| ObjectiveA large number of epidemiological studies suggest that Early life stress(Early life stress,ELS)on human emotion,behavior and cognitive process can produce short or long-term influence.Children in the early exposure to adverse life events are more likely to affect the growth of the brain and the steady state,and even lead to their own cognitive disorder or mental illness.Maternal and child separation(maternal separation,MS)refers to the individual life early birth after separated with his mother for some reason.Under normal circumstances,most of the mammals in a period of time after birth and mother life together,and establish a close contact.Mother to infant touch,breast-feeding,and so on various aspects of behavior for the baby has a very active role in their development of the individual.Maternal separation is a kind of early life stress events,its stress on the individual’s way is deprived of before the baby after birth to weaning and mother close contact.At present,in the maternal separation on children’s behavior and the studies of the effects of neural development,rodent maternal separation model is considered to be one of the most effective model.For now,the clinical acupuncture for prevention and treatment of cognitive dysfunction is more,but less application of moxibustion therapy cognitive dysfunction.But moxibustion for the treatment of psychiatric diseases such as depression,anxiety,vascular dementia cases has gradually increased,Moxibustion and acupuncture as a kind of external treatment,its mild stimulus,the characteristics of adverse reactions,in has its unique advantage and practical value.Therefore,this research adopts randomized controlled animal experiment,establish early maternal separation(maternal separation,MS)model,the selection of "baihui" moxibustion therapy,designed to: 1)to observe the early maternal separation model the influence of cognitive function in rats with early adulthood;2)observation of moxibustion on cognitive behavior to improve maternal separation model rats and the effect on 3)to investigate the moxibustion early maternal separation model rats of life influence cognitive function and mechanism of hippocampal synaptic plasticity effect Method1.Maternal separation model of rats.All the male cub and maternal separation group(control group)randomly assigned the 60 th day after birth,including maternal separation(MS)is divided into model group,Moxibustion group(Moxi)and Electroacupuncture group(EA).Normal group: the animal does not accept any stimulation,free access to food and water.Model group:maternal separation after normal breeding animals,free access to food and water.The 81 th day after birth began experimenting.Separated moxibustion groups: animals from mother to child,on the 61 th day after the birth of moxibustion therapy.Treatment time from 61 days to 81 days after birth,moxibustion treatment once a day,for 3 weeks.With maternal separation after fluoxetine group: animals,fluoxetine treatment in the 61 th day after birth.Treatment time from 61 days to 81 days after birth,irrigation to 1 times a day,for 3 weeks.2.Moxibustion and EA therapyMoxibustion treatment: according to the experimental acupuncture on the definition of "baihui",before treatment with acupuncture point as the center,to shaving the hair area of about 2 cm in diameter,exposed the two points of the skin,with a vertical distance rats "baihui",acupuncture point is about 1.5 cm,lit after moxibustion,suspended moxibustion 20 min,treatment1 times a day,treatment 6 days a week,only one day off,continue treatment for 3 weeks.EA treatment methods: cupping treatment with isoflurane anesthesia system for gas anesthesia in rats,according to the experimental acupuncture on the definition of "baihui",for accurate control of electric acupuncture stimulation output,this study use the Israel A.M.P.I.company produces electrical stimulator for electroacupuncture,stimulation parameters for 10 hz frequency,pulse width 50 us,current 1 ma,duration of 15 min,into the needle depth of about 3 mm.The curative time 20 min,treatment 1 times a day,treatment 6 days a week,only one day off,continue treatment for 3 weeks.3.Maternal separation model rats behavioral tests(1)continue to observe and record maternal separation between groups of rats after model establishment weight change,record and analysis,statistics and analysis between groups of rats after building,body weight before and after treatment.(2)3-Chambered Social Test : assessment of maternal and infant separation model rats ability of Social behavior.(3)Morris water maze: detection of maternal and infant separation model of rat hippocampus dependent changes of spatial learning and memory ability.(4)Fear conditioning test: detection of maternal separation model in rats hippocampus dependent changes of learning and memory ability4.In vivo electrophysiological recordings for long-term potentiation1)Power spectrum analyse of local field potential in CA1 area of hippocampus2)In vivo long-term potentiation recording in SC-CA1 area of hippocampus Results1.The weight index 3 weeks after birth(PND21)on maternal separation,no obvious difference was found between each groups;8 weeks before moxibustion and EA treatment before each weight no statistical difference was found;11 weeks after moxibustion and EA therapy,each weight no statistical difference.2.Behavioural tests: in 3-Chambered Social Test,model group compared with control group,after the stimulation rats into social cage explore a significant reduction in the time;The curative group compared with model group,after the stimulation rats into social cage exploration time without statistical significance;Moxibustion group compared with model group,after the stimulation rats into the exploration of social cage time increased.In water maze experiment,the rats in the navigation training stage,the escape latency period decrease with increasing training time,and the control group,moxibustion and cupping group of rat escape latent period were shortened compared with model group(P<0.05),among them,1,2,3,4 days,there were no statistical difference between each rat escape latent period(P > 0.05);The control group in the 5 days of incubation period to avoid fell significantly compared with model group,the difference statistically significant(p<0.05);Escape latency period in 5 days of moxibustion group was obviously shortened,the difference compared with model group(p<0.05);And escape latency of electric acupuncture group in 5 day fell significantly compared with model group,the difference statistically significant(p<0.01).In the conditioned fear experiments,the model group compared with control group,model group of freeze time decreased significantly;The curative group compared with model group,the curative group of freeze time increased,difference have statistical significance;Moxibustion group compared with model group,moxibustion group froze time increases,difference have statistical significance;Groups of rats in the experiment of voice implied conditions fear froze time no significant differences between groups.3.In the body of the hippocampal synaptic long range enhancement(LTP)recorded in the body the seahorse calendar is recorded in the enhanced LTP,given the high frequency stimulation(HFS),MS group after the excitability of outstanding potential(f EPSP)slope significantly lower compared with control group,moxibustion group compared with model group,no obvious difference compared with drug group.Conclusion1)life early maternal separation stress that offspring rats as adults have a certain degree of social behavior disorders,injury associated with learning and memory and other cognitive behavior at the same time.2)moxibustion therapy can more effectively improve maternal and child separation model of rats induced by social behavior,to improve their ability of social behavior,and cupping can significantly improve social behavior disorders;3)moxibustion and cupping can improve maternal separation model rat hippocampus dependent spatial learning and memory;4)electric acupuncture treatment can improve maternal separation model in different degree in the rat hippocampus dependent fear related learning and memory,5)electric acupuncture and moxibustion can significantly increase the maternal separation model rats impaired SC-CA1 synaptic long range enhancement phenomena,including the improvement of the degree.This study confirmed that the moxibustion and cupping therapy on early life of maternal separation stress lead to abnormal behavior and brain of rats hippocampal synaptic plasticity has improved significantly,but the intervention to the improvement of the different behavioral defects have different effect,so our research will better promote the moxibustion,electric acupuncture and the use of the combination for the treatment of early life together of adverse events of individual creation of modern social barriers and cognitive impairment. |
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