Application of tremella polysaccharide in preparation of product for preventing and treating anxiety caused by high fat diet

Abstract

The invention relates to the technical field of health food, in particular to application of tremella polysaccharide in preparation of a product for preventing and treating anxiety caused by high fat diet. Researches find that tremella polysaccharide can reduce weight gain of mice fed with high-fat diet, reduce liver and fat weight, inhibit microglial cell activation and neuroinflammation in hippocampus and balance intestinal flora, and has an excellent improvement effect on prevention and treatment of anxiety of mice caused by high-fat diet feeding. The application verifies that the tremella polysaccharide has the effect of preventing and treating the anxiety caused by the high-fat diet, can be applied to preparation of products for preventing and treating the anxiety caused by the high-fat diet, serves as a strategy for nutrition intervention on the anxiety disorder caused by the high-fat diet, and has important significance on auxiliary prevention and improvement of the anxiety disorder caused by the high-fat diet.

Description

Technical Field

[0001] The present application belongs to the technical field of health food, and specifically relates to the application of Tremella polysaccharide in the preparation of products for preventing and treating anxiety caused by a high-fat diet. Background Art

[0002] It is well known that a high-fat diet (HFD) causes many diseases, such as obesity, diabetes, and metabolic syndrome, which are associated with an increased risk of neuropsychiatric diseases. An increasing 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 behavior 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 dysbiosis may cause neuroinflammation and brain development disorders by activating microglia. And HFD is associated with anxiety-like behaviors and intestinal flora dysbiosis. The intake of HFD may significantly change the composition and function of the intestinal flora, which is a key regulator of host intestinal homeostasis and immunity.

[0003] 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 can transfer bacterial endotoxins (lipopolysaccharide, LPS) into the systemic blood circulation, which is considered endotoxemia. The increase in systemic LPS may further trigger neuroinflammatory responses. It has been reported that endotoxemia activates microglia and immune cells in the central nervous system and induces inflammatory responses 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.

[0004] Given the popularity of "Western" diets in modern life and the increasing global burden of mood disorders, it is imperative to find effective intervention strategies to combat anxiety-like behavioral disorders and obesity symptoms caused by HFD. Currently, there are no clinical products that can prevent and treat anxiety symptoms caused by a high-fat diet. Therefore, there is an urgent need to provide a product that can prevent and treat anxiety symptoms caused by a high-fat diet. Summary of the invention

[0005] The purpose of the present 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.

[0006] In order to achieve the above application purpose, the technical solution adopted in this application is as follows:

[0007] In a first aspect, the present application provides the use of Tremella polysaccharide in the preparation of a product for preventing and treating anxiety caused by a high-fat diet.

[0008] In some embodiments, the Tremella polysaccharide reduces the activation of microglia, increases the activation of synapses, and improves the expression of brain-derived neurotrophic factor to achieve the effect of preventing and treating anxiety.

[0009] 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, 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.

[0010] In some embodiments, the Tremella polysaccharide reduces the mRNA levels of inflammatory factors TNF-α and MCP-1 in the hippocampus and alleviates the inflammatory response, thereby exerting anti-inflammatory and anxiety-preventing effects.

[0011] 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 improving 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.

[0012] 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.

[0013] In some embodiments, the product includes a medicine containing Tremella polysaccharide, a food containing Tremella polysaccharide, or a food additive containing Tremella polysaccharide.

[0014] In some embodiments, the drug containing Tremella polysaccharide includes a pharmaceutically effective amount of Tremella polysaccharide and a pharmaceutically acceptable carrier.

[0015] 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.

[0016] 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.

[0017] 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.

[0018] In some embodiments, the drug containing Tremella polysaccharide includes at least one of tablets, pills, dispersible tablets, capsules, and oral liquids.

[0019] 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.

[0020] In some embodiments, the product includes a food preparation, a pharmaceutical preparation or a fecal microbiota transplantation preparation containing the intestinal flora improved by Tremella polysaccharide.

[0021] 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.

[0022] 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.

[0023] The first aspect of the present application provides an application of Tremella polysaccharide in the preparation of products 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 anxious 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

[0024] In order to more clearly illustrate the technical solutions in the embodiments of the present application, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. 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 paying any creative work.

[0025] Figure 1 The results of studying the effect of Tremella polysaccharide (TP) intake on metabolic syndrome parameters of HFD-fed mice in Example 1 of this application. A. Experimental design; B. Weight gain; C. Fasting blood glucose; D. Tissue weight. Data are presented as mean ± SEM. Tukey's multiple range test showed that the values ​​of columns with different letters were significantly different (p<0.05).

[0026] Figure 2 The present invention relates to a behavioral test result of studying the effect of Tremella polysaccharide (TP) intake on HFD-induced anxiety-like behavior in mice in Example 1 of the present application, wherein A is a representative track image in OFT; B is a 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 a quantification of the total distance, the time ratio of the open arm travel and the frequency of entry into the open arm; E is a representative image in the marble burying test; F is a 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 per 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 per day; HFD+TPH: mice fed a high-fat diet, gavage of 400 mg / kg TP per day).

[0027] Figure 3 The results of studying the effect of Tremella polysaccharide (TP) intake on brain function of HFD-fed mice in Example 1 of the present application, wherein A is immunofluorescence staining of Iba1 and PSD95 in the hippocampus. Scale: 50 μm; B is immunohistochemical staining of Iba-1 protein in the hippocampus. Scale: 100 μm; C is the protein level of PSD95 and BDNF in the hippocampus; D is the relative mRNA level of Iba-1, PSD95 and BDNF in the hippocampus; E is the relative mRNA level of tight junction proteins Occludin, ZO-1 and Claudin5 in the hippocampus; F is the relative mRNA level of inflammatory factors; G is the level of hippocampal neurotransmitters, 5-hydroxytryptamine (5-HT) and norepinephrine (NE).

[0028] Figure 4These are 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 length of the colon; B is the quantification of the length of the colon; 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 proteins 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.

[0029] Figure 5 These are 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 alpha diversity measured by the Shannon index at week 0 and week 12.

[0030] Figure 6 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.

[0031] Figure 7 These are 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.

[0032] Figure 8These are 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 venue; 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).

[0033] Fig. 9 These are 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 the 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 level of inflammatory factors in the hippocampus; D is the 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: 100 μm.

[0034] Fig.10 These are the results of studying the effect of fecal microbiota transplantation (FMT) on the intestinal flora 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.

[0035] Fig.11 These are the results of studying the effects of fecal microbiota transplantation (FMT) on the intestinal flora 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. DETAILED DESCRIPTION

[0036] In order to make the technical problems, technical solutions and beneficial effects to be solved by the present application more clearly understood, the present application is further described in detail in conjunction with the embodiments below. It should be understood that the specific embodiments described herein are only used to explain the present application and are not used to limit the present application.

[0037] The first aspect of the embodiments of the present application provides the use of Tremella polysaccharide in the preparation of a product for preventing and treating anxiety caused by a high-fat diet.

[0038] The first aspect of the embodiment 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 anxious patients. Therefore, Tremella polysaccharide can be used in the preparation of products for preventing and treating anxiety, as a strategy for intervening in anxiety disorders caused by a high-fat diet, to achieve the effect of preventing and treating anxiety disorders caused by HFD, and has a wide range of application value.

[0039] Tremella has a history of thousands of years in China. Studies have found that Tremella polysaccharide (TP) has hypoglycemic and hypolipidemic effects, anti-obesity effects, and neuroprotective effects. For example, purified polysaccharides from Tremella showed neuroprotective effects in glutamate-induced differentiated PC-12 cell injury. Tremella polysaccharide (TP) can also restore cognitive function by regulating the CREB signaling pathway and cholinergic system in the hippocampus of rats induced by trimethyltin.

[0040] In the study, the applicant found 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 in 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, followed by 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.

[0041] 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 the prevention and treatment of anxiety and obesity.

[0042] In some embodiments, the Tremella polysaccharide reduces the activation of microglia, increases the activation of synapses, and improves the expression of brain-derived neurotrophic factor to achieve the effect of preventing and treating anxiety.

[0043] 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, 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.

[0044] In some embodiments, the Tremella polysaccharide reduces the mRNA levels of inflammatory factors TNF-α and MCP-1 in the hippocampus and alleviates the inflammatory response, thereby exerting anti-inflammatory and anxiety-preventing effects.

[0045] 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 improving 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.

[0046] 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.

[0047] In some embodiments, the product includes a medicine containing Tremella polysaccharide, a food containing Tremella polysaccharide, or a food additive containing Tremella polysaccharide.

[0048] In some embodiments, the drug containing Tremella polysaccharide includes a pharmaceutically effective amount of Tremella polysaccharide and a pharmaceutically acceptable carrier.

[0049] 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.

[0050] 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.

[0051] 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.

[0052] In some embodiments, the drug containing Tremella polysaccharide includes at least one of tablets, pills, dispersible tablets, capsules, and oral liquids.

[0053] 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.

[0054] In some embodiments, the product includes a food preparation, a pharmaceutical preparation or a fecal microbiota transplantation preparation containing the intestinal flora improved by Tremella polysaccharide.

[0055] 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.

[0056] 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.

[0057] The solution of the present application is further described below with reference to specific embodiments.

[0058] Example 1

[0059] 1.1 Experimental animals and treatment

[0060] Male C57BL / 6J mice (7 weeks old) were purchased from Weitong Lihua Co., Ltd. (Beijing, China). Five mice were housed in each cage under standard conditions (12 / 12 light-dark cycle, humidity of 50±15%, temperature of 22±2°C). Mice were randomly divided into 5 groups according to the different treatments they received.

[0061] ND: mice were fed with a normal diet (10% kcal from fat, D12450J, Changzhou Shuyi Shuer Biotechnology Co., Ltd.) and distilled water;

[0062] ND+TPH: mice were fed a normal diet and intragastrically administered with 400 mg / kg of TP (purchased from Bo InnoHealthBiotechnology Company Ltd (Hong Kong, China)) daily;

[0063] HFD: mice were fed a high-fat diet (60% kcal from fat, PD6001, Changzhou Shuyi Shuer Biotechnology Co., Ltd.) and distilled water;

[0064] HFD+TPL: mice were fed a high-fat diet and gavaged with 40 mg / kg TP daily;

[0065] HFD+TPH: Mice were fed a high-fat diet and gavaged with 400 mg / kg TP daily.

[0066] After 12 weeks of intervention, all mice underwent behavioral testing. Subsequently, blood samples were drawn by cardiac puncture, and tissues were collected after euthanasia using carbon dioxide.

[0067] 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).

[0068] 1.2 Provide Tremella polysaccharide (TP) preparations

[0069] Tremella polysaccharide was extracted and supplied by Bo InnoHealth Biotechnology Company Ltd (Hong Kong, China). Briefly, 100 times of pure water was added to Tremella and mixed and boiled at 100°C for 2 h. This extraction process was repeated twice. After filtration and centrifugation, the supernatant was obtained and mixed with 4 times of anhydrous ethanol. Polysaccharide was precipitated by centrifugation. Tremella polysaccharide was dissolved in pure water and protein was removed by TCA method.

[0070] 1.3 Behavioral testing

[0071] Open field test (OFT), elevated plus maze (EPM) and marble burying are methods to evaluate anxiety or depression-like behaviors by assessing spontaneous motor activity in rodents. In this example, behavioral tests including OFT, EPM and marble burying tests were performed to evaluate anxiety-like conditions in mice.

[0072] In OFT and EPM, mice were carefully placed in the center area of ​​the maze and allowed to explore freely for 5 min, and parameters such as total distance, distance 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 with 10 glass beads. The number of beads buried by mice within 30 min was scored.

[0073] 1.4 Alcian blue (AB) staining

[0074] Transverse colon tissues of mice were placed in methanol-Carnoy's fixative (60% methanol, 30% chloroform, 10% glacial acetic acid) for at least 3 hours at room temperature, 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.

[0075] 1.5 Immunohistochemistry and immunofluorescence staining

[0076] The embedded colon and brain were cut into 5 μm and 20 μm sections, respectively. Sodium citrate antigen retrieval solution (PH6.0) was used to repair 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.

[0077] The sections were stained with DAB kit (Servicebio, Wuhan, China) for immunohistochemistry (IHC). Images were observed under an optical microscope. For immunofluorescence (IF) staining, sections were counterstained with 4′6-diamidino-2-phenylindole (DAPI, Beyotime, Shanghai, China) in PBS to observe cell nuclei. Images were taken by a Nikon AX NISElements5.4 confocal microscope (Nikon, Tokyo, Japan).

[0078] 1.6 Western blotting

[0079] 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 supernatant was 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 h at room temperature. The Clarity TMProtein bands were detected using ECL Western Blot Substrate (Bio-Rad, CA, USA) and imaged using the ChemiDoc Touch Imaging System (Bio-Rad, CA, USA).

[0080] 1.7 Reverse transcription qualitative polymerase chain reaction (RT-qPCR)

[0081] Total RNA from colon and brain was extracted 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, SYBR green PCR kit (Biotium Forget-Me-Not TM mRNA expression was determined by qPCR (qPCR Master Mix, Fremont, California, USA). Relative mRNA levels were determined by the 2-ΔΔCt method. Primer sequences are shown in Table 1.

[0082] Table 1 Primer sequences

[0083]

[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 by ELISA kits (Shanghai Xinle Biotechnology Co., Ltd., Shanghai, China).

[0086] 1.9 16S rRNA sequencing analysis

[0087] Fecal samples were collected from the rectum before euthanasia, and fecal microbiota genomic DNA was extracted using the TIANamp Stool DNA Kit (Tiangen, Beijing, China). The extracted DNA was checked on a 1% agarose gel, and the 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 microbiota was amplified using primer pairs 338F (5′-ACTCCTACGGGAGGCAGCAG-3′) and 806R (5′-GGACTACHVGGGTWTCTAAT-3′) at Shanghai Meiji Biopharmaceutical Technology Co., Ltd. (Shanghai, China). The purified amplicons were mixed in equimolar amounts 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, Illinois, USA). All values ​​represent the mean and SEM of six to eight independent experiments. One-way ANOVA and Duncan multiple interval analysis were then used to compare the data. For 16S rRNA gene sequence analysis, all reads were deposited and grouped into operational taxonomic units (OTUs) at 97% sequence identity. Statistical analysis was performed using STAMP, and the functional differences of orthologs between groups were assessed by one-way ANOVA, 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. Figure 1 A. The initial body weights of the groups were at similar levels, and 12 weeks of intervention with high-dose TP (TPH) significantly reduced the body weight of HFD-fed mice ( Figure 1 B). In addition, fasting blood glucose levels were significantly reduced in obese mice supplemented with TPH compared with those in mice fed a HFD ( Figure 1 C). 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 1 D). These results suggest that supplementation with T. fuciformis polysaccharides, especially at high doses, exhibits anti-obesity effects in HFD-fed mice.

[0092] In the OFT test, mice that showed impaired anxiety-like behavior spent less time in the center zone than normal mice. Figure 2 As shown in A and B, HFD intake significantly decreased total distance and speed compared with mice fed ND. In HFD-fed mice, TPH intervention significantly increased these parameters. Meanwhile, HFD mice supplemented with TPH significantly increased the distance and time traveled in the central zone of OFT compared with HFD mice without TP intervention. The results indicate that TPH intervention improves anxiety-like behavioral disorders induced by HFD in mice.

[0093] In the EPM test, anxious mice spent less time in the open arms than normal mice. Figure 2 As shown in C and D, the total distance traveled by HFD-fed mice was significantly reduced, and the distance and time traveled in the open arms were shorter than those of ND mice, while TPH significantly increased the time spent in the open arms by HFD-fed mice. This suggests that TPH intervention alleviated the anxiety tendency of HFD-induced mice.

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

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

[0096] The activation of microglia 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 of Iba-1 and PSD95 was used in all groups to determine the spatial location of microglia and synapses, and the results are shown in Figure 3As shown in Figure 5A, the expression of Iba-1 was increased, while the expression of PSD95 was decreased in the HFD group compared with the ND and TPH groups, indicating that TP supplementation, especially high-dose TP, can attenuate the activation of microglia but increase the activation of synapses. Figure 3 The immunohistochemical staining results in Figure 5B showed that HFD promoted the activation of microglia, while TPH intervention reduced the microglia expression in obese mice. Figure 3 Western blot analysis in Figure C showed that the expression of PSD95 and brain-derived neurotrophic factor (BDNF) was decreased in the HFD and HFD+TPL groups compared with that in ND-fed mice, whereas their expression levels were increased in HFD-fed mice after TPH supplementation. Figure 3 As shown in D, the mRNA levels of Iba-1, PSD95, and BDNF in the five groups of mice in RT-qPCR showed results consistent with immunofluorescence, immunohistochemistry, and western blotting. The results showed that TPH could improve the activation of microglial markers, impaired expression of postsynaptic density protein and brain-derived neurotrophic factor in the hippocampus caused by HFD.

[0098] At the same time, 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. 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), the mRNA levels of key tight junction proteins and inflammatory factors were detected. The results are as follows Figure 3 As shown in E, the test results showed that the transcription levels of occludin, ZO-1 and claudin5 in HFD-fed mice were significantly lower than those in ND-fed control mice. TPL increased the mRNA level of ZO-1, and TPH also increased the transcription levels of occludin, ZO-1 and claudin5 in obese mice. Figure 3 As shown in F, the mRNA levels of inflammatory factors IL-1β, IL-6, and CD68 did not show significant differences among these groups, however, the mRNA levels of inflammatory factors TNF-α and MCP-1 increased in mice in the HFD group, while supplementation with TPH significantly reduced the mRNA levels of inflammatory factors TNF-α and MCP-1. Due to the reduced expression of tight junction proteins, the permeability of the blood-brain barrier in the HFD-fed obese mouse model increased, and the brain was overexposed to inflammatory cytokines, which may aggravate the neuropsychiatric disorders in HFD-induced obese mice. HFD induces early signs of neuropsychiatric disorders in the hippocampus, including reduced levels of tight junction transcripts, systemic and local inflammation. TP intervention can alleviate these adverse reactions, exert anti-inflammatory effects, and restore HFD-induced anxiety-like behavioral disorders.

[0099] This example measures the levels of two neurotransmitters from the intestinal flora, namely 5-hydroxytryptamine (5-HT) and norepinephrine (NE). Figure 3 As shown in G, 5-HT levels were similar in ND and HFD mice, but TP supplementation at both doses, TPH and TPL, increased 5-HT levels in both ND and HFD-fed mice. NE levels were decreased 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 intestinal permeability and immune response of enterocytes in HFD-fed mice, as shown in Figure 4 shown.

[0102] in, Figure 4 The results in Figures A and B showed that HFD feeding resulted in shorter colon length in mice than in ND-fed mice, whereas TPL and TPH interventions increased colon length by 9.64% and 20.25%, respectively, with significant differences.

[0103] like Figure 4 As shown in C, 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] like Figure 4 As shown in DG, both doses of TP increased the expression of occludin and ZO-1 at the protein and transcript levels in the colon compared with HFD-fed mice.

[0105] like Figure 4 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 determined as shown in Figure H. HFD feeding significantly reduced the transcript levels of CLDN7, CLDN12, CLDN15, and VCAM-1 compared with ND-fed mice, while after TPH supplementation, the mRNA levels of HFD-fed mice were partially increased to levels comparable to those of ND-fed mice.

[0106] like Figure 4As shown in Figure 1J, 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] like Figure 4 As shown in Figure K, the mRNA expressions of inflammatory factors TNF-α, IL-1β, IL-6, MCP-1, and CD68 in the colon of obese mice were attenuated by TPL and TPH; Figure 4 As shown in Figure L, the levels of inflammatory factors TNF-α, IL-1β, and IL-6 in the serum of obese mice were attenuated by TPL and TPH. The results showed that TPL and TPH reduced serum inflammatory factors in HFD-induced mice, and TP could protect the colon from HFD-induced deterioration.

[0108] 16S rRNA gene sequencing was used to evaluate the effect of TP on the intestinal flora of HFD-fed mice, and the results were as follows Figure 5 As shown in Figure A, before the intervention in week 0, the bacterial flora composition of all groups was similar at the OTU level, while in the last five weeks, the OTU clusters were clearly separated between the five groups. Figure 5 As shown in Figure 2B, at week 0, alpha diversity based on the Shannon index did not show much difference among mice, whereas at week 12, HFD-fed mice with or without TP treatment showed less diversity than ND-fed mice.

[0109] like Figure 6 As shown in A, at the phylum level, all HFD-fed mice, regardless of TP intervention, showed increased abundance of Firmicutes and decreased abundance of Bacteroidetes. The abundance of Actinobacteria was 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] like Figure 6As shown in Figures B and C, the results showed that many communities at the genus level were significantly changed in ND- and HFD-fed mice. Among them, the abundance of Lactobacillus was significantly reduced in the HFD group compared with the ND group, but TPL and TPH treatment could not rebalance the abundance of Lactobacillus in obese mice. Bifidobacterium, a probiotic, was much less abundant in the HFD group than in ND-fed mice, indicating that HFD was not conducive to the growth of Bifidobacterium, while TPH relatively increased the abundance of Bifidobacterium. And 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] Figure 7 The community heat map analysis shown in A shows that the microbiota of the ND group and the ND+TP group of control mice are closely related to each other, while the intestinal microbiota of the HFD-fed mice, especially the two groups of TP-treated mice, are closely related. Figure 7 As shown in Figure B, PICRUSt-predicted KEGG functional orthologs identified potential functions at level 3. HFD-fed mice, whether or not they received TP treatment, were associated with biosynthesis of secondary metabolites, microbial metabolism in different environments, biosynthesis of amino acids, carbon metabolism, and ABC transporters. Several amino acid metabolites, such as tryptophan and glutamate, are considered to be precursors of neurotransmitters involved in the gut-brain axis. The correlations between gut microbiota and other parameters, including body weight and parameters in behavioral tests, were further investigated. Figure 7 As shown in C, a considerable number of bacterial groups were significantly associated with body weight and marble burying, 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 the parameters in the behavioral tests and TP can improve the anxiety-like behavior of HFD-induced mice by rebalancing the gut microbiota dysbiosis induced by HFD.

[0112] Example 2

[0113] Fecal microbiota transplantation (FMT)

[0114] HFD-fed mice 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 five dietary groups (ND, ND+TPH, HFD, HFD+TPL and HFD+TPH) were collected. Subsequently, fresh fecal samples were resuspended in sterile PBS (15 mL / g feces), vortexed vigorously for 5 minutes, and then left to settle particles for 5 minutes. 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] The behavioral test, Alcian blue (AB) staining, immunohistochemistry and immunofluorescence staining, Western blot, ELISA, 16S rRNA sequencing analysis and data analysis were the same as those used in Example 1.

[0116] Result analysis:

[0117] Figure 8 The behavioral test results shown in AD show that mice in the dHFD→rHFD and dHFD+TPL→rHFD spent more time in the surrounding area or closed arms in the OFT and EPM tests compared to recipient mice transplanted with microbiota from dND, dND+TPH, and dHFD+TPH. Figure 8 As shown in E and F, the microbiota in dHFD+TPH also reduced the number of buried beads in mice on rHFD compared with dHFD→rHFD and dHFD+TPL→rHFD, indicating that the microbiota of obese mice treated with TPH can alleviate the anxiety-like behavior of HFD-induced mice.

[0118] The immune system is considered to be 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 tripartite interaction. The gut microbiota is critical 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 altered brain inflammation, and neuropsychiatric diseases such as anxiety disorders are associated with peripheral inflammation.

[0119] Using double immunofluorescence staining and RT-qPCR, the results were as Fig. 9 As shown in Figures A and B, it was observed that the microbiota from dHFD+TPL and dHFD+TPH improved the HFD-induced expression of Iba-1 and PSD95 at the protein and mRNA levels in the hippocampus of recipient mice. Fig. 9 The results in Figure B showed that the mRNA levels of BDNF were also improved in the mice in the dHFD+TPL→rHFD and dHFD+TPH→rHFD groups. Compared with the dHFD→rHFD mice, the mRNA levels of TNF-α in the hippocampus of mice receiving the dHFD+TPL and dHFD+TPH microbiota were significantly reduced, while only the microbiota from the dHFD+TPH mice significantly attenuated the mRNA levels of IL-1β. Fig. 9 The results in Figure C showed 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. Fig. 9 The results shown in DG in the middle showed 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 in dHFD→rHFD. The results suggest that FMT in mice treated with TP can improve anxiety-like behavioral disorders by alleviating neuroinflammation and protecting colon integrity.

[0120] Changes in intestinal flora of mice receiving fecal flora from different donor groups Figures 10-11 As shown. Fig.10 As shown in Figure A, the bacterial flora in the dND→rHFD and dHFD+TPH→rHFD groups were significantly different from those in the other groups. Fig.10 The results in Figure B show that the α diversity of the bacterial communities in all groups did not show much difference. Fig.11 As shown in A and B, structural analysis of the intestinal microbiota at the phylum and genus levels showed that mice in dHFD+TPH→rHFD showed the lowest abundance of Proteus and its Escherichia coli-Shigella genera, but the abundance of Firmicutes, Blautia, and Romboutsia genera were the highest in mice in the dHFD+TPH→rHFD group, and the abundance of the probiotic Bifidobacterium genus was the lowest in dHFD→rHFD, while fecal microbiota transplantation in other groups rebalanced the levels. The above research results indicate that fecal microbiota transplantation of 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 the weight gain of obese mice fed a high-fat diet (HFD) and reduce liver and fat weight. In the open field test, elevated cross 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 intestinal flora. Fecal flora 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, and 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 substitutions and improvements made within the spirit and principles of the present application should be included in the protection scope 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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