| 1)objective: Parkinson's disease has become the world's second largest neurodegenerative disease affecting the quality of life of the elderly,and its pathogenesis has not been determined.Previously,the research group has demonstrated that electroac upuncture may prevent the destruction of Dopamine energy neurons by regulating the expression of proteins and genes of endoplasmic reticulum stress related signaling pathways,thereby delaying the course of PD.However,few studies have been conducted on the mechanism of inhibiting apoptosis to improve PD related factors and symptoms.Therefore,CHOP induced apoptosis pathway was selected in this study to explore a new target of electroa cupuncture in the treatment of PD,providing experimental and theoretical basis for clinical treatment.2)methods: 60 male SPF-grade SD rats were selected for three behavioral tests after adaptive feeding for one week,and their behavioral performance was not abnormal.They were randomly divided into three groups: normal group(12 rats),sham operation group(15 rats)and model group(33 rats).The PD model(2mg/kg,dissolved in dimethyl sulfoxide at a concentration of 2mg/ml)was replicated with rotenone emulsion subcutaneously injected into the neck and back,once a day for 28 days consecutively.The normal group was not treated,while the sham group was given dimethyl sulfoxide solution with the same volume of unmixed rotenone powder,once a day for 28 days consecutively.After the modeling was completed,the behavior of the rats was scored according to the behavioral scoring criteria of Chen xin.The model rats with scores ranging from 2 to 8 points were deemed as successful replication,and three of the model rats were selected for immunohistochemical detection to evaluate their pathological changes.At the end of modeling,24 rats were ran domly kept in the model group,which were randomly divided into the model group and the electroacupuncture group,with 12 rats in each group.The sham group also kept 12 randomly.In the electroacupuncture treatment group,the acupuncture points "Fengfu" and "Taichong" were selected to be inserted into the electro acupuncture group,with a continuous wave,a frequency of 2Hz and a strength of 1m A,once a day,for 14 d of continuous treatme nt."Fengfu" fixed with a point,"Taichong" alternate alternate days.Rats in the normal group,the sham operation group and the model group were also fixed for fixed time every day,but electroacupuncture was not used.The morphological and behavioral changes of the appearance of rats in each group were observed according to the behavioral scoring criteria of Chen xi.The open field experiment was used toevaluate the changes of the rats' autonomic motor ability and anxiety and depression.Suspension experiment was used to detect the changes of limb muscle strength and brain DA concentration in each group.The rats' midbrain substantia nigra was taken,and half of the experimental animals in each group underwent left ventricular perfusion with normal saline and then underwent paraformaldehyde perfusion for internal fixation of brain tissue.The changes in mean optical density of TH and peg-syn in the dense region of the midbrain substantia nigra were detected by immunohistochemical method.The remaining half of the experimental animals in each group were cut off on the ice surface for brain extraction and homogenized.The changes in protein expression of CHOP,BAX and bcl-2 in the midbrain of rats in each group were detected by Western Blot,and the changes in gene expression of CHOP,BAX and bcl-2 in the midbrain of rats in each group were detected by real-time fluorescence quantitative PCR.3)results: 1.Changes in appearance and behavior score of rats in each group Compared with the normal group and the sham group,the appearance of rats in the model group was significantly changed,and the behavioral score was significantly increased(P<0.01).Compared with the model group,the appearance of the treated rats in the electroacupuncture group was improved,and the behavioral score was significantly reduced(P<0.01).2.Open field experiment of rats in each group Compared with the normal group and the sham group,the total distance,average speed,total exercise time,times of crossing the central grid area and times of vertical lifting in the reaction box of rats in the model group were significantly reduced,and the total rest time was significantly increased(all P<0.01).Compared with the model group,the total distance,average speed,total exercise time,times of crossing the central grid area and times of vertical lifting were significantly increased and the total rest time were significantly reduced in the elec troacupuncture treatment group(all P<0.01).3.Changes in suspension test scores of rats in each group Compared with the normal group and the sham group,the suspensi on test scores of rats in the model group were significantly lower(P<0.01).Compared with the model group,the suspension test scores in the electroacupuncture group were significantly increased(P<0.01).4.Expression changes of TH in the substantia nigra of rats in each group Compared with the normal group and the sham group,the average light density of TH in the dense region of the substantia nigra was significantly reduced in the model group(P<0.01).Compared with the model group,the average optical density of TH in the dense region of the substantia nigra was significantly increased in the electroacupuncture group(P<0.01).5.Expression changes of the midbrain substantia nigra via-syn in rats in each group Compared with the normal group and the sham group,the mean optical density of the midbrain substantia nigra ben-syn was significantly increased in the model group(P<0.01).Compared with the model group,the average optical density of the midbrain substantia nigra bri-syn in the electroacupuncture group was signi ficantly reduced(P<0.01).6.Changes in protein expression of CHOP,BAX and bcl-2 in the midbrain of rats in each group Compared with the normal group and the sham group,the protein expression levels of CHOP and BAX in the midbrain of rats in the model group were significantly increased,and the protein expression levels of bcl-2 were significantly decreased(all P<0.01).Compared with the model group,the protein expression levels of CHOP and BAX in the midbrain of rats in the electroacupunc ture group were significantly reduced,and the protein expressi on levels of bcl-2 were significantly increased(all P<0.01).7.Changes in gene expression of CHOP,BAX and bcl-2 in the midbrain of rats in each group Compared with the normal group and the sham group,the expressi on levels of CHOP and BAX in the midbrain of rats in the model group were significantly increased,and the expression levels of bcl-2 were significantly reduced(all P<0.01).Compared with the model group,the gene expression levels of CHOP and BAX in the midbrain of rats in the electroacupuncture group were significantly reduced,and the gene expression levels of bcl-2 were significantly increased(all P<0.01).4)conclusion: 1.Electroacupuncture points "Fengfu and Taichong" can effectively improve the appearance,morphology and multiple kinds of abnormal PD behaviors of rats with PD disease model.2.The electroacupuncture can degrade the abnormal aggregation of ben-syn,reduce the formation of louisomes in the substantia nigra of PD model rats,and protect the DA energy neurons.3.The mechanism of electroacupuncture intervention in PD modelra ts may be that electroacupuncture can prevent or delay the initiation of CHOP-induced apoptosis signaling pathway,reduce the injury to ERS,protect DA neurons from a large number of in juries,and inhibit the apoptosis pathway,so as to prevent or delay the progression of PD. |
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