Effect Of Acupuncture With "The Method Of Soothing The Liver And Regulating The Spirit" On NE And CORT In CUMS Rats With Hyper-cholinergic Tones

ObjectiveTo observe the effect of acupuncture with "the method of soothing the liver and regulating the spirit" on prefrontal noradrenaline(NE)level and serum cortisol(CORT)level in Chronic Unpredictable Mild Stress(CUMS)rats with hyper-cholinergic tones;explore the mechanism of acupuncture on antidepressant.Methods50 SPF male Sprague-Dawley(SD)rats were employed in the experiment,weight 200±20 g and carried out in accordance with the method of stratified random sucrose preference rate and weight factors of rats group and divided into the control group;model group;electro-acupuncture(EA)group;physostigmine(PHYS)group and EA+physostigmine group.A total of 5 groups,each group of 10rats.21 days chronic unpredictable mild stress was used to build depression model rats,except for the control group.The body weight and sucrose preference test were conducted on the 21st day of modeling,to evaluate the success of the CUMS depression model.The intervention began on the 22 nd day,after successfully modeling to apply electro-acupuncture or drugs were administrated for 3 weeks except for the control group and model group.The acupuncture with "the method of soothing the liver and regulating the spirit"includes baihui GV-20,yintang Ex-HN3,siguan(a combination of hegu LI-4 and taichong LR-3),running the needles were not used after acupuncture,without electricity just retained the needling of baihui,yintang.Electroacupuncture apparatus Hans-LH202H,density wave,frequency 2/100 HZ and current 1～1.2 mA were used acupuncture was performed,every time 20 min,once a day for 3 weeks.The drugs were physostigmine(acetylcholinesterase inhibitor),0.5mg/kg daily,intraperitoneal injection,once a day for 3 weeks.This study is divided into two experiments:Experiment 1:The effects of electro-acupuncture on behavior in CUMS rats with hyper-cholinergic tones.After the intervention,the body weight,sucrose preference test,novelty-suppressed feeding test and open field test were conducted.Differences in each indicator among five groups were compared,to observed the effects of acupuncture with“the method of soothing the liver and regulating the spirit”on the body weight and behavior in CUMS rats with hyper-cholinergic tones.Experiment 2:The effects of electro-acupuncture on prefrontal NE level and serum CORT level in CUMS rats with hyper-cholinergic tones.After the behavior test,six rats were selected from each group,serum and prefrontal tissue were collected.Differences in each indicator among five groups were compared,to explore the effects of acupuncture with“the method of soothing the liver and regulating the spirit”on prefrontal NE level and serum CORT level in CUMS rats with hyper-cholinergic tones were detected by enzyme-linked immunosorbent assay(ELISA).ResultsExperiment 1:The effects of electro-acupuncture on behavior in CUMS rats with hyper-cholinergic tones.The model of chronic unpredictable mild stress(CUMS)depression was successfully established in rats.After the modeling,the body weight and the sucrose preference rate differenced in rats between the five groups(P<0.001).Compared with the control group,the body weight and the sucrose preference rate decreased significantly(P<0.001,P<0.001)in the model group.After the intervention,the body weight,the sucrose preference rate,the latency time to food,level activities,vertical activity times and number of grooming differenced in rats between the five groups(P<0.001).Compared with the control group,the body weight and the sucrose preference rate decreased significantly(P<0.001,P<0.001)and the latency time to food prolonged significantly(P<0.001);level activities,vertical activity times,number of grooming decreased significantly(P<0.001,P<0.001,P<0.001)in the model group.The results showed that the rats developed depressive symptoms after the successful modeling.Compared with the model group,the body weight and the sucrose preference rate increased significantly(P<0.001,P<0.001)and the latency time to food shortened significantly(P<0.001),level activities,vertical activity times,number of grooming increased significantly(P<0.001,P<0.001,P<0.001)in the electro-acupuncture group(EA group).The results showed that electro-acupuncture could improve the depression-like behavior of CUMS-depressed rats and alleviated the depressive symptoms of weight loss,lack of pleasure,lack of interest and reduced activity in rats.Compared with the model group,the depressive behaviors aggravated in the physostigmine group(PHYS group),the body weight and the sucrose preference rate decreased significantly(P<0.001,P<0.001)and the latency time to food prolonged significantly(P<0.001),level activities,vertical activity times decreased significantly(P<0.001,P<0.001)and the results number of grooming were not reliable enough,there was no statistically significant difference(P>0.05)in the PHYS group.Physostigmine group rats in depression behavior were aggravated,may be related to cholinergic excessive activation.Compared with the PHYS group,the depressive behaviors were improved,the body weight and the sucrose preference rate increased significantly(P<0.001,P<0.001);level activities,vertical activity times and number of grooming increased significantly(P<0.001,P<0.001,P<0.001)in the electro-acupuncture group and the electro-acupuncture+physostigmine group.Compared with the PHYS group,the latency time to food was no statistically significant difference(P>0.05)in the electro-acupuncture+physostigmine group;but the latency time to food shortened significantly(P<0.001)in the electro-acupuncture group.Indicating that electro-acupuncture effectively relieved hyper-cholinergic tones to improve the symptoms of depressive rats.Experiment 2:The effects of electro-acupuncture on prefrontal NE level and serum CORT level in CUMS rats with hyper-cholinergic tones[1]We found that after the intervention,the prefrontal NE level and serum CORT level differenced in rats between the five groups(P<0.05).Compared with the control group,the prefrontal NE level decreased significantly(P<0.001)and serem CORT level increased significantly(P<0.05)in the model group.Showing that the modeling had been successfully because under the depression rats show the NE is low and CORT is high.Compared with the model group,the prefrontal NE level increased significantly(P<0.001)and serum CORT level decreased significantly(P<0.05)in the EA group,showing that electro-acupuncture improved the symptoms of depressive rats.Compared with the model group,the prefrontal NE level decreased(P=0.05)and serum CORT level increased significantly(P<0.001)in the PHYS group.It indicates hyper-cholinergic tones lead to the depression was aggravated in rats.Compared with the PHYS group,the prefrontal NE level increased significantly(P<0.001)and serum CORT level decreased significantly(P<0.001)in the electro-acupuncture group and the electro-acupuncture+physostigmine group.It showed that the electro-acupuncture had effected of relieving hyper-cholinergic tones lead to improve the symptoms of depressed rats.ConclusionAcupuncture with "the method of soothing the liver and regulating the spirit" improved the behavior of depression in rats.Its internal mechanism may be that electro-acupuncture related to effectively increase the content of prefrontal noradrenaline,decrease the content of serum cortisol and remove the hyper-cholinergic tones,reduce the susceptibility of rats to stress.