Effect Of Icaritin On Mitochondrial Damage Of MN9D Cells Induced By 6-OHDA

Objective: We investigated the protective effect of Icaritin(ICT)on 6-OHDAinduced mitochondrial damage in MN9 D cells and its possible mechanism by detecting the changes of ICT on 6-OHDA-induced-induced mitochondrial damage.Methods: MN9 D cells were resuscitated and cultured,We selected well-grown MN9 D cells in logarithmic phase and randomly divided them into control group,0.01μM ICT group,50 μM 6-OHDA group,50 μM 6-OHDA + 0.001 μM ICT group and50 μM 6-OHDA + 0.01 μM ICT group.According to the preliminary experimental results of this project,we pre-treated ICT with corresponding concentration for 1 hour and then added 6-OHDA for 24 hours together.ATP content was detected by ATP assay kit,mitochondrial complex Ⅰ activity was detected by mitochondrial complex Ⅰactivity assay kit,MMP was detected by flow cytometry,α-Syn,TH and Cyt C protein levels were detected by western blot.The morphological changes of mitochondria in each group were observed by transmission electron microscopy.Results: Compared with the control group,ATP content,mitochondrial complex I activity,high MMP level and TH protein content were decreased in the 6-OHDA group,while α-Syn and Cyt C protein levels were increased.In the 6-OHDA group,mitochondrial swelling,mitochondrial crest fracture and disappearance,electron density reduction,vacuolation and membrane damage of some mitochondria were observed under electron microscope.After ICT treatment,ATP content,mitochondrial complex I content,MMP level and TH protein content of MN9 D cells increased,whileα-Syn and Cyt C protein levels decreased significantly.It could be observed under electron microscopy that mitochondrial swelling was reduced,and a few mitochondrial crists were still clearly visible.Conclusion: ICT has a protective effect on the cell model of MN9 D cells induced by6-OHDA,which may be related to increase mitochondrial complex I activity,increase the content of ATP,stabilize the mitochondrial membrane potential,and thus alleviate mitochondrial damage.