Composition with depression-relieving effect and application thereof

Abstract

The invention discloses a composition and its application with the effect of relieving depression, and belongs to the field of biomedicine. The raw materials of the composition are measured by weight, including the following components: 7-13 parts of Tremella, 5-12 parts of Dictyophora serrata and 5-13 parts of Morchella solani. The present invention uses Tremella fuciformis, Dictyophora serrata and Morchella solani fruiting bodies of different proportions as raw materials, takes the effect of relieving depression as evaluation criteria, and performs drug efficacy tests by animal experiments and in vitro cell experiments, and is verified by methods such as immunofluorescence staining and immunoblot analysis, and finds that the aqueous extracts of Tremella fuciformis, Dictyophora serrata and Morchella solani fruiting bodies have the effect of relieving depression. The antidepressant component provided by the present invention, the raw material is high-quality edible and medicinal fungi, the toxic and side effects are small, the preparation process is simple, and the treatment effect is unstable due to the difference between different batches of materials, which provides a new direction for the treatment of depression.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine, and in particular to a composition capable of alleviating depression and application thereof. Background Art

[0002] Depressive disorder or depression is a common mental health condition that can happen to anyone. It is characterized by prolonged low mood or loss of pleasure or interest in activities. Depression is different from normal mood changes and feelings about daily life. Depressive mood occurs almost every day, lasts for most of the day, and lasts for at least two weeks. People with depression may experience sleep disorders and changes in appetite. They may feel a lack of self-worth, have suicidal thoughts, and feel hopeless about the future. Physical fatigue and difficulty concentrating are also common. Data show that worldwide, more than 10% of pregnant women and women who have just given birth suffer from depression. Depression can make all aspects of life difficult, affecting schoolwork, work efficiency, and relationships with family, friends and the community. Researchers estimate that 12 billion work days are lost worldwide each year due to depression and anxiety, and productivity losses amount to nearly $1 trillion each year. People with depression have a higher risk of suicide. Studies have also shown a close relationship between depression and physical health problems, including cardiovascular disease, cancer, diabetes and respiratory diseases.

[0003] Depression is the result of a combination of social, psychological and biological factors. Preventive measures have been shown to reduce depression. Psychological therapies and medications are available to effectively treat depression. However, there is not universal access to psychological treatment services or medications for depression. It is estimated that more than 75% of people with depressive disorders in low- and middle-income countries do not receive treatment.

[0004] There are already technologies for treating depression using Chinese medicine compositions or extracts in the prior art, but the preparation process of the composite formula is too many and complicated, and there may be differences between different batches of materials, resulting in unstable treatment effects. Therefore, the current treatment options are not targeted at treating depression, the treatment effect is minimal, and the mechanism of action is unclear. Summary of the invention

[0005] The purpose of the present invention is to provide a composition having the effect of relieving depression and its application, so as to solve the problems existing in the above-mentioned prior art. The present invention uses Tremella fuciformis, Dictyophora short-skirted and Morchella solani fruiting bodies as raw materials to extract effective substances, verifies their anti-depressant efficacy, and has safe raw materials, significant and stable effects, providing a new direction for the treatment of depression.

[0006] To achieve the above object, the present invention provides the following solutions:

[0007] The invention provides a composition with the effect of relieving depression. The raw materials of the composition comprise the following components in parts by weight: 7-13 parts of Tremella fuciformis, 5-12 parts of Dictyophora short-skirted and 5-13 parts of Morchella solani.

[0008] Furthermore, the raw materials of the composition include, by weight, 8 parts of Tremella fuciformis, 12 parts of Dictyophora short-skirted and 5 parts of Morchella solani.

[0009] Furthermore, the preparation method of the composition comprises the following steps: mixing the fruiting bodies of the Tremella fuciformis, the Dictyophora short-skirted and the Morchella solani, adding water to the mixed fruiting bodies and soaking them overnight, then extracting them in a water bath, taking the supernatant after centrifugation, and concentrating them to obtain the composition.

[0010] Furthermore, during soaking, the mass volume ratio of the mixed fruiting body to water is 1 g:50 mL.

[0011] Furthermore, the soaking temperature is 4°C.

[0012] Furthermore, the water bath extraction temperature is 80° C. and the extraction time is 5 h.

[0013] Furthermore, the concentration is to concentrate to 1 / 15 of the original volume.

[0014] The present invention also provides application of the composition in preparing medicine for preventing and treating depression.

[0015] The present invention also provides a medicine for preventing and treating depression, comprising the composition.

[0016] Furthermore, it also includes pharmaceutically acceptable carrier substances and / or adjuvants.

[0017] The present invention discloses the following technical effects:

[0018] The present invention uses different proportions of Tremella fuciformis, Dictyophora short-skirted and Morchella solani fruiting bodies as raw materials, takes the effect of relieving depression as the evaluation standard, conducts efficacy tests through animal experiments and in vitro cell experiments, and verifies through immunofluorescence staining, immunoblot analysis and other methods, and finds that the water extracts of Tremella fuciformis, Dictyophora short-skirted and Morchella solani fruiting bodies have the effect of relieving depression, and further combines the results of metagenomic microbiome and serum metabolomics analysis to explore their mechanism of relieving depression. The results show that the water extracts of the above three high-quality edible and medicinal mushroom fruiting bodies inhibit pro-inflammatory factors in the brain by changing the intestinal flora and serum metabolite composition in animals, thereby exerting a depressive effect.

[0019] The antidepressant component provided by the present invention has high-quality edible and medicinal fungi as raw materials, has little toxic and side effects, has a simple preparation process, and does not cause unstable treatment effects due to differences between different batches of materials. The present invention provides a new direction for the treatment of depression. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0021] Figure 1 The effects of water extracts of Tremella fuciformis, Dictyophora short-skirted and Morchella solani fruiting bodies on the viability of BV2 cells treated with LPS (A) and the amount of LDH released (B), among which, compared with the blank group BV2 cells, ###p<0.001; compared with the model group BV2 cells, ***p<0.001; n=6 / group;

[0022] Figure 2 The effects of water extracts of Tremella fuciformis, Dictyophora short-skirted and Morinda solani fruiting bodies on the expression level of NO in BV2 cells stimulated by LPS, among which, compared with the blank group BV2 cells, ###p<0.01; compared with the model group BV2 cells, **p<0.01; n=6 / group;

[0023] Figure 3 The effects of TDP administration on the behavior of experimental mice, A is the increase in sucrose consumption of CUMS mice in the sucrose preference test, B is the reduction in the immobility time of CUMS mice in the forced swimming test, C is the reduction in the immobility time of CUMS mice in the tail suspension test, D is the increase in the time of CUMS mice crossing the center in the open field test, and E is the open field trajectory diagram;

[0024] Figure 4 This is the OTU Venn diagram of the intestinal flora in the blank group, model group, and TDP2 group;

[0025] Figure 5 This is the taxonomic composition analysis diagram of the intestinal flora in the blank group, model group and TDP2 group;

[0026] Figure 6 Heat map of intestinal flora species composition in the blank group, model group and TDP2 group;

[0027] Figure 7 This is the LEfSe analysis diagram of intestinal flora;

[0028] Figure 8 The Venn diagram of serum metabolic differences between the blank group and the model group, and between the TDP2 group and the model group;

[0029] Fig. 9 Multivariate statistical PLS-DA analysis of serum metabolism in the blank group, model group and TDP2 group;

[0030] Fig.10 This is the heat map of correlation analysis of serum metabolic differential substances in the blank group, model group and TDP2 group;

[0031] Fig.11 KEGG enrichment pathway analysis for serum metabolism;

[0032] Fig.12 This is the heat map of the association between intestinal flora and serum metabolites in the blank group, model group and TDP2 group;

[0033] Fig.13 Immunofluorescence images of SYN and PSD95 in the CA1 and CA3 regions of the mouse hippocampus;

[0034] Fig.14 Quantitative analysis of SYN (A) and PSD95 (B) in the CA1 and CA3 regions of the mouse hippocampus;

[0035] Fig.15 The results of immunoblotting of proinflammatory factors IL-1β and TNF-α in the brains of mice in each group. DETAILED DESCRIPTION

[0036] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0037] It should be understood that the terms described in the present invention are only for describing a particular embodiment and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. The intermediate value in any stated value or stated range, and each smaller range between any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0038] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0039] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present invention description and examples are exemplary only.

[0040] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0041] Example

[0042] 1. Experimental Methods

[0043] 1. Preparation of aqueous extracts (TDP) of Tremella fuciformis, Dictyophora short-skirted and Morchella solani fruiting bodies

[0044] First, the water extracts of Tremella fuciformis, Dictyophora short-skirt and Morchella chuatus were prepared. The three fruiting bodies were first proportioned by weight, respectively, TDP1 (13 parts of Tremella fuciformis, 7 parts of Dictyophora short-skirt and 8 parts of Morchella chuatus); TDP2 (8 parts of Tremella fuciformis, 12 parts of Dictyophora short-skirt and 5 parts of Morchella chuatus); TDP3 (7 parts of Tremella fuciformis, 5 parts of Dictyophora short-skirt and 13 parts of Morchella chuatus), and each group of fruiting bodies with different proportions was added with drinking water at a material-liquid ratio of 1g:50mL, stirred evenly, soaked in a 4℃ refrigerator overnight, and the three were extracted in a constant temperature water bath at 80℃ for 5h. After cooling to room temperature, the three were centrifuged to take the supernatant, and concentrated to 1 / 15 volume by rotary evaporation to obtain three kinds of fruiting body water extracts for subsequent experiments.

[0045] 2. In vitro cell experiment on the relief of depression by aqueous extracts of Tremella fuciformis, Dictyophora short-skirted and Morchella solani fruiting bodies

[0046] (1) Cell culture

[0047] Cell recovery: After the cells are taken out and thawed, they are quickly transferred to a centrifuge tube containing MEM-containing BV2 cell-specific culture medium. A small amount of culture medium is used to repeatedly rinse the cryotube, and then centrifuged at 800 rpm for 5 minutes. Aspirate the supernatant with a pipette, replace a new pipette tip and add 1 mL of new complete culture medium to redisperse the cells. Be gentle and do not collide with the cells. According to the amount of cell precipitation, an appropriate volume of cell suspension is pipetted and added to the adherent culture flask containing complete culture medium. After tightening the lid, shake it crosswise and transfer it horizontally to the incubator to avoid uneven cell growth. The incubator program is set to 37°C and 5% CO2.

[0048] Cell passaging: discard the culture medium in the culture flask, rinse the culture flask with PBS 2-3 times to remove the residual complete culture medium. Add 1mL of pre-melted trypsin, incubate at 37℃ for 1min, observe the degree of cell digestion under a microscope and tap gently. Then add 4mL of BV2 cell-specific culture medium as soon as possible to avoid over-digestion, quickly aspirate the culture medium with an electric pipette to rinse the culture flask to collect cells, centrifuge at 800rpm for 5min, aspirate the supernatant with a pipette, replace the new pipette tip and add 1mL of new complete culture medium to resuspend the cells, be gentle and do not collide with the cells, aspirate an appropriate volume of cell suspension according to the amount of cell sedimentation and transfer it to the adherent culture flask, shake it gently and transfer it horizontally to the incubator, minimize shaking to avoid uneven cell growth, and culture at 37℃ and 5% CO2. After the cells are stably passed to the third generation, they can be used for cell experiments.

[0049] (2) Cell viability assay

[0050] Since LPS can activate inflammation-related pathways and promote the production of inflammatory factors, it is widely used to construct in vitro and in vivo models of inflammation and oxidative stress. One of the classic in vitro models of neuroinflammation is BV2 cells stimulated with LPS. The present invention digests and counts BV2 cells and dilutes the cells to 5×10 4 / mL, take a 96-well plate and add PBS to the outermost circle of wells to prevent the evaporation of the liquid solution, add 100μL of cell suspension to each well in the middle, shake gently and place it horizontally in the incubator to avoid uneven cell growth, and culture overnight. Use the same day to prepare the mother solution of the fruiting body water extract and 1μg / mL LPS. Cell viability detection was performed using a 96-well plate, and all groups kept the total culture volume at each stage at 100μL. BV2 cells in the blank and model groups were not pretreated, and the other groups were pretreated with 100μM fruiting body water extracts of different ratios. After the modeling began, the blank group BV2 cells were not treated, the model group used 1μg / mL LPS to stimulate BV2 cells, and each drug group was modeled with 1μg / mL LPS on the basis of the original drug administration. Add 20μL of sterilized MTT solution to each well in the clean bench in the dark, and incubate in the incubator for 4h under dark conditions. Then, use a syringe needle to carefully suck up the supernatant, use DMSO to dissolve the crystal formazan at the bottom of the well plate, incubate in the incubator for several minutes to completely dissolve the crystals, measure the optical density at a wavelength of 490 nm, and then calculate the cell viability using the following formula.

[0051]

[0052] (3) NO detection

[0053] The gaseous signaling molecule NO has a variety of physiological functions, such as vasodilation, neurotransmitter transmission and host defense. Excessive NO production can also be used as an indicator of inflammation. BV2 cells were digested and diluted to a final cell density of 7.5×10 4 / mL, add 2mL of cell suspension to each well of a 6-well plate, shake it with the cross method and place it horizontally in the incubator to avoid uneven cell growth, and culture it overnight. Use the mother solution of each ratio of fruiting body water extract and 1μg / mL LPS prepared on the same day. All groups maintain the total culture volume of 2mL at each stage. BV2 cells in the blank and model groups were not pretreated, and the other groups were pretreated with 100μM fruiting body water extracts of different ratios. After the modeling began, BV2 cells in the blank group were not treated, and BV2 cells in the model group were stimulated with 1μg / mL LPS. Each drug group was modeled with 1μg / mL LPS on the basis of the original drug administration. After 24h of modeling, the culture medium in the well was discarded, and PBS was gently rinsed twice, and then a total of 500μL of cell lysate was added to each well several times, and the cells were gently scraped off with a cell scraper to collect the sample solution. Collect the supernatant after centrifugation at 13000rpm for 5min. In a new 96-well plate, add 50 μL of sample to each well, then add equal volumes of Griess reagents I and II to each well under light-proof conditions, mix well, and detect the absorbance at 540 nm.

[0054] 3. In vivo animal experiment on the effect of aqueous extracts of Tremella fuciformis, Dictyophora short-skirted and Morchella solani fruiting bodies on relieving depression

[0055] (1) Depression mouse model

[0056] The experimental mice were subjected to the chronic unpredictable animal stress model (CUMS) and the experimental animal species was male C57BL / 6 mice. In the first week, sucrose preference test and cage separation were performed before modeling.

[0057] From the 2nd week to the 7th week, C57BL / 6 mice without chronic unpredictable stimulation were used as the control group in this experiment. The CUMS model was constructed with C57BL / 6 mice subjected to chronic unpredictable stimulation, and used as a group 1 (TDP1, n = 10, ratio: 13 parts of Tremella fuciformis, 7 parts of Dictyophora short-skirted and 8 parts of Morchella solani), a group 2 (TDP2, n = 10, ratio: 8 parts of Tremella fuciformis, 12 parts of Dictyophora short-skirted and 5 parts of Morchella solani), a group 3 (TDP3, n = 10, ratio: 7 parts of Tremella fuciformis, 5 parts of Dictyophora short-skirted and 13 parts of Morchella solani), a positive group (n = 10) and a model group (n = 10). The modeling methods included: restraint stress for 2 hours, forced swimming, wet bedding, fasting for 24 hours, 45°C heat stress for 5 minutes (hot plate), day and night reversal, and 6°C swimming for 5 minutes.

[0058] After 6 weeks of modeling, the modeling effect was tested by sucrose preference experiment. The experimental steps were the same as in the first week. The results of sucrose preference experiment showed that the modeling effect was not obvious, so the modeling time was increased by 2 weeks, and the CUMS model was continued from the 9th to the 10th week. The modeling method was changed to: tail suspension for 10 minutes, forced swimming, wet bedding, fasting for 24 hours, 45℃ heat stress for 8 minutes (hot plate), day and night reversal, and swimming at 4℃ for 6 minutes.

[0059] In the 11th week, the modeling effect was tested by sucrose preference experiment. The experimental steps were the same as in the 1st week. The results of sucrose preference experiment showed that the modeling was successful. In the 12th week, the modeling was continued for one week and then grouped into: blank control group, model group, positive drug group (administered with fluoxetine), TDP1 (administered by weight, the ratio of the prepared fruiting body water extract of 13 parts of Tremella fuciformis, 7 parts of Dictyophora short-skirt and 8 parts of Morchella solani), TDP2 (administered by weight, the ratio of the fruiting body water extract of 8 parts of Tremella fuciformis, 12 parts of Dictyophora short-skirt and 5 parts of Morchella solani), TDP3 (administered by weight, the ratio of the fruiting body water extract of 7 parts of Tremella fuciformis, 5 parts of Dictyophora short-skirt and 13 parts of Morchella solani). Drug administration began after grouping.

[0060] (2) Administration of drugs to mice with depression

[0061] C57BL / 6 mice were modeled and medicated from week 13 to week 17. Mice in the blank control group and model group were given water; mice in the positive drug group were given fluoxetine, with a concentration of 3 mg / kg (prepared into 0.3 mg / mL) based on the human dose; each drug-treated group was given a different ratio of fruiting body water extract, with a concentration of 300 mg / kg (prepared into 30 mg / mL).

[0062] (3) Behavioral experiments on mice with depression

[0063] In the 17th week, behavioral experiments were conducted on depression mice to test the effect of drug administration. The behavioral experiments included forced swimming test, open field test and tail suspension test, and the behavioral results were analyzed and plotted. After the behavioral experiments, sucrose preference test was conducted again.

[0064] (4) Collection of mouse serum and organs

[0065] After the animal experiment, the experimental mice were euthanized. Before the animals were killed, the mice in each group were fasted but not watered for 4 hours, and blood was collected from the tail vein. The blood was left to stand at room temperature for 30 minutes, centrifuged at 3000r / min for 10 minutes, and the supernatant was collected and stored in a -80℃ refrigerator for later use. Subsequently, the experimental animals were euthanized, the contents of the cecum were collected, and the brain, heart, liver, spleen, and kidney were quickly removed on ice and stored in a -80℃ refrigerator for later use.

[0066] (5) Detection of serum metabolic related factors

[0067] Through LC-MS / MS analysis, the levels of various metabolites in the serum of experimental mice were detected, and they were qualitatively and quantitatively analyzed to obtain the quality evaluation of serum metabolite data and analyze them.

[0068] (6) Analysis of intestinal microbial communities

[0069] Fecal samples were randomly selected from the control group, model group, positive drug group, and the group administered with the best fruiting body water extract (n = 4 / group). The bacterial composition of each sample was analyzed by 16S rRNA gene analysis using the Illumina MiSeq platform. Sequences were assigned to operational taxonomic units (OTUs) with >97% pairwise identity. Linear discriminant analysis (LDA) effect size (LEfSe) was used to determine biomarkers for each group (significant when LDA score >2, p≤0.05).

[0070] (7) Immunofluorescence analysis

[0071] The cells were stained with Syn and PSD956 primary antibodies using the TSA method. The nuclei were stained with goat anti-rabbit secondary antibodies and DAPI. The mounted tissue slides were photographed using a Pannoramic SCAN microscope. The intensity profiles were analyzed using ZEN2.6 (Blue Edition) software.

[0072] 2. Experimental Results

[0073] 1. Cytotoxicity of aqueous extracts of Tremella fuciformis, Dictyophora short-skirted and Morchella solani fruiting bodies in different proportions

[0074] The present invention uses the LPS-induced BV2 cell model to explore the anti-inflammatory effect of different ratios of TDP in vitro. First, it was studied whether it has a protective effect on LPS-induced BV2 cells. The results are as follows Figure 1 When cells were stimulated by LPS, cell viability decreased significantly (p<0.001, Figure 1 When the cells stimulated by LPS were treated with different ratios of fruiting body water extracts, the number of BV2 live cells increased significantly (p<0.001, Figure 1 A), which confirmed that TDP could protect BV2 cells induced by LPS, among which the water extract of TDP2 (8 parts of Tremella fuciformis, 12 parts of Dictyophora short-skirted and 5 parts of Morchella solani) had the best effect.

[0075] The large amount of LDH released under cell damage can also be used as one of the indicators to measure the severity of cell damage. After BV2 cells were treated with LPS for 24 hours, the culture medium of all groups was collected to detect the release of LDH by BV2 cells after LPS stimulation to determine whether TDP has an improvement effect. Compared with the blank group BV2 cells, the amount of LDH released in BV2 cells after LPS treatment increased by 17.74% (p<0.001, Figure 1 The water extract of the second ratio (8 parts of Tremella fuciformis, 12 parts of Dictyophora short-skirted and 5 parts of Morchella solani) had the most significant effect on the reduction of LDH release in BV2 cells induced by LPS (p<0.001, Figure 1 B). Therefore, the aqueous extracts of Tremella fuciformis, Dictyophora short-skirted and Morchella solani can protect BV2 cells from LPS-induced inflammatory damage.

[0076] 2. The aqueous extracts of Tremella fuciformis, Dictyophora short-skirted and Morchella solani fruiting bodies can inhibit the production of NO by BV2 cells induced by LPS

[0077] Compared with the blank control group, the NO production in BV2 cells after LPS treatment was significantly increased (p<0.001, Figure 2 ), indicating that LPS induced an inflammatory response in BV2 cells. TDP administration restored NO levels in BV2 cells to normal levels and effectively reduced NO production (p<0.001, Figure 2 ), indicating that the administration of water extract can alleviate the inflammatory response in BV2 cells induced by LPS stimulation.

[0078] 3. Administration of aqueous extracts of Tremella fuciformis, Dictyophora short-skirted and Morchella solani fruiting bodies improves depressive-like behavior in CUMS mice

[0079] SPSS software was used to perform ANOVA analysis on the behavioral data of experimental mice, and Graphpad Pism9.5 was used to plot the analysis results. From the analysis of the experimental data, it can be seen that the administration of TDP to CUMS mice can significantly alleviate their depressive-like behavior. Figure 3 The sucrose preference test data of A showed that after administration of TDP, the percentage of sugar water consumption in the model mice increased significantly, and the anhedonia was alleviated; Figure 3 From B, we can see that in the forced swimming test, compared with the model mice, the immobility time of the mice in the three drug-administered groups was significantly reduced, and the desire to survive increased; Figure 3 It can be seen from the C that in the tail suspension test, after the administration of the fruiting body water extract, the immobility time of CUMS mice in the tail suspension test was significantly reduced; at the same time, the results of the open field test showed that after TDP administration, the number of times the model mice crossed the center of the open field increased significantly, and their desire to explore was stronger than that of the model mice ( Figure 3 D), Figure 3E is the open field trajectory diagram. From the above results, it can be seen that TDP can effectively alleviate the depressive-like behavior of model mice, among which TDP2 (8 parts of Tremella fuciformis, 12 parts of Dictyophora short skirt and 5 parts of Morchella solani) has the best effect.

[0080] 4. Effects of Tremella fuciformis, Dictyophora short-skirted and Morchella solani on intestinal flora and serum metabolome of experimental animals

[0081] The drug administration group with the best efficacy in the behavioral data (TDP2: 8 parts of Tremella fuciformis, 12 parts of Dictyophora short-skirted and 5 parts of Morchella edulis) was selected for intestinal flora analysis and serum metabolome analysis.

[0082] Results of intestinal flora analysis ( Figure 4-Figure 7 ) showed that CUMS modeling changed the abundance of intestinal microorganisms in mice, which was reversed after TDP administration. Compared with the blank group and TDP2 group, the abundance of beneficial bacteria in the Bacteroidetes phylum in CUMS mice decreased, while the abundance of harmful bacteria in the Actinobacteria phylum increased. Studies have shown that patients with NMDAR encephalitis have higher levels of serum inflammatory cytokines, including IL-3, IL-6, and TNF-α, while Prevotella in the Bacteroidetes phylum is negatively correlated with serum inflammatory cytokines, while Bifidobacterium in the Actinobacteria phylum is positively correlated.

[0083] like Figure 8-Figure 12 As shown in the results, serum metabolome analysis revealed that the glutamate content in the CUMS group mice was negatively correlated with that in the two matched groups of mice. Previous studies have shown that glutamate is strongly associated with depression. It can mediate most central nervous excitatory conduction. The main receptor involved in glutamate-mediated neuronal damage is NMDAR, which is essential for neuronal plasticity. The increased activation of glutamate by high concentrations of glutamate plays an important role in the neuronal excitotoxicity of receptor-mediated calcium influx. Studies have shown that NMDAR is related to neuroinflammation, such as the activation of glial cells and the release of inflammatory factors.

[0084] 5. Immunofluorescence results of Tremella fuciformis, Dictyophora short-skirted and Morchella solani on experimental animals

[0085] The best drug-effect group in the behavioral data (TDP2: 8 parts of Tremella fuciformis, 12 parts of Dictyophora short-skirted and 5 parts of Morchella solani) was selected for immunofluorescence staining analysis. The immunofluorescence staining diagram showed that ( Fig.13 and Fig.14 ), after CUMS modeling, the levels of synaptophysin (SYN) and postsynaptic density protein 95 (PSD95) in the CA1 and CA3 regions of the mouse hippocampus were reduced. This situation was reversed in the second-dose group, indicating that TDP administration can play a neuroprotective role.

[0086] 6. Administration of Tremella fuciformis, Dictyophora short-skirted and Morchella solani reduced the content of inflammatory factors in the brain

[0087] The best drug-effect group in the behavioral data (TDP2: 8 parts of Tremella fuciformis, 12 parts of Dictyophora short-skirted, and 5 parts of Morchella solani) was selected for immunoblotting analysis. The results showed that CUMS modeling reduced the levels of pro-inflammatory factors IL-1β and TNF-α in the mouse brain ( Fig.15 ), while TDP administration reduced the levels of these inflammatory factors, indicating that the compound formula can alleviate the inflammatory conditions caused by depression modeling.

[0088] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. A composition having an effect of alleviating depression, characterized in that: The raw materials of the composition are composed of the following components in parts by weight: 7-13 parts of Tremella fuciformis, 5-12 parts of Dictyophora short-skirted and 5-13 parts of Morchella solani; The preparation method of the composition comprises the following steps: The fruiting bodies of Tremella fuciformis, Dictyophora short-skirted and Morchella solani are mixed, water is added to the mixed fruiting bodies to soak overnight, and then water bath extraction is performed, and the supernatant is collected after centrifugation and concentrated to obtain the composition; During immersion, the mass volume ratio of the mixed fruiting body to water is 1 g:50 mL; The soaking temperature is 4°C; The water bath extraction temperature is 80°C and the extraction time is 5h; The concentration is to concentrate to 1 / 15 of the original volume.

2. The composition according to claim 1, characterized in that The raw materials of the composition are composed of 8 parts of Tremella fuciformis, 12 parts of Dictyophora short-skirted and 5 parts of Morchella solani in parts by weight.

3. Use of the composition as claimed in claim 1 or 2 in the preparation of a medicament for preventing and treating depression.

4. A drug for preventing and treating depression, characterized in that: The invention comprises the composition according to claim 1 or 2.

5. The drug according to claim 4, characterized in that It also includes pharmaceutically acceptable carrier substances and / or adjuvants.

Images