Study On Protective Effect And Mechanism Of Ligustrazine Hydrochloride On Iron Overload Induced Cell Injury

OBJECTIVE:1.To study the iron overload model of human umbilical vein endothelial cells by iron ferric ammonium citrate and to study the effect of inflammatory factors on the expression of erythrocytes in endothelial cells under iron overload.2.To investigate the effect of Ligustrazine Hydrochloride Extract on the expression of Hepcidin by inhibiting the expression of inflammatory cytokines,and to reduce the iron overload and inhibit the oxidative stress of endothelial cells and maintain the stability of cell function.METHOD:1.In vitro separation of umbilical vein endothelial cells,identification,culture and passage,with iron ferric ammonium citrate,human umbilical vein endothelial cells to build iron overload model.The experiment was divided into 4 groups: blank group,model group,heparin group and ligustrazine hydrochloride(TMP)group.2.The levels of Hepcidin,ROS,SOD,E-Selection,BMP-6 and IL-6 in the cells or supernatant were detected by enzyme-linked immunosorbent assay(ELISA).The difference of apoptosis rate was detected by flow cytometry.RESULTS:1.To construct the iron overload model of human umbilical vein endothelial cells.After screening,the optimal concentration of ferric citrate was 320 mmol / L.2.The median lethal concentration of ligustrazine hydrochloride and heparin was 4mmol / L and 1*107IU / L respectively in the iron overload model of human umbilical vein endothelial cells.Therefore,1/10 of the lethal concentration was chosen as the concentration of dosing,The concentration of tetramethylpyrazine hydrochloride was 400μmol / L,and the concentration of heparin was 1 *106IU / L.3.The expression of IL-6,BMP-6,Hepcidin,ROS and E-selection in the model group was significantly higher than that in the blank group(P <0.01),Meanwhile,the expression of SOD was significantly increased(P <0.05).The expression of IL-6,BMP-6,Hepcidin,ROS and Eselection was significantly decreased(P <0.01).Meanwhile,the expression of SOD was significantly decreased(P <0.05).4.Compared with the model group,The apoptosis of the model group was significantly increased(P<0.01).Compared with the model group,the apoptosis of ligustrazine hydrochloride group and heparin group was significantly decreased(P <0.01).CONCLUSIONSIron overload increases the oxidative stress of umbilical vein endothelial cells and causes endothelial cell dysfunction.And tetramethylpyrazine hydrochloride inhibits the release of ferritin and reduces intracellular iron overload,to a certain extent,alleviate the damage of cells to oxidative stress.