Study On The Difference And Mechanism Of Different Acupoint Selection Methods In The Treatment Of Primary Insomnia (Spleen Stomach Disharmony Type) Based On The Theory Of "Brain Gut Interaction"

Objective:In this paper,randomized controlled trials were conducted to evaluate the difference in clinical efficacy of acupuncture in the treatment of primary insomnia（spleen-stomach dissonance）with different acupoint selection methods.Meanwhile,ELISA was used to detect the content of brain intestinal peptide in serum of patients and the changes in brain neurotransmitter levels were recorded by brain function fluctuation analysis（ET）.To explore the difference of physiological and neurological effects of different acupoint selection methods in the treatment of primary insomnia（spleen and stomach disharmony type）,in order to find the best acupoint selection scheme for the clinical treatment of primary insomnia（spleen and stomach disharmony type）.The morphological changes of rat hippocampus were observed by electron microscopy in animal experiments,and the content of 5-HT,protein expression of 5-HT_1Aand 5-HT_2A,relative expression of 5-HT_1AmRNA and5-HT_2AmRNA in rat hypothalamus were detected by ELISA combined with Western blot and RT-PCR.To explore the central effect mechanism of different acupoint selection methods in the treatment of primary insomnia（spleen and stomach dissonance type）,and to provide objective basis for the best acupoint selection scheme in clinical treatment of primary insomnia（spleen and stomach dissonance type）.Methods:Clinical trials:96 patients with primary insomnia（spleen and stomach disordered type）were randomly divided into acupoint selection group with symptomatic combination of syndrome differentiation,select acupoint group according to the symptoms,and selection group based on syndrome differentiation.Each group was given acupuncture treatment for a course of 4 weeks.Pittsburgh Sleep Index Scale（PSQI）,TCM Syndrome Scale,Gastrointestinal Symptom Score Scale（GSRS）and clinical effective rate were used for clinical efficacy.ELISA was used to record the content changes of brain and intestinal peptides（5-HT,VIP,SS,GAS,MTL,etc.）in serum before and after treatment.The secretion levels of neurotransmitters（5-HT,GABA,Glu,ACh,NE,DA,etc.）in the brain of patients before and after acupuncture treatment were recorded by brain function fluctuation analysis（ET）.Animal experiment:The model of insomnia rats with discordant spleen and stomach was replicated by iodine acetamide administration combined with intraperitoneal injection of chlorophenylalanine（PCPA）,and the rats were randomly divided into blank group,model group,acupoint selection group with symptomatic combination of syndrome differentiation,select acupoint group according to the symptoms,and selection group based on syndrome differentiation.The general state of the rats was observed and the changes of body weight were recorded.Sleep latency（SL）and Sleep Duration Time（ST）;The morphological differences of hippocampus were observed by electron microscopy,and the content of 5-HT and the relative expression levels of 5-HT_1Aprotein,5-HT_2Aprotein,5-HT_1AmRNA and 5-HT_2AmRNA in hypothalamus of each group were detected by ELISA,Western Blot and RT-PCR.Results:Clinical trial:1.Through the comparison of PSQI,TCM syndrome scale and GSRS,it was found that compared with the select acupoint group according to the symptoms and selection group based on syndrome differentiation,there were significant differences in PSQI,TCM syndrome scale and GSRS in acupoint selection group with symptomatic combination of syndrome differentiation（P<0.05）.2.The clinical effective rate was 87.50%in the acupoint selection group with symptomatic combination of syndrome differentiation,75.00%in the select acupoint group according to the symptoms,and 68.75%in the selection group based on syndrome differentiation.3.ELISA was used to detect the contents of brain and intestinal peptides in serum of patients before and after treatment,and it was found that the contents of5-HT,Gas and MTL in the select acupoint group according to the symptoms were significantly different from those in the acupoint selection group with symptomatic combination of syndrome differentiation（P<0.05）.Compared with the selection group based on syndrome differentiation,the content changes of 5-HT,VIP and SS in the acupoint selection group with symptomatic combination of syndrome differentiation were statistically significant（P<0.05）.4.The comparison of ET recording of neurotransmitter secretion levels in the brain of patients before and after treatment showed that there were significant differences in the levels of Ach,5-HT and DA in select acupoint group according to the symptoms compared with that in the acupoint selection group with symptomatic combination of syndrome differentiation（P<0.05）.Compared with the selection group based on syndrome differentiation,there were significant differences in 5-HT and DA levels in the acupoint selection group with symptomatic combination of syndrome differentiation（P<0.05）.Animal experiment:1.Behavioral observation:compared with the select acupoint group according to the symptoms and selection group based on syndrome differentiation the rats in the acupoint selection group with symptomatic combination of syndrome differentiation had significantly increased body weight,prolonged sleep time,and shortened sleep latency（P<0.05）.2.HE hippocampus staining:compared with the model group,the hippocampal neurons in the acupoint selection group with symptomatic combination of syndrome differentiation,the select acupoint group according to the symptoms and selection group based on syndrome differentiation were arranged relatively neatly,the cell volume was relatively uniform,and the cell contour was relatively clear.3.Electron microscopy observation:compared with the model group,the mitochondrial structure of hippocampal neurons in the acupoint selection group with symptomatic combination of syndrome differentiation,the select acupoint group according to the symptoms and selection group based on syndrome differentiation was relatively orderly,the chromatin distribution was relatively uniform,and a small number of heterochromatin was clustered.4.Hypothalamus ELISA test showed:Compared with the select acupoint group according to the symptoms and selection group based on syndrome differentiation,the content of 5-HT in the hypothalamus of the acupoint selection group with symptomatic combination of syndrome differentiation increased,and the difference was statistically significant（P<0.05）.5.WB showed that the relative expressions of 5-HT_1Aand 5-HT_2Aprotein in the hypothalamus of the acupoint selection group with symptomatic combination of syndrome differentiation were significantly different from those of the select acupoint group according to the symptoms and selection group based on syndrome differentiation（P<0.05）.6.RT-PCR showed that the relative expressions of 5-HT_1AmRNA and5-HT_2AmRNA in the hypothalamus of the acupoint selection group with symptomatic combination of syndrome differentiation were significantly different from those of the select acupoint group according to the symptoms and selection group based on syndrome differentiation（P<0.05）.Conclusion:1.Acupoint selection method with symptomatic combination of syndrome differentiation can improve sleep quality,improve gastrointestinal functional symptoms and clinical effective rate,increase the content of brain intestinal peptide 5-HT,Gas,MTL and VIP in serum,and reduce the content of SS.The level regulation effect of brain neurotransmitters 5-HT,Ach and DA is better than that of Select acupoint method according to the symptoms and Selection method based on syndrome differentiation.2.Based on the difference of clinical effective rate,physiological and neurological effects of the three acupoint selection methods,the best acupoint selection method in clinical acupuncture treatment of primary insomnia（spleen and stomach dissonance）is the acupoint selection method with symptomatic combination of syndrome differentiation.3.Acupoint selection method with symptomatic combination of syndrome differentiation can shorten sleep latency,prolong sleep time,increase weight gain,improve hippocampus and ultrastructure,increase hypothalamus 5-HT content,activate 5-HT_1Aprotein and inhibit 5-HT_2Aprotein expression level in insomnia rat model.It was better to activate 5-HT_1AmRNA and inhibit the relative expression of5-HT_2AmRNA is better than that of Select acupoint method according to the symptoms and Selection method based on syndrome differentiation.4.The mechanism of the central effect of acupuncture in the treatment of primary insomnia（spleen and stomach discordance type）by three acupoint selection methods is to increase the content of 5-HT,activate 5-HT_1Aprotein,inhibit 5-HT_2Aprotein expression level,activate 5-HT_1AmRNA and inhibit the relative expression level of5-HT_2AmRNA in insomnia model rats.