Expression Of Autophagy In Nigra In Rats With Parkinson’s Disease And The Study On Pharmacological Interventions

Objectives By observing the expression of Autophagy in nigra in rats with Parkinson’sdisease and the effects of Eldepryl on that expression, to discuss the relationship betweenthe Autophagy and PD and therapeutic mechanism of Eldepryl.Methods1. All the90health male Sprague-Dawley (SD) rats were divided into thecontrol group, model group and Eldepryl group. Each group was divided into4days and8days subgroups after the success of model preparation. Each group had15rats, amongwhich6rats were used for Immunohistochemistry,6rats were used for western blottingand the let3were used for Transmission Electron Microscope.2. Parkinson’s disease ratmodel were established by injecting rotenone in subcutaneous.3. Gavage Eldepryl(0.5mg/kg)4d and8d after model successfully administered.4. The autophagy in thenigra was observed by transmission electron microscope; The expression of Beclin1andLC3in nigra was detected by Immunohistochemistry and Western blotting.5. Weobserve tissue slices with microscope camera OLYMPUS (400×), then analysis them byMotic Med6.0digital medical image analysis system. The results of Western Blottingwere analyzed with Image J.6. Application SPSS13.0statistical software for dataanalysis at last.Measurement data are presented as mean±SD (x s),using anova designanalysis of variance within the group, P<0.05for the difference was statisticallysignificant.Results1. By TEM we could see: in nigra of control group, the regular karyo-type ofnerve cell, nuclear membrane is complete, chromatin evenly distributed, there are richorganelles in cytoplasm, the structure is normal, there is no autophagosome.Autophagosome appeared in nerve cell of model group in4d, rounded, mostly locatedbeside the nucleus, and the number of lysosomes were increased.Both of them weresignificantly increased in8d. In nerve cell of Eldepryl group, compared with the modelgroup,the injury nerve cell was significantly mitigated.Normal organelles may beseen,the swelling mitochondria and rough endoplasmic reticulum are still visible.Thereare autophagosome in4d,but was significantly decreased compared with model group.There is no autophagosome in8d.2. The expression of Beclin1, LC3and LC3II/LC3I innigra were increased in the model group compared with that in the control group (P<0.05), and there were more expression of Beclin1, LC3and LC3II/LC3I at8dcompared with those at4d, and it has significant difference (P<0.05). The expression ofBeclin1, LC3and LC3II/LC3I in nigra were reduced in the Eldepryl group comparedwith that in the model group (P<0.05), and there were less expression of Beclin1, LC3and LC3II/LC3I at8d compared with those at4d, and it has significant difference(P<0.05).Conclusions1. Autophagy is activated in nigra of the rat model of PD, autophagy hasinvolved in the occurrence and development of PD;2. By reducing the expression ofAutophagy in nigra in rat model of PD, Eldepryl could play a therapeutic role inParkinson’s disease.