CN115501276A

CN115501276A - Traditional Chinese medicine composition for treating stress depression

Info

Publication number: CN115501276A
Application number: CN202211226941.8A
Authority: CN
Inventors: 陈刚; 张海楼; 张煜萱
Original assignee: Jinan University
Current assignee: Jinan University
Priority date: 2022-10-09
Filing date: 2022-10-09
Publication date: 2022-12-23
Anticipated expiration: 2042-10-09
Also published as: CN115501276B

Abstract

The invention relates to a traditional Chinese medicine composition for treating stress depression, which consists of effective components and medically acceptable auxiliary materials, and is characterized in that the effective components are prepared from the following raw material medicines in percentage by weight: is prepared with fried cape jasmine 26-32 wt%, fermented soybean 17-21 wt%, chuanxiong rhizome 17-21 wt%, haw 17-21 wt% and licorice 12-14 wt%. The traditional Chinese medicine composition has the effects of clearing Xuan Yuhuo, relieving restlessness, soothing liver and relieving depression, and not only takes effect quickly, but also has an obvious effect of resisting stress depression and has stable efficacy.

Description

Traditional Chinese medicine composition for treating stress depression

Technical Field

The invention relates to a medical preparation, in particular to a pharmaceutical composition containing an undefined structure from plants, which is suitable for stress depression.

Background

Major Depressive Disorder (MDD), also known as depression, is one of the most serious mental disorders caused by stress, and is ubiquitous in all ages of life. Research studies by the world health organization have shown that depression is one of the most major diseases leading to disability worldwide. Conventional drugs (SSRIs) require several weeks to be taken to achieve efficacy and the patient's symptoms are prone to relapse after withdrawal. In addition, about one third of patients have no significant therapeutic effect after taking such drugs. In addition to this, the disadvantage of slow onset is very dangerous for those with a high suicidal tendency. In recent decades, more and more research has focused on the development and utilization of fast-acting anti-stress, anti-depression drugs. Ketamine is an N-methyl-D-aspartate (NMDA) receptor antagonist whose therapeutic action has been shown to be effective in both stress-induced depression patients and animal models. Within 2 hours, a single dose of ketamine can have a rapid onset of action and its efficacy can last days or even weeks. However, adverse effects such as the potential addiction and neurotoxicity of ketamine greatly limit the wide clinical application and use thereof. Therefore, the development of fast and safe drugs is still urgent and necessary.

Traditional Chinese medicine considers that depression is an emotional disease. It is mainly caused by abnormal emotions resulting from stress. Depression can be caused by frequent mood swings or by stimulation from major events. The traditional Chinese medicine holds that the occurrence of depression is greatly related to factors such as liver qi stagnation, spleen failure in transportation and transportation, heart failure in nourishment, physical weakness and the like. TCM has the theory of "seven emotions", i.e., joy, anger, worry, thinking, sadness, fear and fright. Excessive or insufficient expression of seven emotions can cause adverse effects on the five zang organs of the human body, while deficiency or insufficiency of the five zang organs can also cause emotional changes. Liver qi stagnation is the main pathogenesis of depression syndrome, and the injury of human body by emotion can cause liver failure, qi stagnation is not dispersed, fire is formed by stagnation, and fire inflammation can disturb heart spirit, thus causing the spirit to be unappealing. Adverse emotions caused by various unfortunate events, aspirations and other factors in life, or mental factors such as personal mental stress, worry, sadness and the like are greatly related to the occurrence of malnutrition of mind, so that a series of pathological changes of the human body can be caused, such as vexation, palpitation, shortness of breath, sadness, crying, joy and anger and the like. Kidney governs water, so consumption of kidney yin cannot induce water to heart, resulting in disharmony of kidney water and heart spirit, and restlessness, which can cause depression. Excessive anxiety can injure the heart and spleen, resulting in the consumption of heart blood and insufficiency of spleen-qi, while the deficiency of both heart and spleen can lead to blood deficiency and inability to nourish the heart, so that the heart fails to nourish and the mind is restless, thereby causing emotions such as depression and sadness. Therefore, the traditional Chinese medicine treatment starts with the aspects of soothing liver-qi stagnation, strengthening spleen and eliminating phlegm, activating blood and nourishing heart, tonifying qi and deficiency and the like, and mainly regulates the qi activity of the five internal organs of the human body.

The classic famous book "Shanghai treatise" discloses a cape jasmine and liquorice fermented soybean decoction, which comprises the following components: fourteen (kohlrabi) (10 g), two (roasted) liquorice (6 g) and four (30 g) fermented soybeans (fermented soybean) (sponge dressing). The formula of the gardenia, liquorice and fermented soybean soup is as follows: (1) the prescription is prepared by adding liquorice in the gardenia and fermented soybean decoction, and is a medicament for clearing heat, relieving restlessness and tonifying deficiency. The decoction of cape jasmine and fermented soya beans in the recipe has the functions of clearing dysphoria with smothery sensation in chest and diaphragm, and licorice root, which is sweet and warm, tonifying middle-jiao and Qi, and strengthening body resistance. (2) Gardenia jasminoides is reddish in color and looks at the heart, bitter in taste belongs to fire, cold in nature and down-flow of fire-heat, kidney is like a bean, black in color enters the kidney, the raw materials are made into fermented beans, light and superficial priming fluid rises, yin and yang and fire-water are complemented, and the symptoms of heat worry, cheerful and pain are relieved . Zhi Zi and Chi Zhi are listed above. Licorice root, radix Glycyrrhizae is added to tonify the middle-jiao, because it fails to transport the body upward and downward due to the deficiency of middle-jiao during sweating, vomiting and descent. The gardenia licorice fermented soybean soup has the following functions: to open the diaphragm and clear heat. Tranquilizing, antipyretic, anti-inflammatory promoting bile flow, stopping bleeding, and inducing diuresis. Clear heat and relieve restlessness, replenish qi and calm middle energizer. It is indicated for qi deficiency with symptoms of both Zhi and Qiao. Restlessness due to deficiency, insomnia, feeling of restlessness, or repeated reversal of heart-mind, restlessness, fever, and lack of qi. Exogenous febrile disease, warm disease, restlessness and insomnia due to sweating and descending, and shortness of breath.

However, the gardenia, liquorice and fermented soybean soup for treating stress depression has the defects of slow response and unsatisfactory effect. Therefore, the finding of a traditional Chinese medicine compound for treating stress depression with quick response and obvious curative effect has important significance.

Disclosure of Invention

The invention aims to solve the technical problem of providing a traditional Chinese medicine composition for treating stress depression, which has the advantages of quick response and obvious curative effect.

The technical scheme for solving the problems comprises the following steps:

the invention relates to a traditional Chinese medicine composition for treating stress depression, which consists of effective components and medically acceptable auxiliary materials, and is characterized in that the effective components are prepared from the following raw material medicines in percentage by weight:

26 to 32 percent of fried gardenia, 17 to 21 percent of fermented soybean, 17 to 21 percent of Szechuan lovage rhizome, 17 to 21 percent of hawthorn and 12 to 14 percent of liquorice.

In the scheme, the optimal weight percentage of the raw material medicines is 30 percent of fried gardenia, 19 percent of fermented soybean, 19 percent of ligusticum wallichii, 19 percent of hawthorn and 13 percent of honey-fried licorice root.

The traditional Chinese medicine composition is prepared from the following effective components by the following method:

(1) The raw materials are taken according to the proportion, the raw materials are placed in boiling water of which the amount is 10 times of that of the raw materials for distillation, distillate is collected, and dregs of a decoction are stored in another container;

(2) Soaking the residue in water for 0.5 hr, decocting twice with 10 times of water for 1.5 hr, adding 8 times of water for 1hr, filtering, and mixing filtrates;

(3) Mixing the distillate of step (1) and the filtrate of step (2), concentrating under reduced pressure at 60 deg.C to relative density of 1.05-1.20, spray drying, and pulverizing into fine powder to obtain the effective component.

The traditional Chinese medicine composition can be prepared into oral preparations such as oral liquid, granules or pills, and the oral preparations are prepared from the effective components and medically acceptable auxiliary materials according to a conventional method.

The formula of the traditional Chinese medicine composition consists of five medicines of fried gardenia, fermented soybean, ligusticum wallichii, hawthorn and honey-fried licorice root, wherein the gardenia is bitter and cold, enters heart, lung and triple energizer meridians, is good at clearing triple energizer fire, has special effects of relieving restlessness and purging fire, and cooling blood and resolving depression, and is a monarch drug; the fermented soybean is pungent in taste, has the effect of dispersing, is slightly bitter and cold, has slight heat clearing effect, has the function of relieving restlessness, and is a ministerial medicament, the monarch medicament and the ministerial medicament are combined, so that the fermented soybean is capable of clearing and ventilating, pathogenic heat is clear, and the restlessness is eliminated; the combination of the hemlock parsley and the hawthorn is an adjuvant, the hemlock parsley is pungent and warm and can promote qi, the rangoon medicine used slightly is bitter and cold without stagnation, and can enhance the dispersing power of ministerial medicines, and can also dredge wood to achieve depression as the chuanxiong medicine enters the liver channel; hawthorn is sour and sweet in taste, slightly warm and not hot, has good effect of invigorating spleen and stomach, enters liver and benefits liver; prepared licorice root, radix Glycyrrhizae Praeparata, sweet and mild in flavor, is used as a guiding drug for harmonizing the effects of the other drugs in the recipe. The whole formula is used for both cold and heat, and has the effects of clearing Xuan Chufan and soothing liver to achieve the effect of depression.

Compared with the meridian formula gardenia, liquorice and fermented soybean soup, the formula not only increases the ligusticum wallichii and the hawthorn, but also changes the original formula from the fermented soybean to the gardenia, so that the effect of resisting depression can be achieved after the formula is given for 30 minutes once.

To facilitate a better understanding of the present invention to the public, the following further illustrate the advantageous effects of the present invention through experiments on pharmaceutical effects and specific embodiments.

Drawings

FIG. 1 is a graph of a factorial design interaction visual analysis of the interaction between the drug used in control group 1 and the drug used in control group 2 in the tail-overhang experiment described below.

FIG. 2 is an electrophoretogram showing the change in PACAP expression in mouse hippocampus, wherein the groups from left to right are: blank group, model group, control group 1, control group 2, yang ginseng group and experimental group.

Detailed Description

Example 1 (oral liquid)

1. Prescription: 90g of fried gardenia, 60g of fermented soybean, 60g of ligusticum wallichii, 60g of hawthorn and 40g of liquorice.

2. The preparation method comprises the following steps:

(3) Mixing the distillate of step (1) and the filtrate of step (2), concentrating under reduced pressure at 60 deg.C until the relative density is 1.05-1.20, spray drying, and pulverizing into fine powder to obtain effective component;

(4) And (4) taking the effective components prepared in the step (3), adding 80g of cane sugar, and sterilizing by flowing steam at 100 ℃ for 30 minutes to prepare 100ml of oral liquid.

3. Method of administration and dosage

Is administered 1 time a day, 1ml each time, and 7 days as a treatment course.

Example 2 (pill)

1. Prescription: 98g of fried gardenia, 54g of fermented soybean, 56g of ligusticum wallichii, 59g of hawthorn and 43g of liquorice.

2. The preparation method comprises the following steps:

the preparation method of the effective components in this example is the same as that of example 1; 40g of refined honey is added into the prepared active ingredients to prepare 100 pills.

3. Method of administration and dosage

The preparation is administered 1 time per day, 10 granules each time, and 7 days as a treatment course.

Example 3 (granules)

1. Prescription: 83g of fried gardenia, 58g of fermented soybean, 61g of ligusticum wallichii, 64g of hawthorn and 44g of liquorice.

2. The preparation method comprises the following steps:

the preparation method of the effective components in this example is the same as that of example 1; adding a proper amount of dextrin into the prepared effective components, placing the effective components in a fluidized bed, setting the air inlet temperature to be 100 ℃, starting to feed liquid when the temperature of the materials rises to 70 ℃, and controlling the liquid feeding speed to be 80-150 drops/min, and the atomization pressure to be 0.2MPa outside and 0.15MPa inside. After the spraying was finished, the drying was continued at 70 ℃ for 1 hour to obtain 100 g of granules.

3. Method of administration and dosage

The medicine is taken 1 time a day, 10g each time, and 7 days as a treatment course.

Example 4 (efficacy test)

1. Laboratory animals and groups thereof

Adult male ICR mice 126, aged 1-2 months, weighed 18-22 grams. Experimental mice were purchased from Shanghai family planning science institute (SCXK (Shanghai) -2018-0006); the animals are all bred in an animal house with SPF grade, the temperature of the animal house is kept at 22 +/-2 ℃, and the humidity is controlled at 50% +/-10%. Normal feeding acclimation was performed one week before the start of the experiment and all animals were fed ad libitum. Experiments were performed with approval by the ethical committee of animal experiments. All the behavior experiments were carried out for 8a.m. -5p.m. 126 adult male ICR mice were divided into a blank group, a model group, a control group 1, a control group 2, an experimental group and a positive ginseng group, 21 mice per group, and randomly assigned, 8 mice for the anti-stress experiment, 8 mice for the anti-depression experiment and 5 mice for the anti-depression molecular experiment.

2. Test drugs and methods and dosages of administration

Blank group: the administration was performed by gavage with 0.1ml/10g of 0.9% physiological saline according to the body weight of the mice.

Model group: the administration was performed by gavage with 0.1ml/10g of 0.9% physiological saline according to the body weight of the mice.

Control group 1: the tested drugs are: taking 90g of fried gardenia, 40g of liquorice and 60g of fermented soybean, preparing the mixture into oral liquid according to the method of the embodiment 1, and adding 0.9% of normal saline to dilute the oral liquid to be equivalent to 0.5g/ml of crude drug; the administration method and dosage are as follows: the administration was performed by gavage at 0.1ml/10g based on the body weight of the mice.

Control group 2: the tested drugs are: taking 60g of hawthorn and ligusticum wallichii respectively, preparing into oral liquid according to the method in the embodiment 1, and adding 0.9% of normal saline to dilute the oral liquid to be equivalent to 0.182g/ml of crude drug (bulk drug); the administration method and dosage are as follows: the administration was performed by gavage at 0.1ml/10g based on the body weight of the mice.

Experimental groups: the oral liquid of example 1 is diluted to 0.592g/ml crude drug (bulk drug) by adding 0.9% physiological saline, and the administration method and the dosage are as follows: the administration was performed by gavage at 0.1ml/10g based on the body weight of the mice.

Yang ginseng group: fluoxetine, 0.9% physiological saline diluted to the concentration of 2mg/ml, according to the body weight of mice according to 0.1ml/10g intraperitoneal injection administration.

3. Establishing a chronic stress depression model:

the establishment of the chronic stress model is to simulate the disease state of depression patients under the long-term stress state on animals, and can be used for screening anti-stress antidepressant drugs and experimental research on the pathophysiological mechanism of depression. The anti-stress drug can reverse the behavioral changes of stressed mice. The types of stress mice received are as follows: keeping water for 24hr, fasting for 24hr, reversing day and night for 12hr, binding for 6hr, inclining squirrel cage at 45 deg.C for 18hr, wetting cage for 24hr, and strangling articles for 24hr. The stress cycle was 4 weeks; during this period, the blank mice did not receive any stress, were fed water ad libitum, and were fed normally. At the end of molding, mice were randomly assigned to the blank group, model group, control group 1, control group 2, experimental group and positive control group for further drug administration and behavior detection.

4. Anti-stress efficacy test

4.1 anti-stress behavioural assay

Tail suspension test: tail suspension behavior detection is widely used in the basic research anti-stress drug testing process. The principle is that the mouse can often try to escape after being hung, but cannot escape, struggle is given up after a long time, and the mouse enters a unique state of depression and quiescence and mental power weakening to the environmental stress which cannot escape. During the course of the experiment, the immobility time (rest time) of the mice was recorded to assess whether the mice were in a depressed or excited state. Anti-stress drugs can greatly shorten the time to change their immobility state, from which they can be generally predicted to have antidepressant effects. Briefly, tail overhang testing was performed using a computer device, which simultaneously tested 6 animals at a time. Mice were placed in a sealed sound-proof box, both acoustically and visually isolated, taped and hung 50cm from the ground with a hook at about 1cm from the tip of the mouse tail, while the animal's activity was recorded and analyzed for total immobility over the last 4 minutes of the 6 minute test period using ANY-maze software (stowing co. The anti-stress drug can obviously reduce the immobility time of tail suspension of the mice.

Forced swimming test: forced swim testing is one of the techniques used to evaluate the anti-stress effects of drugs during the course of stress animal model studies, an experimental test of behavioral despair. When the animal is placed in water with proper temperature, the struggle is one of the states which the animal usually shows under the environment, the other performance is escape and often escape failure or no escape, the inevitable stress environment is generated by the struggle, and the animal shows typical movement and static state and is the performance of mental power reduction. A series of parameters are observed and recorded during the process of the movement and floating of the experimental animal, and can be used for evaluating the pharmacodynamic action. The mice were removed from their cages and placed in a transparent glass jar (40 cm in height, 20 cm in diameter) filled with 30 cm of water (22-24 ℃) and forced to swim for 6 minutes. At the end of the experiment, the mice were removed from the water, dried with a hair dryer, and returned to their cages after drying. The mouse is considered to be in a stationary state when the mouse keeps its head above the water surface for breathing and floats in the water without struggling and with only minute movements of the paw. The total immobility time within the last 4 minutes of the 6 minute test period was recorded and analyzed by the ANY-maze software. The anti-stress drug can obviously reduce the immobility time of forced swimming of the mouse.

4.2 statistical methods

All data were analyzed by SPSS 21.0 software (SPSS, armonk, USA). To compare two or more groups, one-way analysis of variance was used, followed by post-hoc analysis of LSD. All data are expressed as mean ± SD, with P <0.05 considered statistically significant. 2 x 2 factorial design analysis of variance analysis the interaction effect of two drugs in combination. P <0.05 indicates significant difference.

4.3 results

4.3.1 the results show: in the tail suspension experiment, compared with a blank group, after 4 weeks of stress, the tail suspension immobility time of the model mouse is obviously increased (P < 0.05), which shows that the stress model is successful; in comparison with the model group, the immobility time of mice could not be significantly reduced after a single administration of either control group 1 or control group 2hr after 4 weeks of molding (P > 0.05). However, compared with the model group, the immobility time of the mouse tail suspension can be significantly reduced after a single administration of the experimental group for 2hr (P < 0.05). Meanwhile, after the positive reference drug fluoxetine is given for 2 hours, the immobility time of tail suspension of the mouse cannot be improved (P > 0.05). In addition, compared with the control group 1 and the control group 2, the test group can obviously reduce the immobility time (P < 0.05) of the tail suspension of the mice, and the test group is prompted to have obvious rapid anti-stress effect. The results are shown in Table 1.

Table 1 immobility time of mice tail suspension detection 2 hours after single administration of each group of drugs: (

n＝8)

Group of	Time of immobility(s)
		Blank group	107.66±18.56*
Model set	154.49±11.69
		Control group 1	144.32±11.54
Control group 2	152.76±13.84
		Experimental group	110.95±11.29* ^&#
Yang ginseng group	153.82±14.77

(note: in comparison to the model set, ^* P<0.001, compared with the control group 1, ^& P<0.001, compared with the control group 2, ^# P<0.001)

4.3.2 the results show: in forced swim experiments, the time to immobility for forced swim was significantly increased (P < 0.05) in model mice after 4 weeks of stress compared to the blank group; in contrast to the model group, the immobility time of the mice could not be significantly reduced after 4 weeks of molding after a single administration of either control group 1 or control group for 2 days (P > 0.05). However, compared to the model group, the immobility time of the mouse tail suspension can be significantly reduced after 1 day of single administration to the experimental group (P < 0.05). Meanwhile, the immobility time of forced swimming of mice cannot be improved after 1 day of administration of the positive reference drug fluoxetine (P > 0.05). In addition, compared with the control group 1 and the control group 2, the test group can obviously reduce the immobility time (P < 0.05) of forced swimming of the mice, and the test group is prompted to have obvious anti-stress effect and stable curative effect. The results are shown in Table 2.

TABLE 2 forced swim test of mice 1 day after single administration of each group of drugsDynamic time (

n＝8)

Group of	Time of immobility(s)
		Blank group	106.79±12.69*
Model set	177.26±16.51
		Control group 1	158.96±24.44
Control group 2	180.43±16.74
		Experimental group	108.10±16.17* ^&#
Yang ginseng group	165.29±17.88

4.4 analysis of cause of disease results

The experiment was designed with two factors, each at two levels. Factor 1 is the Gardenia jasminoides Ellis and Glycyrrhiza uralensis fermented soybean group (the tested medicine used in the control group 1 in the experiment is factor A), and the level is divided into use and non-use; factor 2 is the hawthorn and ligusticum wallichii group (the tested medicine used in the control group 2 in the experiment, factor B), the level is also divided into use and non-use, the interaction is gardenia, liquorice, fermented soya beans and hawthorn and ligusticum wallichii (the tested medicine used in the experiment group in the experiment), and 4 combinations are provided, namely: blank group (A0B 0), gardenia Glycyrrhiza fermented soya bean group (A1B 0), hawthorn Szechuan lovage rhizome group (A0B 1), gardenia Glycyrrhiza fermented soya bean and Hawthorn Szechuan lovage rhizome group (A1B 1) which have no two factors.

Observation indexes are as follows: immobility time of tail suspension and immobility time of forced swimming

The results are shown in tables 3 and 4.

TABLE 3 influence of the combination of Gardenia jasminoides, glycyrrhiza uralensis, fermented soya beans, hawthorn fruit and Ligusticum wallichii on the immobility time of the suspended tail

TABLE 4 analysis of the Effect of the combination of Gardenia jasminoides, glycyrrhiza uralensis and fermented soya bean soup and Hawthorn fruit and Ligusticum wallichii on the immobility time in forced swimming

4.4.1 factorial design analysis of variance

The data in tables 3 and 4 are subjected to statistical analysis of main effects and interactive effects of various factors, and the results are shown in tables 5 and 6, wherein P of the two main effects is less than 0.01, P of the interactive effects is less than 0.05, and the differences have statistical significance, so that the interaction between the gardenia liquorice fermented soya beans and the hawthorn and ligusticum wallichii is shown, and the stress can be obviously improved. The figure 1 shows the analysis of the interaction between the drug used in the tail-suspended control group 1 and the drug used in the control group 2.

TABLE 5 analysis of variance analysis for analysis of hanging tail results

TABLE 6 analysis of variance analysis for analysis of forced swimming results

5. Antidepressant effect test

5.1 antidepressant behavioural test

Novel environmental suppression feeding experiments the resolution of stress-induced depressive states by animals was assessed by the behavioral approach of a novel environmental suppression feeding experiment (NSF) in which animals respond to seeking appetite stimulation. In the NSF test, after food deprivation for 18 hours, mice were placed in an open field (15cm × 15cm) with a single grain placed in the center of the field (weighed). Each animal was initially placed in a corner of the field and allowed to explore for 10 minutes, ending when the rat chews the rat chow or 10 minutes. At the same time, the food intake in the cages was measured as an index for appetite control. The time from the start of the recording to the feeding period is defined as a latency period, and the mice actively gnaw the food or the mice hold the food with both claws to feed for the first feeding, but the latency period is not counted when the food is just hit to gnaw or smell the food. The formula for calculating the food intake per unit weight is as follows: the total food intake (g)/mouse body weight (g) is the unit body weight food intake (g/kg). Depressed mice show a significant increase in feeding latency and a significant decrease in food intake per unit body weight, and administration of antidepressant drugs can significantly reverse these indicators.

And (4) testing syrup preference: the carbohydrate preference test (SPT) is an evaluation index for simulating anhedonia caused by depression due to stress. Briefly, mice were housed individually, two bottles of 1% sucrose solution were placed on either side of the cage to acclimate to the sweetness of sugar water for 72 hours, and then the water was removed for 18 hours. After the water exclusion was complete, the test was performed by giving two bottles of solution for 2 hours (one bottle filled with pure water and the other one of them with 1% sucrose solution). After the end of the 2 hours of the test, the consumption of the sucrose solution or water was measured. Sugar water preference is defined as the ratio of the mass of sucrose to the total mass of water consumed and sucrose consumed (sucrose + water) in a 2 hour test, normalized to the body weight of each animal. Depressed mice show a significant reduction in carbohydrate preference, which can be significantly reversed after administration of antidepressant drugs.

5.2 statistical methods

5.3 results

5.3.1 the results show: in the experiment of inhibiting food intake in a novel environment, compared with a blank group, after 4 weeks of stress, the food intake latency of a model mouse is remarkably increased (P < 0.05), and the food intake per unit body weight is remarkably reduced (P < 0.05); compared with the model group, after 4 weeks of modeling, the food intake latency period of the control group 1 or the control group 2 30min can not be obviously reduced after a single administration (P > 0.05) and the food intake per unit body weight is obviously reduced (P > 0.05). However, compared with the model group, 30min after single administration of the experimental group, the food intake latency period can be remarkably reduced (P < 0.05), and the food intake per unit body weight is remarkably reduced (P < 0.05). Meanwhile, after the positive reference medicament fluoxetine is given for 30min, the feeding latency period cannot be obviously reduced, and the feeding amount per unit body weight cannot be obviously increased (P > 0.05) and the feeding amount per unit body weight cannot be obviously reduced (P > 0.05). In addition, compared with the control group 1 and the control group 2, the experimental group can obviously reduce the obvious increase of the intake incubation period (P < 0.05) and the obvious decrease of the intake of unit weight (P < 0.05), and the Chinese herbal compound formed by the gardenia, the liquorice, the fermented soya beans, the ligusticum wallichii and the hawthorn is prompted to have obvious quick antidepressant effect. The results are given in Table 7,8.

TABLE 7 feeding latency time for the novel environmental feeding assay in mice 30 minutes after single administration of each group of drugs: (

n＝8)

Group of	Incubation period of food intake(s)
		Blank group	29.38±8.45*
Model set	80.50±11.30
		Control group 1	77.88±11.45
Control group 2	78.63±10.54
		Experiment of group of	29.75±5.75* ^&#
Yang ginseng group	82.38±11.00

table 8:unit food intake for a novelty ambient food intake test 30 minutes after a single administration of each group of drugs: (

n＝8)

Group of	Food intake per unit weight (g/kg)
		Blank group	0.024±0.002*
Model set	0.009±0.001
		Control group 1	0.010±003
Control group 2	0.010±0.001
		Experimental group	0.020±0.006* ^&#
Yang ginseng group	0.008±0.002

5.3.2 the results show: in the sugar water preference experiment, compared to the blank group, the sugar water preference of the model mice was significantly reduced after 4 weeks of stress (P < 0.05); in contrast to the model group, the sugar water preference of mice could not be significantly increased after 4 weeks of molding, either after a single administration of control group 1 or control group for 2 days (P > 0.05). However, compared to the model group, 1 day after a single administration to the experimental group, the sugar water preference of the mice could be significantly increased (P < 0.05). At the same time, the glucose preference of mice was not significantly increased (P > 0.05) 1 day after the administration of the positive reference drug fluoxetine. In addition, compared with the control group 1 and the control group 2, the sugar water preference (P < 0.05) of the mice can be obviously improved in the experimental group, and the Chinese herbal compound formed by combining gardenia liquorice, fermented soya beans, ligusticum wallichii and hawthorn fruits is prompted to have an obvious anti-depression effect and a stable curative effect. The results are shown in Table 9.

TABLE 9 detection of change in sugar Water preference of mice 1 day after single administration of each group of drugs: (

n＝8)

Group of	Sweet water preference (g/g + g)
		Blank group	0.867±0.036*
Model set	0.602±0.072
		Control group 1	0.657±0.042
Control group 2	0.616±0.056
		Experimental group	0.906±0.020* ^&#
Yang ginseng group	0.618±0.074

5.4 analysis of cause of analysis results

The experiment was designed with two factors, each at two levels. Factor 1 is the Gardenia jasminoides ellis licorice fermented soybean group (the tested medicine used in the control group 1 in the experiment), and the level is divided into use and non-use; factor 2 is the hawthorn chuanxiong rhizome group (the test drug used in the control group 2 in the above experiment), the level also is divided into use and non-use, the interaction is the gardenia liquorice fermented soya beans and the hawthorn chuanxiong rhizome (the test drug used in the experiment group in the above experiment), there are 4 combinations, namely: blank group (A0B 0), gardenia Glycyrrhiza fermented soya bean group (A1B 0), hawthorn Szechuan lovage rhizome group (A0B 1), gardenia Glycyrrhiza fermented soya bean and Hawthorn Szechuan lovage rhizome group (A1B 1) which have no two factors.

Observation indexes are as follows: latency to feed in the novel environment, feed intake per unit weight and sugar water preference

The results are reported in tables 10, 11 and 12.

TABLE 10 analysis of the effect of the combination of Gardenia jasminoides, glycyrrhiza uralensis, fermented soya beans and Crataegus pinnatifida and Ligusticum wallichii on the incubation period of food intake

TABLE 11 analysis of the influence of the combination of Gardenia jasminoides, glycyrrhiza uralensis, fermented soya beans and Hawthorn fruit and Ligusticum wallichii on the food intake per unit weight

TABLE 12 analysis of the effect of Gardenia jasminoides Ellis, glycyrrhiza uralensis and Crataegus pinnatifida on sugar water preference

5.4.1 factorial design analysis of variance

The data in tables 10, 11 and 12 were analyzed statistically for the main effect and the interaction effect of each factor, and the results are shown in tables 13, 14 and 15, where P of the two main effects is less than 0.01, P of the interaction effect is less than 0.05, and the difference is statistically significant, indicating that there is interaction between gardenia glycyrrhiza fermented soya bean and hawthorn ligusticum wallichii.

TABLE 13 analysis of factorial design of feeding latency results analysis of variance

TABLE 14 analysis of the cause design analysis of food intake results per unit weight analysis of variance

TABLE 15 analysis of sugar Water preference results factoring design analysis of variance

6. Molecular experiments against depressive disorders

6.1 molecular detection

The pituitary gland adenosine activating enzyme (PACAP) is closely related to the occurrence and development of stress and depression. In order to detect the protein level expression amount of PACAP in the hippocampus of the mouse, total protein required for the experiment is extracted from the hippocampus of the mouse, the concentration of the protein contained in each tissue sample is measured by an instrument (Nanodrop), PBS and a Loading Buffer solution are added according to the protein concentration in each sample in proportion, then the sample is boiled for 5min in an incubator at 100 ℃ and cooled to room temperature, and then the sample is subpackaged and stored in a refrigerator at-80 ℃. SDS-PAGE gels (15% gels were used in each case) were selected at the corresponding concentrations according to the molecular weight of PACAP, followed by separation of the proteins by electrophoresis, transfer to polyvinylidene fluoride (PVDF) membranes after successful separation, 5% blocking of the PVDF membranes in BSA solution at room temperature for 1hr, addition of antibodies containing primary antibodies (BDNF, PAC-AP, tubulin) respectively, and subsequent incubation at 4 ℃ in a refrigerator for 12 hr. Rinsing in TBST buffer at room temperature for 3 times, adding goat anti-rabbit secondary antibody, incubating at room temperature for 1hr, and developing with ECL. And acquiring the gray values of the strips of PACAP and Tubulin, and then counting the gray ratio of each sample PACAP to Tubulin.

6.2 statistical methods

All data were analyzed by SPSS 21.0 software (SPSS, armonk, USA). To compare two or more groups, one-way analysis of variance was used, followed by post-hoc analysis of LSD. All data are expressed as mean ± SEM, P <0.05 was considered statistically significant. 2 x 2 factorial design analysis of variance analysis the interaction effect of two drugs in combination. P <0.05 indicates significant difference.

6.3 results

The results show that: in a protein detection experiment, compared with a blank group, after 4 weeks of stress, the PACAP protein expression level of the hippocampus of a model mouse is obviously reduced (P < 0.05); compared with the model group, after 4 weeks of modeling, the PACAP protein expression level of the hippocampus of the mice cannot be obviously increased after a single administration of the control group 1 or the control group 2 30min (P is more than 0.05). However, compared with the model group, the PACAP protein expression level of the mouse hippocampus can be obviously increased after a single administration of the experimental group for 30min (P < 0.05); whereas there was no significant change in PACAP in hippocampus 30min after a single administration of fluoxetine (yang ginseng). In addition, the experimental group can obviously increase the PACAP protein expression level (P < 0.05) of mouse hippocampus by respectively comparing with the control group 1 and the control group 2, and the Chinese herbal compound formed by combining the gardenia, liquorice and fermented soybean soup, the ligusticum wallichii and the hawthorn has obvious quick antidepressant effect. The results are shown in Table 16 and FIG. 2.

TABLE 16 expression of PACAP in hippocampal brain region of mice 30 minutes after single administration of each group of drugs: (

n＝5)

Group of	PACAP/Tubulin
		Blank group	0.73±0.23*
Model set	0.44±0.15
		Control group 1	0.51±0.21
Control group 2	0.54±0.16
		Yang ginseng group	0.48±0.17
Experimental group	0.82±0.20* ^&#

6.4 analysis of cause of analysis results

The experiment was designed with two factors, each at two levels. Factor 1 is the Gardenia jasminoides Ellis and Glycyrrhiza uralensis fermented soybean group (the tested medicine used in the control group 1 in the experiment), and the level is divided into use and non-use; factor 2 is the hawthorn and ligusticum wallichii group (the tested medicine used in the control group 2 in the experiment), the level is also divided into use and non-use, the interaction is gardenia, liquorice, fermented soya beans and hawthorn and ligusticum wallichii (the tested medicine used in the experiment group in the experiment), and 4 combinations are provided, namely: blank group (A0B 0), fructus Gardeniae and Glycyrrhrizae radix semen Sojae Preparatum group (A1B 0), fructus crataegi and rhizoma Ligustici Chuanxiong group (A0B 1), fructus Gardeniae and Glycyrrhrizae radix semen Sojae Preparatum + fructus crataegi and rhizoma Ligustici Chuanxiong group (A1B 1) with both factors being not used.

Observation indexes are as follows: expression of PACAP in mouse hippocampus

The results of the experiment are reported in table 17.

TABLE 17 analysis of the Effect of Gardenia jasminoides, glycyrrhiza uralensis and fermented soya bean decoction and Hawthorn fruit and Ligusticum wallichii on Hippocampus PACAP protein

6.4.1 factorial design analysis of variance

The data in table 17 were analyzed statistically for major effects and interaction effects of each factor, and the results are shown in table 18, where P of the two major effects is less than 0.01, P of the interaction effect is less than 0.05, and the differences are both statistically significant, indicating that there is an interaction between gardenia glycyrrhiza fermented soya beans and hawthorn ligusticum wallichii.

TABLE 18 Hippocampus PACAP protein expression results factorial design analysis of variance

Conclusion

The experimental result shows that the traditional Chinese medicine compound formed by using the gardenia, liquorice, fermented soya beans and the ligusticum wallichii and hawthorn in a combined way for one time can obviously and rapidly shorten the tail suspension immobility time and the feeding latency of a mouse and rapidly increase the food intake of unit weight, has stable effect, and can significantly reduce the immobility time of forced swimming of the mouse and increase the sweet water preference of the mouse after 1 day of single use. The traditional Chinese medicine compound formed by combining the gardenia, liquorice, fermented soya beans and the ligusticum wallichii and hawthorn can quickly resist stress and depression, has stable and obvious curative effect on stress-induced depression.

Claims

1. The invention relates to a traditional Chinese medicine composition for treating stress depression, which consists of effective components and medically acceptable auxiliary materials, and is characterized in that the effective components are prepared from the following raw material medicines in percentage by weight:

2. The traditional Chinese medicine composition for treating stress depression according to claim 1, wherein the effective components are prepared from the following raw material medicines in percentage by weight:

30% of fried gardenia, 19% of fermented soybean, 19% of ligusticum wallichii, 19% of hawthorn and 13% of honey-fried licorice root.

3. The traditional Chinese medicine composition for treating stress depression according to claim 1 or 2, wherein the effective components are prepared by the following method:

(2) Soaking the residue in water for 0.5 hr, decocting twice, adding 10 times of water for the first time, decocting for 1.5 hr, adding 8 times of water for the second time, decocting for 1hr, filtering, and mixing filtrates;

4. The traditional Chinese medicine composition for treating stress depression according to claim 1 or 2, wherein the pharmaceutical composition is oral liquid, pills or granules.