Study In Optimized Human α-Synuclein DNA Vaccine Immunize Rotenone Inducing On Chronic Mouse Model Of Parkinson’s Disease

Objiective1. To study the immuotherapeutic effects of the optimized humanα-synuclein DNA vaccine(pVAX1-IL4/SYN-B) immunize rotenoneinducing on chronic mouse of Parkinson’s disease.2. To explore the neuroprotective effects of the optimized humanα-synuclein DNA vaccine(pVAX1-IL4/SYN-B) on mouse after chronicrotenone damage.Methods1. At the first, those mouse were treated with rotenone(1mg/kg)through subcutaneous injection for40days. Then those mouse wererandomly divided into three groups:the optimized human α-synuclein DNAvaccine group(pVAX1-IL4/SYN-B)、the empty plasmid group and thePBS control group.The anterior tibial muscles of those mouse hind legwere injected with the pVAX1-IL4/SYN-B,vacant vector plasmid andPBS(100μl),one time every3weeks, total3times.After the lastimmunization for2weeks,all of the mouse were observed the behavioral changes. The expression of α-syn mRNA was detected by RT-PCR.Thechanges of TH and α-synuclein in substantia nigra(SN) was assayed byimmunohistochemistry.2. Firstly,those mouse were randomly divided into three groups:theoptimized human α-synuclein DNA vaccine group(pVAX1-IL4/SYN-B)、the empty plasmid group(pVAX1) and the PBS control group.Secondly, theanterior tibial muscles of those mouse hind leg were injected with thepVAX1-IL4/SYN-B,vacant vector plasmid and PBS(100μl), total3timesand one time every3weeks,.Three weeks after the last immunization,allmouse were treated with rotenone (1mg/kg)through subcutaneous injectionfor40days.After the last rotenone injection,all mouse were observed thebehavioral changes. The expression of α-syn mRNA was detected byRT-PCR.The changes of TH and α-synuclein in substantia nigra(SN) wasassayed by immunohistochemistry.Results1.significant differences were found between the DNA vaccine groupand the control group in the average ambulation grids（P＜0.05P)andfrequency of rearing（P＜0.05). Compared with the empty plasmid groupthe DNA vaccine group can decrease the α-syn expression in substantianigra45.64%(P＜0.05),increase the TH positive cells51.43%(P＜0.05).Compared with the PBS control group the DNA vaccine group candecrease the α-syn expression in substantia nigra43.28%(P＜0.05),increase the TH positive cells59.00%(P＜0.05).Compared to the empty plasmidgroup and the PBS control group,the α-syn mRNA of the DNA vaccinegroup was decreased significantly(P＜0.05).2.There are Significant differences between the DNA vaccine groupand the control group in the average ambulation grids （P<0.05)andfrequency of rearing（P<0.05).Compared with the empty plasmid groupthe DNA vaccine group can decrease the α-syn expression in substantianigra55.68%(P<0.05),increase the TH positive cells63.27%(P<0.05).Compared with the PBS control group the DNA vaccinegroup can decrease the α-syn expression in substantia nigra55.34%(P<0.05),increase the TH positive cells55.34%(P<0.05).Comparedto the empty plasmid group and the PBS control group,the α-syn mRNAof the DNA vaccine group was decreased significantly(P<0.05).Conclusion1. In the study,we conclude that the optimized human α-synucleinDNA vaccine(pVAX1-IL4/SYN-B) has much effects to improve the micebehavior of Parkinson’s disease and has a better therapeutic effect on micewith Parkinson’s disease.2. We concluded that the optimized human α-synuclein DNAvaccine(pVAX1-IL4/SYN-B)can improve the behavior of mice withParkinson’s disease and exerting neuroprotective effects on dopaminergicneuron aganist rotenone neurotoxin damage.