Mechanism Of Inhibition Of Lipid Peroxidation By Lon Protease 1 In Alleviating Ferroptosis In Dopaminergic Nerve Cells

Objective: To investigate the mechanism of ferroptosis and lipid peroxidation damage in the cell model of Parkinson’s disease(PD),and to investigate the possible mechanism of inhibition of lipid peroxidation by Lon Protease 1(LONP1)to alleviate ferroptosis in dopaminergic nerve cells.Methods:(1)Using 6-hydroxydopamine Human neuroblastoma(SH-SY5Y)cells were induced by 6-OHDA to establish a PD cell model,and Ferrostatin-1(Fer-1)was used to intervene and detect the improvement of lipid peroxidation.Cell morphology was observed using an inverted microscope,cell survival rate was measured using CCK-8 method,and the levels of iron,malondialdehyde(MDA),and reduced glutathione(GSH)were measured using a kit to detect ferrous ions,lipid peroxidation products MDA,and GSH;RT-q PCR was used to detect the m RNA expression levels of Keap1,Nrf2,and GPX4;Western blot was used to detect the protein expression levels of Keap1,Nrf2,and GPX4.(2)Using plasmid transfection technology to overexpress the Homo LONP1 gene to intervene in a PD cell model.The expression level of LONP1 was detected by ELISA assay;The kit detects the levels of ferrous ions,lipid peroxidation products MDA,and GSH;RT-q PCR was used to detect the m RNA expression levels of LONP1,Keap1,Nrf2,and GPX4;Western blot was used to detect the protein expression levels of LONP1,Keap1,Nrf2,and GPX4.Result:(1)○1 SH-SY5 Y cells exposed to After 24 hours in 6-OHDA of 80μM,most of the cell bodies became rounded,and the synaptic structure decreased or even disappeared.○2 Compared with the normal control group,the levels of ferrous ions and MDA in the PD model group increased,while the levels of GSH decreased(P<0.01);○3 Compared with the normal control group,the level of Keap1 m RNA in the PD model group increased,while the levels of Nrf2 and GPX4 m RNA decreased(P<0.05).The Western blot results suggested that the protein expression level was consistent with the above results;○4 Compared with the PD model group,the survival rate of DA neurons increased to 89.50±3.27% after using Fer-1,and the levels of ferrous ions and MDA decreased with the increase of GSH levels(P<0.01);The level of Keap1 m RNA decreased,while the levels of Nrf2 and GPX4 m RNA increased(P<0.05).The protein expression level was consistent with the above results.(2)○1 Compared with the PD model group,the levels of iron ions and MDA in the LONP1 overexpression+PD model group decreased,while the levels of GSH increased(P<0.01).○2 Compared with the PD model group,the level of Keap1 m RNA in the LONP1 overexpression+PD model group decreased,while the levels of Nrf2 and GPX4 m RNA increased(P<0.05).The protein expression level was consistent with the above results.Conclusion: On the basis of 6-OHDA induced SH-SY5 Y cells,ferroptosis lipid peroxidation participates in the damage process of PD cell model,and Fer-1 can reduce the level of lipid peroxidation and alleviate cell damage;LONP1 can improve cell damage by reducing lipid peroxidation levels and inhibiting ferroptosis in DA neurons.