The Preliminary Study Of Anti-angiogenesis Of Sanguinarine

ObjectiveMacleaya is kind of chinese traditional medicine,its main components are sanguinarine(SA)and chelerythrine.The recent years studies show that Macleaya is found with anti-tumor effect.According to the previous experimental studies,we through the anti-angiogenesis direction to study the anti-tumor effect of sanguinarine,which may provides the theory basis for the Chinese medicine of Macleaya or sanguinarine.Methods and Results1.In vitro experimentMTT method was used to study the inhibition rate of CT26.WT cell and HUVECs cell treated with sanguinarine in different concentration and time.The IC50 of each cell was calculated to evaluate the anti-proliferative effect of SA.The results showed that SA could play anti-proliferative effect on both cells.The IC50 is between 1.08 ?mol/L and 1.30 ?mol/L.CoCl2 was used to build up the model of hypoxia.MTT method was used to observe the best concentration and time of CoCl2,The results showed that the best conditions to establish the hypoxia model were:CT26.WT cell worked with the 100 ?mol/L CoCl2 for 6h;HUVECs cell worked with the 150 ?mol/L CoCl2 for 4h.Beside,MTT method was used to study the inhibition rate of CT26.WT cell and HUVECs cell treated with SA in different concentration and time under the condition of hypoxia.Results showed that SA still played anti-proliferative effect on both cells. cell scratch test was used to study the migration of SA under the condition of hypoxia.Results showed that SA could prevent migration of CT26.WT cell and HUVECs cell.2.In vivo experimentTumor-bearing mice model was established by injecting of CT26.WT cells.We took the inhibition rate and relative tumor volume as the index of antitumor effect.The results indicated SA significantly inhibited the growth of CT26.WT tumor-bearing mice.The tumor inhibition rate results showed that the inhibition rates of 5mg/kg SA group,10mg/kg SA group,20mg/kg SA group, 40mg/kg SA group mice were 19.11%,19.96%,36.76%,49.37%.Expression of VEGF and CD31 from tumor tissues were tested by immunohistochemistry.The experimental results showed that the VEGF and CD31 expression in tumor tissue of SA drug group were lower.Further research on the mechanism of anti-angiogenesis was the detection of PI3K,AKT,HIF-1 ?,VEGF protein expression which were from the HIF-1 signaling pathway by western blotting.The experimental results suggest that the anti-angiogenesis effect of SA may be related to HIF-1 pathwayConclusionThe antitumor activity of SA which was extracted from Macleaya was confirmed by in vivo and in vitro experiments.SA might through inhibition angiogenesis play its anti tumor effect.But its role in inhibiting angiogenesis was something relation to do with HIF-1 signaling pathway.