Expression Changes Of Vascular Endothelial Growth Factor And Epithelial Cell Kinase A2 During The Apoptosis Of Human Umbilical Vein Endothelial Cells Induced By Strychnine

Background and purpose:Chinese traditional medicine strychnix(also known as brucine),Brucine is the mature dried seeds of strychnix brucellae,with anti-inflammatory and analgesic,soft and firm knot,detumescence and blood circulation,activating channels and collaterals,etc.The indole-type structure alkaloids presented from strychnix Brucine are called strychnix,that also one of the important components of composition and efficacy.Clinically,strychnine is widely used in the treatment of pain and rheumatism,tumor inhibition and pain relief.Clinical studies have found that brucine in vitro can not only significantly inhibit the proliferation of a variety of cells,induce multiple myeloma,breast cancer,colon cancer,liver cancer and other cancer cells apoptosis.Human umbilical vein endothelial cells(human umbilical vein endothelial cells,HUVECs)is a common vascular endothelial cell function experiments of cell model.VEGF and Eph A2 are two important markers of vascular endothelial cells.As survival factor and specific mitogen of vascular endothelial cells,VEGF is also an important regulator of angiogenesis.Eph A2 is a member of the receptor tyrosine kinase family,but is highly expressed in malignant tumors.Neovascularization plays an important role in tumor development and metastasis.The process of endothelial cells forming new blood vessels from existing blood vessels and establishing new blood vessel network is very complicated,among which the migration of endothelial cells and the formation of tubes are the key links.The experiment through exploring strychnine HUVECs proliferation activity and cell apoptosis induction effect,and vascular endothelial growth factor(VEGF)and epithelial cell kinase A2(epithelial cell kinase A2,Eph A2)expression changes,and explore the mechanism of action.To investigate the changes in expression of vascular endothelial growth factor(VEGF)and epithelial kinase A2(Eph A2)during brucine induced apoptosis of human umbilical vein endothelial cells(HUVECs).Methods HUVECs were routinely cultured and divided into blank control group,low(10 ?mol/L),medium(20 ?mol/L),and high(40 ?mol/L)doses of strychnine.CCK-8 method was used to detect horses of different concentrations.Comparison of chrysophylline on the viability of HUVECs cells.The migration changes of HUVECs were detected by scratch test to observe the inhibition of cell proliferation.Hoechst33258 cell immunofluorescence,flow cytometry Annexin V/PI double staining method to detect the effects of brucine on HUVECs apoptosis changes,Western blot method to detect the concentration dependence of strychnine on HUVECs cells Changes in VEGF and Eph A2 protein expression.Results Zygospermine could inhibit the proliferation of HUVECs in a concentration-dependent manner.Scratch experiments showed that zygostigmine could inhibit the migration of HUVECs.Hoechst 33258 cell immunofluorescence and flow cytometry showed that styrenine promoted HUVECs in a concentration-dependent manner Apoptosis,Western blot results showed that strychnine reduced the expression of VEGF and Eph A2 in HUVECs in a concentration-dependent manner.Image analysis showed that compared with the blank control,as the concentration increased,the expressions of VEGF / GAPDH and Eph A2 / GAPDH protein in the treatment group of strychnine(10,20,40 ?mol / L)significantly decreased,and the difference was statistically significant(P < 0.05,P < 0.01),the high concentration of strychnine group has significant difference in VEGF and Eph A2 protein expression compared with the low concentration group and the blank group(P < 0.05,P < 0.01).Conclusion Brucine can inhibit the proliferation and migration of HUVECs,and induce apoptosis,which may be related to the down-regulated expression of VEGF and Eph A2.