The Screening And Mechanism Study Of Phyllanthus Urinaria Which Have Anti-angiogenic Effect

Part OneScreen Traditional Chinese Medicine Which Have Anti-angiogenic EffectObjectives To screen Traditional Chinese Medicine composition of 10 alcohol extracts of Traditional Chinese Medicine, Phyllanthus urinaria, obviously difference cyotoxic on human lung cancer A549 cells, human hepatoma HepG2 cells, human colon carcinoma Lovo cells, human gastric carcinoma MKN-28 cells, human breast cancer MCF-7 cells and human umbilical vein endothelial cell (HUVEC) of human umbilical vein in vitro.Methods The MTT assay had been developed for quantitative evaluation of the inhibitory effect of 10 alcohol extracts of Traditional Chinese Medicine on proliferation of human cell lines of lung cancer,hepatoma,colon carcinoma,gastric carcinoma,breast cancer and HUVEC of human umbilical vein. Growth curve assay was used to detect the variability cell toxic action to tumour cell and HUVEC which was cultured in vitro by 10 alcohol extracts of Traditional Chinese Medicine. Screened the most sensitive cell to the Traditional Chinese Medicine and the Traditional Chinese Medicine which had different cytotoxic effect on HUVEC.Results The most sensitive cell to the medicine was human colon carcinoma Lovo cells. Alcohol extract of Phyllanthus urinaria could obviously inhibit the proliferation of HUVEC in deuto-cytotoxic concentration(P=0.001).Effect was most obviously at 48h, and had fine dose-dependent relationship.Conclusion Alcohol extract of Phyllanthus urinaria in 10-320μg/ml,had obviously variability cell toxic action to tumour cell and HUVEC, the inhibitory effect to HUVEC proliferation was most obviously at 48h, and had fine dose-dependent relationship.Part TwoThe Effect of Alcohol Extract of Phyllanthus Urinaria on The Biological Behaviour of Human Umbilical Vein Endothelial Cell in VitroObjectives To investigate the effect of alcohol extract of Phyllanthus urinaria on human umbilical vein endothelial cell migration and tube formation. Methods Transwell cabin and out-body canaliculization test were used to observe the action of alcohol extract of Phyllanthus urinaria on the migrational and vaso-formed ability of HUVEC.Results The alcohol extract of Phyllanthu s urinaria at the doses of 10-80μg/ml inhibited migration of HUVEC in vitro.The number of the migrated HUVEC can be obviously reduced, alcohol extract of Phyllanthus urinaria (10-80μg/ml) inhibited the migration of HUVECs(14.76%-69.85%)after medicine effected for 12 hours.While the alcohol extract of Phyllanthus urinaria in 10-80μg/ml effected at 24 hours to the HUVEC, the number of the tubule was reduced and the lumen structure which is closely knitted capillary-like plexus was destroyed. Conclusion The alcohol extract of Phyllanthus urinaria could actively suppress angiogenesis in vitro and the mechanism may related to block the migration and canaliculization of the HUVEC.Part ThreeResearch The Mechanism of Alcohol Extract of Phyllanthus Urinaria to Anti-angiogenesis of The Chick Embryo in VivoObjectives To investigate the effect of anti-angiogenic of the alcohol extract of Phyllan thus urinaria in vivo. Methods Chickchorio allantoic membrane (CAM) model was used to observe the effect of anti-angiogenic of the alcohol extract of Phyllanthus urinaria. Results Surrounding the carrying agent of CAM the blood vessel was suppressed.There was inhibitory action on the CAM angiogenesis when the concentration of the alcohol extract of Phyllanthus urinaria is between 10-80μg/ml,inhibited the angiogenesis of CAM (21.60%-63.56%) after added medicine for 72h.Conclusion The alcohol extract of Phyllanthus urinaria could inhibit CAM angiogenesis in vivo.