Improvement Of PCBP1on PD Model Rats

Objective:For making sure whether poly (rC) binding protein1(PCBP1) took part in the pathology progress of PD, this study discussed the possibly existed relationship with PCBP1in the the occurrence of Parkinson’s disease(PD) and PCBPl’s modulatory and protective action, using the PD model of intrastriatal6-Hydroxydopamine of rat through adenoassociated virus transfecting recombinant plasmid into neuronal cells to express. Aim at providing reliable theoretical basis for clinically using virus to treat PD, and discovering potential target spots and new thoughts for PD’s treatment.Materials and Methods:SD male rats(200—250g), using PD rats model by6-OHDA injected to bilateral striatums damage, randomly divided into the model group and the virus-treated group and control group. The treated group was injected rAAV2-PCBP1-EGFP in the left part of striatums one week before the surgery of6-OHDA injected to bilateral striatums. A week later,start the behavior test once a week, lasting for6weeks. The animals were sacrificed quickly at the7th week after6-OHDA injured, and the control group’s behaviors were tested and the animals were sacrificed at the same time.1. The rotational behavior test was performed to observe the changes of the behavior induced by6-OHDA.2. The survival ratio of doperminergic neuron fibers in the ST was tested by immuno-histochemical staining to detect different dosages of6-OHDA’s change to rats.3. Confocal microscopy was performed to detect the expression of rAAV2-PCBP1-EGFP in neuronal cells.4. Local motion test experiment was used to detect the rats’behavior changes of the control group, the model group and the virus-treated group.5.The expression content of PCBP1was detected by Immunohistochemical stainingin the neurons of the control group, the model group and the virus-treated group.6. Western blot was used to detect the content of TA and HSP70in the neurons of the control group, the model group and the virus-treated group.Results1. After4weeks of the damage, all group of the rats lesioned by6-OHDA had appeared unusual rotation behavior induced by Apomorphine(Apo); As increasing the dosages of6-OHDA, the unusual rotation times increased and progressively aggravated as time went by. 24ug group have significant differences compared with the control group(p<0.01); Immunohistochemical staining also showed that the TH positive Neuron fiber in the Striatum appeared different levels of death, and the survival neuron percentage at the8week was79%and75%.2. Different dosages of rAAV2-PCBP1-EGFP were injected into rats’striatums and observed its expressions in neurons at different time by Laser Scanning Confocal Microscope. The result showed that rAAV2-PCBP1-EGFP began to express1week after the injection into rats’ striatums, lasting to the eighth week.3. Local motion test experiment results showed that the virus-treated group had significant improvements compared with the model group (p<0.01). Immunohistochemical staining results also showed that the expression content of PCBP1in the virus-treated group increased compared with in the control group, and the model group.4. Western blot detected the expression condition of TH and HSP70in the6-OHDA rats’ midbrain substantia nigra, discovered that PCBP1against the decreasing trend of6-OHDA rat substantia nigra TH protein expression and PCBP1improve the substantia nigra expression of HSP70in a growing trend.Conclusions1. PD rat model made by6-OHDA injection into striatums caused dopaminergic neuron in SNc region to emerge progressive, recessive regression and was a suitable animal model for PD early treatment;2. Firstly demonstrated in PD model that PCBP1had clearly neuronal protective action to dopaminergic neurons;3. In the PD rat model made by6-OHDA injection into striatums, demonstrated4ul rAAV2-PCBP1-EGFP(Genome particles drops of degrees2.5×1011vg/ml) treatment could improve rats movement ability.4. rAAV2-PCBP1-EGFP treatment had clearly neuronal protective action to dopaminergic neuron in SNc region of PD rat model.5. rAAV2-PCBP1-EGFP could increase TH expression in substantia nigra and decrease substantia nigra neuron’s loss to improve PD’s movement behavior by increasing the HSP70content in midbrain substantia nigra.