Effect Of The Three Roots Balanced Acupuncture Of Mongolian Medicine On Clock Gene Expression In Suprachiasmatic Nucleus Of Sleep Disorder Model Of Depressd Rats

Objective:1.Establish a rat model of depression sleep disorder(SDD),intervene with three roots balanced acupuncture Mongolian medicine,and observe thethree roots balanced acupuncture Mongolian medicine to improve sleep through behavioral detection method.2.Observe the regulation effect of the protein expression of Block,Bmal 1,Per 1 and Per 3 of rats(Suprachiasmatic nucleus,SCN),explore the possible mechanism of the treatment of sleep disorders in depression,and provide a scientific theoretical basis for the clinical application of the three roots balanced needles in the treatment of depression.Methods:24 adult male healthy SD rats,with weight(180 m 20)g,were kept in natural light,well ventilated,temperature(22 ± 2)℃ and humidity(35 m 2%),and adapted for 7 days.One week later,the experimental rats were divided into blank group,model group,western medicine group and Mongolian medicine group by random number table method.Except for the blank group,the other three groups developed a depression sleep disorder(SDD)model using chronic mild and unpredictable stress(chronic unpredictable mild stress,CUMS)combined with the sleep deprivation method of isolation plus rapid eye movement(REM).Model group:1 animal per cage alone,received CUMS+REM sleep deprivation stimulation for 28d,to establish the SDD model.Western medicine group:1 person per cage was kept alone,received CUMS+REM sleep deprivation stimulation for 28d,SDD model was established,1 hour after stress stimulation every day,and agomelatine was prepared into lmg/1 mL at 2mg/kg body weight and normal saline,and administered by gavage.Mongolian doctor group:received CUMS+REM sleep deprivation stimulation for 28d,and received three balanced injections for 1 hour daily after stress stimulation.After 28d,the model was evaluated by behavioral detection of general situation,body weight within 24 hours,sugar water consumption experiment,Open-Field open box experiment,and Morris water maze experiment.Rats were killed on ice and the expression of Clock,Bmal 1,Per 1 and Per 3 genes in SCN by Western-blot and Immunohistochemistry.Results:1.Weight and behavioral experiments:The results of weight gain,sugar water preference,Open-Field open box experiment and Morris water maze experiment of rats before SDD modeling were not statistically significant(p>0.05).After SDD modeling,the weight gain,sugar water preference,and Open-Field Open box scores decreased(p<0.05).Morris Water maze experiment,the platform search time is longer(p<0.05).Compared with the model group,the body weight,sugar and water consumption,and Open-Field open box experimental scores of the western medicine group and the Mongolian medicine group were significantly higher,and the difference was statistically significant(p<0.05).Morris Water maze experiment,the platform search time is shorter(p<0.05).2.Western-blot test results:high expression of Block,Bmal 1 and Per 1,and Per 1 in SCN of blank group,Block,Bmal 1 and Per 1 in SCN of model group(P<0.05),and Block,Bmal 1 and Per 1 in SCN of western medicine and Mongolian medicine group after intervention(P<0.05).The expression of Per 3 protein in SCN was low,while the expression of Per 3 protein in SCN of model group increased(P<0.05),and the expression of Per 3 protein in SCN of Western medicine group and Mongolian medicine group decreased P<0.05).3.Immunohistochemical test results:high expression of Block,Bmal 1 and Per 1,Per 1 in SCN,Bmal 1 and Per 1 in SCN of model group(P<0.05),and Block,Bmal 1 and Per 1 in SCN of western medicine group and Mongolian medicine group after intervention(P<0.05).The expression of Per 3 protein in SCN was low,while the expression of Per 3 protein in SCN of model group increased(P<0.05),and the expression of Per 3 protein in SCN of Western medicine group and Mongolian medicine group decreased P<0.05).Conclusion:1.The results of weight and behavioral tests showed that the weight,absent field level score and vertical score,sugar and water consumption were all significantly increased and the platform finding time became shorter.It shows that the three roots balanced acupuncture Mongolian medicine can significantly improve the sleep disorder of depression,significantly improve the autonomic activity,spatial exploration,learning and memory ability of depression sleep disorder(SDD)model rats,and reduce the level of anhedonia.2.The protein expression trend of Clock,Bmal 1,Per 1 and Per 3 genes in SCN was regulated.According to the results of this study,the course of thethree roots balanced acupuncture Mongolian medicine may change the depression state by affecting the protein expression of Clock,Bmal 1,Per 1 and Per 3 in the brain in depressed sleep disorder rats.In general,the three balance needles can improve the depression state of depression sleep disorder(SDD)model rats,and the mechanism may be related to the expression of Clock,Bmal 1,Per 1,Per 3 and Per 3 in SCN.