| Chapter 1 Literature ResearchObjective: Parkinson’s disease is currently the second largest senile dege nerative disease.Acupuncture treatment of Parkinson’s disease has always been a research hotspot in this field.In this study,by searching relevant literatures on acupuncture treatment of Parkinson’s disease at home and abroad,the current research status of acupuncture treatment of Parkin son’s disease is statistically provided to provide a literature basis for the research of this subject.Methods: The search database for this study was CNKI,CBM,Weipu,Wanfang,Pubmed.The search formula "Parkinson" AND "Acupuncture" was developed and a professional search was conducted.The search was conducted since the establishment of the database until February 2020.According to the inclusion criteria and exclusion criteria,the bibliography is imported into Note Express3.2.0 software,read the document title and abstract,delete the documents that do not match the theme,download the full text of the filtered documents,and integrate the documents according to the inclusion and exclusion criteria for subsequent screening.The final screening literature was analyzed for its basic characteristics,disease model,acupuncture points,research level,and Excel was used to organize,summarize,and analyze the data.Results: Through analysis,a total of 1,654 papers were obtained.According to the inclusion criteria and exclusion criteria,excluding repeated reports finally determined that the number of rat experiment literature was 147.44 academic dissertations,103 journal articles,the earliest year 1996,the most recent year 2020.It is concluded that the mice currently used for modeling Parkinson’s disease mainly include C57 BL / 6 male mice,SD rats,and Wistar rats;the modeling reagents mainly include rotenone,6-OHDA,MPTP,LPS and other biochemical methods,and there are also Physical methods such as unilateral MFB damage and mechanical damage MFB.The research level covers molecular biology such as genomics,transcriptomics,proteomics,and metabolomics.Commonly use dacupoints are Fengfu,Taichong,Zusanli and other nerve points and brain reflex zones,which lay the literature for subsequent research.basis.Discussion: 1.Parkinson’s disease model strains mainly include SD rats,C57 BL / 6 mice,Wistar rats,etc.;2.Parkinson’s disease modeling methods include MPTP intraperitoneal injection model,6-hydroxydopamine(6-OHDA)intracerebral injection model,rotenone subcutaneous injection model of neck and back,etc.;3.The research fields of acupuncture treatment of Parkinson’s disease include transcriptomics,proteomics,metabolomics,etc.Chapter 2 Experimental ResearchObjective: Parkinson’s disease is a common central nervous system degenerative disease that affects the physical and mental health of the elderly.Acupuncture has a wide range of indications and has achieved significant results in the treatment of Parkinson’s disease.Previous research by this research group shows that electroacupuncture can effectively prevent the occurrence and development of Parkinson’s disease,which may be related to its regulation of endoplasmic reticulum stress and protection of dopaminergic neurons.However,there are few studies on how electroacupuncture can treat Parkinson’s disease by regulating the ATF4 / Parkin signaling pathway.In this experiment,a rat model of Parkinson’s was replicated by subcutaneous injection of rotenone in the back of the neck.The effects of electroacupuncture at "Fengfu"(GV16)and "Taichong"(LR3)on the mesencephalic substantia nigra in rats with Parkinson’s disease were observed.mechanism.Methods: Sixty SPF male healthy SD rats were used.Three open field experiments were performed to ensure that the rats had no spontaneous behavioral abnormalities and were fed adaptively for one week.Grouped according to the random number table,they were randomly divided into normal group(12 animals),sham operation group(15 animals),and model(33 animals).The rat model of Parkinson’s disease was replicated by subcutaneous injection of rotenone on the back of the neck,and arotenone-dimethylsulfoxide emulsion(rotenone: dimethylsulfoxide: physiological saline solution = 1g: 3.5L: 0.5L)was injected subcutaneously into the model.,Dosed once a day according to the body weight of rats,2mg / kg,for 28 consecutive days;The sham operation group was given rotenone-free dimethyl sulfoxide physiological saline mixed solution(dimethyl sulfoxide: physiological saline solution = 3.5L: 0.5L)subcutaneously at the neck and back,and was administered once a day according to the body weight of rats.2mg / kg for 28 consecutive days;Normal group: Grasp and weigh daily without any other treatment.After modeling,observe the behavioral changes and score the rats according to the behavioral scoring system established by Chen Xin et al.Rats with model behavior scores ranging from 2 to 8 were regarded a s successful models,27 successful models were randomly selected,and 3 successful model rats were randomly selected for left ventricular paraformaldehyde internal fixation to fix brain tissue.,Immunohistochemical method was used to detect the expression level of TH in rat midbrain to make a histological identification model.After successful modeling,there were 12 in the normal group and 12 in the sham operation group.The model was randomly divided into 12 in the model group and 12 in the electroacupuncture group.The open field experiments were used to observe the behavioral changes of the rats in each group.Electroacupuncture group electroacupuncture points "Fengfu" and "Taichong",positive pole-"Fengfu",negative pole-"Taichong"(double-sided "Taichong" exchanges electric needles every other day),waveform-contin uous wave,frequency-2Hz,Current-1m A,20 min / time,once / day,continuous treatment for 14 days.Normal group,sham operation group and model group: the same grasping and fixing methods as the electroa cupuncture group were adopted daily,and no treatment was performed.After treatment,observe the behavioral changes and scores of the rats in each group according to the behavioral scoring system established by Chen Xin et al.Use open field experiments to observe the behavioral changes in the autonomous activities of the rats in each group.After the behavioral tests of rats in each group were completed,6 rats in each group were randomly selected for left ventricular paraformaldehyde internal fixation,and the expression changes of TH and α-syn in the sub stantia nigra of rats were detected by immunohistochemistry;the remaining in each group Six rats were decapitated quickly on ice to remove fresh brain tissue.Western blot analysis was used to detect changes in ATF4 and Parkin protein levels in the rat mesencephalic substantia nigra.Real-time fluorescence quantitative-PCR was used to detect rat substant ianigra ATF4 m RNA,Parkin m RNA gene expression changes.Result: 1.Observation of behavior changes in experimental groups of rats After modeling,compared with the normal group and the sham operation group,the behavioral observation scores of the model group and the electroacupuncture group were significantly increased(P <0.01).After treatment,compared with the normal group and the sham operation group,the behavioral observation score of the model group was significantly increased(P <0.01);compared with the model group,the behavioral observation score of the electroacupuncture group was significantly decreased(P <0.01).2.Changes in behavior of rats in experimental groups After modeling,compared with the normal group and the sham operation group,the total distance of autonomous movement,the total time of autonomous movement,the average speed of autonomous movement,and the frequency of vertical movement were significantly reduced in the model group and the electroacupuncture group(P <0.01).The total rest time was significantly increased(P <0.01).After treatment,compared with the normal group and the sham operation group,the total distance of autonomous movement,the total time of autonomous movement,the average speed of autonomous movement,and the frequency of vertical movement were significantly reduced in the model group(P <0.01),and the total rest time increased significantly.High(P <0.01);compared with the model group,the total distance of autonomous movement,the total time of autonomous movement,the average speed of autonomous movement,and the frequency of vertical movement significantly increased in the electroacupuncture group(P <0.01),and the total rest time Significantly reduced(P <0.01).3.Changes of TH expression in the substantia nigra of rats in each experimental group Compared with the normal group and the sham operation group,the average absorbance of TH in rats in the model group was significantly reduced(P <0.01);compared with the model group,the average absorbance of TH in rats was significantly increased in the electroacupunctur e group(P <0.01).4.Changes of α-syn expression in the substantia nigra of rats in each experimental group Compared with the normal group and the sham operation group,the average absorbance of mesencephalic α-syn in the model group was significantly increased(P <0.01).Compared with the model group,the mesencephalic α-syn in the electroacupuncture group was significantly higher.The average absorbance was significantly reduced(P <0.01).5.Changes in ATF4 and Parkin protein expression in the mesencephalic substantia nigra of each group of rats Compared with the normal group and sham operation group,the ex pression of ATF4 protein in the mesencephalic substantia nigra of the model group was significantly increased(P <0.01),and the expression of Parkin protein was significantly reduced(P <0.01).Compared with the model group,the expression of ATF4 protein in the substantia nigra of the electroacupuncture group was significantly reduced(P <0.01),and the expression of Parkin protein was significantly increased(P <0.01).6.Experimental expression of ATF4 m RNA and Parkinm RNA in the substantia nigra of rats in each group Compared with the normal group and the sham operation group,the expression level of ATF4 m RNA gene in the substantia nigra of the model group was significantly increased(P <0.01),and the expression level of Parkin m RNA gene was significantly decreased(P <0.01).Compared with the model group,the expression level of ATF4 mRNA gene in the substantia nigra of the electroacupuncture group was significantl yreduced(P <0.01),and the expression level of Parkin mRNA gene was significantly increased(P <0.01).Conclusion: 1.Rotenone can well induce a rat model of Parkinson’s disease.2.Electroacupuncture can effectively improve the behavioral abnormalities of rats with Parkinson’s disease.3.Electroacupuncture can effectively clear or degrade α-syn over-expressed or misfolded,clear neurotoxicity,improve dopaminergic neuron activity,and protect dopaminergic neurons.4.The effect mechanism of electroacupuncture on Parkinson’s disease model rats may be similar to that of electroacupuncture at "Fengfu"(GV16)and "Taichong"(LR3)points,which can effectively activate the ATF4/Parkin signaling pathway,relieve endoplasmic reticulum stress,protect DA neurons,and prevent Or delay the progression of Parkinson’s disease. |
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