An antidepressant polysaccharide soluble fiber preparation and a preparation method thereof

Abstract

This invention provides an antidepressant polysaccharide soluble fiber preparation and its preparation method, belonging to the field of functional food preparation technology. The preparation consists of the following raw materials in parts by weight: 30-60 parts of shiitake mushroom soluble dietary fiber, 2-8 parts of γ-aminobutyric acid (GABA), 3-10 parts of Dendrobium officinale extract, 5-15 parts of Poria cocos water extract, and 20-40 parts of maltodextrin. The preparation method of the shiitake mushroom soluble dietary fiber includes: pulverizing the shiitake mushroom stems and subjecting them to high-temperature and high-pressure steam treatment, followed by hot water extraction and filtration. The filtrates are then combined and sequentially enzymatically hydrolyzed with alkaline protease, α-amylase, and cellulase. After enzyme inactivation and filtration, the hydrolysate is concentrated and precipitated with ethanol to obtain the final product. This invention's preparation, through the synergistic effect of shiitake mushroom dietary fiber with GABA, Dendrobium officinale, and Poria cocos, can effectively improve depressive symptoms and can be used to prepare antidepressant drugs or functional foods. It has the advantages of natural raw materials, clear efficacy, and stable processing.

Description

Technical Field

[0001] This invention belongs to the field of functional food formulation technology, specifically an antidepressant polysaccharide soluble fiber formulation and its preparation method. Background Technology

[0002] Depression is a common and serious mental disorder characterized by persistent low mood, loss of interest, impaired cognitive function, and autonomic dysfunction, severely impacting patients' quality of life and social functioning. Currently, drug treatment for depression primarily relies on first-line antidepressants such as selective serotonin reuptake inhibitors (SSRIs) and serotonin-norepinephrine reuptake inhibitors (SNRIs). However, existing antidepressants have clinical limitations, including slow onset of action and significant side effects. Therefore, developing alternative or adjunctive antidepressant regimens with faster onset of action, higher safety profiles, and novel mechanisms of action is of significant clinical importance and has broad market prospects.

[0003] In recent years, the gut-brain axis theory has provided a new perspective for the treatment of depression. The gut-brain axis is a complex network of bidirectional communication between the gut and the central nervous system, encompassing multiple pathways including the immune, neuroendocrine, and vagus nerve pathways. The gut microbiota plays a crucial role in the occurrence and development of depression by influencing neurotransmitter metabolism, immune system regulation, and the function of the hypothalamus-pituitary-adrenal axis. Dietary fiber, as an important substrate for the gut microbiota, is fermented by microorganisms in the gut to produce short-chain fatty acids, which can affect the function of the central nervous system through various pathways, including regulating neurotransmitter levels, reducing neuroinflammatory responses, and enhancing gut-brain communication. Dietary interventions rich in dietary fiber can improve depressive symptoms by regulating gut microbiota composition and increasing microbial diversity. Functional foods and nutritional intervention strategies based on the gut-brain axis have become a research hotspot in the field of adjunctive treatment for depression.

[0004] Shiitake mushrooms (Lentinus edodes) are rich in dietary fiber and bioactive components such as β-glucan. Shiitake mushroom soluble dietary fiber exhibits excellent prebiotic activity, promoting the proliferation of beneficial bacteria such as Bifidobacteria and Lactobacillus, repairing the intestinal barrier, and indirectly influencing neurotransmitter secretion and mood through the gut-brain axis. Animal experiments have shown that shiitake mushroom dietary fiber can exert neuroprotective effects by regulating gut microbiota and short-chain fatty acid metabolism. It is noteworthy that the shiitake mushroom stem is a major byproduct of shiitake mushroom processing. Due to its high fiber content and poor taste, shiitake mushroom stems were often treated as waste in the past, causing resource waste and environmental pollution. However, studies have shown that the total dietary fiber content of shiitake mushroom stems is much higher than that of the cap, making them a high-quality raw material for extracting soluble dietary fiber. Currently, there are no reports of using shiitake mushroom soluble dietary fiber as a main raw material for the development of antidepressant preparations. Summary of the Invention

[0005] In view of this, the purpose of the present invention is to provide a polysaccharide soluble fiber preparation that can effectively improve depressive symptoms and has high safety.

[0006] To achieve the above-mentioned objectives, the present invention provides the following technical solution: This invention provides a polysaccharide soluble fiber preparation comprising the following raw materials in parts by weight: 30-60 parts of soluble dietary fiber from shiitake mushrooms, 2-8 parts of γ-aminobutyric acid, 3-10 parts of Dendrobium officinale extract, 5-15 parts of Poria cocos water extract, and 20-40 parts of maltodextrin.

[0007] Preferably, the method for preparing the soluble dietary fiber of shiitake mushrooms includes the following steps: shredding the shiitake mushroom stems and subjecting them to high-temperature and high-pressure steam treatment to obtain shiitake mushroom material; adding water to the shiitake mushroom material and extracting it with high-temperature water, filtering, and combining the filtrates; sequentially hydrolyzing the filtrate with alkaline protease, α-amylase, and cellulase, inactivating the enzymes, filtering, and obtaining an enzymatic hydrolysate; concentrating the enzymatic hydrolysate and adding ethanol for precipitation to obtain soluble dietary fiber of shiitake mushrooms.

[0008] More preferably, the conditions for the high-temperature and high-pressure steam treatment are as follows: the treatment temperature is 119~123℃, the treatment pressure is 0.1~0.15MPa, and the treatment time is 25~40min.

[0009] More preferably, the amount of water added during water extraction is 20 to 25 times the mass of the shiitake mushroom material.

[0010] More preferably, the water extraction conditions are as follows: water extraction temperature is 98~100℃, water extraction time is 1.5~2.5h, and water extraction is performed 2~3 times.

[0011] More preferably, the amount of alkaline protease added is 0.15%~0.35% of the mass of shiitake mushroom material, the enzymatic hydrolysis pH is 8.5~10.5, the enzymatic hydrolysis temperature is 53~58℃, and the enzymatic hydrolysis time is 25~45 min; the amount of α-amylase added is 2.0%~3.0% of the mass of shiitake mushroom material, the enzymatic hydrolysis pH is 6.0~7.0, the enzymatic hydrolysis temperature is 50~55℃, and the enzymatic hydrolysis time is 25~45 min; the amount of cellulase added is 0.5%~1.0% of the mass of shiitake mushroom material, the enzymatic hydrolysis pH is 4.5~6.0, the enzymatic hydrolysis temperature is 50~60℃, and the enzymatic hydrolysis time is 0.5~5 h.

[0012] More preferably, the enzymatic hydrolysate is concentrated to 1 / 8 to 1 / 12 of its original volume.

[0013] More preferably, the final volume concentration of the ethanol is 60% to 80%.

[0014] The present invention also provides a method for preparing the polysaccharide soluble fiber preparation, comprising the following steps: weighing each raw material according to the ratio and passing it through an 80-120 mesh sieve; mixing at 10-25℃ and 30-60 r / min for 20-40 min to obtain a uniform mixture; and sterilizing by dry granulation or direct packaging to obtain the final product.

[0015] The present invention also provides the application of the polysaccharide soluble fiber preparation, or the polysaccharide soluble fiber preparation prepared by the preparation method, in the preparation of drugs for treating depression.

[0016] Compared with the prior art, the present invention has the following advantages: (1) This invention uses shiitake mushroom stems as raw materials to extract soluble dietary fiber, turning waste into treasure and reducing raw material costs. At the same time, the dietary fiber from shiitake mushroom stems has a high molecular weight (degree of polymerization DP≥60), which can be accurately detected by the enzyme gravimetric method in GB 5009.88-2023, solving the problem of low detection results of existing dietary fiber products.

[0017] (2) This invention combines soluble dietary fiber from shiitake mushrooms with γ-aminobutyric acid, dendrobium, and poria cocos to exert its effects through multiple targets on the gut-brain axis. Animal experiments have shown that it can significantly increase the levels of 5-HT and BDNF in the hippocampus and increase the production of short-chain fatty acids in the intestine, thus having a clear antidepressant effect.

[0018] (3) The preparation process of this invention is stable, easy to operate, and suitable for industrial production. The resulting preparation can be used directly as a functional food or for the preparation of antidepressant drugs. Attached Figure Description

[0019] Figure 1 The figure shows the comparison of 5-HT content in the hippocampus of mice in different treatment groups; in the figure, ## represents p<0.01 compared with the blank control group, and p<0.01 compared with the model control group. This means p < 0.05. This means p < 0.01.

[0020] Figure 2 The figure shows the comparison of BDNF content in the hippocampus of mice in different treatment groups; in the figure, ## represents p<0.01 compared with the blank control group, and p<0.01 compared with the model control group. This means p < 0.05. This means p < 0.01.

[0021] Figure 3 The figure shows the comparison of fecal acetic acid content in mice of different treatment groups; in the figure, ## represents p<0.01 compared with the blank control group, and p<0.01 compared with the model control group. This means p < 0.05. This means p < 0.01.

[0022] Figure 4 The figure shows the comparison of fecal propionic acid content in mice of different treatment groups; in the figure, ## represents p<0.01 compared with the blank control group, and p<0.01 compared with the model control group. This means p < 0.05. This means p < 0.01.

[0023] Figure 5 The figure shows the comparison of fecal butyrate content in mice of different treatment groups; in the figure, ## represents p<0.01 compared with the blank control group, and p<0.01 compared with the model control group. This means p < 0.05. This means p < 0.01. Detailed Implementation

[0024] This invention provides a polysaccharide soluble fiber preparation, wherein the preparation preferably comprises the following raw materials in parts by weight: 30-60 parts of soluble dietary fiber from shiitake mushrooms, 2-8 parts of γ-aminobutyric acid (GABA), 3-10 parts of Dendrobium officinale extract, 5-15 parts of Poria cocos water extract, and 20-40 parts of maltodextrin; more preferably, the preparation preferably comprises the following raw materials in parts by weight: 35-50 parts of soluble dietary fiber from shiitake mushrooms, 3-6 parts of γ-aminobutyric acid (GABA), 5-8 parts of Dendrobium officinale extract, 8-12 parts of Poria cocos water extract, and 25-35 parts of maltodextrin; even more preferably, the preparation preferably comprises the following raw materials in parts by weight: 45 parts of soluble dietary fiber from shiitake mushrooms, 8 parts of γ-aminobutyric acid (GABA), 6 parts of Dendrobium officinale extract, 10 parts of Poria cocos water extract, and 34 parts of maltodextrin. In a specific embodiment of the present invention, the soluble dietary fiber from shiitake mushrooms is a dried powder obtained from the stems of shiitake mushrooms through high-temperature and high-pressure steam treatment, hot water extraction, compound enzymatic hydrolysis, and ethanol precipitation, with an average molecular weight of not less than 1.0 × 10⁻⁶. 4 Da, degree of polymerization DP≥60, can be completely precipitated by 78% ethanol, and the soluble dietary fiber content, as determined by the enzyme gravimetric method in GB 5009.88-2023, is not less than 90%. γ-aminobutyric acid (GABA) is preferably a food-grade high-purity product prepared by microbial fermentation, with a purity ≥98%. Dendrobium extract is preferably obtained from the dried stems of Dendrobium officinale, after water extraction, concentration, and drying, with its dendrobine content controlled between 0.1% and 0.5%. Poria cocos water extract is a powder obtained from Poria cocos sclerotia through water extraction, filtration, concentration, and spray drying, with a Poria cocos polysaccharide content of not less than 30%. Maltodextrin is used as an excipient and diluent.

[0025] In this invention, the method for preparing soluble dietary fiber from shiitake mushrooms includes the following steps: shiitake mushroom stems are crushed and subjected to high-temperature and high-pressure steam treatment to obtain shiitake mushroom material; the shiitake mushroom material is added to water and extracted at high temperature, filtered, and the filtrates are combined; the filtrate is sequentially hydrolyzed by alkaline protease, α-amylase, and cellulase, the enzymes are inactivated, and the filtrate is filtered to obtain an enzymatic hydrolysate; the enzymatic hydrolysate is concentrated and then precipitated with ethanol to obtain soluble dietary fiber from shiitake mushrooms.

[0026] In a specific embodiment of the present invention, fresh or dried shiitake mushroom stems are preferably pulverized to 40-80 mesh using a pulverizer to obtain coarse shiitake mushroom powder. The coarse shiitake mushroom powder is then placed in a sealed high-pressure reactor, and deionized water is added at a mass ratio of 1:0.2-0.4 to control the moisture content of the material between 20% and 30%, ensuring uniform steam penetration and preventing excessive wetting of the material. After closing the reactor, saturated steam is introduced, rapidly raising the temperature inside the reactor to 119-123°C and the pressure to 0.1-0.15 MPa. This high-temperature, high-pressure treatment is maintained for 25-40 minutes. During this time, the cellulose crystallization zone in the material is destroyed, hydrogen bonds and some covalent bonds in the lignocellulose complex break, and the cell wall structure becomes loose, thereby transforming the originally tightly bound insoluble fiber into more easily soluble dietary fiber. After the treatment, the steam inside the reactor is slowly released to atmospheric pressure, and the material is removed, yielding the shiitake mushroom material treated with high-temperature, high-pressure steam. The preferred processing conditions for the high-temperature and high-pressure steam are a processing temperature of 120°C, a pressure of 0.12 MPa, and a processing time of 30 minutes.

[0027] In a specific embodiment of the present invention, the shiitake mushroom material after high-temperature and high-pressure steam treatment needs to be extracted with hot water to dissolve soluble dietary fiber. The preferred steps are as follows: The treated shiitake mushroom material is transferred to a stainless steel extraction tank equipped with a stirring device, and deionized water is added at a ratio of 20-25 times the mass of the shiitake mushroom material, i.e., 20-25L of water is added per 1kg of shiitake mushroom material. After the material and water are mixed evenly, the temperature is raised to 98-100℃ and maintained at a slight boiling state for 1.5-2.5 hours. During the extraction process, the high temperature enhances the hydration of cellulose and hemicellulose, and soluble dietary fiber is released from the loose cell wall structure and dissolves in the water. After extraction, solid-liquid separation is performed while hot using a 200-300 mesh filter cloth or an industrial centrifuge, and the filtrate is collected. To ensure that the soluble dietary fiber is extracted as fully as possible, the separated solid residue is extracted 2-3 times under the above conditions, i.e., 20-25 times the mass of deionized water is added again, and extraction is carried out at 98-100℃ for 1.5-2.5 hours. The filtrate is then collected after filtration. The filtrates obtained from multiple extractions are combined and used for subsequent enzymatic hydrolysis. Using a multiple extraction method can effectively increase the total yield of soluble dietary fiber and avoid raw material waste caused by insufficient extraction in a single step.

[0028] In a specific embodiment of the present invention, the combined filtrate is sequentially subjected to enzymatic hydrolysis with alkaline protease, α-amylase, and cellulase to remove residual protein and starch, and further degrade cellulose components in the cell wall, thereby purifying soluble dietary fiber. The preferred steps are as follows: the pH of the filtrate is adjusted to 8.5-10.5, preferably 9.0-9.5, and then 0.15%-0.35% (by weight of the initial shiitake mushroom material) of alkaline protease (enzyme activity ≥100000 U / g) is added. The mixture is then hydrolyzed at a constant temperature in a water bath at 53-58°C for 25-45 minutes. This step hydrolyzes the residual protein in the filtrate into small peptides and amino acids. After alkaline protease hydrolysis, enzyme inactivation is not required; α-amylase hydrolysis is then performed directly. Adjust the pH of the above enzymatic hydrolysate to 6.0-7.0, preferably 6.5, and add 2.0%-3.0% α-amylase (enzyme activity ≥4000U / g) based on the initial mass of shiitake mushroom material. Hydrolyze at a constant temperature of 50-55℃ for 25-45 minutes. This step decomposes the residual starch in the filtrate into oligosaccharides such as dextrin and maltose. Then, perform cellulase hydrolysis, further adjusting the pH to 4.5-6.0, preferably 5.0-5.5, and add 0.5%-1.0% cellulase (enzyme activity ≥10000U / g) based on the initial mass of shiitake mushroom material. Hydrolyze at a constant temperature of 50-60℃ for 0.5-5 hours. This step further degrades residual cellulose and hemicellulose in the cell walls, releasing more high-molecular-weight soluble dietary fiber and allowing it to remain stably in the solution. The amounts of the three enzymes added are calculated based on the mass of the shiitake mushroom material before the initial high-temperature and high-pressure steam treatment. During enzymatic hydrolysis, proper stirring should be maintained to ensure sufficient contact between the enzyme and the substrate. After all enzymatic hydrolysis steps are completed, the hydrolysate is heated to 90-100°C and held for 10-15 minutes to inactivate the enzyme protein by denaturation. After inactivation, the hydrolysate is cooled to room temperature and filtered through a 200-300 mesh filter cloth or a plate and frame filter to remove insoluble residues and denatured enzyme proteins. The clear filtrate is collected as the hydrolysate. This hydrolysate contains high-purity soluble dietary fiber while removing most proteins, starches, and other impurities, providing a good foundation for subsequent concentration and ethanol precipitation.

[0029] In a specific embodiment of the present invention, after enzyme inactivation and filtration, the enzymatic hydrolysate needs to be concentrated and precipitated with ethanol to separate the soluble dietary fiber from shiitake mushrooms. The preferred steps are as follows: The clarified enzymatic hydrolysate obtained from filtration is transferred to a rotary evaporator or a vacuum concentration tank and concentrated under reduced pressure at 55-65°C and a vacuum degree of -0.08 to -0.1 MPa until the volume of the concentrated solution is 1 / 8 to 1 / 12 of the original volume of the enzymatic hydrolysate. At this point, the concentration of soluble dietary fiber is significantly increased, which is beneficial for subsequent ethanol precipitation. After concentration, the concentrated solution is cooled to room temperature, and anhydrous ethanol is slowly added under stirring to achieve a final volume concentration of ethanol in the mixture of 60-80%, preferably 75%. Stirring should be continuous during the addition of ethanol to ensure thorough mixing of the concentrated solution and ethanol and prevent localized supersaturation. After the addition is complete, the mixture is transferred to a refrigerator at 4°C and allowed to stand for 8-12 hours to allow the soluble dietary fiber to fully flocculate and precipitate. After settling, centrifuge at 3000-4000 rpm for 15-25 minutes, discard the supernatant, and collect the precipitate. To further remove pigments and low-molecular-weight impurities, wash the precipitate 2-3 times with a 60%-80% (v / v) ethanol aqueous solution, adding an appropriate amount of washing solution each time, stirring well, and then centrifuging again to collect the precipitate. The washed precipitate is pale yellow to off-white and has a loose texture. Finally, place the washed precipitate in a vacuum drying oven and dry it at 50-60℃ and a vacuum of -0.08 to -0.1 MPa for 12-24 hours, or freeze-dry it until constant weight is obtained, thus obtaining the soluble dietary fiber product from shiitake mushrooms. Analysis shows that the fiber is in powder form with an average molecular weight of not less than 1.0 × 10⁻⁶. 4 The degree of polymerization (DP) is ≥60, and the soluble dietary fiber content, as measured by the enzyme gravimetric method according to GB5009.88-2023, can reach over 90%. The soluble dietary fiber of shiitake mushrooms prepared by this invention can be directly used to compound with raw materials such as γ-aminobutyric acid, dendrobium extract, poria cocos water extract, and maltodextrin to prepare polysaccharide soluble fiber preparations with antidepressant effects.

[0030] The present invention also provides a method for preparing the polysaccharide soluble fiber preparation, comprising the following steps: weighing each raw material according to the ratio and passing it through an 80-120 mesh sieve; mixing at 10-25℃ and 30-60 r / min for 20-40 min to obtain a uniform mixture; and sterilizing by dry granulation or direct packaging to obtain the final product.

[0031] In a specific embodiment of the present invention, the preferred method for preparing the polysaccharide soluble fiber preparation is as follows: Accurately weigh 30-60 parts of shiitake mushroom soluble dietary fiber, 2-8 parts of γ-aminobutyric acid (GABA), 3-10 parts of Dendrobium officinale extract, 5-15 parts of Poria cocos water extract, and 20-40 parts of maltodextrin according to the aforementioned weight ratio. Pass all raw materials through an 80-120 mesh vibrating sieve to remove any possible lumps or coarse particles, ensuring uniform powder particle size. After sieving, add a smaller amount of GABA, Dendrobium officinale extract, and Poria cocos water extract to an equal amount of maltodextrin in a mixing bag for incremental premixing. Specifically, first, take a small amount of the raw material and mix it manually with an equal amount of maltodextrin until homogeneous. Then, add an equal amount of maltodextrin to the mixture and mix again. Repeat this process until all the maltodextrin is added to obtain the premix. Then, the premix, along with the remaining soluble dietary fiber from shiitake mushrooms and the remaining maltodextrin, are added to a three-dimensional motion mixer or a V-type mixer. At room temperature (10-25°C), the mixing speed is set to 30-60 rpm, and continuous mixing is performed for 20-40 minutes until all components are visibly homogeneous, uniform in color, and free of discoloration. The resulting powder is the formulation intermediate. Depending on the target dosage form, granules can be prepared using a dry granulator: the mixed powder is fed into a dry granulator, where it is pressed into thin sheets by roller pressure, then crushed, sized, and passed through a 14-20 mesh sieve to obtain uniformly sized granules; alternatively, the mixed powder can be directly packaged as a powder. For packaging, an automatic packaging machine is used to fill granules or powder into food-grade aluminum-plastic composite film bags or paper-aluminum-plastic composite bags at 3-5 grams per bag, and then heat-sealed. Finally, the packaged products are sterilized, preferably by cobalt-60 irradiation at a dose of 5-10 kGy. Microwave or electron beam sterilization can also be used. After sterilization, the products pass inspection to obtain the finished product. Throughout the preparation process, the relative humidity is controlled at 40%-60% to prevent the powder from absorbing moisture and clumping. Those skilled in the art can adjust the mixer's loading capacity and mixing time according to the actual production scale, as long as uniform mixing is achieved.

[0032] The present invention also provides the application of the polysaccharide soluble fiber preparation, or the polysaccharide soluble fiber preparation prepared by the preparation method, in the preparation of drugs for treating depression.

[0033] Unless otherwise specified, the experimental methods used in the following embodiments are conventional methods. Unless otherwise specified, the experimental materials used in the following embodiments are commercially available products.

[0034] Example 1: Preparation of soluble dietary fiber from shiitake mushrooms Take dried shiitake mushroom stems and crush them through a 40-mesh sieve. Mix the shiitake mushroom powder with deionized water at a mass ratio of 1:0.3 until the moisture content is approximately 23%. Place the mixture in a high-pressure reactor and treat it with high-temperature and high-pressure steam at 120℃ and 0.12MPa for 30 minutes to obtain the treated shiitake mushroom material.

[0035] The processed shiitake mushroom material was added to 22 times its weight of deionized water and heated at 100℃ for 2 hours. The mixture was then filtered and the filtrate was collected. The residue was extracted twice more under the same conditions, and the filtrates were combined.

[0036] Adjust the pH of the combined filtrate to 9.5, add 0.25% alkaline protease (by weight of the shiitake mushroom material), and hydrolyze in a 55°C water bath for 35 min. Then adjust the pH to 6.5, add 2.5% α-amylase (by weight of the shiitake mushroom material), and hydrolyze in a 52°C water bath for 35 min. Finally, adjust the pH to 5.2, add 0.8% cellulase (by weight of the shiitake mushroom material), and hydrolyze in a 55°C water bath for 2 h. After hydrolysis, boil for 10 min to inactivate the enzyme, cool to room temperature, filter, and collect the hydrolysate.

[0037] The enzymatic hydrolysate was concentrated under reduced pressure to 1 / 10 of its original volume to obtain a concentrated solution. Anhydrous ethanol was slowly added to the concentrated solution to bring the final ethanol concentration to 75%, and the solution was allowed to stand at 4°C for 12 hours. The solution was then centrifuged at 3000 rpm for 20 minutes, and the precipitate was collected. The precipitate was washed twice with 75% ethanol and freeze-dried to obtain soluble dietary fiber from shiitake mushrooms.

[0038] The average molecular weight of the soluble dietary fiber obtained from shiitake mushrooms was determined to be 2.3 × 10⁻⁶. 5 Da, with a degree of polymerization of approximately 1400, can be completely precipitated by 78% ethanol. The soluble dietary fiber content was determined to be 91.2% according to the enzymatic gravimetric method in GB 5009.88-2023.

[0039] Example 2 A polysaccharide soluble fiber preparation comprises the following raw materials in parts by weight: 30 parts of shiitake mushroom soluble dietary fiber prepared in Example 1, 2 parts of γ-aminobutyric acid, 3 parts of Dendrobium officinale extract (water extract of Dendrobium officinale, dry powder), 5 parts of Poria cocos water extract (dry powder of Poria cocos water extract), and 20 parts of maltodextrin.

[0040] The preparation method of the above-mentioned polysaccharide soluble fiber preparation is as follows: All raw materials are passed through a 100-mesh sieve. γ-aminobutyric acid, Dendrobium officinale extract, Poria cocos water extract, and an equal amount of maltodextrin are added and mixed in equal increments to prepare a premix. The premix, along with the remaining lentinan soluble dietary fiber and maltodextrin, is added to a three-dimensional motion mixer and mixed at 20℃ and 45 r / min for 30 min to obtain a homogeneous mixture. The mixture is then dry-granulated (using a 14-mesh sieve), sterilized by irradiation (6 kGy), and packaged to obtain granules.

[0041] The polysaccharide soluble fiber preparation prepared in this embodiment was tested according to the method in GB 5009.88-2023. The total dietary fiber content of the preparation prepared in this embodiment was determined to be 26.8% (of which 24.1% was precipitable soluble dietary fiber determined by enzyme gravimetric method and 2.7% was non-precipitable soluble dietary fiber determined by liquid chromatography), meeting the requirements of the national food safety standards.

[0042] Example 3 A polysaccharide soluble fiber preparation comprises the following raw materials in parts by weight: 45 parts of shiitake mushroom soluble dietary fiber prepared in Example 1, 5 parts of γ-aminobutyric acid, 6 parts of Dendrobium officinale extract (Dendrobium officinale water extract, dry powder), 10 parts of Poria cocos water extract (Poria cocos water extract dry powder), and 34 parts of maltodextrin.

[0043] The preparation method is the same as in Example 2.

[0044] The polysaccharide soluble fiber preparation prepared in this embodiment was tested according to the method in GB 5009.88-2023. The total dietary fiber content of the preparation prepared in this embodiment was determined to be 40.5% (35.8% by enzyme gravimetric method and 4.7% by liquid chromatography).

[0045] Example 4 A polysaccharide soluble fiber preparation comprises the following raw materials in parts by weight: 60 parts of shiitake mushroom soluble dietary fiber prepared in Example 1, 8 parts of γ-aminobutyric acid, 10 parts of Dendrobium officinale extract (water extract of Dendrobium officinale, dry powder), 15 parts of Poria cocos water extract (dry powder of Poria cocos water extract), and 40 parts of maltodextrin.

[0046] The preparation method is the same as in Example 2.

[0047] The polysaccharide soluble fiber preparation prepared in this embodiment was tested according to the method in GB 5009.88-2023. The total dietary fiber content of the preparation prepared in this embodiment was determined to be 53.2% (47.6% by enzyme gravimetric method and 5.6% by liquid chromatography).

[0048] Example 5: Verification of the antidepressant efficacy of polysaccharide soluble fiber preparations 1. Experimental Grouping SPF-grade male C57BL / 6J mice, 6-8 weeks old and weighing 20-22g, were randomly divided into 6 groups of 12 mice each after one week of acclimatization. The grouping results are shown in Table 1.

[0049] Table 1 Experimental Groups

[0050] 2. Establishment of a chronic unpredictable mild stress (CUMS) model Except for the blank control group, the mice in the other groups received CUMS stimulation for 6 weeks. The stimuli included: fasting for 24 hours, water deprivation for 24 hours, 45° tilted cage for 12 hours, day-night reversal (12 hours of light / 12 hours of darkness reversed), tail clamping for 1 minute, swimming in 4℃ cold water for 5 minutes, moist bedding for 12 hours, restraint for 2 hours, noise stimulation for 2 hours, and day-night lighting for 30 minutes. One or two stimuli were randomly selected each day, and the same stimuli did not appear consecutively.

[0051] 3. Dosing regimen Administered via gavage starting in week 5, once daily for two consecutive weeks. The blank control group and the model control group received an equal volume of distilled water. During the administration period, all groups except the blank control group continued to receive CUMS stimulation.

[0052] 4. Behavioral tests (1) On the first day after the administration of the drug, a sucrose preference test (SPT) was conducted: Mice were housed in single cages and deprived of water for 24 hours. Then, they were given one bottle each of 1% sucrose solution and pure water. After 2 hours, they were swapped and placed for another 2 hours. The total fluid consumption and sucrose preference index were calculated (sucrose preference % = sucrose consumption / total fluid consumption × 100%).

[0053] (2) On the second day after the administration of the drug, the tail suspension test (TST) was performed: the tip of the mouse tail was fixed to a horizontal bar with tape at 2 cm, and the mouse was hung upside down. After 1 minute of adaptation, the immobility time within the next 4 minutes was recorded.

[0054] (3) Forced swimming test (FST) was performed on the 3rd day after the drug administration: the mice were placed in a transparent cylinder with a diameter of 15cm, a water depth of 15cm and a water temperature of 25±1℃. After acclimatization for 1 minute, the immobility time within the next 4 minutes was recorded.

[0055] 5. Biochemical indicator testing Mice were euthanized after behavioral testing, and hippocampal tissue was isolated on ice. The levels of 5-HT and BDNF in the hippocampal tissue were detected using ELISA. Fecal samples were collected, and the content of short-chain fatty acids (acetic acid, propionic acid, and butyric acid) was detected using GC-FID.

[0056] 6. Experimental Results (1) Behavioral results As shown in Table 2, compared with the blank control group, the sucrose preference index of mice in the model control group was significantly reduced (p<0.01), and the immobility time in the tail suspension test and forced swimming test was significantly prolonged, indicating successful CUMS modeling. Compared with the model control group, the sucrose preference index of mice in the medium and high dose groups of the formulation was significantly increased (p<0.05, p<0.01), and the immobility time in the tail suspension test and forced swimming test was significantly shortened (p<0.05, p<0.01). Although the low dose group showed an improvement trend, the difference was not statistically significant compared with the model control group. The behavioral improvement effect of the positive control group (fluoxetine group) was comparable to that of the medium dose group of the formulation.

[0057] Table 2 Comparison of behavioral test results of mice in each group ( (n=12)

[0058] Note: ##p<0.01 vs. blank control group; p<0.05, p<0.01 vs model control group.

[0059] (2) Biochemical index results according to Figures 1-2 The results showed that, compared with the blank control group, the levels of 5-HT and BDNF in the hippocampus of mice in the CUMS model control group were significantly reduced (p<0.01), indicating that the depression model was successfully established. After intervention with the formulation of this invention, both the medium-dose and high-dose groups significantly reversed the above changes. Specifically, the levels of 5-HT and BDNF in both the medium-dose and high-dose groups were significantly increased, with the high-dose group showing a better effect than the medium-dose group. The two indicators of the positive control, fluoxetine, were also significantly increased. These results indicate that the formulation of this invention exerts its antidepressant effect by upregulating the levels of 5-HT and BDNF in the hippocampus.

[0060] (3) Results of fecal short-chain fatty acids according to Figures 3-5 The results showed that after CUMS modeling, the levels of acetic acid, propionic acid, and butyric acid in the feces of mice in the model control group were significantly lower than those in the blank control group, suggesting that depressive state is accompanied by intestinal flora metabolic disorder. After intervention with the formulation of this invention, the levels of the three short-chain fatty acids in the medium-dose and high-dose groups all rebounded in a dose-dependent manner, with butyric acid showing the most significant recovery effect. Fluoxetine had no significant effect on short-chain fatty acids. The results indicate that the formulation of this invention can improve depressive symptoms through the gut-brain axis pathway by regulating intestinal flora metabolism and increasing the production of short-chain fatty acids.

[0061] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A polysaccharide soluble fiber preparation, characterized in that, The ingredients include the following parts by weight: 30-60 parts of soluble dietary fiber from shiitake mushrooms, 2-8 parts of γ-aminobutyric acid, 3-10 parts of Dendrobium officinale extract, 5-15 parts of Poria cocos water extract, and 20-40 parts of maltodextrin.

2. The polysaccharide soluble fiber preparation according to claim 1, characterized in that, The method for preparing soluble dietary fiber from shiitake mushrooms includes the following steps: crushing the stems of shiitake mushrooms and then subjecting them to high-temperature and high-pressure steam treatment to obtain shiitake mushroom material; adding water to the shiitake mushroom material and extracting it with high-temperature water, filtering, and combining the filtrates; sequentially subjecting the filtrate to enzymatic hydrolysis by alkaline protease, α-amylase, and cellulase, inactivating the enzymes, filtering, and obtaining an enzymatic hydrolysate; concentrating the enzymatic hydrolysate and adding ethanol for precipitation to obtain soluble dietary fiber from shiitake mushrooms.

3. The formulation according to claim 2, characterized in that, The conditions for the high-temperature and high-pressure steam treatment are as follows: the treatment temperature is 119~123℃, the treatment pressure is 0.1~0.15MPa, and the treatment time is 25~40min.

4. The formulation according to claim 2, characterized in that, The amount of water added during water extraction is 20 to 25 times the mass of the shiitake mushroom material.

5. The formulation according to claim 2 or 4, characterized in that, The conditions for water extraction are as follows: water extraction temperature is 98~100℃, water extraction time is 1.5~2.5h, and water extraction is performed 2~3 times.

6. The formulation according to claim 2, characterized in that, The alkaline protease is added at 0.15%~0.35% of the mass of the shiitake mushroom material, with an enzymatic hydrolysis pH of 8.5~10.5, an enzymatic hydrolysis temperature of 53~58℃, and an enzymatic hydrolysis time of 25~45 min; the α-amylase is added at 2.0%~3.0% of the mass of the shiitake mushroom material, with an enzymatic hydrolysis pH of 6.0~7.0, an enzymatic hydrolysis temperature of 50~55℃, and an enzymatic hydrolysis time of 25~45 min; the cellulase is added at 0.5%~1.0% of the mass of the shiitake mushroom material, with an enzymatic hydrolysis pH of 4.5~6.0, an enzymatic hydrolysis temperature of 50~60℃, and an enzymatic hydrolysis time of 0.5~5 h.

7. The formulation according to claim 2, characterized in that, The enzymatic hydrolysate is concentrated to 1 / 8 to 1 / 12 of its original volume.

8. The formulation according to claim 2, characterized in that, The final volume concentration of the ethanol is 60% to 80%.

9. A method for preparing the polysaccharide soluble fiber formulation according to any one of claims 1 to 8, characterized in that, The process includes the following steps: weigh each raw material according to the formula and pass it through an 80-120 mesh sieve; mix it at 10-25℃ and 30-60r / min for 20-40 minutes to obtain a homogeneous mixture; then granulate it by dry method or directly package it and sterilize it to obtain the final product.

10. The use of the polysaccharide soluble fiber preparation according to any one of claims 1 to 8, or the polysaccharide soluble fiber preparation prepared by the preparation method according to claim 9, in the preparation of a drug for treating depression.

Images