Research On Part Of Neuro-immunological Mechanism Of Anti-chronic Psycho Logical Stress Of "Regulating The Liver Function Formula"

Part one: Literature summary and Theory approchPsychological stress is the course of psychological and physiological change resulted by organism that perceive and surge the threats of stress causing psychology through knowing appraise. With the development of society and the increase of pressure from every facet, psychological stress threaten the human heath, as an important fact which can not be ignored, How the psychological and socialological stimulation and the psychological action convert into physiological change, ie from concept to substance, is the key of the psychological stress mechanism.From the perspective of pathological physiology, psychological stress is a series of neuro-endocrine change which caused by intense stimulation. Sympathesis nerve excitation and pituitary-adrenal cortex secretion are the main mechanism. The stress system contain central and external parts. The former mainly include hypothalamic and hippocampus, however, the latter are consist of hypothalamic- pituitary-adrenal cortex axis(HPA) and sympathesis/adrenal system and sub sympathesis system. HPA is the main channel which play a key role in the psychological stress mechanism.Immune system is the out-receptor of HPA, so the infuence on it become more popular. Immunologycal inhibition is the main response. T cell cluster's disorder in quantity and function leads to the disorder in Immune system. Research indicats that HPA is the main channel which play a key role in the psychological stress mechanism. As complement, CRH and ACTH took part in immunoregulation except glucocorticoid. On the other hand, hippocampus becomes the central immunoregulational one. Studies found that normal hippocampus can constrain the excessive response of HPA and lower it to normal level. Furthermore, hippocampus is a sensitive and easy be hurt area in stress. In a word, hippocampus's hurt in stress and HPA's excessive response are the results of infernal circle.The traditional Chinese medicine does not have the concept called "psychological stress", but the bad circumstances and mental stimulation have close relationship with the body disease in state of viscera and seven sentiments theories. Stress is nonspecificity systemic reaction, thus the study of the Chinese medicine for anti-stress must involve many organs. At present, many researches are base on the emotion theory, involving the liver, the kidney, the heart, and the spleen, especially the liver. The detective indexes primarily involve praxiology and neuro-endocrine-immune system. But the conclusion does not sufficient in the holistic level.The studies of our working team indicats that there are some concrete central nerve mechanism in adtion to the whole regulation of the "liver"function (liver controlling dispersion, adjusting emotion)in TCM on hypothalamic-pituitary-adrenal cortex system. Meanwhile there are unity between part and whole in mechanism of action. We take JiaWeiSiNiSan (JWSNS) as a representative regulation liver formula to regulate the psychological stress reaction and have done some relative researches which have tentatively confirmed our investing hypothesis.To make animal model in psychological stress is the foundation, but there are still some vital segments which can't be resolved. For example, the somatic stress factor in the course of previous model-making can't be avoid, which infuenced the experimental results in some extent. Willner p. who established the chronic mild unpredictable stress model (CMUS) solve the problem.Given to the importance in the NIM net, the HPAA and hippocampus become more significant in the regulation of the 'liver'. And with the improvement of animal model, we will get close to the essence of the regulation of the 'liver'. Base on that, we will study further the mchanism of HPAA in chronic psychological stress and the action of Chinese compouned medcine. In order to interpret the mchanism of psychological stress and Chinese compouned medcine in holistic level.Part two: Study target and contentTarget: according to study the mechanism of action of jiaweisinisan (JWSNS) against psychological stress on HPAA and the chang on neuro-endocrine-immune system, we will try to interpret the the mchanism of regulation "liver" theory against psychological stress on neuro-endocrine-immune level, and finally develop a way against psychological stress with TCM. Furthermore, we can reveal the substance foundation and the mechanism of regulation NIM net on multistrata and multitarget from cell and even molecule level.Content: to establish the CUMS animal model and detect the chang of neuro-endoc rine-immune system, so as to observe the regulation action of TCM and discuss the mechanism.Composition and dosage of JWSNS: Bupleurum root 15 g, White peony root 15g, Bitter orange 6g, Wolfberry fruit 15g, Capejasmine fruit 5g, Rehmannia root 18g, Concha haliotidis 30g. The formula are bought from the first affiliated hospital pharmacy of our school which concidered as pure crude herbs by relative department. Finally, we preserved it in refrigerator at 4 centigrade after being prepared. We should provide 2 ml per animal each treatment group for intragastric administration before building mode.Model building and grouping: we take the modified CUMS animal model which to be cancel lightning stroke installation and to be lighten intensity. Model rats were placed in one independent room and accepted different stress stimulus within 21 days. The stressors consist of restraint for 1h, 45°cage tilted for 7h, cage moistened (padding: water=200g:100ml), new intruder added (two cages combined ) for 23 days, persistent illumination for 36h, bottle free lh, fasting diet and water 23h.Section one: The establish of CMUS modelThe target of this experiment is to observe the characters of different ages and sex according to establish of CMUS, and then evaluate it through three facet of face, structure and predictive validity, we saw that, in the course of building model, the model group decline in whole level. Show that, anxiety, dysphoria, increased defecation, reject in some course, shortness of breath, color pattern get darker and even become yellow, the modified behavior decrease even disappear. As compared, the treatment group was between model and normal one, normal group shows best. As to sacchar-concume, the result are similar, although there are transient increase because of baby rats' grouth. But three weeks later it come down as others. Results also similar on weight. Hence, we can come to conclude that JWSNS can relieve stress state effectively, but it can' t make them recovered as long as the stressor exist. The sex is also an important factor. In our experiment, the differences comes from behavior, neuro-endocrine-immune index ect. Compared with male rats, the female one's sacchar-concume present fluctuation state. Say it specificially, that is, decline on first week and then increase in some extent, finally low down again on third week. Base on that, we deduce female' s reaction are more intensity on acute stress and then change into chronic stress state.Section two: JWSNS regulate CUMS on neuro-endocrine-immune systemIn this section, we will observe the change and the way to act of adrenal hormone, thyroid hormone and hebin. This finding show that, female adult rats' T3, T4 of every group don't have obvious chang except the hebin and androgen of adult group. Say it specificially, in male model group, androgen increase obviously(p＜0.01), and then decrease after be given JWSNS. Compared with normal group, there isn't obvious difference. Cue that psychological stress can infuence the periphery sex gland of adult male rats, and can be lower down by JWSNS effectively. However, the female group' androgen didn't lower dow later, which need study further. We have known about the HPAA is the main chanel for chronic psychological stress, the result displayed the certain change of it, and even in different sexes.From the whole level, the result thoroughly displayed the interaction among CRH,ACTH and CORT. At the same time, it also explain the unobvious but changeable result. For example, the general tendency is the activation and accentuation on HPAA, but the situation on female group only showed the tendency without obvious sense. Taking the degression of ATCH as reference, we can deduce the regulation action of it on female' s CORT, ie the elevated tendency get gently because of the decrease of ATCH. The same situation happened to the male group. However, the CRH in hypothalamus are higher than control group obviously, maybe it' s caused by the feedback action for ATCH and CORT in blood plasma.Section three: the regulation of JWSNS on immune function of CMUS modelIn this section we will approach the mutual relationship between immune function and neuroendocrine, and try to identify the action chanel of HPAA. Results show that each model group obviously decrease on thymus gland index, after being given JWSNS both of them rise with the former obviously. It displayed that JWSNS could effectively recover the chang on immune function. From the shape change of thymus gland to improve, we can also found the obvious effect of JWSNS on immunodepression. T cell is the most important immunocell, so we take it for example. Resuts show that, in female model group, CD3+, CD4+ and CD8+ all decrease, especially CD3+ (p＜0.05), and then increase after be given JWSNS, especially CD3+ and CD4+, the three are higher than normal one. We can deduce that the female are more sensitive for stress and medicine; In male group, the three indexes have the same change but without statistical significance.. CD4+ and CDS+ decrease in model group and increase in medicine group both without statistical significance, maybe have relative with the number, which need study further. Compared with male group, the female one's expresion are better which is equal to previous report.The change of adrenal cortex shape, compared with normal group, displayed the excited state of endocrine function of adrenal gland. After be given JWSNS, corticohyp erplassia were restrained and glucocorticosteroid secreted become slow. Combined with the obvious degrade on immune function, we can get the causal relation between the two.Section four: the protection of JWSNS on hippocampus neurone of CMUS modelIn this section we will approach the mutual relationship between the change of hippocampus neurone and the regulation of periphery neuroendocrine, and try to identify the action chanel of HPAA. Hippocampus is not only the senior centers of accommodation but also the area of sensitivity coping with stress hurt. In this experiment, compared with normal group, apoptosis rate of model group both male and female increase obviously, hippocampus optical density value obviously lower than normal group; Compared with model group, hippocampus neurone apoptosis rate of medicine group both male and female decrease obviously, hippocampus optical density value obviously increase. Show that JWSNS can protect hippocampus neurone of CMUS model.Part three: ConclusionFirst, the duplicate and modify of CMUS model are successful, and then evaluate it through three facets of face, structure and predictive validity, we can say there are some similarity between praxiology appearance of rat and emotional expression of human; It is reflected that there are some essential character of emotional disturbance on praxiology and physiology, and collective neurobiology mechanism——the regulation of HPAA etc. Moreover, JWSNS works effectively.Second, the main active path of neuroendocrine against chronic psychological stress is HPA axis. The activation of it is physiological foundation of periphery body reaction and immune function disorder. However, JWSNS can regulate HPA axis effectively, so we could think the mechanism of it have close relationship with HPA axis.Third, chronic psychological stress displayed the obvious change in different sexes. the hypothalamus-pituitarium-thyroid gland axis and hypothalamus-pituitarium-sex gland axis were involved, but it need to be study further.Fourth, immune function were restrained obviously in state of chronic psychological stress, and JWSNS can improve it effectively.Last, hippocampus neurone apoptosis rate of chronic psychological stress rats increase obviously, we can think the regulation of HPAA were influenced by the increase death of hippocampus neurone, and JWSNS can improve it effectively, the mechanism of it have close relationship with hippocampus.Part Four Deficiency and Prospect Because of the limitation of time and fund, the model's duplication and evaluation as well as the whole regulation mechanism on neuro-endocrine-immune system need to be consummated, especially the relative pathes. Another, hippocampus neurone apoptosis is only the start of hippocampus hurt. In the future, we will focus on the validation section of JWSNS against chronic psychological stress.