| ObjectiveDepression is a common mental disorder with complicated pathogenesis, it has a high prevalence rate, high recurrence rate, high disability rate and high cost of medical treatment.Depression is closely related to stress and with the change of the social economic framework, the psychological stress from all aspects gradually occupy the leading position in the occurrence and development of depression. Studies have indicated that chronic psychological stress can cause degenerative changes in the nervous system, which not only can cause significant physical symptoms, but also can bring about cognitive impairment, which can lead to depression.The core of psychological stress induced depression is to make the neuron apoptosis in the hippocampus of the brain and the key intermediate mechanism may involved in the reactive oxygen species,which had induced oxidative stress. It directly damaged protein, DNA and biological membrane and activated the mitochondrial apoptosis pathway to promote the apoptosis of hippocampal neurons. Because the hippocampus is the important brain area in the pathogenesis of depression, how to inhibit the apoptosis of hippocampal neurons become the key to the treatment of depression.Due to the early symptoms of psychological stress only in inattention and insomnia and other similar neuro endocrine disorders, long-term psychological stress will cause unimaginable consequences when these mild symptoms are often ignored by patients and can not be treated in a timely manner. In recent years, due to the shortage of anti-depression drugs, such as toxic and side effects, depression has gradually become one of the advantages diseases of acupuncture treatment. which has obvious clinical curative effect, little side effect, and has obvious advantages in improving the early stage of psychological stress, anxiety, work and interest and sleep and other symptoms. In addition it can inhibit the apoptosis of hippocampal neurons in order to maintain the survival of neurons in the hippocampus, so as to play an anti-depression effect.In the previous study, our team found that acupuncture can improve the apoptosis of hippocampal neurons from the signal pathway and related factors.But there is little research about the relationship with psychological stress-- oxidative stress--mitochondrial apoptosis pathway--depression. Related report that whether acupuncture can improve the mechanism of psychological stress induced apoptosis is involved in the regulation of oxidative stress and inhibition of apoptosis pathway is also very rare.Using fluorescence probe technique and Western blotting to detect content of reactive oxygen species and protein expression levels of cytochrome c, caspase-3 and AIF.In this paper, we continue the study of acupuncture on apoptosis,and to establish a rat model of chronic psychological stress by binding method,which can simulate the process of psychological stress in the pathogenesis of human depression, through investigating behavior, open field test and sucrose preference test evaluation of acupuncture on chronic stress rat model of depression anhedonia behavior. We selected mitochondrial apoptosis pathway and oxidative stress, which is closely related to psychological stress, as the research focus. And to used acupuncture on "Baihui" and "Yintang" and "Sanyinjiao" points as the intervention measures.In addition, fluoxetine was selected as a positive control drug. To study the regulating effect of acupuncture on the content of reactive oxygen species induced by psychological stress and whether the anti-apoptotic mechanism of acupuncture is involved in the inhibition of mitochondrial apoptosis pathway. The aim of the study was to establish the anti-apoptotic network of acupuncture, to improve the anti-depression mechanism of acupuncture, and to provide experimental evidence for the prevention and treatment of depression induced by psychological stress.Methods1.Experimental groups:32 male SD rats were divided into 4 groups:control group, model group, acupuncture group and fluoxetine group according to the random number table. The rats in each group were fed for one week before the experiment.2. Modeling methods:In this study, we established the animal model of chronic psychological stress by using the internationally recognized chronic restraint stress rat model. The specific method:using barbed wire, Both ends of whicn are fixed by a clamp. The aim was to keep the breathing unobstructed, and not to oppress the rats. The rats were only subjected to psychological stress, which basically controlled the effect of physiological stress. The results mainly reflected the effect of psychological and emotional factors on the rats. Restraint stress time for the daily 9:00-15:00, during the period of restraint, intake and water deprivation. After the end of the shackles, the rats were put into mouse cage, free food intake.28 days in a row.3. Intervention methods:(1) control group:conventional breeding for 28 days, only in the 9:00-15:00 daily intake and water deprivation, the rest of the time free feeding water and food.(2) model group:chronic restraint combined with isolated for 28 days, restraint stress time for the daily 9:00-15:00, during the period of restraint, intake and water deprivation, the rest of the time free feeding water and food.(3) acupuncture group:chronic restraint combined with isolated for 28 days, restraint stress time for the daily 9:00-15:00, during the period of restraint stress, intake and water deprivation, the rest of the time free feeding water and food. At the same time, selecting the "Baihui, Yintang" and "Sanyinjiao" points to acupuncture intervention,1 hour before restraint stress,retaining needle for 20 minutes, once a day, continuous 28 days.(4) fluoxetine group:chronic restraint combined with isolated for 28 days, restraint stress time for the daily 9:00-15:00, during the period of restraint stress, intake and water deprivation, the rest of the time free feeding water and food. At the same time,useing fluoxetine administered intervention 30 minutes before restraint stress, once a day, continuous 28 days.4 Detection index and methods:Using open-field test, body weight and sucrose preference test to evaluate change of depression model rats and using fluorescence probe technique and Western blotting to detect content of reactive oxygen species and protein expression levels of cytochrome c, caspase-3 and AIF in the hippocampus of rats.Results1. behavioral results:Before the experiment, there was no significant difference among groups (all P>0.05). After 28-day stress procedure, the model group showed a significant decrease of the number of crossing and rearing, sucrose consumption, sucrose preference index and the body weight compared to the control group (P<0.01); compared with the model group, in the open field test,acupuncture group and fluoxetine group showed a significant increase of the number of crossing and rearing (all P<0.01),and the improvement effect of acupuncture was significantly better than that of fluoxetine (P<0.01, P<0.05). in the body weight, acupuncture group and fluoxetine group showed a significant increase(P<0.01, P<0.05); in the sucrose preference test, acupuncture group and fluoxetine group showed a significant increase of sucrose consumption and sucrose preference index (all P<0.01).2.The content of reactive oxygen species (ROS) in the hippocampus:After 28-day stress procedure, the model group showed a significant increase of the content of reactive oxygen species in the hippocampus compared to the controll group (P<0.01); compared with the model group, acupuncture group and fluoxetine group showed a significant decrease of the content of reactive oxygen species (ROS) in the hippocampus (all P<0.01).3.The protein expression level of cytochrome C and caspase-3 in the hippocampus:After 28-day stress procedure, the model group showed a significant increase of the protein expression level of cytochrome C and caspase-3 in the hippocampus compared to the control group (all P<0.01); compared with the model group, acupuncture group and fluoxetine group showed a significant decrease of the protein expression level of cytochrome C and caspase-3 in the hippocampus(all P<0.01). But in the down-regulation of caspase-3 protein expression level, the improvement of acupuncture has obvious advantages (P<0.01).4. The protein expression level of AIF in the hippocampus:After 28-day stress procedure, the model group showed a significant increase of the protein expression level of AIF in the hippocampus compared to the controll group (P<0.01); compared with the model group, acupuncture group and fluoxetine group showed a significant decrease of the protein expression level of AIF in the hippocampus(all P<0.01).Conclusions1. Acupuncture and fluoxetine intervention can improve the expression level of behavioral in rats with depression. Acupuncture was superior to fluoxetine in the improvement of exploratory behavior and physical activity status.2. The combination of solitary and chronic restraint stress stimulation can brings about oxidative stress, leading to excessive amounts of reactive oxygen species in the rat hippocampus. Acupuncture intervention can effectively reduce the content of reactive oxygen species in the hippocampus of rats, and inhibit the series of adverse effects caused by oxidative stress.3. Acupuncture intervention can significantly reduce the protein expression level of cytochrome c and caspase-3,AIF in the hippocampal of model rats which inhibit hippocampal mitochondria apoptosis pathway activation may be related to the down-regulation of the content of reactive oxygen species.4. According to the relationship between psychological stress, oxidative stress, mitochondrial, it is suggested that acupuncture may be regulating chronic psychological stress--oxidative stress-- mitochondrial apoptotic pathway this special pathway on hippocampal neurons apoptosis to improve the antidepressant effect. |
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