Experimental Study Of The Rhizoma Paridis Combined With Oxaliplatin Inhibiting Angiogenesis

[Background]Angiogenesis means forming of neovessels from preexisting blood vessels in the form of sprout.Doctor Folkman have raised the hypothesis that tumor growth depends on the growth of neovessels, because the author observed that the domain of human body that lacking of blood vessels has less opportunity with malignancy.Neovessels may provide the tumor growth with oxygen and nutrient substance and induce the Neoplasm Metastasis more quickly.Inhibiting the formation of neovessels may inhibite the growth of tumor.When some chemotherapeutics are used in low-dose and high-frequency model,they could inhibit the growth of tumor by inhibitting the angiogenesis of tissue of tumor selectively.In this therapy model,the target point of chemotherapeutics is vascular endothelial cell but not tumor cell.Low-dose chemotherapy is also named antiangiogenic chemotherapy.Now,the antiangiogenesis are so expensive that the economy of China can't afford it.The traditional Chinese drug is a treasurehouse of China.So,searching for the antiangiogenesis in traditional chinese drug which would be excellent in quality and reasonable in price is especially important. The traditional chinese drug have abundant experience.For the past few years,along with lucubrate of antiangiogenic - therapy,more and more traditional Chinese medicine and it's active component may have antiangiogenic affect.The traditional Chinese medicine is cheaper and has less toxic and side-effect.So, antiangiogenic effect of traditional Chinese drugs become warm spots of the dom research.The antitumor effect of the Rhizoma Paridis and it's chemical composition have been proved in vitro and in vivo.We need to lucubrate antiangiogenic affect of the Rhizoma Paridis and the combination of chemotherapeutics.[Objective] To investigate the posssible effect of the alcohol extracts of Rhizoma Paridis,oxaliplatin,and the combination of the two drugs on anti-angiogenesis in vitro and in vivo.[Methods]We used MTT method to observe the influence on proliferation of the alcohol extracts of Rhizoma Paridis,oxaliplatin and the combination of two drugs on human umbilical vein endothelial cells(HUVEC) or intestinal cancer cell line LOVO.The influence of drugs on HUVEC migration was evaluated by Transwell model.We also observed the morphologic change of formation of anastomosing tubes on triaxial Matrigel model.Annexinâ…¤fluorescein stain and cell cycle test kit and flow cytometry were used to detect the development of apoptosis and cell cycle of HUVEC after treated with drugs.Chick chorioallantoic membrane(CAM) model was used to observe the effect of anti-angiogenesis of the drugs in vivo.[Results]The survival rate of the alcohol extracts of Rhizoma Paridis on HUVEC is dose-dependent within the dose of 7.5μg/mL～240μg/mL and the 50%inhibiting concentration(IC50) of the alcohol extracts of Rhizoma Paridis on HUVEC is 36.83μg/ml.The survival rate of the alcohol extracts of Rhizoma Paridis on LOVO is also dose-dependent and the IC50 of the alcohol extracts of Rhizoma Paridis on LOVO is 267.54μg/ml.The survival rate of oxaliplatin on HUVEC is dose-dependent within the dose of 0.5μg/mL～16μg/mL and the 50%inhibiting concentration(IC₅₀) of oxaliplatin on HUVEC is 3.52μg/ml.The survival rate of oxaliplatin on LOVO is dose-dependent and the IC₅₀ of oxaliplatin on LOVO is 8.20μg/ml.The alcohol extracts of Rhizoma Paridis and oxaliplatin has cytotoxic variability between HUVEC and LOVO.The combination of the alcohol extracts of Rhizoma Paridis and oxaliplatin have a synergistic interaction or subadditive interaction.The alcohol extracts of Rhizoma Paridis within the dose of 7.5μg/mL～60μg/mL and oxaliplatin within the dose of 0.5μg/mL～4μg/mL inhibited migration of HUVEC in vitro.The inhibition ratios of the alcohol extracts of Rhizoma Paridis on migration are 14.80%,25.73%,35.57%,70.12%at the dose of 7.5μg/ml,15μg/ml,30μg/ml,60μg/ml.The inhibition ratios of the oxaliplatin on migration are 17.45%,31.23%,45.45%,59.04%at the dose of 0.5μg/ml,1μg/ml,2μg/ml,4μg/ml. The number of the migrated HUVEC can be obviously reduced by the two drugs.The combination of the two drugs have a synergistic interaction or subadditive interaction.The alcohol extracts of Rhizoma Paridis and oxaliplatin can also destroy lumen structure which is closely knitted capillary-like plexus and the combination of the two drugs may have a synergistic interaction.The alcohol extracts of Rhizoma Paridis and oxaliplatin may induce apoptosis of the HUVEC cell cycle after 48h.The combination of the two drugs may have a synergistic interaction.The alcohol extracts of Rhizoma Paridis and oxaliplatin may block the HUVEC cell cycle and arrest at G0-G1 phase after 24h.The combination of the two drugs may have a synergistic interaction.The alcohol extracts of Rhizoma Paridis and oxaliplatin suppress angiogenesis of CAM in vivo and reduces the number of neovessels.The combination of the two drugs have a subadditive interaction. [Conclusion]This experiment shows that the alcohol extracts of Rhizoma Paridis and oxaliplatin may have anti-angiogenic effect in vitro and in vivo,and the combination of the two drugs may have a synergistic interaction or subadditive interaction.