| BackgroundRecently, depression syndrome model, as empirical research has been widely adopted by others. The animal model is on the basis of the experimental study of depression syndrome, is also the key of the success or failure of the reaserch. Depression syndrome model simulate specific TCM (Traditional Chinese medicine) syndrome on base of traditional Chinese medicine diagnosis and treatment theory by modern technology to offer the clinical ideal experimental observation object.TCM syndrome model must be base on TCM etiology theory.To take into account the critical role of emotional disorders in liver depression, the pathogenesis have appeared a variety of models both at home and abroad, mainly two aspects: First, using excess or toxic drugs to induce acute and chronic liver poisoning injury to simulate depression model; Secondly, to change the physiological status of the animal to replicate the model by emotional stimulation.Drug modeling, acute and chronic toxicity can cause liver substantial damage,TCM depression Syndrome does not mean to substantial damage to the liver, and drugs also affect the animal multiple organ system function. Drug modeling had a significant difference from TCM theory, it has been an unapproved by others, phased out gradually.The kernel institutions to regulate the body’s is five organs. In the psychological aspects:the liver plays a decisive role in this procedure.To adjust the Qi caused by a variety of external stimulation, mainly regulated by the liver. Liver have a catharsis function, control the blood circulation. Liver regulate the catharsis that is it have the physiological characteristics of dispersion, catharsis. The normal function of liver catharsis can circulate systemic blood smoothly, regulate the emotions, digestion and excretion of dietary cereal better, semen excretion and help to reach to whole body. Therefore, once liver fail up to charge of Qi, will lead to liver dysfunction of catharsis, air-block pathological changes will happen first, then hepatic depression syndrome will be formed. Hepatic depression is an extensive syndrome, common disease in clinic and common syndromes in TCM viscera syndrome, also is one of the major aspects of liver disease pathogenesis in TCM. Therefore, explore the substance of hepatic depression syndrome can provide new ideas and methods for clinical diagnosis and prevention of many diseases.Generally, liver depression disease and syndrome produce many unmoral emotions due to a variety of factors result from diet disorder, maladjustment, drug misconduct.Liver emotional disorders, such as depression, mood restless due to liver catharsis is not sufficient. From the pathological mechanisms of psychological stress and the specific features of various organs in Chinese medicine," liver" is the core which the body regulate the psychological stress response.Under abnormal emotional (psychological stress), Qi of body will be affected firstly, it can cause circulation and functional abnormalities of Qi and blood for further development. From the pathological mechanisms of psychological stress and the specific features of various organs in Chinese medicine," liver" is the core which the body regulate the psychological stress response. Under abnormal emotional (psychological stress), Qi of body will bet firstly affected, it can cause circulation and functional abnormalities of Qi and blood for further development.Recently, from the physiological pathology of the liver research, the changes of the liver physiological function and pathological involve in the multiple systems, and the nervous,endocrine, immune etc. For example,When stagnation,the central neurotransmitter norepinephrine, epinephrine, serotonin amine,dopamine, skin potential, several catecholamines in the blood, adrenal organizational structure, and CD4+/CD8+in immune system will be changed; and more associated with sympathetic abnormalism.This show that liver function in TCM closely relate to Western medicine neuro-endocrine-immune network. Research on liver further show that the physiological function and pathological changes of the liver is involved in many systems, the nervous, endocrine, immune, digestive, cardiovascular system diseases are be observed frequently, suggesting that TCM liver function are closely related neuroendocrine disorders, immune dysfunction caused by poor emotional state and psychological stress.Studies suggest that hepatic depression syndrome generalize about the nervous, endocrine, immune, circulatory, digestive, sensory and other systems of certain pathological changes depression, anxiety, grief, stress, premising on nerve center of the adjustment disorder. Although there is no stress formulation in Chinese medicine classical medical books, the ancient physicians had early recognized that a variety of adverse stimuli effects on the body, they attached great importance to the influence of emotional factors on the body. According to the theory of Zang organs,Liver control dispersion is one of a liver basic physiological functions, it plays a crucial role in regulating entire body, the liver of the physiological, psychological functions relate with the flow of Qi. In recent years, studies of the liver parenchyma further show that the physiological functions and pathological changes of the liver relate to many of the body system and the nervous, endocrine, immune, digestive, cardiovascular system diseases, which indicates that liver function are closely related with neuroendocrine disorders, immune dysfunction caused by bad emotional and psychological stress.The rearch of stress involved in the hypothalamic-pituitary-adrenal cortex system (HPA), the locus coeruleus-sympathetic-adrenal medullary system (LC-NE) and so more comprehensive in abroad than our country. In our country, although the research focus on the relationship of the liver and neuroendocrine system is popular, the research focus on the HPA axis, the hypothalamic-pituitary-thyroid axis (HPT) or the hypothalamic-pituitary gonadal axis (HPGA).The material basis of hepatic depression syndrome relevance of neurotransmitters of LC-NE secretion under chronic restraint stress has not been reported.Thereby,we try to explore liver function of controlling dispersion is of great significance base on the Chinese medicine theory.Research purposesTo observe the general state of rats in the liver depression model, body weight, behavior, serological NE level and histological evaluation model to validate the model.Detected protein and gene expression of the synthetic enzyme TH and DBH in LC-NE system and neuropeptides CRF, and observing variation level after Xiaoyao powder and fluoxetine intervention. Explore the liver substantiality in the nervous system in LC-NE system, and offer a evidence for liver substantiality research.Research Methods(1) Establishment of chronic restraint stress liver depression rats modelGroups:5-weeks-old72SD rats(150±20) g were randomly divided into four groups with18rats in each group after one-week’s adaptation living:group A(control group), group B(model group), group C(Xiaoyao Powder treatment group),fluoxetine treatment group (D),The rats from the group B,C and D were housed individually in cages while rats from group A were be housed in two cage.Model establishment:Rats from group B, C and D were be bound in a specific bound frame, fixed body and tail. Restraint time were be randomly and gradually durated1day1h to6h at day21th.Drug Intervention:Rats from group C, D were deprived from diet and water during chronic immobilization stress. The rats in group A did not give any other intervention.At16:00pm daily, Rats from group A, B were given2ml physiological saline by intragastric administration. Rats from group C were given Xiaoyao powder with the same ways according to body surface area converted into rat dosage(10g/kg.d), Group D rats were given fluoxetine (5mg/kg.d).Model evaluation:mode reliability will be evaluated by the observation of behavior, fluid consumption test, serum NE level, drug intervention efficacy.2. H&E staining to observe the morphological changes in rat LC and the changes of Nissl bodies by Nissl staining.3. Expression of TH, DBH in LC and neuropeptides CRF detected by immunohistochemical method and analyzed with Imagepro plus6.0.4. Expression of TH, DBH in LC and neuropeptides CRF detected by Western blotting.5. Expression of TH, DBH in LC and neuropeptides CRF detected by Real-time fluorescence quantitative (real-time PCR).6. All data were be statistically analyzed using SPSS17.0software package. The measurement data results show in x±s.Statistics of comparison between the two groups were be used t-test;Statistical comparison among multiple groups will be chose One way ANOVA;The first test for homogeneity of variance of the data, homogeneity of variance with LSD method; if the data is the heterogeneity of variance, the Welch method of variance analysis;Comparison between each two by Dunnett T3method;Hierarchical data using Kruskal-Wallis H nonparametric test analysis.P<0.05was considered significant difference.Results1. Group B rat model manifested irritability, struggle, and then manifested as fatigue, lack of exercise preferences adherent sounds weak and thin, unresponsive, eating less, accompanied by the coat dull yellow stool thin pond Syndrome of depressed performance. C and D rats were mild, while no abnormal behavior state in group A. The rats’ weight increased and significantly different from other groups (P<0.01). It showed rats in group B grew the slowest, while rats in group C, D has been rectified after durg intervention. In open field test,The erect time,grooming time and traversing grid also reduced in group B(P<0.01).Fluid consumption test revealed that group B lost appetite for sweet water during the whole procession. Statistics showed there were significant differences between these four groups (P<0.01).2. The serum level of NE was also increased in rat model(P<0.01).3. H&E staining showed that the LC morphology in rats are normal. The Nissl visible neurons and the number. of Nissl body enhanced, but model group has no obvious morphological abnormalities.4. Immunohistochemical staining showed positive neurons of the TH, DBH and neuropeptides of CRF in group B were greater than group A (P<0.05), the staining intensity was significantly enhanced, the average optical density values significantly higher than group A (P<0.05). Treatment group showed a downward expression.5. Western blot showed TH, DBH and neuropeptides of CRF in LC. Compared with group A, TH, DBH and CRF protein expression was significantly higher (P <0.05) in Group B, Group C and D significantly lower than group B (P<0.01).6. Real-time fluorescence quantitative (real-time PCR) detection TH, DBH and neuropeptides of CRF mRNA in LC. Compared with group A, TH, DBH and neuropeptides of CRF mRNA in group B significantly higher (P<0.05), Group C and D significantly lower than group B (P<0.01).Conclusion1.The rat model of liver depression syndrome from chronic constraint stress is reliable and credible.2. In chronic restraint stress process, no obvious abnormalities in the morphology of the rat LC, no obvious abnormalities; Nissl staining revealed that the LC neurons Nissl body staining increased.But there were no obvious morphological abnormalities in model group.This showed that chronic stress(21days) does not cause morphological changes in rat LC tissue.3. This project firstly explored the hepatic depression syndrome in Locus coeruleus-noradrenergic system, and provides a theoretical basis for the further study of the molecular mechanisms of hepatic depression syndrome, and also give a beneficial trial for deeper research of the essence of liver depression syndrome.4. Chronic restraint stress activated rat LC-NE system to release catecholamines NE, thereby TH and DBH level was increased in synthase NE neurons; at the same time, stress activating the neuropeptide CRF promoted LC-NE neurons activity, with the formation of LC-NE system interaction, causing the body’s excitement, alertness, tension and anxiety and other emotional responses.However, Xiaoyao powder treatment can effectively reverse the effect. |
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