| Objective: Acupuncture and moxibustion is effective and safe in the prevention and treatment of Parkinson's disease.Studies at home and abroad have shown that endoplasmic reticulum stress isinvolved in the pathogenesis of Parkinson's disease.Previousstudies showed that there gulation mechanism of electro-acupuncturein the prevention and treatment of Parkinson's disease may be related to the inhibition of protein and gene expression of endoplasmic reticulum stress pathway-related factors and the protection of dopaminergic neurons from apoptosis.However,the effect of electroacupuncture on the control of endoplasmic reticulum stress in the treatment of Parkinson's disease by influencing the PI3K/Akt signaling pathway is rarely studied.In this experiment,rats with Parkinson's disease induced by subcutaneous injection of rotenone into the neck and back were used as the model,and electro-acupuncture at Fengfu,Taichong point was used to intervene,the expression level of TH,?-syn and the expression levels of ATF6,PI3 K,Akt,Caspase-3 protein and gene in the midbrain of rat model rats were observed,to explore the mechanism of electro-acupuncture in the treatment of Parkinson's disease(PD)model rats,in order to provide experimental basis for the clinical application of EA in the prevention and treatment of Parkinson's disease and the improvement of its related clinical symptoms.Methods: 47 healthy male SD rats of SPF grade were selected and purchased.After 3 behavioral tests,no self-issued learning abnormality was observed.After one week of adaptive feeding,they were randomly divided into two groups: control group(12 rats)and model group(35 rats).The rat model of Parkinson's disease was made by subcutaneous injection of rotenone into the neck and back.The rats in the model group were injected subcutaneously with rotenone(2 mg / kg,dissolved in sunflower oil,concentration of 2mg/mL)into the neck and back.The control group was injected with the same dose of sunflower oil emulsion without rotenone,once a dayfor 28 d,with the same dose of rotenone-free sunflower oilemulsion.Referring to the behavioral scoring standard established by Chen Xin and others,the group regarded the rats with a score of 2 to 8 as a successful model,the rats who are not in the scoring range are excluded,27 successful model rats were randomly retained,and 3 rats were randomly selected for left ventricular paraformaldehyde internal fixation.The pathological changes of successful PD model rats were observed by histochemical identification.After successful modeling of Parkinson's disease rats,the model group was randomly divided into two groups: modelgroup(n=12)and electroacupuncture group(n=12),rats in electroacupuncture group were followed by electroacupuncture after acupuncture at Fengfu,Taichong points,Fengfu is a fixed point,Taichong alternates around the other day,continuous wave,frequency 2Hz,intensity 1mA,once a day,continuous treatment for 14 days;The rats in the control group andthe modelgroup were fixed with electric acupuncture every day and fed synchronously,but they were not treated with electro-acupuncture.The behavioral symptoms of rats in each group were observedby the behavioral scoring standard established by ChenXin et al,and the behavioral changes of rats in each groupwere observed by the open field test;Sampleswere taken from the substantia nigra of the midbrain.Half of the rats in each group were treated with left ventricular paraformaldehyde perfusion and internal fixation of brain tissue,the expression of TH,?-syn in substantia nigra was detected by immunohistochemistry;The fresh brain tissue homogenate wastaken from the other half of the rats in each group,the protein expression of ATF6,PI3 K,Akt,Caspase-3 in the substantia nigra of each group was detected by Western Blot,The expression of ATF6,PI3 K,Akt,Caspase-3 gene in substantia nigra was detected by real-time fluorescence quantitative-PCR.Results:1.Changes of behavioral symptom scores in all groups of ratsCompared with the control group,the behavioral score in the model group was significantly higher than that in thecontrol group(P<0.01);Compared with the model group,the behavioral score of EA group was significantly lower(P<0.01).2.Behavioral changes of autonomic exercise in rats of each groupCompared with the control group,the total distance,average speed,total time of exercise,the distance of movement in the central area and the exercise time in the centralarea of the model group were significantly decreased,and the total rest time was significantly increased(all P<0.01);Compared with the model group,the total distance,average velocity,total time of exercise,the distance of movement in the central area and the exercise time in the central area of the electroacupuncturegroup were significantly higher than those in the model group,while the total rest time was significantly decreased(all P<0.01).3.Changes of TH expression in substantia nigra of rats in each groupCompared with the control group,the mean optical density of the substantia nigra TH in the model group was significantlylower than that in the control group(P<0.01);Compared with the model group,the mean optical density of the substantia nigra TH in the electroacupuncture group wassignificantly higher than that in the model group(P<0.01).4.Changes of ?-syn expression in substantia nigra of all groups of ratsCompared with the control group,the mean optical density of ?-syn in the substantia nigra of the model group wassignificantly higher than that of the control group(P<0.01);Compared with the modelgroup,the mean optical density of ?-syn in the substantia nigra of the midbrain of the electroacupuncture group was significantly lower than that of the model group(P<0.01).5.Changes of ATF6,PI3 K,Akt,Caspase-3 protein expression in substantia nigra of all groups of ratsCompared with the control group,the protein expression level of ATF6,Caspase-3 in the substantia nigra of the model group was significantly higher,while the protein expression of PI3 K,Akt was significantly decreased in the model group(all P<0.01);Compared with the model group,the expression level of ATF6,Caspase-3 protein in substantia nigra decreased significantly and the protein expression level of PI3 K,Akt increased significantly in EA group(all P<0.01).6.Changes of gene expression of ATF6,PI3 K,Akt,Caspase-3 in substantianigra of rat mesencephalon in each groupCompared with the control group,the expression level of ATF6,Caspase-3 gene in the substantia nigra of the model group was significantly higher,while the gene expression level of PI3 K,Akt was significantly decreased in the model group(all P<0.01);Compared with the model group,the expression level of ATF6,Caspase-3 gene in substantia nigra decreased significantly and the gene expression level of PI3 K,Akt increased significantly in EA group(all P<0.01).Conclusion:1.Electroacupuncture can effectively improve the typical behavioral symptomsand abnormal behavioral manifestations in Parkinson's disease model rats.2.Electroacupuncture can degrade or clear the unfolded or misfolded ?-syn,in the substantia nigra of the midbrain of Parkinson's disease model rats to protect dopaminergic neurons from injury or apoptosis.3.The mechanism of the effect of electroacupuncture on Parkinson's disease model rats may be related to the activation of PI3K/Akt signaling pathway to inhibit ERS,to play an anti-apoptotic role and to prevent or delay the progressive course of PD. |
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