Therapeutic Effect Of Human Olfactory Mucosa Mesenchymal Stem Cells On Alzheimer's Disease Model Cells

Objective: This study induced hyperphosphorylation of TAU protein in human neuroblastoma cell line(SH-SY5Y)by Okadaic acid(OA),and established the pathological mechanism of Alzheimer's disease(AD)in vitro cell model.On this basis,the therapeutic effect of human Olfactory Mucosa mesenchymal stem cells(hOM-MSCs)co-cultured with AD cell model was studied,providing theoretical and experimental basis for the application of hOM-MSCs in AD treatment.Method:1.SH-SY5 Y cells were resuscitated,and subculture was carried out with good cell condition and satisfactory density;2.SH-SY5 Y cells was treated with different concentration(0,10,20,30,40 nmol/L)of OA treatment SH SY5 Y cells in different time(8 h,24 h),by observing the cell morphology,detection cell apoptosis rate,cell microfilament,microtubule structure and using method of western blot(WB)to detect intracellular phosphorylation TAU(P-TAU)protein content changes to determine the optimal concentration of OA building and processing time;3.Obtain human olfactory mucosa tissue with nasal endoscope,cut the tissue into pieces and stick to the wall,conduct h OM-MSCs primary culture,and subculture was carried out with good cell condition and satisfactory density;4.Randomized modeling was conducted according to the determined OA treatment time and concentration.hOM-MSCs were transplanted into the upper chamber of transwell device in the treatment group and co-cultured with the successful modeling AD cells and then compare with the control group and the modeling group;5.The therapeutic effect of OM-MSCs on the cells after modeling was analyzed by observing the cell morphology,immunofluorescence results and the changes of P-TAU amount in cells detected by western blot.Result:1.According to the observation of cell morphology,detection of apoptosis rate,immunofluorescence results and analysis of WB results,there were significant differences between the cells and the control group after OA treatment.Among them,the cell apoptosis rate increased,cell morphology changed,cells became round,dendrites shortened,microtubule fiber bundles became thinner,and the amount of intracellular P-TAU increased significantly.2.After OM-MSCs were co-cultured with SH-SY5 Y cells after modeling,cell morphology began to restore to normal,dendrites began to elongate,microtubule fiber bundles began to thicken,and compared with the non-cocultured model group,the amount of P-TAU in cells significantly decreased.Conclusion:1.SH-SY5 Y cells were induced by OA to conduct AD modeling cells.The optimal concentration of OA should be 30nmol/L and 24 h is the appropriate processing time.2.Co-culture with hOM-MSCs has a therapeutic effect on the AD modeling cells which is established by OA induced injury of SH-SY5 Y cells.