Adenovirus-mediated therapeutic effect of Parkin on Parkinson's model rats
Objective To explore the therapeutic effect of adeno-associated virus-mediated Parkin on Parkinson's model rats.Methods A total of 60 SD rats were randomly divided into a control group,a model group,and a Parkin group.Parkinson's disease(PD)model was established by subcutaneous injec-tion of fisetin emulsion into the neck and back in the model group and Parkin group.After modeling,the rats in the Parkin group were injected with adeno-associated viruses carrying the Parkin gene in the left sub-stantia nigra.The rats in the control and model groups were injected with negative adeno-associated viruses carrying empty vectors in the left substantia nigra.The behavioral analysis was done on three groups of rats after 21 days of injection.The rats were euthanized,and the apoptosis index of substantia nigra cells in the three groups was analyzed using terminal-deoxynucleotidyl transferase mediated nick end labeling(TUNEL).The superoxide dismutase(SOD)and glutathione(GSH)-px levels were analyzed using SOD and GSH-px assay kits.The levels of apoptosis related proteins[cysteinyl aspartate-specific protease-3(Caspase-3),B cell lymphoma/leukemia-2(bcl-2),and bcl-2 associated X protein(bax)]were detected by Western blotting.Different treatments were compared using analysis of variance,and inter-group com-parisons were made using LSD.Results The expression level of Parkin protein in the substantia nigra of rats in the Parkin group(1.54±0.18)was significantly higher than that in the control group and model group(0.81±0.11,0.89±0.12,t=9.257,8.014,P＜0.05).The number of rotations within 10 min in the Parkin group[(4.87±0.85)circles]was significantly less than that in the model group[(12.25±3.21)circles,t=4.264,P＜0.05].The apoptosis index of substantia nigra cells in the Parkin group[(10.67± 4.91)％]was significantly less than that in the model group[(34.59±8.32)％,t=3.698,P＜0.05].The peripheral serum SOD and GSH-px enzyme activities in the Parkin group[(3.87±0.61),(7.98± 1.27)U/ml]were significantly higher than those in the model group[(2.18±0.51),(4.10±1.09)U/ml,t=6.594,7.251,P＜0.05).The levels of Caspase-3 and bax in the brain tissue of Parkin group rats(1.12±0.08,1.23±0.06)were significantly lower than those in the model group rats(1.58±0.08,1.69±0.11,t=8.174,9.241,P＜0.05).The level of bcl-2 in the brain tissue of Parkin group rats(1.38±0.10)was higher than that of the model group(1.02±0.08,t=5.141,P＜0.05).Conclusion Adenovirus-mediated expression of Parkin protein in the substantia nigra of Parkinson's rats can significantly inhibit cell apoptosis,and significantly improve rat behavior.
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