The Mechanism Of Regulating Liver Formula-JWSNS Against Stress Induced Depression

Part One Theory Research1. The relationship between stress and depressionStress was Put forward by Han Salye in 1930',ti is a kind if syndrome reaction state which including nerve, endocrinology and Immunity when our bodies facing strongly stimulus from outside world . When facing stressors, the HPA axis(hypothalamic- pituitaryadrenal axis) is excited, the adrenal endocrines much more GC (Glucocorticoids) ,at the same time, the Glucose level in blood is also higher, the immunity function is depressed. Proper stress reaction is good to the body spend short term hard environment, keep living ,but if the body is under stress state for a long time, the HPA axis is over excited, the GC level is too high, the body will be hurt. Research shows: long time strongly stress influence human being's health, different kinds of disease including depression will appear, and the diseases all connected with the exciting of HPA axis and the increasing of GC level in blood. Chronic stress can induce depression is widely accepted now, the behaviors of suicide patients who has been depressed is the same as the symptom induced by chronic stress, the mechanism of which is still unknown, the depression patients has not only the inturbulence of mood, but also hurt of nervous system. Li yunfeng made a hypothesis for the relationship of stress and depression that: chronic stress→, the level of GC is higher→, the GR in hippocampus is not sensitive→the rate of MR/GR is changed→the HPA axis highly excited→the conductive function of NA is decreased→set off EAAs,→the level of Ca²⁺ imbalanced→the express of BDNF mRNA decreased→the hippocampal neurons damaged→depression.2. The mechanism of depression2.1. The traditional hypothesis of monoamine transmitter2.2. The hippocampal neuron damage of depression.3. The mechanism of antidepressant medicine.4. The theory analysis of treating depression by regulating Liver.Part Two the Research Reports Section One The pharmacological research on the antidepressant effect of JWSNSKM mouse, male, 17-22g in weight, providing by animals' center of Guangong Chinese traditional university, Grade SPF.The mouse were raised in quiet environment ,eating and drinking freely, the room temperature was 20-23·C.Composition and dosage of JWSNS: Bupleurum Root 15g,White Peony Root 15g,Bitter Orange 6g,Wolfberry Fruit 15g,Capejasmine Fruit 5g,Rehmannia Root 18g,Concha haliotidis 30g.The herbs are bought from the medical material company of Guangzhou and appraised as pure crude herbs by the drug department, The concentration of crude herbs is 0.294g/ml 0.588g/ml 1.176g/ml after being cooled under the common temperature.1. Tail suspending test 1 : All mouse were divided into 5 groups at random according to the weight: Normal group, applied with saline; JWSNS low dose group ; JWSNS middle dose group; JWSNS middle dose group ; fluoxetine group. Gave them medicine for 7 days. The morning of the 7th day, pended the mouse tail on the oar for 6 minutes, recorded the still time of the last 5 minutes.2. Tail suspending test 2: the method is the same as experiment 1,the time of giving JWSNS prolonged to 14 days.3. Tail suspending test 2: the method is the same as experiment 1, the time of giving JWSNS prolonged to 21 days.Result1. The influence of giving JWSNS 7 days on still time of TST: There are no significant difference between the low, middle, high dose of JWSNS group and fluoxetine group. There are significant difference between fluoxetine group and normal group. (P＜0.05).2. The influence of giving JWSNS 14 days on still time of TST: There are no significant difference between the low, middle, high dose of JWSNS group and fluoxetine group. There are significant difference between fluoxetine group and normal group.(P＜0.05).3. The influence of giving JWSNS 21 days on still time of TST: There are no significant difference between the high dose of JWSNS group and fluoxetine group. There are significant difference between the low, middle dose group and normal group. (P＜0,05). There are significant difference between fluoxetine group and normal group ( P＜0.05 ).All the data were expressed with X+S and analyzed by t-test and ANOVA with SPSS statistics software.Section Two: The influence of regulating "liver" formula-JWSNS on weight and sucrose preference of CMUS model.Establish the modified chronic mild unpredictable stress model (CMUS) according to Wilier P. SD rats, grade SPF, male,180-220g in weight. Offered by animal's center in Guangzhou university of TCM. Model rats were placed in one independent room and accepted different stress stimuli within 21 days. The stressors consist of restraint for 1 hour,45℃cage tilt for 7h,soiled cage (300ml water in sawdust bedding)for 17h,new invader (two cages combined) for 23h,persistent illumination for 36h,empty bottle for 1 h,food and water deprivation(23h each time).All rats were divided into 3 groups according to the weight and base Sucrose Preference: Normal group, without any stimulus; CMUS group, applied stimulus without formula; JWSSN group, offered 2ml JWSNS concentration every day at AM 8:30.Normal group and Model group were given 2ml distilled water. The Normal group was placed in different room compared with others.After food and water deprivation for 23h, the Sucrose Preference test was taken on the 7th, 14th, 21th day of the experiment. There's no significant difference between different groups in weight and sucrose preference (p＞0.05) before CUMS procedure. After 7 days of model-making, the weight of model group JWSNS group was significantly lower than the Normal group (P＜0.01).there was no significant difference between model group and JWSNS group. After 14 day of model-making ,the weight of model group JWSNS group was significantly lower than the Normal group (P＜0.01).there was significant difference between model group and JWSNS group (P＜0.01); After 21 day of model-making ,the weight of model group JWSNS group was significantly lower than the Normal group (P＜0.01).there was significant difference between model group and JWSNS group (P＜0.01).It indicated that CMUS can decrease the body weight of rats, while JWSNS can reverse the tendency deduced by. The sucrose preference of model group is significantly lower than the normal group (P＜0.01), indicated that CMUS can decrease the sucrose preference, while JWSNS can reverse the tendency deduced by CMUS. through the sucrose preference test on CMUS model, we can find that JWSNS has effects of anti-depression.Section Three: The influence of regulating "liver" formula-JWSNS on monoamine neurotransmitters of central nervous system.Wistar rats, SPF grade, 180-220g,herb and administration is basically the same as section two.From the result we found , after 21 days of CMUS, the content of 5-HT,NE,DA of model group is significantly lower than the normal group (P＜0.01); the content of 5-HT,DA of JWSNS group is significantly higher than the model group (P＜0.01).NE is also increased, but nit significantly. CMUS can decrease the content of 5-HT,NE,DA in central nervous system (p＜0.05); JWSNS can increase the content of 5-HT,DA, NE also has the tendency of increasing, so we conclude up-regulating the content of central nervous system monoamine neurotransmitter is one of the anti-depression mechanism of JWSNS.Section Four: The protective effects of regulating "liver" formula on hippocampus neuron damage of CMUS ratsThe subgroup is the same as section two, herb and administration is basically the same as section two.The apoptosis rates of hippocampus neurons are detected by flow cytometry. The results indicate that there exist enormous apoptosis in model group. The normal group and JWSNS group has little percent of subG1 period, indicated that there were small amount of hippocampus neuron apoptosis ; Compare with the normal group , the apoptosis rate of model group increased significantly, indicated that 21days of CMUS can deduce the apoptosis of hippocampus neurons.JWSNS has protective effects on hippocampus neurons of CMUS rats. But the mechanism is still needs to be researched.Part Three : Conclusion1. JWSNS can significantly increase the sucrose preference of CUMS rats, which indicate that JWSNS has anti-depressive effect.2. JWSNS can significantly up-regulate the DANE content of central nervous system, and it probably exert the antidepressant effect by up-regulate part of the monoamine neurotransmitter content of central nervous system.3. JWSNS can decrease the hippocampus neuron's apoptosis rate of CUMS group, which indicate it has the effect of protecting hippocampus neurons, it also maybe one of the antidepressant mechanism of JWSNS.