Construction and identification method of liver depression and spleen deficiency type generalized anxiety disorder animal model

Abstract

The invention relates to the field of traditional Chinese medicine, in particular to a construction and identification method of a liver depression and spleen deficiency type generalized anxiety disorder animal model. The method mainly comprises the following steps: establishing a liver depression and spleen deficiency type generalized anxiety disorder animal model based on a chronic constraint stress method, and identifying the liver depression and spleen deficiency type generalized anxiety disorder rat model by adopting a combined multi-dimensional evaluation mode and combining general evaluation of general conditions, weight change and the like of rats and behavioral tests of sweet water consumption and the like. The constructed liver depression and spleen deficiency type wide anxiety syndrome combined rat model provides treatment suggestions of traditional Chinese medicine dialectical treatment, and has important contributions to clear model construction and optimization model evaluation.

Description

Technical Field

[0001] The present invention relates to the field of traditional Chinese medicine, and particularly to a method for constructing and identifying an animal model of generalized anxiety disorder with liver depression and spleen deficiency. Background Art

[0002] In modern society, the competition is becoming increasingly fierce, and the emergency factors and pressures faced by people are gradually increasing, seriously affecting their physical and mental health and posing a serious threat. Chronic anxiety, also known as generalized anxiety disorder, mostly occurs due to long-term exposure to stress. The existing clinical treatment plans for chronic anxiety often deviate from the expected curative effects. In traditional Chinese medicine, anxiety disorder belongs to the category of emotional diseases, mostly caused by emotional discomfort, liver qi stagnation, and spleen dysfunction. Based on the two major principles of the overall concept and syndrome differentiation and treatment, traditional Chinese medicine treatment has unique advantages. However, the existing mechanism research mostly focuses on the simple generalized anxiety disorder model and cannot demonstrate the advantages of traditional Chinese medicine in preventing and treating diseases in combination with syndromes. Summary of the Invention

[0003] In view of this, the present invention provides a method for establishing and identifying an animal model of generalized anxiety disorder with liver depression and spleen deficiency induced by chronic restraint stress, adopting a combined multi-dimensional evaluation method, and combining the overall evaluation of the general conditions and weight changes of rats, etc., and behavioral tests such as sucrose consumption, etc., to identify the rat model of generalized anxiety disorder with liver depression and spleen deficiency. Based on the constructed disease-syndrome combination rat model of generalized anxiety disorder with liver depression and spleen deficiency, the present invention provides treatment suggestions for traditional Chinese medicine syndrome differentiation and treatment (such as Xiaoyao Powder). The present invention makes important contributions to clarifying model construction and optimizing model evaluation, and is conducive to the scientific dissemination of the model construction and identification methods involved in the present invention.

[0004] In order to achieve the above invention purposes, the present invention provides the following technical solutions:

[0005] The present invention provides a method for constructing an animal model of generalized anxiety disorder with liver depression and spleen deficiency, comprising the following steps:

[0006] Step (1): Divide rats into a control group, a model group 1, a model group 2, and a model group 3 according to the random block method;

[0007] Step (2): Record the status of rats every day, administer Xiaoyao Powder to the rats in the model group 1, administer rolipram to the rats in the model group 2, and perform chronic restraint stress on the rats in the model group 1, the model group 2, and the model group 3 for 3 h after 30 min, and repeat for 21 days. During this period, record the weight, sucrose consumption, number of entries into the open arms, and residence time in the open arms of the rats to obtain an animal model of generalized anxiety disorder with liver depression and spleen deficiency;

[0008] The operating steps of the chronic restraint stress include: restraining the rats in a rat restraint bag, taking two adjustable adhesive soft bands to fix the chest and abdomen of the rats on a plane respectively, making the heads of the rats protrude from the notch of the restraint bag, adjusting the tightness of the adhesive soft bands to ensure the normal breathing of the rats. During the whole restraint process, all the rats are in the same environment and are not given food or water.

[0009] In some specific embodiments of the present invention, the model group 3 in the above construction method is given the test drug or normal saline.

[0010] In some specific embodiments of the present invention, step (2) of the above construction method is: recording the state of the rats every day, giving Xiaoyao Powder to the rats in model group 1, giving rolipram to the rats in model group 2, giving the test drug to the rats in model group 3. 30 minutes later, chronic restraint stress is applied to the rats in model group 1, model group 2, and model group 3 for 3 hours, and this is repeated for 21 days. During this period, the sugar water consumption and the body weight of the rats are recorded to obtain an animal model of generalized anxiety disorder with liver depression and spleen deficiency.

[0011] In some specific embodiments of the present invention, the rats in the above construction method are male SD rats weighing 220 - 250 g.

[0012] In some specific embodiments of the present invention, the administration method of the above Xiaoyao Powder is intragastric administration.

[0013] In some specific embodiments of the present invention, the dosage of the above Xiaoyao Powder is 3.854 g·kg -1 ·d -1 。

[0014] In some specific embodiments of the present invention, the above Xiaoyao Powder is Xiaoyao Powder suspension.

[0015] In some specific embodiments of the present invention, the above Xiaoyao Powder suspension is obtained by dissolving the dry extract in water. The preparation method of the dry extract can be: Poria cocos, Paeonia lactiflora, Glycyrrhiza uralensis, extracted by boiling water 3 times, and the water addition amounts are 10 times, 8 times, and 8 times the amount of the medicinal materials respectively, and the extraction times are 2 hours, 1 hour, and 1 hour in sequence to obtain the water extract 1. Bupleurum chinense, Angelica sinensis, Atractylodes macrocephala, Mentha haplocalyx, and Zingiber officinale are first soaked in 10 times the amount of water for 2 hours to extract the volatile oil to obtain the medicinal liquid and the medicinal residues; the medicinal liquid and the water extract 1 are combined to obtain the water extract 2. The medicinal residues are decocted with water 2 times, and the water addition amount is 8 times the amount of the medicinal materials each time, and each decoction is for 1 hour. The decoction is combined with the water extract 2 to obtain the water extract 3, which is filtered, centrifuged, concentrated into a thick extract, and vacuum dried at 70°C to obtain the dry extract.

[0016] In some specific embodiments of the present invention, the way of administering the rolipram in the above construction method is intraperitoneal injection.

[0017] In some specific embodiments of the present invention, the rolipram in the above construction method is an injection solution with a content of 0.3 mg / mL.

[0018] In some specific embodiments of the present invention, the consumption of the sugar water in the above construction method is obtained according to the sucrose preference experiment; the number of entries into the open arms and the residence time in the open arms are obtained according to the elevated plus-maze experiment.

[0019] In some specific embodiments of the present invention, the steps of the sucrose preference experiment in the above construction method include: after the rats are fasted for 24 h, pure water and 1% sucrose water are provided, and after 1 h, the consumption of the sugar water is calculated according to the remaining pure water and the 1% sucrose water.

[0020] In some specific embodiments of the present invention, the above-mentioned state includes at least one of the mental state, body posture, fur color and luster, activity level, degree of response to restraint, color of the palpebral fissure mucosa, color of the auricle, and fecal form of the rats.

[0021] In some specific embodiments of the present invention, the body weight and the consumption of the sugar water of the above-mentioned rats are recorded on the 0th day, the 7th day, the 14th day, and the 21st day; the number of entries into the open arms and the residence time in the open arms are recorded on the 21st day.

[0022] In some specific embodiments of the present invention, the above construction method further includes the step of judging whether the animal model of generalized anxiety disorder with liver depression and spleen deficiency is successfully constructed according to the above-mentioned state, body weight, and consumption of the sugar water.

[0023] The present invention also provides a drug evaluation method, which is based on the animal model of generalized anxiety disorder with liver depression and spleen deficiency obtained by the above construction method for evaluation.

[0024] The present invention has the following effects:

[0025] The present invention uses the method of chronic restraint stress to establish a disease-syndrome combined animal model. This model can be evaluated and identified from multiple dimensions such as characterization and molecular level testing, and is mainly applied to the research on the pathological mechanism and efficacy evaluation of the rat model of generalized anxiety disorder with liver depression and spleen deficiency. The present invention simplifies the modeling operation of the disease-syndrome combined animal model of the rat model of generalized anxiety disorder with liver depression and spleen deficiency, enhances the replicability of the model; multiple evaluation means highlight the disease-syndrome characteristics, enhance the scientific nature of the model, and provide a simple, scientific, and traditional Chinese medicine-characteristic operation plan for establishing the disease-syndrome combined animal model of generalized anxiety disorder with liver depression and spleen deficiency at present. Description of the Drawings

[0026] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required for the description of the embodiments or the prior art.

[0027] Figure 1 Showing the technological process of Xiaoyao Powder

[0028] Figure 2 Showing the changes in the body weights of rats in each group. Among them, compared with the normal group, ***P < 0.001, and compared with the model group, #P < 0.05;

[0029] Figure 3 Showing the consumption of sugar water by rats in each group on the 14th day of stress. Among them, compared with the normal group, ***P < 0.001, and compared with the model group, P < 0.001, #P < 0.05;

[0030] Figure 4 Showing the consumption of sugar water by rats in each group on the 21st day of stress. Among them, compared with the normal group, ***P < 0.001, and compared with the model group, ##P < 0.01;

[0031] Figure 5 Showing the number of times rats in each group entered the bilateral open arms on the 21st day of stress. Among them, compared with the normal group, **P < 0.01, and compared with the model group, ##P < 0.01;

[0032] Figure 6 Showing the staying time of rats in each group entering the bilateral open arms on the 21st day of stress. Among them, compared with the normal group, **P < 0.01, and compared with the model group, #P < 0.05. Detailed implementation manners

[0033] The present invention discloses a method for establishing a rat model of generalized anxiety disorder with liver depression and spleen deficiency by chronic restraint stress. Those skilled in the art can draw on the content of this article and appropriately improve the process parameters to achieve it. It should be particularly noted that all similar substitutions and modifications are obvious to those skilled in the art, and they are all regarded as included in the present invention. The methods and applications of the present invention have been described through preferred embodiments, and those related can obviously make changes or appropriate alterations and combinations to the methods and applications described herein without departing from the content, spirit and scope of the present invention to implement and apply the technology of the present invention.

[0034] The present invention provides a method for constructing and identifying an animal model of generalized anxiety disorder with liver depression and spleen deficiency, as follows.

[0035] S1. Animal Housing: Male Sprague-Dawley rats, weighing 220–250 g, were housed in a room maintained at a constant temperature and humidity, with a 12-h light / 12-h dark cycle. All animals were acclimated to the feeding regimen for one week before formal experiments.

[0036] S2. Animal grouping: The animals were divided into four groups using the randomized block method: normal control group, model group (S), Xiaoyaosan + model group (X), and rolipram + model group (R).

[0037] S3. Modeling Method: Rats in the model group (S), Xiaoyaosan + model group (X), and rolipram + model group (R) were subjected to chronic restraint stress for 3 hours daily at randomized times for 21 consecutive days. Rats in the normal control group were housed in their own cages and did not receive any modeling treatment. During the modeling period, all animals in all groups were deprived of food and water.

[0038] S4. Treatment methods: 30 minutes before restraint, the Xiaoyaosan + model group was given Xiaoyaosan suspension; the model group was given an equal volume of normal saline; and the rolipram + model group was given rolipram injection.

[0039] S5. Evaluation indicators: Before daily administration, observe the rats' mental state, body posture, fur color, and other general conditions; record the body weight of each group of rats at regular intervals; and monitor the animals' sugar water consumption at regular intervals.

[0040] In some embodiments, the method for establishing the S3 model of generalized anxiety disorder of the liver depression and spleen deficiency type is as follows.

[0041] S31. After drug administration, the animals were allowed to rest for 30 minutes before being restrained. Rats in the rolipram, model, and Xiaoyaosan groups were restrained in rat restraint bags. Two adjustable adhesive straps were attached to the rat's chest and abdomen, respectively, on the wire mesh at the top of the cage. The bag opening was trimmed to allow the rat's head to protrude, and the tightness of the straps was adjusted to ensure normal breathing. The animals remained in the same environment throughout the restraint process and were not given food or water. Restraint was performed for 3 hours daily, at randomized times, for 21 consecutive days.

[0042] S32. The rats in the normal group were kept in their own cages and no modeling was performed.

[0043] In some embodiments, the treatment method of S4 is as follows.

[0044] S41. 30 minutes before restraint, the Xiaoyaosan + model group was given Xiaoyaosan suspension by gavage (the rat equivalent dose was converted from the human dose, and the rat dose was 3.854 g·kg). -1 ·d -1 ) treatment, with an oral gavage volume of 10 mL / kg.

[0045] S42. The model group was intragastrically administered with an equal volume of normal saline;

[0046] S43. The rolipram + model group was intraperitoneally injected with the rolipram group injection solution (the dosage was 0.3 mg / mL) 30 min before restraint daily.

[0047] In some embodiments, the evaluation method of S5 is specifically as follows.

[0048] S51. General condition

[0049] Before drug administration daily, carefully observe the mental state, body posture, fur color and luster, activity level, response degree to restraint, color of palpebral fissure mucosa, color of auricle, and feces and other general conditions of the rats.

[0050] S52. Change in body weight of rats

[0051] After the experiment started, weigh the rats with an electronic scale before gavage every other day, and adjust the corresponding gavage and injection dosages according to the body weight. Record the body weights of rats in each group at regular intervals and make a dynamic comparison.

[0052] S53. Sucrose preference experiment

[0053] Observe the sugar water consumption of animals at regular intervals. After 24 h of fasting and water deprivation, give each rat 2 pre - quantified bottles of water: 1 bottle of 1% sucrose water and 1 bottle of pure water. After 1 h, take away the 2 bottles and weigh them. When conducting the test, all animals are measured individually in a single cage. The sugar water consumption reflects the response degree of animals to rewards.

[0054] S54. Elevated plus - maze experiment

[0055] The test was conducted on the 21st day of the experiment. Move all animals into the operation room 1 h in advance. The operation room should be quiet, and the light source should be blocked with a black light - proof cloth. Before the test, place the rats to be tested in a separate rat cage. After adapting for 5 min, quickly place them on the central platform of the elevated plus - maze. Ensure that the head direction is the same each time. Observe each rat for 5 min. After taking out the rat, clean the maze with 75% alcohol and then water in sequence. After the smell dissipates, conduct the next test. Try to ensure that the shooting position is the same for each experiment.

[0056] It should be understood that the expression "one or more of..." individually includes each of the objects recited after the expression and various different combinations of two or more of the recited objects, unless otherwise understood from the context and usage. The expression "and / or" in combination with three or more recited objects should be understood to have the same meaning, unless otherwise understood from the context.

[0057] The terms "comprising", "having", or "including", including the use of their grammatical synonyms, should generally be understood as open-ended and non-limiting, e.g., not excluding other unrecited elements or steps, unless specifically stated otherwise or understood from the context otherwise.

[0058] It should be understood that the order of steps or the order of performing certain actions is not important as long as the present application remains operable. In addition, two or more steps or actions can be carried out simultaneously.

[0059] The use of any and all examples or exemplary language herein, such as "for example" or "including", is merely intended to better illustrate the present application and does not limit the scope of the present application. No language in this specification should be construed as indicating that any unclaimed element is essential for the practice of the present application.

[0060] In addition, the numerical ranges and parameters used to define the present application are approximate values. The relevant values in the specific embodiments have been presented as precisely as possible herein. However, any numerical value inherently and inevitably contains standard deviations due to individual testing methods. Therefore, unless otherwise clearly stated, it should be understood that all ranges, amounts, numerical values, and percentages used in this disclosure are modified by "about". Herein, "about" generally means that the actual value is within plus or minus 10%, 5%, 1%, or 0.5% of a specific value or range.

[0061] Unless otherwise specified, the raw materials, reagents, consumables, and instruments involved in the present invention are all ordinary commercially available products and can be purchased from the market.

[0062] The present invention will be further described below in conjunction with embodiments.

[0063] Embodiment

[0064] (1) Animal feeding environment

[0065] Forty-eight male Sprague-Dawley rats, weighing 220 - 250 g, were housed in a room with constant temperature and humidity, with a light cycle of 12 h light / 12 h dark. Before the start of the experiment, all animals were fed adaptively for one week before the formal experiment. All experimental operations followed the NIH Guide for the Care and Use of Laboratory Animals (NIH publications No. 80 - 23, 1996). The experimental protocol was approved by the Animal Use Committee of the West Virginia University Health Sciences Center.

[0066] (2) Animal grouping

[0067] Using the random block method, the animals were equally divided into 4 groups: normal control group, model group (S), Xiaoyaosan + model group (X), and rolipram + model group (R), with 2 rats in each cage.

[0068] (3) Modeling method

[0069] The rat model was established using chronic restraint stress in three groups: the model group (S), the Xiaoyaosan + model group (X), and the rolipram + model group (R). The stress was applied 3 hours daily at randomized times for 21 consecutive days. The normal group was housed in their own cages and did not receive any modeling treatment. During the modeling period, all animals in the groups were deprived of food or water.

[0070] Specifically, the modeling method is as follows.

[0071] a) After drug administration, the animals were allowed to rest for 30 minutes before being restrained. Rats in the rolipram, model, and Xiaoyaosan groups were restrained in rat restraint bags. Two adjustable adhesive straps were attached to the rat's chest and abdomen, respectively, on the wire mesh at the top of the cage. The bag opening was trimmed to allow the rat's head to protrude, and the tightness of the straps was adjusted to ensure normal breathing. The animals remained in the same environment throughout the restraint process and were deprived of food and water. Restraint was performed for 3 hours daily, at randomized times, for 21 consecutive days.

[0072] b) The mice in the normal group were kept in their own cages without any modeling treatment.

[0073] (4) Treatment methods

[0074] 30 minutes before restraint, the Xiaoyaosan+model group was given Xiaoyaosan suspension; the model group was given an equal volume of normal saline; and the rolipram+model group was given rolipram injection.

[0075] Specifically, the treatment procedure is as follows.

[0076] a) 30 min before restraint, the Xiaoyaosan + model group was given Xiaoyaosan suspension by gavage (the rat equivalent dose was converted from the human dose, and the dose was 3.854 g·kg -1 ·d -1 ) treatment, with an oral gavage volume of 10 mL / kg. The Xiaoyao Powder used in the experiment originated from the Taiping Huimin Hejijufang, and the prescription composition is: 30 g of Bupleurum chinense, 30 g of Angelica sinensis, 30 g of White Peony Root, 30 g of Atractylodes macrocephala, 30 g of Poria cocos, 15 g of Roasted Licorice Root, 10 g of Ginger, and 10 g of Menthol. It was purchased from Beijing Tongrentang (Bozhou) Pieces Co., Ltd. (the processing standard is the "Beijing Traditional Chinese Medicine Processing Standard"; production license number: (Anhui) Y20030002). It was identified as a pure medicinal material by the National Medical Hall Pharmacy of Beijing University of Chinese Medicine, and was made into a dry extract in strict accordance with the process flow shown in the figure below for use. When used, it was dissolved and prepared with distilled water and an appropriate container as needed (the process flow is as shown in the figure below). Figure 1 ).

[0077] b) The model group was given an equal volume of normal saline;

[0078] c) In the rolipram + model group, rats were intraperitoneally injected with rolipram injection (dosage: 0.3 mg / mL, injection volume: 1 mL / kg) 30 minutes before restraint daily.

[0079] (5) Evaluation indicators

[0080] Before drug administration daily, observe the general conditions of the rats such as mental state, body posture, fur color, etc.; record the body weights of rats in each group on days 0, 7, 14, and 21; detect the sugar water consumption of the animals on days 0, 7, 14, and 21. Specifically, the evaluation methods are as follows.

[0081] a) General conditions

[0082] Before drug administration daily, carefully observe the general conditions of the rats including mental state, body posture, fur color and luster, activity level, reaction degree to restraint, color of the palpebral fissure mucosa, color of the auricle, and feces, etc.

[0083] b) Changes in rat body weight

[0084] After the start of the experiment, weigh the rats with an electronic scale before gavage every other day, and adjust the corresponding gavage and injection dosages according to the body weight. Record the body weights of rats in each group on days 0, 7, 14, and 21 and make a dynamic comparison.

[0085] c) Sucrose preference experiment

[0086] At four time points on days 0, 7, 14, and 21, observe the sugar water consumption of the animals. After 24 hours of fasting and water deprivation, give each rat two pre - quantified bottles of water: one bottle of 1% sucrose water and one bottle of pure water. After 1 hour, take away the two bottles and weigh them. When conducting the test, all animals are measured individually in cages. The sugar water consumption reflects the reaction degree of the animals to rewards.

[0087] d) Elevated plus - maze experiment

[0088] Conduct on day 21 to evaluate the anxiety level of the rats. The inner wall and bottom surface of the elevated plus - maze are dark gray, 50 cm high from the bottom surface, and it consists of two opposite open arms (length × width = 45 cm × 15 cm), two opposite closed arms (length × width × height = 45 cm × 15 cm × 30 cm), and a central platform (15 cm × 15 cm) connecting the four arms, in a cross shape. Place a camera directly above the central platform and connect it to a computer, and adjust the focal length. Evaluate the anxiety of the rats by recording the number of entries into the open arms and the residence time.

[0089] (6) Experimental results

[0090] As shown in Tables 1 to 4, Figures 2 to 6As shown. Table 1 records the states of each group of rats during the experiment.

[0091] Table 1: Observation of the general states of each group of rats

[0092]

[0093] Figure 2 (The corresponding data are shown in Table 2) shows the weight changes of each group of rats, Figure 3 (The corresponding data are shown in Table 3) shows the sugar water consumption of each group of rats on the 14th day of stress, Figure 4 shows (the corresponding data are shown in Table 3) the sugar water consumption of each group of rats on the 21st day of stress, Figure 5 (The corresponding data are shown in Table 4) shows the number of times each group of rats entered the bilateral open arms on the 21st day of stress, Figure 6 (The corresponding data are shown in Table 4) shows the staying time of each group of rats in the bilateral open arms on the 21st day of stress. Compared with the model group, the weight loss of rats in the Rolipram + model group was significantly reduced, the sugar water consumption increased, and the number of times and staying time of entering the bilateral open arms increased. Since Rolipram is known to have the effect of reducing anxiety, combined with the above description of the general state, it can be seen that Rolipram reduced the anxiety behavior of rats, indicating that the model group successfully induced anxiety disorder. Compared with the model group, the weight loss of rats in the Xiaoyaosan + model group was significantly reduced, the sugar water consumption increased, and the number of times and staying time of entering the bilateral open arms increased. Since Xiaoyaosan is known to be mainly used for treating liver depression and spleen weakness syndrome, combined with the above description of the general state, it can be seen that Xiaoyaosan improved the anxiety behavior of rats, indicating that liver depression and spleen weakness syndrome was successfully induced.

[0094] Table 2: Weight changes of each group of rats (g)

[0095]

[0096] Note: Compared with the normal group *** P<0.001; compared with the model group # P<0.05.

[0097] Table 3: Sugar water consumption of each group of rats on the 14th and 21st days of stress (g)

[0098]

[0099] Note: Compared with the normal group *** P<0.001, ** P<0.01; compared with the model group ### P <0.001, ## P <0.01, # P<0.05.

[0100] Table 4: Number of times and staying time of each group of rats entering the bilateral open arms on the 21st day of stress

[0101]

[0102] Note: Compared with the normal group ** P < 0.01; compared with the model group ## P < 0.01, # P < 0.05.

[0103] The above are only the preferred embodiments of the present invention. It should be pointed out that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and modifications can be made, and these improvements and modifications should also be regarded as the protection scope of the present invention.

Claims

1. A method for constructing an animal model of generalized anxiety disorder with liver depression and spleen deficiency type, characterized in that, It includes the following steps: Step (1): Divide the rats into a control group, model group 1, model group 2, and model group 3 according to the randomized block method; Step (2): Record the status of the rats every day. Administer Xiaoyao Powder to the rats in model group 1, and administer rolipram to the rats in model group 2. After 30 min, perform chronic restraint stress on the rats in model group 1, model group 2, and model group 3 for 3 h, and repeat for 21 days. During this period, record the body weight, sugar water consumption, number of entries into the open arms, and residence time in the open arms of the rats to obtain an animal model of generalized anxiety disorder with liver depression and spleen deficiency; The operation steps of the chronic restraint stress include: Restrain the rats in a rat restraint bag. Take two adjustable adhesive soft bands to fix the chest and abdomen of the rats on a plane respectively, so that the heads of the rats protrude from the notch of the restraint bag. Adjust the tightness of the adhesive soft bands to ensure normal breathing of the rats. All rats are in the same environment during the whole restraint process and are not given food or water.

2. The construction method according to claim 1, wherein The rats are male SD rats weighing 220 - 250 g.

3. The construction method according to claim 1 or 2, characterized in that, The administration method of Xiaoyao Powder is gavage.

4. The construction method according to claim 3, characterized in that The dosage of the Xiaoyao Powder is 3.854 g·kg -1 ·d -1 .

5. The construction method according to any one of claims 1 to 4, characterized in that, The administration method of rolipram is intraperitoneal injection.

6. The construction method according to claim 5, characterized in that, The rolipram is an injection with a content of 0.3 mg / mL.

7. The construction method according to any one of claims 1 to 6, characterized in that The sugar water consumption is obtained according to the sucrose preference experiment; the number of entries into the open arms and the residence time in the open arms are obtained according to the elevated plus maze experiment.

8. The construction method according to claim 7, characterized in that, The steps of the sucrose preference experiment include: After the rats are fasted for 24 h, provide pure water and 1% sucrose water. After 1 h, calculate the sugar water consumption based on the remaining pure water and 1% sucrose water.

9. The construction method according to any one of claims 1 to 8, characterized in that, The status includes at least one of the mental state, body posture, fur color and luster, activity level, reaction degree to restraint, color of the fissure mucosa of the eyes, color of the auricles, and fecal form of the rats.

10. The construction method according to any one of claims 1 to 9, characterized in that, The body weight and sugar water consumption of the rats are recorded on the 0th day, 7th day, 14th day, and 21st day; the number of entries into the open arms and the residence time in the open arms are recorded on the 21st day.

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