Protective Effect Of Ndfip1 Overexpression On Apoptosis Of Nerve Cells Induced By Iron Overload

ObjectiveTo observe the effect of Ndfip1 overexpression on the survival rate of nerve cells under the conditions of iron overload,in order to clarify the protective effect of Ndfip1 on nerve cells;and to detect the level of ROS,MDA and the expression of Bcl-2,Bax and Caspase-3,in order to explore the protective mechanism of Ndfip1 on nerve cells.MethodsSH-SY5Y cells in the experimental group were transfected with Ndfip1plasmid,and SH-SY5Y cells in the control group were transfected with empty plasmid.First,the expressions of Ndfip1 protein in the cells of experimental group and control group after transfected for 48h were detected by Western blot to determine the effect of transfection.MTT methods were used to detect the survival rate of SH-SY5Y cells under the action of different concentrations of FeSO₄（0,50,100,200,500μM）to determine the concentration of iron overload.Second,MTT methods were used to determine the survival rate of SH-SY5Y cells in the experimental group and the control group under the condition of iron overload,and the effect of Ndfip1 overexpression on the survival rate of nerve cells was analyzed.Finally,under conditions of iron overload,the levels of ROS and the content of MDA in the cells of experimental group and control group were detected by kits to analyze the effect of Ndfip1 overexpression on oxidative stress responses in nerve cells.The expressions of Bcl-2,Bax and Caspase-3 protein were detected by Western blot to analyze the effect of Ndfip1 overexpression on apoptosis and explore the protective mechanism of Ndfip1 on nerve cells.ResultsAfter plasmid transfected,Western blot results showed that the expression of Ndfip1 in the experimental group increased compared with the control group,indicating that the plasmid were successfully transfected and the cell models were successfully constructed,which can be used in subsequent experiments.The MTT results showed that with the increase of FeSO₄ concentration,the cell survival rates gradually decreased.Among them,the cell survival rate was significantly reduced after treatment with 200μM FeSO₄（P<0.01）,and this concentration can be used as an experimental concentration of iron overload for subsequent experiment.After treated with 200μM FeSO₄,the survival rate of cells in the experimental group increased compared with the control group（P<0.05）,indicating that Ndfip1 can resist the apoptosis of nerve cells induced by iron overload and has protective effect on nerve cells.The ROS results showed that compared with the control group,the fluorescences of cells were significantly weakened in the experimental group,indicating that ROS were reduced.The MDA results showed that the contents of MDA were significantly reduced in the experimental group compared with the control group,indicating that the oxidative stress responses in the experimental group were reduced.Western blot results showed that the expressions of Bcl-2 in the experimental group were increased compared with the control group,indicating that the inhibition of apoptosis was enhanced.The expressions of Bax in the experimental group were decreased compared with the control group,indicating that the role of promoting apoptosis was weakened.The expressions of Caspase-3 in the experimental group were decreased compared with the control group,indicating that the expression of apoptotic enzymes were decreased.Thus,the apoptosis in the experimental group was inhibited.Conclusions1.Ndfip1 could reduce the apoptosis that induced by iron overload and improve the survival rate of nerve cells.2.The protective effect of Ndfip1 on nerve cells may be realized by reducing the oxidative stress response and inhibiting the mitochondrial apoptosis pathway.