Study On The Effect Of Acupuncture On GABA In Rats With Insomnia Model Based On ERK Regulating MAP-2 Pathway

Objective:Using the ERK signaling pathway and the skeletal protein map-2 in the cell,in order to discuss whether the effect of acupuncture on GABA in rats with insomnia model was transmitted through a series of signal transduction,from stimulated,the cytoskeleton protein transports neurotransmitter,step by step,to the relevant target cells to produce the neurotransmitter which show the therapeutic effect of acupuncture on the disease.In addition,acupuncture treatment affect the release of neurotransmitter by regulating the cytoskeleton proteins of neurons,which can regulate insomnia.Methods:In the experiment,selected 60 male healthy clean grade SD rats(200-220g),after adaptation for a week,50 rats in normal physiological state were screened for activities,diet,sleep,hair color,eyes and so on.The 50 rats were divided into 5 groups by the random method of digital tables.In the blank group 10,intraperitoneal injection of saline injection the proportion according to body weight 100g/mL.Groups that need to build modules include model group 10,acupuncture group 10,inhibitor group 10,and inhibitor+acupuncture group 10,the model was used for the intraperitoneal injection of chlorphenylalanine(PCPA),and injection of 100g/mL of weight for two consecutive days.After modeling,the model was judged to be successful according to the behavioral characteristics of rats.Acupuncture group 10,after two days of modeling,the acupuncture treatment was performed for 7 days consecutively;The inhibitor group 10,were injected with ERK inhibitor at 30mg/kg each day on the 3rd day,7 days in total;Inhibitor+acupuncture group 10,injection ERK inhibitor after acupuncture treatment,total 7 days.After experiment,the rats brain tissues were taken,anesthesia in rats after disinfection in each group of 5 groups,part of the rats were selected for beheaded processing,open the skull with surgical instuments on a plate of dry ice,stripping brain tissues,and remove the hypothalamus organizations carefully.In addition,the remaining rats were anesthetized.Supine fixed on the plate,with surgical scissors to open chest skin and subcutaneous tissue,exposed chest,ribs cut open to expose the heart,after using a syringe needle from the left ventricle after intubation immediately cut to the right atrial appendage.Fixed treatment on rat brain tissue,finally fixed brain tissue is taken out for detection.The protein expression was detected by Western-blot and immunohistochemistry,and the number of synaptic vesicles in GABA neurons was observed by electron microscopy.Using SPSS16.0 software for statistical analysis,one-way analysis of variance and the normal distribution test was performed.Results:1.Building after 24-48h,a module in the rat began to appear manic,part of an erect posture,increased food intake,activities during the day,and the time of sleep was reduced.There was a significant difference between the model group and the model group,which proves that the model is successful.2.According to the results of ERK in rats hypothalamus the blank control group to compare model group was significantly higher(P<0.05);Compared with the model group,the acupuncture group was significantly lower(P<0.01),the inhibitor group was significantly decreased(P<0.01),and the inhibitor+acupuncture group had no significant changes(P>0.05);Compared with the acupuncture group the inhibitor group and the inhibitor+acupuncture group(P>0.05)did not change significantly;the inhibitor group was compared with the inhibitor+acupuncture group(P>0.05).3.The results of MAP-2 in the hypothalamus of rats showed that in the model group there was significantly lower than that in the blank control group(P<0.05).Compared with model group the relative content of MAP-2 in the acupuncture group increased significantly(P<0.01),the inhibitor group increased(P<0.01),and the inhibitor+acupuncture group increased significantly(P<0.01).Compared with the acupuncture group there were no obvious change in the content of MAP-2 in the inhibitor group and the inhibitor+acupuncture group(P>0.05).The content of the inhibitor+acupuncture group increased compared with the inhibitor group(P>0.05).4.The results of GABA in the hypothalamus of rats showed a significant decrease in the comparison model group with the blank control group(P<0.05).Compared with the model group,the acupuncture group,inhibitor group,inhibitor+acupuncture group was significantly increased(P<0.01).Compared with the acupuncture group,the inhibitor group and the inhibitor+acupuncture group had no statistical significance(P>0.05).The inhibitor+acupuncture group had no statistical significance compared with the inhibitor group(P>0.05).Conclusions:1.Acupuncture does participate in ERK1/2 signal regulation mechanism,and acupuncture has inhibitory effect on ERK expression.2.Acupuncture and ERK inhibitors can increase the content of MAP-2 and GABA in hypothalamus of insomnia model rats.3.Acupuncture may inhibit the expression of ERK and increase the expression of MAP-2,and eventually increase the content of GABA.