Use of tremella polysaccharides in preparing products for preventing and treating high-fat diet induced anxiety

Abstract

The present application relates to the technical field of health-care foods and particularly relates to the use of tremella polysaccharides in preparing products for preventing and treating high-fat diet induced anxiety. It is found by researches in the present application that tremella polysaccharides can relieve the weight gaining of mice fed by high-fat diet, reduce the weights of the liver and fat, inhibit microglial cell activation and neuritis in the hippocampus, balance the intestinal flora, and have an excellent improvement effect on preventing and treating anxiety of mice caused by high-fat diet feeding. The present application verifies that tremella polysaccharides have the effect of preventing and treating high-fat diet induced anxiety and thus can be used to prepare products for preventing and treating high-fat diet induced anxiety. Therefore, as a nutritional intervention strategy for high-fat diet induced anxiety disorders, tremella polysaccharides are of great significance for assisting in preventing and relieving high-fat diet induced anxiety disorders.

Description

Application of Tremella polysaccharide in preparing products for preventing and treating anxiety caused by high-fat diet

[0001] This application claims priority to the Chinese patent application filed with the China Patent Office on August 14, 2023, with application number 202311020498.3 and invention name “Application of Tremella Polysaccharide in the Preparation of Products for Preventing and Treating Anxiety Caused by a High-Fat Diet”, the entire contents of which are incorporated by reference into this application. Technical Field

[0002] The present application belongs to the technical field of health foods, and specifically relates to the use of Tremella polysaccharide in the preparation of products for preventing and treating anxiety caused by a high-fat diet. Background Art

[0003] It is well known that a high-fat diet (HFD) can lead to many diseases, such as obesity, diabetes, and metabolic syndrome, which are associated with an increased risk of neuropsychiatric disorders. A growing number of studies have shown that HFD significantly increases anxiety-like behaviors, and long-term HFD intake affects the morphology and activation of microglia. Microglia, as mononuclear phagocytes, play a vital role in maintaining the homeostasis of the central nervous system and inducing its pathological processes. Therefore, the anxiety-like behaviors of HFD-fed mice may be caused by changes in microglia. Neuroinflammation is also a hallmark of several neuropsychiatric diseases associated with HFD intake. Studies have reported that intestinal flora imbalance may cause neuroinflammation and brain development disorders by activating microglia. HFD is associated with anxiety-like behaviors and intestinal flora imbalance. The intake of HFD may significantly alter the composition and function of the intestinal flora, which is a key regulator of host intestinal homeostasis and immunity.

[0004] The epithelial barrier and mucus layer in the intestine can prevent external antigens from entering the host. The intake of HFD reduces the thickness of the mucus layer and the expression of epithelial tight junction proteins, thereby exacerbating local and systemic immune responses. In addition, changes in intestinal mucus and epithelial barriers lead to increased intestinal permeability, which allows bacterial endotoxins (lipopolysaccharide, LPS) to be transferred into the systemic blood circulation, which is considered endotoxemia. The increase in systemic LPS may further trigger a neuroinflammatory response. It has been reported that endotoxemia activates microglia and immune cells in the central nervous system and induces an inflammatory response in the mouse hippocampus. Therefore, the dysregulation of the microbiota-gut-brain axis caused by the intake of HFD can lead to neuroinflammation, microglial and synaptic damage, and subsequently lead to decreased brain function.

[0005] Given the prevalence of Western-style diets in modern life and the increasing global burden of mood disorders, it is imperative to find effective intervention strategies to combat the anxiety-like behavioral disorders and obesity symptoms caused by high-fat diets. Currently, there are no clinical products that can prevent and treat anxiety symptoms caused by high-fat diets. Therefore, there is an urgent need to provide a product that can prevent and treat anxiety symptoms caused by high-fat diets. Technical issues

[0006] The purpose of this application is to provide the use of Tremella polysaccharide in the preparation of a product for preventing and treating anxiety caused by a high-fat diet, aiming to solve the problem that there is no product in the prior art for preventing and treating anxiety symptoms caused by a high-fat diet. Technical Solutions

[0007] To achieve the above application objectives, the technical solutions adopted in this application are as follows:

[0008] In a first aspect, the present application provides the use of Tremella fuciformis polysaccharide in the preparation of a product for preventing and treating anxiety caused by a high-fat diet.

[0009] In some embodiments, the Tremella polysaccharide reduces the activation of microglia, increases the activation of synapses, and enhances the expression of brain-derived neurotrophic factor, thereby achieving the effect of preventing and treating anxiety.

[0010] In some embodiments, the Tremella polysaccharide improves the transcription levels of the three genes occludin, ZO-1 and claudin5 in the hippocampus and increases the expression of tight junction proteins to improve the blood-brain barrier and prevent the brain from being overly exposed to inflammatory cytokines that pass through the blood-brain barrier, thereby achieving the effect of preventing and treating anxiety.

[0011] In some embodiments, the Tremella polysaccharide reduces the mRNA levels of inflammatory factors TNF-α and MCP-1 in the hippocampus, thereby alleviating the inflammatory response and exerting anti-inflammatory and anxiety-preventing effects.

[0012] In some embodiments, the Tremella polysaccharide enhances the intestinal barrier by increasing the thickness of intestinal mucus, improving the expression of intestinal tight junction proteins, and increasing the expression of the antimicrobial peptide Reg3γ, thereby reducing the mRNA expression of inflammatory factors TNF-α, IL-1β, IL-6, MCP-1 and CD68 in the colon, thereby achieving anti-inflammatory and immune regulatory effects.

[0013] In some embodiments, the Tremella polysaccharide can achieve the effect of preventing and treating anxiety by directly improving the intestinal flora; and the intestinal flora improved by the Tremella polysaccharide can also reduce the inflammatory response in the brain and intestines induced by a high-fat diet after transplantation, thereby achieving the effect of preventing and treating anxiety.

[0014] In some embodiments, the product includes a medicine containing Tremella polysaccharide, a food containing Tremella polysaccharide, or a food additive containing Tremella polysaccharide.

[0015] In some embodiments, the drug containing Tremella fuciformis polysaccharide comprises a pharmaceutically effective amount of Tremella fuciformis polysaccharide and a pharmaceutically acceptable carrier.

[0016] In some embodiments, the food containing Tremella polysaccharide includes at least one of a functional food containing Tremella polysaccharide, a health product containing Tremella polysaccharide, a dietary supplement containing Tremella polysaccharide, and a common food containing Tremella polysaccharide.

[0017] In some embodiments, the food containing Tremella polysaccharide includes at least one of Tremella soup, Tremella slurry, Tremella paste, Tremella soup, dry Tremella soup, Tremella raw powder, and Tremella dry powder.

[0018] In some embodiments, the food additive containing Tremella polysaccharide includes at least one of Tremella soup, Tremella slurry, Tremella paste, Tremella soup, dry Tremella soup, Tremella raw powder, and Tremella dry powder.

[0019] In some embodiments, the medicine containing Tremella polysaccharide includes at least one of tablets, pills, dispersible tablets, capsules, and oral liquids.

[0020] In some embodiments, the pharmaceutically acceptable carrier includes at least one of a diluent, a filler, a binder, a preservative, a lubricant, a dispersant, a wetting agent, a sweetener, a flavoring agent, an emulsifier, a suspending agent, a preservative, an antioxidant, a colorant, and a stabilizer.

[0021] In some embodiments, the product includes a food preparation, a pharmaceutical preparation, or a fecal microbiota transplantation preparation containing intestinal flora improved by Tremella polysaccharide.

[0022] In some embodiments, the food preparation containing the intestinal flora improved by Tremella fuciformis polysaccharide includes at least one of a functional food containing the intestinal flora improved by Tremella fuciformis polysaccharide, a health product containing the intestinal flora improved by Tremella fuciformis polysaccharide, a dietary supplement containing the intestinal flora improved by Tremella fuciformis polysaccharide, and a common food containing the intestinal flora improved by Tremella fuciformis polysaccharide.

[0023] In some embodiments, the pharmaceutical preparation or fecal microbiota transplantation preparation containing the intestinal flora improved by Tremella polysaccharide includes at least one of tablets, pills, dispersible tablets, capsules, and oral liquids. Beneficial effects

[0024] The first aspect of the present application provides an application of Tremella polysaccharide in the preparation of a product for preventing and treating anxiety caused by a high-fat diet. Tremella polysaccharide has an effective preventive and therapeutic effect on anxiety-like behavioral disorders caused by the intake of a high-fat diet (HFD). Tremella is economical and affordable, and Tremella polysaccharide is a natural extract with few side effects. It has the advantages of safety, convenience and low cost, and can meet the needs of anxiety patients. Therefore, Tremella polysaccharide can be used in the preparation of products for preventing and treating anxiety caused by a high-fat diet, intervene in anxiety symptoms caused by a high-fat diet, and achieve the effect of preventing and treating anxiety symptoms caused by HFD, which has a wide range of application value. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] In order to more clearly illustrate the technical solutions in the embodiments of the present application, the following briefly introduces the drawings required for use in the embodiments or descriptions of the prior art. Obviously, the drawings described below are only some embodiments of the present application. For ordinary technicians in this field, other drawings can be obtained based on these drawings without any creative work.

[0026] Figure 1 shows the results of the study in Example 1 of this application on the effects of Tremella fuciformis polysaccharide (TP) intake on metabolic syndrome parameters in HFD-fed mice. A. Experimental design; B. Weight gain; C. Fasting blood glucose; D. Tissue weight. Data are presented as mean ± SEM. Values ​​in columns with different letters were significantly different (p < 0.05) as shown by Tukey's multiple range test.

[0027] Figure 2 is the behavioral test results of Example 1 of the present application to study the effect of Tremella polysaccharide (TP) intake on HFD-induced anxiety-like behavior in mice, wherein A is a representative track image in OFT; B is the quantification of the total distance, speed, distance and time in the internal area of ​​the venue; C is a representative track image and heat map image in EPM; D is the quantification of the total distance, time ratio of open arm travel and frequency of entry into the open arm; E is a representative image in the marble burying test; F is the quantification of the number of buried beads; (ND: mice fed a normal diet and distilled water; ND+TPH: mice fed a normal diet, gavage of 400 mg / kg TP and distilled water every day; HFD: mice fed a high-fat diet and distilled water; HFD+TPL: mice fed a high-fat diet, gavage of 40 mg / kg TP every day; HFD+TPH: mice fed a high-fat diet, gavage of 400 mg / kg TP every day).

[0028] Figure 3 shows the results of studying the effects of Tremella polysaccharide (TP) intake on brain function in HFD-fed mice in Example 1 of the present application, wherein A is immunofluorescence staining of Iba1 and PSD95 in the hippocampus. Scale bar: 50 μm; B is immunohistochemical staining of Iba-1 protein in the hippocampus. Scale bar: 100 μm; C is the protein levels of PSD95 and BDNF in the hippocampus; D is the relative mRNA levels of Iba-1, PSD95, and BDNF in the hippocampus; E is the relative mRNA levels of tight junction proteins Occludin, ZO-1, and Claudin 5 in the hippocampus; F is the relative mRNA levels of inflammatory factors; G is the levels of neurotransmitters, serotonin (5-HT), and norepinephrine (NE) in the hippocampus.

[0029] Figure 4 shows the results of studying the effects of Tremella polysaccharide (TP) intake on colon structure and function changes in HFD-fed mice in Example 1 of the present application, wherein A is the colon length; B is the quantification of colon length; C is the Alcian blue staining and immunofluorescence staining of the mucus layer; D is the immunohistochemical staining of the protein occludin; E is the relative mRNA level of occludin; F is the immunohistochemical staining of the ZO-1 protein; G is the relative mRNA level of ZO-1; H is the relative mRNA level of tight junction protein in the colon; I is the relative mRNA level of Reg3γ; J is the serum LPS level; K is the relative mRNA level of inflammatory factors in the colon; L is the level of inflammatory factors in serum, scale: 100 μm.

[0030] Figure 5 shows the results of studying the effect of Tremella polysaccharide (TP) intake on the changes in the intestinal flora structure of HFD-fed mice in Example 1 of the present application, wherein A is the principal coordinate analysis diagram of the unweighted UniFrac distance (PCoA) at week 0 and week 12; B is the α diversity measured by the Shannon index at week 0 and week 12.

[0031] Figure 6 shows the results of studying the effect of Tremella polysaccharide (TP) intake on the changes in the intestinal flora structure of HFD-fed mice in Example 1 of the present application, A is the community changes at the phylum level; B is the community changes at the genus level; C is the differential abundance analysis at the genus level, using one-way ANOVA with post hoc Kruskal-Wallis H test, *p<0.05; **p<0.01; ***p<0.001.

[0032] Figure 7 shows the results of studying the effects of Tremella polysaccharide (TP) intake on functional changes in intestinal flora in HFD-fed mice in Example 1 of the present application, wherein A is a community heat map analysis at the mouse genus level; B is the prediction of KEGG functional pathway differences using PICRUSt2; and C is a Spearman rank correlation heat map showing the correlation between neurobehavioral changes and specific flora.

[0033] Figure 8 is the behavioral test results of studying the effect of fecal microbiota transplantation (FMT) on HFD-induced anxiety-like behavior in mice in Example 2 of the present application, wherein A is a representative trajectory image in OFT; B is the quantification of the distance and time of the inner circle of the field; C is a representative trajectory image in EPM; D is the quantification of the open arm time ratio and entry frequency; E is a representative image in the marble burying test; F is the quantification of the number of buried beads; (dND: ND-fed mice; dND+TPH: ND-fed TPH-intervention mice; dHFD: HFD-fed mice; dHFD+TPL: HFD-fed TPL-intervention mice; dHFD+TPH: HFD-fed TPH-intervention mice, rHFD: HFD-fed mice are recipients of fecal microbiota transplantation).

[0034] Figure 9 shows the results of studying the effects of fecal microbiota transplantation (FMT) on the brain and colon of HFD-fed mice in Example 2 of the present application, wherein A is immunofluorescence staining of Iba1 and PSD95 in the hippocampus; B is the relative mRNA levels of Iba-1, PSD95 and BDNF in the hippocampus; C is the relative mRNA levels of inflammatory factors in the hippocampus; D is immunofluorescence staining of the mucus layer; E is the relative mRNA level of ZO-1 in the colon; F is the relative mRNA level of Occludin; G is the relative mRNA level of Reg3γ, scale bar: 100 μm.

[0035] Figure 10 shows the results of studying the effect of fecal microbiota transplantation (FMT) on the intestinal microbiota of HFD-fed mice in Example 2 of the present application, A is the principal coordinate analysis diagram of the unweighted UniFrac distance (PCoA); B is the alpha diversity measured by the Shannon index.

[0036] Figure 11 shows the results of studying the effect of fecal microbiota transplantation (FMT) on the intestinal microbiota of HFD-fed mice in Example 2 of the present application, A is the comparison of dominant phyla; B is the comparison of dominant genera, wherein, one-way ANOVA with post hoc Kruskal-Wallis H test was used, *p<0.05; **p<0.01; ***p<0.001. Modes for Carrying Out the Invention

[0037] In order to make the technical problems, technical solutions and beneficial effects to be solved by this application more clearly understood, this application is further described in detail below in conjunction with the embodiments. It should be understood that the specific embodiments described herein are only used to explain this application and are not intended to limit this application.

[0038] In a first aspect, the embodiments of the present application provide the use of Tremella fuciformis polysaccharide in the preparation of a product for preventing and treating anxiety caused by a high-fat diet.

[0039] The first aspect of the present application provides an application of Tremella polysaccharide in the preparation of a product for preventing and treating anxiety caused by a high-fat diet. Tremella polysaccharide has an effective preventive and therapeutic effect on anxiety-like behavioral disorders caused by the intake of a high-fat diet (HFD). Tremella is economical and affordable, and Tremella polysaccharide is a natural extract with few side effects. It has the advantages of safety, convenience and low cost, which can meet the needs of anxiety patients. Therefore, Tremella polysaccharide can be used in the preparation of products for preventing and treating anxiety as a strategy for intervening in anxiety symptoms caused by a high-fat diet, achieving the effect of preventing and treating anxiety symptoms caused by HFD, and has wide application value.

[0040] Tremella fuciformis (Tremella) has a history of thousands of years in China. Studies have shown that its polysaccharides (TP) have hypoglycemic and hypolipidemic effects, anti-obesity properties, and neuroprotective properties. For example, purified polysaccharides from Tremella fuciformis exhibit neuroprotective effects against glutamate-induced injury in differentiated PC-12 cells. TP has also been shown to restore cognitive function by modulating the CREB signaling pathway and cholinergic system in the rat hippocampus induced by trimethyltin.

[0041] The applicant found in the study that HFD-fed mice with anxiety-like behavioral disorders showed lower intestinal flora diversity (Shannon index) and structural microecological imbalance, and the transcription level of tight junction proteins in the colon of HFD-induced mice was significantly reduced, and the LPS concentration in the serum was increased, indicating that the intestinal barrier function was impaired, that is, HFD may destroy the intestinal barrier function by causing imbalance of the intestinal flora of mice. The imbalance of the flora-gut-brain axis caused by the intake of HFD can lead to neuroinflammation, microglial and synaptic damage, and subsequently lead to decreased brain function. The flora-gut-brain axis, that is, the two-way communication between the intestinal flora and the brain, is important for maintaining the homeostasis of the gastrointestinal tract, central nervous system and flora systems in animals and humans. The intestinal flora is essential to the basic structure and function of the brain.

[0042] It was also found that TP intervention can inhibit neuroinflammation, microglial activation and synaptic damage, and TP intake increased the mRNA level of tight junction proteins and neurotransmitter levels in the blood-brain barrier of HFD-fed mice. Moreover, TP intake can prevent the dysbiosis of intestinal flora, and rebalance the intestinal flora by fecal flora transplantation (FMT) to improve the intestinal flora of HFD-fed mice, further achieving the effect of directly improving their anxiety-like behavior. Therefore, the applicant verified that TP has a beneficial effect on HFD-induced anxiety-like behavior from the perspective of the flora-gut-brain axis. This shows that Tremella polysaccharide has an anti-anxiety effect and can be used in the preparation of products for preventing and treating anxiety and obesity.

[0043] In some embodiments, the Tremella polysaccharide reduces the activation of microglia, increases the activation of synapses, and enhances the expression of brain-derived neurotrophic factor, thereby achieving the effect of preventing and treating anxiety.

[0044] In some embodiments, the Tremella polysaccharide improves the transcription levels of the three genes occludin, ZO-1 and claudin5 in the hippocampus and increases the expression of tight junction proteins to improve the blood-brain barrier and prevent the brain from being overly exposed to inflammatory cytokines that pass through the blood-brain barrier, thereby achieving the effect of preventing and treating anxiety.

[0045] In some embodiments, the Tremella polysaccharide reduces the mRNA levels of inflammatory factors TNF-α and MCP-1 in the hippocampus, thereby alleviating the inflammatory response and exerting anti-inflammatory and anxiety-preventing effects.

[0046] In some embodiments, the Tremella polysaccharide enhances the intestinal barrier by increasing the thickness of intestinal mucus, improving the expression of intestinal tight junction proteins, and increasing the expression of the antimicrobial peptide Reg3γ, thereby reducing the mRNA expression of inflammatory factors TNF-α, IL-1β, IL-6, MCP-1 and CD68 in the colon, thereby achieving anti-inflammatory and immune regulatory effects.

[0047] In some embodiments, the Tremella polysaccharide can achieve the effect of preventing and treating anxiety by directly improving the intestinal flora; and the intestinal flora improved by the Tremella polysaccharide can reduce the inflammatory response in the brain and intestines, thereby achieving the effect of preventing and treating anxiety.

[0048] In some embodiments, the product includes a medicine containing Tremella polysaccharide, a food containing Tremella polysaccharide, or a food additive containing Tremella polysaccharide.

[0049] In some embodiments, the drug containing Tremella fuciformis polysaccharide comprises a pharmaceutically effective amount of Tremella fuciformis polysaccharide and a pharmaceutically acceptable carrier.

[0050] In some embodiments, the food containing Tremella polysaccharide includes at least one of a functional food containing Tremella polysaccharide, a health product containing Tremella polysaccharide, a dietary supplement containing Tremella polysaccharide, and a common food containing Tremella polysaccharide.

[0051] In some embodiments, the food containing Tremella polysaccharide includes at least one of Tremella soup, Tremella slurry, Tremella paste, Tremella soup, dry Tremella soup, Tremella raw powder, and Tremella dry powder.

[0052] In some embodiments, the food additive containing Tremella polysaccharide includes at least one of Tremella soup, Tremella slurry, Tremella paste, Tremella soup, dry Tremella soup, Tremella raw powder, and Tremella dry powder.

[0053] In some embodiments, the medicine containing Tremella polysaccharide includes at least one of tablets, pills, dispersible tablets, capsules, and oral liquids.

[0054] In some embodiments, the pharmaceutically acceptable carrier includes at least one of a diluent, a filler, a binder, a preservative, a lubricant, a dispersant, a wetting agent, a sweetener, a flavoring agent, an emulsifier, a suspending agent, a preservative, an antioxidant, a colorant, and a stabilizer.

[0055] In some embodiments, the product includes a food preparation, a pharmaceutical preparation, or a fecal microbiota transplantation preparation containing intestinal flora improved by Tremella polysaccharide.

[0056] In some embodiments, the food preparation containing the intestinal flora improved by Tremella fuciformis polysaccharide includes at least one of a functional food containing the intestinal flora improved by Tremella fuciformis polysaccharide, a health product containing the intestinal flora improved by Tremella fuciformis polysaccharide, a dietary supplement containing the intestinal flora improved by Tremella fuciformis polysaccharide, and a common food containing the intestinal flora improved by Tremella fuciformis polysaccharide.

[0057] In some embodiments, the pharmaceutical preparation or fecal microbiota transplantation preparation containing the intestinal flora improved by Tremella polysaccharide includes at least one of tablets, pills, dispersible tablets, capsules, and oral liquids.

[0058] The solution of this application is further described below with reference to specific embodiments.

[0059] Example 1

[0060] 1.1 Experimental animals and treatment

[0061] Male C57BL / 6J mice (7 weeks old) were purchased from Weitonglihua Co., Ltd. (Beijing, China). Five mice were housed per cage under standard conditions (12 / 12 light / dark cycle, humidity 50 ± 15%, and temperature 22 ± 2°C). Mice were randomly divided into five groups according to the different treatments they received.

[0062] ND: mice were fed a normal diet (10% kcal from fat, D12450J, Changzhou Shuyishuer Biotechnology Co., Ltd.) and distilled water;

[0063] ND+TPH: mice were fed a normal diet and gavaged with 400 mg / kg of TP (purchased from Bo InnoHealth Biotechnology Company Ltd (Hong Kong, China)) daily;

[0064] HFD: mice were fed a high-fat diet (60% kcal from fat, PD6001, Changzhou Shuyi Shuer Biotechnology Co., Ltd.) and distilled water;

[0065] HFD+TPL: mice were fed a high-fat diet and gavaged with 40 mg / kg TP daily;

[0066] HFD+TPH: Mice were fed a high-fat diet and gavaged with 400 mg / kg of TP daily.

[0067] After 12 weeks of intervention, all mice underwent behavioral testing, followed by blood samples drawn via cardiac puncture, and tissue collection after euthanasia using carbon dioxide.

[0068] The experimental procedures used were approved by the Animal Subjects Ethics Committee of the Hong Kong Polytechnic University (ASESC No. 21-22 / 87-ABCT-R-OTHERS).

[0069] 1.2 Provide Tremella polysaccharide (TP) preparations

[0070] Tremella polysaccharides were extracted and supplied by Bo InnoHealth Biotechnology Company Ltd (Hong Kong, China). Briefly, Tremella was mixed with 100% pure water and boiled at 100°C for 2 hours. This extraction process was repeated twice. After filtration and centrifugation, the supernatant was obtained and mixed with 4% anhydrous ethanol. The polysaccharides were precipitated by centrifugation. Tremella polysaccharides were dissolved in pure water, and proteins were removed using the TCA method.

[0071] 1.3 Behavioral testing

[0072] The open field test (OFT), elevated plus maze (EPM), and marble burying are methods for evaluating anxiety or depression-like behaviors by assessing the spontaneous locomotor activity of rodents. In this example, behavioral tests including the OFT, EPM, and marble burying tests were performed to assess anxiety-like conditions in mice.

[0073] In both the OFT and EPM, mice were carefully placed in the center area of ​​the maze and allowed to explore freely for 5 minutes. Parameters such as total distance traveled, distance traveled in the center or open arms, and time were recorded using the EthoVisionXT video imaging system. In the marble burying test, mice were placed in a cage containing 10 glass beads. The number of beads buried by the mice within 30 minutes was scored.

[0074] 1.4 Alcian blue (AB) staining

[0075] Transverse colon tissue of mice was placed in methanol-Carnoy's fixative (60% methanol, 30% chloroform, 10% glacial acetic acid) at room temperature for at least 3 hours, then dehydrated and embedded in paraffin. To measure the thickness of the colonic mucus layer, the sectioned colon samples were dewaxed and hydrated in distilled water, then placed in 3% acetic acid solution for 5 minutes, and then stained in Alcian blue solution. The samples were rinsed in distilled water and counterstained with filtered nuclear fast red solution.

[0076] 1.5 Immunohistochemistry and immunofluorescence staining

[0077] The embedded colon and brain were cut into 5 μm and 20 μm sections, respectively. Sodium citrate antigen retrieval solution (pH 6.0) was used to retrieve antigens, and 3% methanol peroxide was used to quench the samples. After washing with PBS, the sections were incubated with primary antibodies against Iba1 (Servicebio, Wuhan, China), Occludin (Abcam, Cambridge, UK), ZO1 (Santa Cruz, Heidelberg, Germany), PSD95 (Cell Signaling Technology, Boston, USA), and MUC2 (ABclone, Wuhan, China) at 4°C overnight. The samples were incubated with the corresponding secondary antibodies at room temperature for 30 minutes.

[0078] Immunohistochemistry (IHC) staining was performed using a DAB kit (Servicebio, Wuhan, China). Images were observed under a light microscope. For immunofluorescence (IF) staining, sections were counterstained with 4′6-diamidino-2-phenylindole (DAPI, Beyotime, Shanghai, China) in PBS to visualize cell nuclei. Images were captured using a Nikon AX NIS Elements 5.4 confocal microscope (Nikon, Tokyo, Japan).

[0079] 1.6 Western blotting

[0080] Hippocampal tissue was mixed with grinding beads and homogenized in pre-cooled RIPA lysis buffer containing EDTA-free protease inhibitor cocktail and PhosSTOP phosphatase inhibitor. The collected supernatants were quantified using the BCA assay. The obtained protein samples were loaded and separated in SDS-PAGE and transferred to PVDF membranes, which were blocked and incubated with primary antibodies against PSD95 (Cell Signaling Technology, Boston, USA) and BDNF (Santa Cruz, Heidelberg, Germany). Subsequently, the membranes were washed in TBST and incubated with HRP-linked anti-mouse IgG secondary antibodies (Cell Signaling Technology, Boston, USA) for 2 hours at room temperature. The membranes were visualized using Clarity ELISA kit (Sigma-Aldrich, Sigma-Aldrich). TMProtein bands were detected using ECL Western Blot Substrate (Bio-Rad, CA, USA) and images were obtained using the ChemiDoc Touch Imaging System (Bio-Rad, CA, USA).

[0081] 1.7 Reverse transcription qualitative polymerase chain reaction (RT-qPCR)

[0082] Total RNA was extracted from the colon and brain using Trizol reagent (Thermo Fisher Scientific, Waltham, MA, USA). Up to 1 μg of total RNA was reverse transcribed into cDNA using a reverse transcription kit (Takara PrimeScript RT Master Mix, Dalian, China). Subsequently, the cDNA was detected using a SYBR green PCR kit (Biotium Forget-Me-Not) in a real-time PCR detection system (Bio-Rad, Hercules, CA, USA). TM mRNA expression was determined by qPCR using qPCR Master Mix (Fremont, CA, USA). Relative mRNA levels were determined by the 2-ΔΔCt method. Primer sequences are shown in Table 1.

[0083] Table 1 Primer sequences

[0084] 1.8 ELISA

[0085] TNF-α, IL-6, IL-1β, and lipopolysaccharide (LPS) levels were determined using ELISA kits (Jiangsu Enzyme Immunity Industry Co., Ltd., Yancheng, China). 5-hydroxytryptamine (5-HT) and norepinephrine (NE) levels in the hippocampus were measured using ELISA kits (Shanghai Xinle Biotechnology Co., Ltd., Shanghai, China).

[0086] 1.9 16S rRNA sequencing analysis

[0087] Fecal samples were collected rectally before euthanasia, and fecal microbial genomic DNA was extracted using the TIANamp Stool DNA Kit (Tiangen, Beijing, China). Extracted DNA was checked on 1% agarose gels, and DNA concentration and purity were determined using a NanoDrop UV-Vis spectrophotometer (Thermo Scientific, Wilmington, USA). The hypervariable region V3-V4 of the 16S rRNA gene of the microbial community was amplified using primer pairs 338F (5'-ACTCCTACGGGAGGCAGCAG-3') and 806R (5'-GGACTACHVGGGTWTCTAAT-3') at Shanghai Meiji Biopharmaceutical Technology Co., Ltd. (Shanghai, China). Purified amplicons were mixed in equal moles and subjected to paired-end sequencing on an Illumina MiSeq PE300 platform / NovaSeq PE250 platform (Illumina, San Diego, USA) according to the standard protocol of Shanghai Meiji Biopharmaceutical Technology Co., Ltd. (Shanghai, China).

[0088] 2.0 Data Analysis

[0089] Data were analyzed using the SPSS statistical software package (version 20, IBM Corporation, Chicago, IL, USA). All values ​​represent the mean and SEM of six to eight independent experiments. The data were then compared using one-way analysis of variance and Duncan multiple interval analysis. For 16S rRNA gene sequence analysis, all reads were deposited at 97% sequence identity and grouped into operational taxonomic units (OTUs). Statistical analysis was performed using STAMP, and functional differences between orthologs were assessed by one-way analysis of variance, followed by post hoc Tukey Kramer or Kruskal-Wallis tests for multiple comparisons.

[0090] Result analysis:

[0091] Tremella polysaccharides improved metabolic syndrome parameters in HFD-fed mice, which can induce obesity and its associated metabolic syndrome. In the study, all mice were weighed before the start of their assigned diet and at the end of the experiment. The experimental design is shown in Figure 1A. The initial body weight of each group was at similar levels, and high-dose TP (TPH) significantly reduced the body weight of HFD-fed mice after 12 weeks of intervention (Figure 1B). In addition, fasting blood glucose levels in obese mice supplemented with TPH were significantly reduced compared with HFD-fed mice (Figure 1C). Long-term HFD feeding increases the weight of multiple tissues, including liver, subcutaneous fat, and epididymal fat. However, TPH intervention significantly reduced the increase in the weight of these tissues in HFD-fed mice (Figure 1D). These results indicate that supplementation with Tremella polysaccharides, especially high-dose Tremella polysaccharides, exhibits anti-obesity effects in HFD-fed mice.

[0092] In the OFT test, mice exhibiting anxiety-like behavioral disorders spent less time in the central area than normal mice. As shown in Figure 2A and B, HFD intake significantly reduced total distance and speed compared with mice fed ND. In HFD-fed mice, TPH intervention significantly increased these parameters: total distance and speed. At the same time, compared with HFD mice without TP intervention, HFD mice supplemented with TPH significantly increased the distance and time traveled in the OFT central area. The results indicate that TPH intervention improved HFD-induced anxiety-like behavioral disorders in mice.

[0093] In the EPM test, anxious mice spend less time in the open arms than normal mice. In the study of this example, as shown in Figure 2C and D, the total distance traveled by HFD-fed mice was significantly reduced compared to the ND group, with a shorter distance and time spent in the open arms. However, TPH significantly increased the time spent in the open arms by HFD-fed mice. This suggests that TPH intervention alleviates HFD-induced anxiety in mice.

[0094] In the marble burying test, mice with poor sociability buried more beads than normal mice. As shown in Figure 2, E and F, HFD-fed mice buried more beads than ND-fed mice, while TPH intervention significantly reduced the number of beads buried. Therefore, TPH intervention improved HFD-induced sociability in mice, alleviating anxiety-like behavioral impairments.

[0095] In the open field test, elevated plus maze and marble burying behavioral tests, the results of the three tests all showed that lower doses of TP had limited effects on improving anxiety-like behavioral disorders in mice, while high doses of TP significantly improved HFD-induced anxiety-like behaviors in mice.

[0096] Microglial activation is crucial in the pathogenesis of brain diseases and neuroinflammation. Microglia play an important role in cognitive dysfunction through excessive synaptic pruning. A high-fat and fiber-deficient diet can induce microglial activation in the hippocampus.

[0097] Double immunofluorescence staining for Iba-1 and PSD95 was used in all groups to determine the spatial location of microglia and synapses. The results, shown in Figure 3A, indicate that compared with the ND and TPH groups, the HFD group showed increased Iba-1 expression and decreased PSD95 expression, suggesting that TP supplementation, especially high-dose TP, attenuates microglial activation but increases synaptic activation. Immunohistochemical staining results in Figure 3B demonstrate that the HFD promotes microglial activation, while TPH treatment reduces microglial expression in obese mice. Western blot analysis in Figure 3C demonstrates that compared with ND-fed mice, the HFD and HFD+TPL groups have decreased PSD95 and brain-derived neurotrophic factor (BDNF) expression, while TPH supplementation increases their expression in HFD-fed mice. As shown in Figure 3D, RT-qPCR analysis of Iba-1, PSD95, and BDNF mRNA levels in the five groups of mice showed consistent results with immunofluorescence, immunohistochemistry, and Western blot. The results showed that TPH could improve the impaired activation of microglial markers, postsynaptic density protein, and brain-derived neurotrophic factor expression in the hippocampus caused by HFD.

[0098] At the same time, changes in circulating cytokines in systemic immunity lead to altered brain inflammation, and neuropsychiatric disorders such as anxiety disorders are associated with peripheral inflammation. Circulating cytokines and chemokines can be directly transported across the blood-brain barrier and enter the central nervous system. To further examine the integrity of the hippocampal blood-brain barrier (BBB), mRNA levels of key tight junction proteins and inflammatory factors were measured. The results are shown in Figure 3E. The results showed that the transcript levels of occludin, ZO-1, and claudin5 were significantly lower in HFD-fed mice than in ND-fed control mice. TPL increased the mRNA level of ZO-1, and TPH increased the transcript levels of all three genes in obese mice. As shown in Figure 3F, the mRNA levels of the inflammatory factors IL-1β, IL-6, and CD68 did not show significant differences between the groups. However, the mRNA levels of the inflammatory factors TNF-α and MCP-1 increased in the HFD group, while TPH supplementation significantly reduced these mRNA levels. Due to reduced expression of tight junction proteins, blood-brain barrier permeability is increased in HFD-fed obese mice, leading to overexposure of the brain to inflammatory cytokines, which may aggravate neuropsychiatric disorders in HFD-induced obese mice. HFD induces early signs of neuropsychiatric disorders in the hippocampus, including decreased levels of tight junction transcripts and systemic and local inflammation. TP intervention can alleviate these adverse effects, exert anti-inflammatory effects, and restore HFD-induced anxiety-like behavioral disorders.

[0099] This example measured the levels of two neurotransmitters from the gut microbiota, serotonin (5-HT) and norepinephrine (NE). As shown in Figure 3G, 5-HT levels were similar in ND and HFD mice, but in mice fed ND and HFD, TP supplementation at two doses, TPH and TPL, increased 5-HT levels. NE levels were reduced in HFD-induced obese mice, but TPH and TPL significantly improved NE levels in HFD-fed mice.

[0100] These results suggest that TP improves the blood-brain barrier by increasing the expression of tight junction proteins, attenuating inflammatory responses, and regulating neurotransmitter pathways.

[0101] Meanwhile, TP supplementation improved the intestinal permeability and immune response of enterocytes in HFD-fed mice, and the results are shown in Figure 4 .

[0102] Among them, the results in Figure 4A and B showed that HFD feeding caused the colon length of mice to be shorter than that of ND-fed mice, while TPL and TPH interventions increased the colon length by 9.64% and 20.25%, respectively, and the differences were significant.

[0103] As shown in Figure 4C, HFD feeding reduced the thickness of colonic mucus in mice compared with ND-fed mice, as evidenced by mucus staining and MUC2 immunofluorescence staining of enterocytes. TPL and TPH intervention increased the thickness of the mucus layer in HFD-induced mice.

[0104] As shown in Figure 4, DG, both doses of TP increased the expression of occludin and ZO-1 in the colon at both the protein and transcript levels compared with those in HFD-fed mice.

[0105] As shown in Figure 4H, in addition to occludin and ZO-1, the mRNA levels of other colon-specific tight junction genes, including CLDN2, CLDN4, CLDN7, CLDN12, CLDN15, ICAM-1, and VCAM-1, were also measured. Compared with ND-fed mice, HFD feeding significantly reduced the transcript levels of CLDN7, CLDN12, CLDN15, and VCAM-1, while after TPH supplementation, the mRNA levels in HFD-fed mice were partially increased to levels comparable to those of ND-fed mice.

[0106] As shown in Figure 4, IJ, the deterioration of the colonic mucus layer and tight junction proteins may affect the expression of the antimicrobial peptide Reg3γ and inflammatory factors. It can be observed that TPH increased the mRNA expression of Reg3γ and significantly reduced endotoxemia in the serum compared with the HFD group mice.

[0107] As shown in Figure 4(K), TPL and TPH attenuated the mRNA expression of inflammatory factors TNF-α, IL-1β, IL-6, MCP-1, and CD68 in the colon of obese mice. As shown in Figure 4(L), TPL and TPH attenuated the levels of inflammatory factors TNF-α, IL-1β, and IL-6 in the serum of obese mice. These results suggest that TPL and TPH reduce serum inflammatory factors in HFD-induced mice and that TP protects the colon from HFD-induced exacerbations.

[0108] 16S rRNA gene sequencing was used to evaluate the effects of TP on the intestinal microbiota of HFD-fed mice. The results are shown in Figure 5A. Before the intervention at week 0, the microbiota composition of all groups was similar at the OTU level, while OTU clusters were clearly separated between the five groups in the final week. As shown in Figure 5B, at week 0, alpha diversity based on the Shannon index did not show much difference among mice, while at week 12, HFD-fed mice with or without TP treatment showed less diversity than mice fed ND.

[0109] As shown in Figure 6A, at the phylum level, all HFD-fed mice, regardless of whether they received TP intervention, showed increased abundance of Firmicutes and decreased abundance of Bacteroidetes. The abundance of Actinobacteria decreased in the HFD group, while TPH intervention increased the abundance of Actinobacteria in HFD-fed mice. Proteobacteria, which contain various pathogenic bacteria, were enriched in HFD-fed mice without TP treatment, and TPH could rebalance their abundance.

[0110] As shown in Figure 6B and C, the results showed that many communities at the genus level in ND- and HFD-fed mice underwent significant changes. Among them, the abundance of Lactobacillus in the HFD group was significantly reduced compared with the ND group, but TPL and TPH treatment could not rebalance the abundance of Lactobacillus in obese mice. Bifidobacterium is a probiotic, and its abundance in the HFD group was much lower than that in ND-fed mice, indicating that HFD is not conducive to the growth of Bifidobacterium, while TPH relatively increased the abundance of Bifidobacterium. It can be seen that TPH also rebalanced the abundance of Allobaculum. In addition, both TPL and TPH reduced the abundance of Escherichia-Shigella, which was enriched in the HFD group, indicating that TP has a beneficial effect on maintaining a healthy intestinal flora.

[0111] Community heatmap analysis (Figure 7A) demonstrated that the microbiota of control mice (ND and ND+TP groups) were closely related to each other, while the gut microbiota of HFD-fed mice, particularly those treated with TP, were closely related. As shown in Figure 7B, PICRUSt-predicted KEGG functional orthologs identified three potential functions. HFD-fed mice, regardless of TP treatment, showed similarities in secondary metabolite biosynthesis, microbial metabolism in diverse environments, amino acid biosynthesis, carbon metabolism, and ABC transporters. Several amino acid metabolites, such as tryptophan and glutamate, are thought to be precursors of neurotransmitters involved in the gut-brain axis. Correlations between the gut microbiota and other parameters, including body weight and behavioral tests, were further investigated. As shown in Figure 7C, a significant number of bacterial groups were significantly associated with body weight and marble burial, while only two genera were associated with OFT and one genus was significantly associated with EPM. This suggests that the gut microbiota is associated with behavioral test parameters and that TP can ameliorate HFD-induced anxiety-like behavior in mice by rebalancing the HFD-induced gut microbiota imbalance.

[0112] Example 2

[0113] Fecal microbiota transplantation (FMT)

[0114] Mice fed with a HFD were treated with an antibiotic solution (ATB) containing 0.5 g / L vancomycin hydrochloride, 1 g / L neomycin sulfate, 1 g / L ampicillin sulfate, and 1 g / L metronidazole to eradicate intestinal flora. The ATB solution was added to the sterile drinking water of HFD-fed mice for 10 days. Fecal samples from mice in the five dietary groups (ND, ND+TPH, HFD, HFD+TPL, and HFD+TPH) were collected. Subsequently, the fresh fecal samples were resuspended in sterile PBS (15 mL / g feces), vortexed vigorously for 5 minutes, and then left to settle for 5 minutes to precipitate particles. The supernatant was used to colonize mice treated with ATB by oral gavage. Fecal microbiota from ND-fed mice (dND), ND-fed TPH-intervention mice (dND+TPH), HFD-fed mice (dHFD), and HFD-fed TPL (dHFD+TPL) and TPH (dHFD+TPH)-intervention mice were transplanted into HFD-fed recipient mice (rHFD) to further evaluate the effects of the microbiota on HFD-induced mice.

[0115] Behavioral testing, Alcian blue (AB) staining, immunohistochemistry and immunofluorescence staining, Western blotting, ELISA, 16S rRNA sequencing analysis and data analysis were the same as those used in Example 1.

[0116] Result analysis:

[0117] The behavioral test results shown in Figures 8A to D show that compared with recipient mice transplanted with microbiota from dND, dND+TPH, and dHFD+TPH, mice on the dHFD→rHFD and dHFD+TPL→rHFD spent more time in the surrounding area or closed arms during the OFT and EPM tests. In the marble burying test, as shown in Figures 8E and F, the microbiota from the dHFD+TPH group also reduced the number of beads buried by mice on the rHFD group compared with those from the dHFD→rHFD and dHFD+TPL→rHFD groups. This suggests that the microbiota from obese mice treated with TPH can alleviate the anxiety-like behavior induced by HFD in mice.

[0118] The immune system is considered one of the communication pathways in the microbiota-gut-brain axis. The gut microbiota, various immune cells, and nerves are closely connected to the intestinal lining and influence the immune response through a three-way interaction. The gut microbiota is crucial in regulating the development and function of innate immune cells in the brain and is required for the maturation and activation of microglia in the brain. The gut microbiota also interacts with the brain through the systemic immune system. Changes in circulating cytokines in systemic immunity lead to changes in brain inflammation, and neuropsychiatric diseases such as anxiety disorders are associated with peripheral inflammation.

[0119] Using double immunofluorescence staining and RT-qPCR, the results, shown in Figures 9A and 9B, indicate that the microbiota from dHFD+TPL and dHFD+TPH improved HFD-induced expression of Iba-1 and PSD95 at both the protein and mRNA levels in the hippocampus of recipient mice. As shown in Figure 9B, BDNF mRNA levels were also improved in mice in the dHFD+TPL→rHFD and dHFD+TPH→rHFD groups. Compared to dHFD→rHFD mice, TNF-α mRNA levels in the hippocampus of mice receiving the dHFD+TPL and dHFD+TPH microbiota were significantly reduced, whereas only the microbiota from dHFD+TPH mice significantly attenuated IL-1β mRNA levels. The results in Figure 9C demonstrate that the microbiota from dHFD+TPL and dHFD+TPH had limited effects on reducing IL-6 mRNA levels in rHFD mice, with no significant differences observed. The results shown in Figure 9 (DG) show that in the colon, fecal microbiota transplantation of dHFD+TPH significantly increased the thickness of the mucus layer, tight junction proteins, and the expression of the antimicrobial peptide Reg3γ compared with mice on dHFD→rHFD. These results suggest that FMT in mice treated with TP can improve anxiety-like behavioral disorders by alleviating neuroinflammation and protecting colon integrity.

[0120] Figures 10 and 11 show changes in the intestinal microbiota of mice receiving fecal microbiota from different donor groups. As shown in Figure 10A, the microbiota in the dND→rHFD and dHFD+TPH→rHFD groups differed significantly from those in the other groups. However, the results in Figure 10B show that the α-diversity of the microbiota did not differ significantly across all groups. As shown in Figures 11A and 11B, structural analysis of the intestinal microbial community at the phylum and genus levels revealed that mice on the dHFD+TPH→rHFD diet had the lowest abundance of Proteobacteria and their Escherichia coli-Shigella genera. However, the abundance of Firmicutes, Blautia, and Romboutsia genera was highest in the dHFD+TPH→rHFD group. The abundance of the probiotic Bifidobacterium was lowest in the dHFD→rHFD group, while fecal microbiota transplantation in the other groups rebalanced these levels. The above research results indicate that fecal microbiota transplantation from mice receiving TP intervention can reconstruct or improve the intestinal microbiota of HFD-fed recipient mice, thereby improving the anxiety-like behavior of HFD-induced mice.

[0121] In summary, the present application proposes the use of Tremella polysaccharide in the preparation of products for preventing and treating anxiety caused by a high-fat diet. Tremella polysaccharide can reduce weight gain in obese mice fed a high-fat diet (HFD) and reduce liver and fat weight. In the open field experiment, elevated plus maze and marble burying behavioral tests, the intake of Tremella polysaccharide (TP) can prevent and improve anxiety-like behavioral disorders caused by the intake of a high-fat diet (HFD), and TP intervention can inhibit neuroinflammation, microglial activation and synaptic damage. The intake of TP increases the mRNA level of tight junction proteins and neurotransmitter levels in the blood-brain barrier of HFD-fed mice, and can prevent the dysbiosis of the intestinal flora. Fecal microbiota transplantation (FMT) can rebalance the intestinal flora to improve the intestinal flora of HFD-fed mice, further achieving the effect of directly improving their anxiety-like behavior and preventing obesity. Tremella polysaccharide has the effect of preventing and treating anxiety caused by a high-fat diet. It can be used in the preparation of products for preventing and treating anxiety caused by obesity and / or intestinal flora imbalance induced by a high-fat diet. As a strategy for nutritional intervention in anxiety disorders caused by a high-fat diet, it is of great significance for preventing and improving anxiety disorders caused by a high-fat diet.

[0122] The above description is only a preferred embodiment of the present application and is not intended to limit the present application. Any modifications, equivalent replacements and improvements made within the spirit and principles of the present application should be included in the scope of protection of the present application.

Claims

1. Application of Tremella polysaccharide in the preparation of products for preventing and treating anxiety caused by high-fat diet.

2. The use according to claim 1, characterized in that: The tremella polysaccharide reduces the activation of microglia, increases the activation of synapses and improves the expression of brain-derived neurotrophic factor, so as to achieve the effect of preventing and treating anxiety.

3. The use according to claim 1, characterized in that: The tremella polysaccharide improves the transcription levels of the three genes occludin, ZO-1 and claudin5 in the hippocampus and increases the expression of tight junction proteins to improve the blood-brain barrier, prevent the brain from being overly exposed to inflammatory cytokines that pass through the blood-brain barrier, and achieve the effect of preventing and treating anxiety.

4. The use according to claim 1, characterized in that: The Tremella polysaccharide reduces the mRNA levels of inflammatory factors TNF-α and MCP-1 in the hippocampus and alleviates the inflammatory response, thereby playing an anti-inflammatory and anxiety-preventing role.

5. The use according to claim 1, characterized in that: The tremella polysaccharide increases the thickness of intestinal mucus, improves the expression of intestinal tight junction proteins, and improves the expression of the antimicrobial peptide Reg3γ, thereby enhancing the intestinal barrier, and further reduces the mRNA expression of inflammatory factors TNF-α, IL-1β, IL-6, MCP-1 and CD68 in the colon, thereby achieving anti-inflammatory and immune-regulating effects.

6. The use according to claim 1, characterized in that: The Tremella polysaccharide can achieve the effect of preventing and treating anxiety by directly improving the intestinal flora; and the intestinal flora improved by the Tremella polysaccharide can also reduce the inflammatory response in the brain and intestines induced by a high-fat diet after transplantation, thereby achieving the effect of preventing and treating anxiety.

7. The use according to claim 1, characterized in that: The products include medicines containing tremella polysaccharides, foods containing tremella polysaccharides or food additives containing tremella polysaccharides.

8. The use according to claim 7, characterized in that: The drug containing Tremella polysaccharide comprises a pharmaceutically effective amount of Tremella polysaccharide and a pharmaceutically acceptable carrier; and / or, The food containing Tremella polysaccharide includes at least one of a functional food containing Tremella polysaccharide, a health product containing Tremella polysaccharide, a dietary supplement containing Tremella polysaccharide, and a common food containing Tremella polysaccharide.

9. The use according to claim 7, characterized in that: The food containing Tremella polysaccharide includes at least one of Tremella soup, Tremella slurry, Tremella paste, Tremella soup, dry Tremella soup, Tremella original powder, and Tremella dry powder; and / or, The food additive containing Tremella polysaccharide includes at least one of Tremella soup, Tremella slurry, Tremella paste, Tremella soup, dry Tremella soup, Tremella original powder, and Tremella dry powder; and / or, The medicine containing Tremella polysaccharide includes at least one of tablets, dropping pills, dispersible tablets, capsules, and oral liquid; and / or, The pharmaceutically acceptable carrier includes at least one of a diluent, a filler, a binder, a preservative, a lubricant, a dispersant, a wetting agent, a sweetener, a flavoring agent, an emulsifier, a suspending agent, a preservative, an antioxidant, a colorant, and a stabilizer.

10. The use according to claim 1, characterized in that: The products include food preparations, pharmaceutical preparations or fecal microbiota transplantation preparations containing intestinal flora improved by Tremella polysaccharide.

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