Edgeworthia chrysantha polysaccharide as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of biological medicine and natural product preparation, and relates to edgeworthia chrysantha polysaccharide as well as a preparation method and application thereof. Edgeworthia chrysantha flower buds are used as a raw material, a water extraction and alcohol precipitation method is adopted, further purification is performed to prepare the Edgeworthia chrysantha polysaccharide, and experiments prove that the Edgeworthia chrysantha (EGM) polysaccharide has anti-inflammatory and anti-oxidation effects, can achieve a good liver protection function and remarkably improves liver injury and intestinal flora disorder caused by CCl4. The extraction and preparation method of the edgeworthia chrysantha polysaccharide is simple and very suitable for industrial implementation, so that the edgeworthia chrysantha polysaccharide has good practical application value.

Description

Technical Field

[0001] The invention belongs to the technical field of biomedicine and natural product preparation, and relates to a dianthus polysaccharide and a preparation method and application thereof. Background Art

[0002] The information disclosed in this background technology section is only intended to enhance the understanding of the overall background of the invention, and should not necessarily be regarded as an admission or any form of suggestion that the information constitutes the prior art already known to a person skilled in the art.

[0003] As the core digestive and metabolic organ of the human body, the liver plays a vital role in the decomposition of endogenous and exogenous toxins. It is not only responsible for synthesizing bile and metabolizing substances such as fat and protein, but also plays a key role in the detoxification process. However, with the rapid development of industrial technology and the significant improvement of people's living standards, the challenges faced by the liver have also increased. Damage from toxic substances and their metabolites, deterioration of air quality, increased environmental pollution, excessive drinking and drug abuse all increase the risk of chemical liver damage. Long-term or acute liver damage may further evolve into fatty liver, hepatitis, cirrhosis, and even liver cancer.

[0004] Most of the liver protection drugs currently available on the market have certain limitations, such as strong side effects and relatively single efficacy. Therefore, the development of liver protection drugs or natural products with low toxicity and side effects and significant protective effects is of vital importance for preventing and treating liver damage and reducing the morbidity and mortality of liver disease, and has also become one of the hot topics in the global scientific research field. In particular, natural products are considered to be a powerful supplement to chemical synthetic drugs due to their advantages of low toxicity and side effects, multiple targets, and multiple pathways. These natural products effectively alleviate liver damage and protect liver tissue by improving the liver tissue's anti-oxidative stress, anti-inflammatory response, and regulating intestinal microorganisms.

[0005] Edgeworthia gardneri (Wall.) Meisn is a small tree of the family Thymelaeaceae and the genus Edgeworthia. It is widely used in Tibetan areas to prevent and treat "three highs" (high blood pressure, high blood sugar, and high blood lipids) and cardiovascular and cerebrovascular diseases. Its flower buds have the effect of clearing the liver and improving eyesight. In recent years, a large number of researchers have studied the chemical composition and pharmacological effects of Edgeworthia gardneri and found that it is rich in flavonoids, coumarins, phenylpropanoids and other ingredients, and has biological activities such as lowering blood sugar, lowering blood lipids, anti-oxidation, anti-tumor and liver protection. However, there are few reports on the biological activity of its polysaccharide components. Summary of the invention

[0006] In view of the deficiencies of the prior art, the present invention provides a polysaccharide of Dianjiexiang and a preparation method and application thereof. Specifically, the present invention extracts Dianjiexiang polysaccharide from Dianjiexiang and verifies through experiments that it has many pharmaceutical activities such as liver protection, anti-inflammatory, anti-oxidation and intestinal flora regulation. Based on the above research results, the present invention is completed.

[0007] In the present invention, E. yunnanensis polysaccharide may also be referred to as EGM polysaccharide.

[0008] In order to achieve the above technical objectives, the technical solution adopted by the present invention is as follows:

[0009] The first aspect of the present invention provides the use of Dianjiexiang polysaccharide in any one or more of the following:

[0010] (a) Anti-oxidation or preparation of anti-oxidation products;

[0011] (b) Anti-inflammatory or preparation of anti-inflammatory products;

[0012] (c) liver protection or products for preparing liver protection;

[0013] (d) Products for regulating intestinal flora or preparing intestinal flora conditions.

[0014] Among them, in (a), the anti-oxidation is specifically manifested as: reducing the levels of SOD, MDA, T-AOC and GSH-px.

[0015] In the above (b), the anti-inflammatory effect is specifically manifested as: increasing the expression level of IL-10 and reducing the expression levels of TNF-α, IL-6, and IL-1β.

[0016] In (c), liver protection is specifically manifested as: improving symptoms related to liver damage; further, the liver damage is caused by CCl 4 induced;

[0017] Furthermore, the improvement of liver injury-related symptoms is specifically manifested as: inhibiting the subject's weight loss, improving CCl 4 Pathological damage to liver tissue caused by leukemia reduces the expression levels of ALT and AST.

[0018] In (d), the intestinal flora regulation is specifically manifested as: at the phylum level, the abundance of Firmicutes is reduced, and the abundance of Bacteroidetes, Actinobacteria and Proteobacteria is increased. At the genus level, Lactobacillus, Faecalibacterium prausnitzii and Lactobacillus reuteri are increased, and the number of Brevibacterium, Lactobacillus mucosa and Deinococcus is reduced. At the same time, the functional pathways of the microbial community were analyzed through KEGG enrichment, which proved that the Yunnan knot fragrance polysaccharide can increase nucleotide metabolism, replication and repair, and carbohydrate metabolism.

[0019] The second aspect of the present invention provides a method for preparing Dianjiexiang polysaccharide, the preparation method comprising:

[0020] (1) extracting the buds of the osmanthus fragrans with ethanol by refluxing to remove fat-soluble substances to obtain a residue;

[0021] (2) boiling the residue obtained in step (1) with water to obtain a water extract, and collecting the supernatant after centrifugation;

[0022] (3) subjecting the supernatant obtained in step (2) to alcohol precipitation, and collecting the precipitate to obtain crude polysaccharide;

[0023] (4) The crude polysaccharide obtained in step (3) is obtained by removing the protein.

[0024] Wherein, in the step (1), the specific conditions of the ethanol reflux extraction are: using high concentration ethanol (such as 95% ethanol) for reflux extraction 1 to 5 times (preferably 3 times), each time for 1 to 5 hours (preferably 2 hours); further, before ethanol reflux extraction, the Yunnan knotwood is dried, such as drying at 40-60°C (preferably 50°C).

[0025] In the step (2), the specific conditions for boiling with water are: the material-liquid ratio is 1:1-10 (kg / L), preferably 1:5, the boiling treatment time is 1-3 hours, preferably 2 hours, and the number of times of boiling with water is 1-5 times (preferably 3 times).

[0026] In the step (3), the alcohol precipitation treatment method comprises: adding 95% ethanol to the supernatant, letting it stand and collecting the precipitate; the standing treatment time is 10-40 hours (preferably 24 hours).

[0027] In step (4), the protein removal can be performed by Sevage method, specifically, Sevage reagent (CH 3 Cl:BuOH=4:1, v / v) to remove protein 5 to 8 times.

[0028] Furthermore, the preparation method also includes freeze-drying the Dianjiexiang polysaccharide obtained in step (4).

[0029] The third aspect of the present invention provides a product, which at least contains the Yunnan knotweed polysaccharide obtained by the above preparation method.

[0030] The product has at least the following effects:

[0031] (a) Antioxidant;

[0032] (b) anti-inflammatory;

[0033] (c) liver protection;

[0034] (d) Regulation of intestinal flora.

[0035] The product may be food or medicine, which is not specifically limited here.

[0036] A fourth aspect of the present invention provides a method for preventing and treating CCl 4 A method for treating induced liver damage and intestinal flora disorder, comprising administering the above-mentioned Dianjiexiang polysaccharide or the above-mentioned product to a subject.

[0037] Beneficial technical effects of one or more of the above technical solutions:

[0038] In the above technical scheme, the buds of Eryngium yunnanensis are used as raw materials, and the Eryngium yunnanensis polysaccharide is prepared by water extraction and alcohol precipitation method and further purification. Experiments have shown that the Eryngium yunnanensis (EGM) polysaccharide has anti-inflammatory and antioxidant effects, and can also play a good role in liver protection and significantly improve CCl 4 The extraction and preparation method of Dianjiexiang polysaccharide in the above technical solution is simple and very suitable for industrial implementation, so it has good practical application value. BRIEF DESCRIPTION OF THE DRAWINGS

[0039] The accompanying drawings in the specification, which constitute a part of the present invention, are used to provide a further understanding of the present invention. The exemplary embodiments of the present invention and their descriptions are used to explain the present invention and do not constitute improper limitations on the present invention.

[0040] Figure 1 The EGM polysaccharide of the present invention regulates CCl 4 Body weight, liver index and pathological sections of induced mice; (a) body weight data, (b) liver index, (c) mouse liver H&E sections and Sirius red staining.

[0041] Figure 2 The present invention is a determination of the effects of EGM polysaccharide of the present invention on the inflammation level, liver function-related indicators and antioxidant capacity of each group; wherein (A) IL-6, (B) IL-10, (C) IL-1β, (D) TNF-α, (E) ALT, (F) AST, (G) SOD, (H) MDA, (I) T-AOC and (J) GSH-px.

[0042] Figure 3 The effect of EGM polysaccharide of the present invention on the richness of intestinal microorganisms. Among them, (A) ASV Venn diagram, (B) PCA analysis.

[0043] Figure 4 Alpha diversity analysis of EGM polysaccharides of the present invention; (A) diversity analysis, (B) rarefaction curve, (C) rank abundance curve.

[0044] Figure 5The effects of the EGM polysaccharide of the present invention on the microbial flora of different groups of mice; wherein, (A) phylum, (B) class, (C) order, (D) family, and (E) genus.

[0045] Figure 6 The variance analysis of the present invention showed the differences in the intestinal microbiota of different groups of mice.

[0046] Figure 7 Functional analysis of mouse microbiota for the present KEGG analysis. DETAILED DESCRIPTION

[0047] It should be noted that the following detailed descriptions are all illustrative and intended to provide further explanation of the present invention. Unless otherwise specified, all technical and scientific terms used herein have the same meanings as those commonly understood by those skilled in the art to which the present invention belongs.

[0048] It should be noted that the terms used herein are only for describing specific embodiments and are not intended to limit exemplary embodiments according to the present invention. As used herein, unless the context clearly indicates otherwise, the singular form is also intended to include the plural form. In addition, it should be understood that when the terms "comprising" and / or "including" are used in this specification, they indicate the presence of features, steps, operations, devices, components and / or combinations thereof. It should be understood that the scope of protection of the present invention is not limited to the specific embodiments described below; it should also be understood that the terms used in the embodiments of the present invention are intended to describe specific embodiments, rather than to limit the scope of protection of the present invention.

[0049] In a typical embodiment of the present invention, the use of Dianjiexiang polysaccharide in any one or more of the following is provided:

[0050] (a) Anti-oxidation or preparation of anti-oxidation products;

[0051] (b) Anti-inflammatory or preparation of anti-inflammatory products;

[0052] (c) liver protection or products for preparing liver protection;

[0053] (d) Products for regulating intestinal flora or preparing intestinal flora conditions.

[0054] in,

[0055] In the application (a), the anti-oxidation is specifically manifested as: reducing the levels of SOD, MDA, T-AOC and GSH-px.

[0056] In the application (b), the anti-inflammatory effect is specifically manifested as: increasing the expression level of IL-10 and reducing the expression levels of TNF-α, IL-6, and IL-1β.

[0057] In the application (c), liver protection is specifically embodied in: improving symptoms related to liver damage; further, the liver damage is caused by CCl 4 induced;

[0058] Furthermore, the improvement of liver injury-related symptoms is specifically manifested as: inhibiting the subject's weight loss, improving CCl 4 Pathological damage to liver tissue caused by leukemia reduces the expression levels of ALT and AST.

[0059] In the application (d), the intestinal flora regulation is specifically manifested as: at the phylum level, the abundance of Firmicutes is reduced, and the abundance of Bacteroidetes, Actinobacteria and Proteobacteria is increased. At the genus level, Lactobacillus, Faecalibacterium prausnitzii and Lactobacillus reuteri are increased, and the number of Brevibacterium, Lactobacillus mucosa and Deinococcus is reduced. At the same time, the functional pathways of the microbial community were analyzed through KEGG enrichment, which proved that the Yunnan knot fragrance polysaccharide can increase nucleotide metabolism, replication and repair, and carbohydrate metabolism.

[0060] In another specific embodiment of the present invention, a method for preparing Dianjiexiang polysaccharide is provided, and the preparation method comprises:

[0061] (1) extracting the buds of the osmanthus fragrans with ethanol by refluxing to remove fat-soluble substances to obtain a residue;

[0062] (2) boiling the residue obtained in step (1) with water to obtain a water extract, and collecting the supernatant after centrifugation;

[0063] (3) subjecting the supernatant obtained in step (2) to alcohol precipitation, and collecting the precipitate to obtain crude polysaccharide;

[0064] (4) The crude polysaccharide obtained in step (3) is obtained by removing the protein.

[0065] Wherein, in the step (1), the specific conditions of the ethanol reflux extraction are: using high concentration ethanol (such as 95% ethanol) to perform reflux extraction 1 to 5 times (preferably 3 times), each time for 1 to 5 hours (preferably 2 hours); further, before ethanol reflux extraction, the Yunnan knot wood is dried, such as drying at 40-60°C (preferably 50°C).

[0066] In the step (2), the specific conditions for boiling with water are: the material-liquid ratio is 1:1-10 (kg / L), preferably 1:5, the boiling treatment time is 1-3 hours, preferably 2 hours, and the number of times of boiling with water is 1-5 times (preferably 3 times).

[0067] In the step (3), the alcohol precipitation treatment method comprises: adding 95% ethanol to the supernatant, letting it stand and collecting the precipitate; the standing treatment time is 10-40 hours (preferably 24 hours).

[0068] In step (4), the protein removal can be performed by Sevage method, specifically, Sevage reagent (CH 3 Cl:BuOH=4:1, v / v) to remove protein 5 to 8 times.

[0069] Furthermore, the preparation method also includes freeze-drying the Dianjiexiang polysaccharide obtained in step (4).

[0070] In another specific embodiment of the present invention, a product is provided, wherein the product at least comprises the Dianjiexiang polysaccharide obtained by the above-mentioned preparation method.

[0071] The product has at least the following effects:

[0072] (a) Antioxidant;

[0073] (b) anti-inflammatory;

[0074] (c) liver protection;

[0075] (d) Regulation of intestinal flora.

[0076] The product may be food or medicine, which is not specifically limited here.

[0077] The food is understood in a broad sense in the present invention, and can be understood as any form that can be eaten. Therefore, the food in the present invention includes ordinary food and special food. The special food in the present invention includes health food and special medical formula food; and ordinary food is relative to special food, which is food suitable for everyone.

[0078] The drug can be administered in a unit dosage form, and the route of administration can be enteral or parenteral, such as oral, intramuscular, subcutaneous, nasal, oral mucosa, skin, peritoneum or rectum. The dosage form can be a liquid dosage form or a solid dosage form. For example, the liquid dosage form can be a true solution, a colloid, a microparticle dosage form, an emulsion dosage form, or a suspension dosage form. Other dosage forms such as tablets, capsules, dripping pills, aerosols, pills, powders, solutions, suspensions, emulsions, granules, suppositories, freeze-dried powder injections, etc. are not specifically limited here.

[0079] In the present invention, the medicine can be used for humans and non-human animals. The non-human animals can be non-human mammals, such as mice, rats, guinea pigs, pigs, monkeys, orangutans, etc., which are not specifically limited here.

[0080] In another specific embodiment of the present invention, a method for preventing and treating CCl is provided. 4 A method for treating induced liver damage and intestinal flora disorder, comprising administering the above-mentioned Dianjiexiang polysaccharide or the above-mentioned product to a subject.

[0081] The subject may be a human or a non-human animal, and the non-human animal is further a non-human mammal (such as mouse, rat, guinea pig, pig, monkey, orangutan, etc.), but humans are preferred.

[0082] The present invention is further explained by the following examples, but they are not intended to limit the present invention. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention.

[0083] Example 1: Preparation of EGM polysaccharide

[0084] EGM polysaccharide was prepared by the following steps: extracting the 50°C dried buds of Dian Jie Xiang with 4 times the mass of 95% ethanol (i.e., the mass ratio of Dian Jie Xiang buds to 95% ethanol was 1:4) at 65°C for 3 times, each time for 2 hours, to remove fat-soluble substances, and merging to obtain residues; adding water to the obtained residue at a solid-liquid ratio of 1:5 (kg / L), boiling for 2 hours, boiling for 3 times, extracting, merging to obtain water extracts, centrifuging at 5000rpm for 10min to remove insoluble components, and collecting supernatants; adding 95% ethanol to the supernatant, standing and collecting precipitates; standing for 24 hours, collecting the precipitates, i.e., crude polysaccharides, and using the Sevage method (using Sevage reagent (CH 3 Cl:BuOH=4:1,v / v) to remove protein 7 times) and then freeze-dried to obtain the product.

[0085] The sugar content of the EGM polysaccharide was determined to be 87.06% by the phenol-sulfuric acid method.

[0086] EGM polysaccharide was used in subsequent Examples 2-5.

[0087] Example 2: EGM polysaccharide treatment by CCl 4 Induced liver injury in mice

[0088] Specifically, in order to explore the liver protective effect of EGM polysaccharide, 45 4-week-old ICR mice (24.56±0.75g) were purchased and randomly divided into 3 groups. They were control group (IC), model group (IM), and treatment group (IT). The IT group was given EGM solution (50mg / kg) every day, and the IC group and IM group were given an equal amount of normal saline every day. The IT group and IM group were intraperitoneally injected with CCl every 3 days. 4 A mixture of 1:1 (v / v) (2 mL / kg) with olive oil was administered intraperitoneally to the IC group. On day 32, all mice were euthanized, and blood, liver samples, feces, and intestinal contents were collected.

[0089] Body weight and liver index were analyzed; liver tissue was collected and stored in 4% paraformaldehyde fixative for more than 48 hours. The fixed tissue was dehydrated in a gradient manner, embedded in paraffin, sliced ​​into 5 μm thick slices, stained with H&E and Sirius red, and the morphological changes of liver tissue were observed under a microscope ( Figure 1 The results of mouse body weight measurement showed that EGM polysaccharide slowed down the CCl 4 The trend of mouse body weight loss and mouse liver index after induction ( Figure 1 ab). H&E staining and Sirius red staining results showed that the hepatic cords in the IM group were irregular in many places, and some cells were lighter in color. A large number of red blood cells were seen in the central veins, with light plasma staining, balloon-like degeneration, and inflammatory cell infiltration. Vacuoles of varying sizes were seen in the hepatocytes, and there was obvious fatty degeneration. The hepatic cords in the IT group were irregularly arranged, and the cytoplasmic staining was darker than that in the IM group. Balloons and lipid droplets appeared in some areas, and the pathological damage was alleviated compared with the IM group.

[0090] Determination of liver function indexes: After blood collection, let stand at 4°C for 2 hours and then centrifuge (3500rpm, 15min) to collect the supernatant. Then, perform the test according to the instructions of the ALT and AST kits ( Figure 2 E, F). ELISA results showed that EGM polysaccharide could reduce the levels of ALT and AST ( Figure 2 AD).

[0091] Example 3: Anti-inflammatory effect of EGM polysaccharide in vivo

[0092] Experimental methods and animal models: Same as in Example 2.

[0093] Index determination: After blood collection, let stand for 2 hours at 4°C and then centrifuge (3500rpm, 15min) to collect the supernatant. Then, test according to the instructions of IL-6, IL-10, IL-1β, and TNF-α kits. ELISA results showed that EGM polysaccharide can increase the expression level of IL-10 and reduce the levels of TNF-α, IL-6, and IL-1β ( Figure 2 AD).

[0094] Example 4: Antioxidant effect of EGM polysaccharide in vivo

[0095] Experimental methods and animal models: Same as in Example 2.

[0096] Index determination: After blood collection, let stand at 4℃ for 2h and then centrifuge (3500rpm, 15min), collect the supernatant. Then, test according to the instructions of SOD, MDA, T-AOC and GSH-px kits. ELISA results showed that EGM polysaccharide can increase and decrease the levels of SOD, MDA, T-AOC and GSH-px ( Figure 2 GJ).

[0097] Example 5: EGM polysaccharides can improve CCl 4 Induced intestinal microbial disturbance in mice

[0098] Experimental methods and animal models: Same as in Example 2.

[0099] By sequencing 16S rRNA in mouse colonic feces, CCl 4 After 32 days of induction of liver injury, the rectums of mice in the IC, IM, and IT groups were harvested to study the microbial regulatory effects of EGM polysaccharides. Rectal microbial DNA was extracted according to the kit instructions, the V3-V4 region was amplified, the library was constructed, and sequencing was performed on the Illumina NovaSeq platform. QIIME2 was used to analyze the alpha diversity of intestinal microorganisms. The distinguishing species between different ICR animal groups were revealed by the analysis of LEfSe and DEseq2. Non-metric multidimensional scaling, partial least squares discriminant, and principal coordinates were used to analyze the microbiota structure. Microbiome evolution analysis was performed using ggtree in the R package. MetaCyc and enzyme databases were used to analyze the functional differences of the mouse microbiota for targeted KEGG analysis.

[0100] The sequencing results showed that a total of 481 ASVs were obtained in the three groups. There were 2361 ASVs in the IC group, 1559 ASVs in the IM group, and 1218 ASVs in the IT group. Among them, there were 102 ASVs in the three groups, 156 ASVs in the IC group and the IM group, and 131 ASVs in the IC group and the IT group ( Figure 3 A). Alpha diversity analysis was used to calculate the sample species richness and species diversity within each sample. β-diversity analysis was used to describe the microbial differences between groups. The scatter plots between groups showed that there were differences in the microbial composition among the three groups ( Figure 3 B).

[0101] Used(521.2±289.6985, 284.4±39.5576, 360.2±150.3669), ACEFeature(559.526±314.571, 325 .4394±55.1975, 417.0117±147.5698), Chao1 (557.4736±320.4449, 308.9692±47.4726, 407.204 9±151.4841), Simpson (0.9269±0.0384, 0.8427±0.0684), Chao1 (557.4736±320.444, 9308.969 2±47.4726, 407.2049±151.4841), Shannon (5.5523±0.9531, 4.0392±05308, 4.1187±0.2585), PD The whole tree (54.0727±37.4249, 48.7306±18.0363, 77.086±33.5954) and Coverage (99.38±99.97%, 99.72±99.90%, 99.57±99.83%) were used to describe the species abundance and diversity of groups IC, IT and IM. Figure 4 A). The results of the rarefaction curve and abundance level curve show that the curve tends to be flat, indicating that the sequencing depth is sufficient ( Figure 4 BC).

[0102] A detailed comparison of bacteria with high abundance among the three groups was conducted at the phylum and genus levels. At the phylum level, Firmicutes (IC = 63.08%, IT = 79.11%, IM = 85.66%), Bacteroidetes (IC = 12.53%, IT = 13.44%, IM = 7.18%), Actinobacteria (IC = 7.44%, IT = 2.66%, IM = 2.65%), and Proteobacteria (IC = 3.65%, IT = 3.97%, IM = 1.34%) were the bacteria with high abundance in the three groups (IC, IT, and IM). Figure 5 ).

[0103] In addition, Desulfobacterota also accounted for a certain proportion in the IC group (7.40%), while its abundance was lower in the IM group (0.13%) and the IT group (0.11%). At the genus level, the marker bacteria of the IC group, IT group and IM group were all Lactobacillus (IC = 11.0%, IT group = 41.87%, IM group = 41.88%). Faecalibaculum (10.27%) and Limosilactobacillus (11.95%) were abundant in the IT group, and Candidatus-Arthromitus (16.52%) and Limosilactobacillus (16.26%) were the dominant genera in the IM group. Other marker bacteria in the IC group included atopstipes (9.33%), Mammaliicoccus (9.18%) and Desulfovibrio (7.31%) ( Figure 6 ).

[0104] KEGG analysis of the mouse microbiota showed that it could increase nucleotide metabolism, replication and repair, and carbohydrate metabolism ( Figure 7 ).

[0105] It should be noted that the above examples are only used to illustrate the technical solution of the present invention rather than to limit it. Although the present invention is described in detail with reference to the given examples, those skilled in the art may modify or replace the technical solution of the present invention as needed without departing from the spirit and scope of the technical solution of the present invention.

Claims

1. Application of Yunnan Jiexiang polysaccharide in any one or more of the following: (a) Anti-oxidation or preparation of anti-oxidation products; (b) Anti-inflammatory or preparation of anti-inflammatory products; (c) liver protection or products for preparing liver protection; (d) Products for regulating intestinal flora or preparing intestinal flora conditions.

2. The use according to claim 1, characterized in that In the application (a), the anti-oxidation is specifically manifested as: reducing the levels of SOD, MDA, T-AOC and GSH-px.

3. The use according to claim 1, characterized in that In the application (b), the anti-inflammatory effect is specifically manifested as: increasing the expression level of IL-10 and reducing the expression levels of TNF-α, IL-6, and IL-1β.

4. The use according to claim 1, characterized in that In the application (c), liver protection is specifically manifested as: improving symptoms related to liver damage; further, the liver damage is induced by CCl4; Furthermore, the improvement of liver injury-related symptoms is specifically manifested in: inhibiting the subject's weight loss, improving liver tissue pathological damage caused by CCl4, and reducing ALT and AST expression levels.

5. The use according to claim 1, characterized in that In the application (d), the intestinal flora regulation is specifically manifested as: at the phylum level, the abundance of Firmicutes is reduced, and the abundance of Bacteroidetes, Actinobacteria and Proteobacteria is increased. At the genus level, Lactobacillus, Faecalibacterium prausnitzii and Lactobacillus reuteri are increased, and the number of Brevibacterium, Lactobacillus mucosus and Deinococcus is reduced.

6. A method for preparing Dianjiexiang polysaccharide, characterized in that: The preparation method comprises: (1) extracting the buds of the osmanthus fragrans with ethanol by refluxing to remove fat-soluble substances to obtain a residue; (2) boiling the residue obtained in step (1) with water to obtain a water extract, and collecting the supernatant after centrifugation; (3) subjecting the supernatant obtained in step (2) to alcohol precipitation, and collecting the precipitate to obtain crude polysaccharide; (4) The crude polysaccharide obtained in step (3) is obtained by removing the protein.

7. The preparation method according to claim 6, characterized in that: In the step (1), the specific conditions of the ethanol reflux extraction are: using high concentration ethanol for reflux extraction 1 to 5 times, each time for 1 to 5 hours; further, before the ethanol reflux extraction, the Yunnan knot wood is dried at 40-60°C; In the step (2), the specific conditions for boiling with water are: the material-liquid ratio is 1:1-10 (kg / L), preferably 1:5, the boiling treatment time is 1-3 hours, and the number of times of boiling with water is 1-5 times; In the step (3), the alcohol precipitation treatment method comprises: adding 95% ethanol to the supernatant, letting it stand and collecting the precipitate; the standing treatment time is 10-40 hours; In the step (4), the protein removal is performed using the Sevage method.

8. The preparation method according to claim 6, characterized in that: The preparation method further comprises freeze-drying the Dianjiexiang polysaccharide obtained in step (4).

9. A product, characterized in that The product at least comprises the Dianjiexiang polysaccharide obtained by the preparation method according to any one of claims 6 to 8.

10. The product according to claim 9, characterized in that The product has at least the following effects: (a) Antioxidant; (b) anti-inflammatory; (c) liver protection; (d) Regulation of intestinal flora; Wherein, the product is food or medicine.

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