Influence Of Angiogenesis Inhibitors, Angiostatin And Thalidomide, On The Lymphagiogenesis

Objective To study the effect and mechanism of angiostatin and thalidomide on proliferation and migration of lymphatic endothelial cells (LEC) and further to find the safe,effective and practical inhibitors of lymphangiogenesis.Material and Methods LECs from pig thoracic duct were cultured. 1. Inhibitory assay:Two groups,angiostatin and thalidomide were set.In each group,two methods were used to observe the proliferation and migration of LECs.(1)Method of scraping line:In angiostatin group and thalidomide group,each included control group and 3 experimental groups of different concentration.(2)MTT:In angiostatin group,the control group and 4 experimental groups of different doses were set.In thalidomide group,it included control group and 5 experimental groups of different doses.2. Apoptotic assay:(l)Hoechst staining. (2) Flow cytometer detection:Different concentrations of angiostatin and thalidomide were used on cultured LECs to detect apoptosis.3.Electron microscope observation.Results 1.Method of scraping line:Comparing with the control group,when dose of angiostatin and thalidomide was 0.5 μg/ml,both could inhibit the proliferation and migration of LECs (P<0.01).2.MTT:When dose of angiostatin and thalidomide reached 0.75ug/ml and 0.5ug/ml respectively,a strong inhibitive effect can be found (P<0.01).3.Apoptotic assay:(l)Hoechst staining proved that the apoptotic body surrounding the nucleus of LECs which were treated by angiostatin and thalidomide could be observed.(2)Flow cytometer detection:Lots of sub-diploid nuclear peak appeared (apoptotic peak) and diploid cells peak reduced.Conclusion 1 .Angiostatin and thalidomide could evidently inhibit the proliferation and migration of LECs and were dose-dependent.2.Angiostatin and thalidomide could induce apoptosis of LECs.