THSWD + CTX group regressed significantly the volume ,weight of tumor in bearing-B16 mice compared to saline group (P<0.01) . In addition ,KDR/flk-1 and MVD were deduced in experimental groups. Conclusions: 1. The promoting blood flow prescriptions and herbs of THSWD have potent activity to inhibit the CAM agiogenisis,whereas, the invigorationg blood prescriptions and herbs of THSWD can obviously stimulate the CAM agiogenisis. 2. THSWD has activity against the proliferation of ECV304. 3. THSWD can inhibit the growth of B16 melanoma in C57BL/6J mice, and can inhibit the expression of KDR/flk-1 and MVD, which suggest that THSWD has the effects of antiagiogenisis might responsible for the antitumor effect. Innovation and signification: It is discovered that the promoting blood flow prescriptions and herbs in THSWD can inhibit the CAM agiogenisis ,on the contrary, the invigorationg blood prescriptions and herbs in THSWD have angiostimulatory effects on CAM. THSWD has activity against the proliferation of ECV304. THSWD can inhibit the growth of B16 melanoma in C57BL/6J mice, and can inhibit the expression of KDR/flk-1 and MVD, which suggest that THSWD has the effects of antiagiogenisis might responsible for the antitumor effect. Antiagiogenisis may be one of the possible pharmacological mechanism which was responsible for THSWD treating blood stasis syndrome angiogenesis diseases.