| BackgroundVascular anomaly (VA) is a group of congenital diseases including hemangiomas and vascular malformations, which incidence rate is 0.3%~1% in the young. Based on the hemodynamic and biological characteristics, VA is consisted of infantile hemangiomas, capillary malformations (CMs), venous malformations (VMs), lymphatic malformations and arteriovenous malformations (AVMs). Vascular malformations, especially extensive and deep complicated malformations, would destroy patient's appearance and function due to untimely and improper treatment. Resection of lesions and/or vessel deligation might bring about severe local hemodynamic disorder and worsen the state of illness. Therefore, the theraputics of vascular malformations are most challenging to clinical scenarios.Pingyangmycin (PYM, bleomycin AS), is an antitumour antibiotic. Since the 1980', PYM has been used for treating infantile lymphangioma and hemangioma and the effects were satisfying. Then it was used in the area of interventional radiology by domestic radiologists from 1990'. Several benigh vascular diseases such as hysteromyoma and huge hemangioma of liver, were cured well after arterial embolization with pingyangmycin lipiodol emulsion (PLE). Because of fewer complications and less injuries than surgical treatment, the theraputics received more and more attention of related clinical department. It has a tendency to take the place of the routine treatment in the suitable indications.Preliminary study revealed the devascular effect of PYM correlated with the inhibitory of vascular endothelial cells (ECs). The effects of PLE arterial perfusion on vessels and surrounding histology havn't been realized until now. Otherwise, some clinical problems including the possibility of devascular orsclerotic treatment of PYM on vascular malformations are worthy of investigating totally.Section I In vitro study of PYM on growth inhibition of human umbilicalvein endothelial cells. ObjectiveTo confirm inhibitory effect of PYM on vascular endothelial cell, we investigated the effect of PYM on growth and proliferation of EVC304 cells in vitro, a cell strain of human umbilical vein endothelial cells. Marterials & MethodsThe growth inhibition of ECV304 cells was evaluated by MTT assay after 24-72 hours' incubation with PYM and BLM in different concentration gradient (Ixl0"2~lxl03ug/ml). The effects of PYM on cell cycle and apoptosis were measured by flow cytometry and fluorescence microscope. The patterns of ECV304 cell death were also analysed after annexinV-FTTC/PI double-variance flow cytometry assay. ResultsPYM and BLM can effectively inhibit the growth of ECV304 cells. After cultured with 10ng/ml PYM for 24, 48 and 72 hours, inhibitory rates of ECV304 cells were 44.7%, 59.7% and 74.4% respectively. The inhibitory effects of the drugs were time-dependent and dose-dependent. ICso of PYM and BLM treatment for 24, 48, 72 hours were 32.94, 2.56, 0.75ug/ml and 125.75,6.59,2.46ug/ml respectively.Flow cytometry analysis showed that ratio of Go-Gi phase cells decreased from (45.38±1.09)% to (44.72±1.44)% by lug/ml PYM treatment for 24 hours, with that of S phase cells significantiy(P<0.01) decreasing from (44.91±1.76)% to (16.24±1.40)% and of G2-M phase cells significantly (P<0.01) increased from (9.72±0.77)% to (39.04±2.40)%. Meanwhile, typical changes of cell apoptosis were observed under fluorescent microscope. Apoptosis rates of ECV304 cells by lug/ml PYM treatment for 24,48 and 72hours were respectively 6.6%, 15.3%, 19.3% and 11.7%,16.8%, 22.7%(P<0.01 within group) by 100ng/ml PYM (P<0.05 between two groups). Furthermore, necrosis rate of 100ug/ml concentration displayed stronger cytotoxic effect than that of lug/ml with 48.7%~60.3% vs 16.8%~23.9%Conclusion1. PYM and BLM could effectively inhibit the growth and proliferation of ECV304 cells and the effects were time-dependent and dose-dependent. Inhibitory effect of PYM on ECV304 cells was stronger than that of BLM.2. The cycle of ECV304 cells was sele... |
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