Effect Of Electroacupuncture On Expression Of Autophagy Related Proteins LC3-? And P62 In Substantia Nigra Of Rats With Parkinson's Disease

Objective: The Parkinson's Disease model was prepared by subcutaneous injection of rotenone.To observe the effect of electroacupuncture on the expression of autophagy-related proteins LC3-? and P62 in PD rats and to investigate the role of autophagy in acupuncture and moxibustion for the prevention and treatment of PD.Methods: A total of 120 healthy male SD rats were selected,and the rats were divided into eight groups after one week of adaptive feeding.The eight groups were normal group,model group,sham operation group,four EA groups(treatment for 28 days,21 days,14 days,7 days)and pretreatment group,15 rats in each group.PD rat model was induced by a subcutaneous injection of rotenone at the rat's nape,and rats in the sham operation group were injected with sun-flower oil that did not include rotenone.Rats in the EA groups were treated by EA at“Fengfu”(GV16)and“Taichong”(LR3)While the normal group,sham operation group and model group were not treated,Thetreatment was given once a day for EA groups of 7 days,14 days,21 days and 28 days.Rats in the pretreatment group received the same treatment as the EA groups for 7 days and then established the model.(1)To observe the behavioral changes of all rats and conduct a behavioral score,The horizontal movementand vertical movement of rats in each group were detected by open box test.(2)Immunohistochemical staining was used to detect the expression of TH and ?-syn in the substantia nigra of each group.(3)The expression of LC3-?m RNA and P62 m RNA in the substantia nigra of the rats were detected by RT-PCR.(4)Western blot was used to detect the expression of LC3-? and P62 protein in the substantia nigra of rats.Results:(1)Behavioral scores showed that compared with the normal group and the sham operation group,the behavioral scores of the model group increased significantly(P<0.01);The EA groups and the pretreatment group were significantly lower than that in the model group(P<0.01);Open field test results showed that compared with the normal group and sham operation group,horizontal motion and vertical motion of model group decreased significantly(P<0.01);horizontal motion and vertical motion of the EA groups and the pretreatment group was obviously improved compared with the model group(P<0.01).(2)Immunohistochemical detection showed that compared with the normal group and the sham operation group,the positive expression of TH in the substantia nigra decreased significantly and the positive expression of a-syn was significantly higher in the model group.Compared with the model group,the positive expression of TH in the EA groups and thepretreatment group were significantly increased,the positive expression of a-syn were decreased significantly(P<0.01).There were no significant difference between the positive expression of TH and a-syn in the normal group and the sham operation group rats in substantia nigra(P>0.05).(3)RT-PCR showed that compared with the normal group and sham operation group,the expression of LC3-? m RNA and P62 m RNA in model group were significantly increased(P<0.01).Compared with the model group,the expression of LC3-?m RNA in the EA groups of 28 days,21 days,14 days were significantly higher,P62 m RNA decreased significantly(P<0.01),There were no signnificant difference between the positive expression of LC3-? m RNA and P62 m RNA in the EA group of 7 days and the pretreatment group.(P>0.05)(4)The results of Western blot showed that compared with the normal group and the sham operation group,the expression of LC3-? and P62 were highter in the model group.(both P<0.01).Compared with the model group,the expression level of LC3-?was significantly increased(P<0.01)and the expression of P62 was significantly decreased in the EA group and the pretreatment group(P<0.01).Conclusion: Autophagy activation and well function can effectively remove deposition of ?-syn,delay the process of PD.Electroacupuncture treatment can improve autophagy dysfunction induce by the cytotoxicity of rotenone,and promote the degradation of cytotoxic ?-syn,which can play a protective role against DA neurons.