Tupistra chinensis polysaccharide with antibacterial and anti-inflammatory effects as well as preparation method and application of tupistra chinensis polysaccharide

Abstract

The invention discloses tupistra chinensis polysaccharide with antibacterial and anti-inflammatory effects as well as a preparation method and application thereof, and belongs to the technical field of biotechnology and medicine. The preparation method comprises the following steps: (1) selecting tupistra chinensis rhizomes, removing impurities, drying, grinding into fine powder, wrapping the fine powder with gauze, putting into a Soxhlet extractor, continuously and circularly extracting until an extracting solution becomes colorless, collecting medicine residues, transferring the medicine residues into a flask, refluxing, and drying; (2) putting the dried medicine residues into a flask, carrying out reflux extraction in a constant-temperature water bath, merging filtrate, and concentrating under reduced pressure until the filtrate is thick, so as to obtain concentrated filtrate; and (3) carrying out alcohol precipitation treatment on the concentrated filtrate, carrying out suction filtration separation to obtain a precipitate, and washing, drying and grinding the precipitate to obtain the tupistra chinensis polysaccharide. The tupistra chinensis polysaccharide prepared by the invention can recover a damaged intestinal flora structure and increase the diversity of intestinal microorganisms, and is also beneficial to relieving body inflammation caused by LPS and escherichia coli treatment and relieving symptoms of inflammatory bowel diseases.

Description

Technical Field

[0001] The present invention belongs to the fields of biotechnology and medicine, and particularly relates to a Tupistra chinensis polysaccharide with antibacterial and anti-inflammatory effects, a preparation method thereof, and an application thereof. Background Art

[0002] The intestine is not only the core organ for human digestion and absorption, but also harbors a complex microbial community - the gut microbiota, which plays an important role in maintaining the body's immune balance, metabolic regulation, and overall health. However, intestinal inflammatory diseases (such as inflammatory bowel disease, colitis) and related diarrhea problems not only seriously affect the quality of human life, but also restrict the development of the livestock industry. Especially in the context of "reducing and banning antibiotics", finding safe and effective natural antibacterial and anti-inflammatory drugs has become an urgent need.

[0003] Tupistra chinensis Baker, a perennial herb of the Liliaceae family, has attracted attention due to its significant antibacterial, anti-inflammatory, antioxidant, and immunomodulatory effects. Traditional Chinese medicine commonly uses Tupistra chinensis to treat various diseases such as sore throat, stomachache, cancer, and snake bites, and it is recorded in important literature such as "Chinese Herbal Medicine Dictionary" and "Compendium of Chinese Herbal Medicines across the Country". Modern pharmacological research shows that Tupistra chinensis is rich in various bioactive components, including steroid compounds (such as spirosteroids, furosteroids), saponins, polysaccharides, and flavonoid compounds, which endow Tupistra chinensis with significant antibacterial, anti-inflammatory, antioxidant, and immunomodulatory effects.

[0004] In particular, the polysaccharide component in Tupistra chinensis can not only reduce inflammation by directly inhibiting the growth of pathogenic microorganisms or regulating the host immune response, but also act as a prebiotic to promote the proliferation of beneficial bacteria and help restore and maintain the balance of the gut microbiota. This dual mechanism of action makes Tupistra chinensis and its extracts have great potential in the development of drugs for intestinal inflammatory diseases and the improvement of gut health. Therefore, further studying the specific mechanism of action of Tupistra chinensis and exploring its application prospects in the fields of medicine and animal health are of great significance for improving human health levels. Summary of the Invention

[0005] In view of this, the present invention discloses a Tupistra chinensis polysaccharide with antibacterial and anti-inflammatory effects, a preparation method thereof, and an application thereof. The Tupistra chinensis polysaccharide prepared by the present invention can reduce the body inflammation caused by LPS (lipopolysaccharide) and pathogenic Escherichia coli treatment, relieve inflammatory bowel disease, and reduce the diarrhea caused by pathogenic Escherichia coli by restoring the structure of the damaged gut microbiota, increasing the gut microbiota diversity, and changing the metabolic pathway.

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

[0007] The present invention provides a preparation method of tupistra chinensis bak. polysaccharide with antibacterial and anti-inflammatory effects, comprising the following steps:

[0008] (1) Defatting and impurity removal of tupistra chinensis bak.: Select an appropriate amount of tupistra chinensis bak. rhizomes, sieve to remove impurities, dry the screened rhizomes and crush them into fine powder. Wrap the crushed rhizome fine powder with gauze and put it into a Soxhlet extractor. Use ethanol solution with a concentration of 95% as the solvent for continuous circulation extraction until the extract becomes colorless. Collect the medicinal residues defatted through the above process. Then transfer the defatted medicinal residues to a flask, reflux with ethanol solution with a concentration of 95% at 60 °C for 2 times, 1 h each time. After reflux, take out the medicinal residues and dry them.

[0009] (2) Extraction of tupistra chinensis bak. polysaccharide: Put the dried medicinal residues in step (1) into a flask, add distilled water, reflux and extract in a constant temperature water bath at 90 °C for 3 times, 2 h each time. Filter after each extraction, collect the filtrate, and combine all the filtrates. Use a vacuum concentrator to concentrate the combined filtrate to 1 / 10 of the original volume to form a concentrated filtrate.

[0010] (3) Preparation of tupistra chinensis bak. polysaccharide: Slowly add ethanol solution to the concentrated filtrate in step (2), while constantly stirring, for alcohol precipitation treatment. Use a suction filtration device to separate the precipitate and the supernatant. Wash the precipitate with absolute ethanol and dry it in a drying oven at 50 °C to constant weight. Crush the dried precipitate to obtain light brown particles, that is, obtain tupistra chinensis bak. polysaccharide.

[0011] Preferably, in step (1), the mass-volume ratio of the tupistra chinensis bak. rhizome fine powder to the ethanol solution with a concentration of 95% during the defatting process is 1 g:(2 - 5) mL; the mass-volume ratio of the defatted medicinal residues to the ethanol solution with a concentration of 95% during the reflux process is 1 g:(2 - 5) mL.

[0012] Preferably, the mass-volume ratio of the tupistra chinensis bak. rhizome fine powder to the ethanol solution with a concentration of 95% during the defatting process is 1 g:4 mL; the mass-volume ratio of the defatted medicinal residues to the ethanol solution with a concentration of 95% during the reflux process is 1 g:3 mL.

[0013] Preferably, in step (2), the mass-volume ratio of the dried medicinal residues to distilled water is 1 g:(3 - 10) mL.

[0014] Preferably, the mass-volume ratio of the dried medicinal residues to distilled water is 1 g:6 mL.

[0015] Preferably, in step (3), the concentration of the ethanol solution is 30 - 95%; the volume ratio of the concentrated filtrate to the ethanol solution is 1:(0.8 - 1.5).

[0016] Preferably, the specific steps of step (3) are as follows:

[0017] (ⅰ) 40% ethanol precipitation: Slowly add a 40% ethanol solution to the concentrated filtrate while continuously stirring. After standing for 12 - 24 h, a precipitate is obtained. Use a suction filtration device to separate the 40% ethanol precipitate and the supernatant, and wash with absolute ethanol.

[0018] (ⅱ) 60% ethanol precipitation: Retain the supernatant from step (ⅰ), slowly add a 60% ethanol solution to it while continuously stirring. After standing for 12 - 24 h, a precipitate is obtained. Use a suction filtration device to separate the 60% ethanol precipitate and the supernatant, and wash with absolute ethanol.

[0019] (ⅲ) 80% ethanol precipitation: Retain the supernatant from step (ⅱ), slowly add an 80% ethanol solution to it while continuously stirring and standing for 12 - 24 h to obtain a precipitate. Use a suction filtration device to separate the 60% ethanol precipitate and the supernatant, and wash with absolute ethanol.

[0020] (ⅳ) Place the precipitates of 40% ethanol, 60% ethanol, and 80% ethanol above in a drying oven at 50 °C and dry to a constant weight, then mix them evenly according to a mass ratio of 1:1:1. Then crush the evenly mixed precipitate to finally obtain the tupistra root polysaccharide.

[0021] The present invention also discloses the tupistra root polysaccharide prepared by the above preparation method.

[0022] The present invention also discloses the application of the above tupistra root polysaccharide in pharmaceutical products with antibacterial and anti-inflammatory effects.

[0023] Preferably, the pharmaceutical product with antibacterial and anti-inflammatory effects is a pharmaceutical product for preventing or treating diarrhea or inflammatory bowel disease.

[0024] Compared with the prior art, the present invention has the following beneficial effects:

[0025] (1) The tupistra root polysaccharide provided by the present invention can significantly improve the total antioxidant capacity (T-AOC), superoxide dismutase (SOD) activity, and glutathione peroxidase (GSH-Px) level in animals, while reducing the content of malondialdehyde (MDA), which helps to enhance the body's ability to resist oxidative stress and protect cells from free radical damage; and by regulating the levels of inflammatory factors, including increasing the concentration of anti-inflammatory factors such as interleukin 10 (IL-10), and controlling the production of pro-inflammatory factors such as interleukin 1β (IL-1β), interleukin 6 (IL-6), and tumor necrosis factor α (TNF-α), it effectively reduces the body's inflammatory response caused by LPS (lipopolysaccharide) and Escherichia coli, helps to reduce the impact of inflammatory bowel disease on the body, and relieve symptoms.

[0026] (2) The tupistra polysaccharide provided by the present invention can regulate the intestinal microbial community structure and effectively reverse the intestinal flora changes caused by pathogenic Escherichia coli. For example, it can reduce the abnormal increase of genera such as Alloprevotella, UBA7173, UBA3263, and Muribaculum, and restore the abundances of various beneficial genera such as UBA3282, Desulfovibrio_R_446353, and Lawsonibacter. At the same time, the tupistra polysaccharide helps to increase or maintain the abundances of butyrate-producing bacteria (such as Lawsonibacter, Eubacterium_J, and Dysosmobacter), and these bacteria support the health of intestinal epithelial cells by producing butyrate, promote intestinal villus proliferation, and enhance nutrient absorption.

[0027] The tupistra polysaccharide provided by the present invention can restore the damaged intestinal flora structure, increase intestinal microbial diversity, and maintain the intestinal barrier function by changing specific metabolic pathways. Moreover, it also helps to reduce the body inflammation caused by LPS and Escherichia coli treatment and further relieve the symptoms of inflammatory bowel disease. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 is a flow chart for the preparation and efficacy verification of the tupistra polysaccharide provided by the present invention;

[0029] Figure 2 is a microscopic examination stained image of the effects of the tupistra polysaccharide provided by the present invention on the intestinal and spleen tissue morphology of mice with LPS-induced intestinal inflammation (groups C, M, and P are the control group, model group, and test group, respectively);

[0030] Figure 3 is a graph showing the effects of the tupistra polysaccharide provided by the present invention on serum interleukin and antioxidant enzymes in mice with LPS-induced intestinal inflammation (groups C, M, and P are the control group, model group, and test group, respectively);

[0031] Figure 4 is a graph showing the effects of the tupistra polysaccharide provided by the present invention on the intestinal flora structure of mice with LPS-induced intestinal inflammation (groups C, M, and P are the control group, model group, and test group, respectively);

[0032] Figure 5 is a graph showing the possible metabolic pathways of the intestinal flora in mice with LPS-induced intestinal inflammation affected by the tupistra polysaccharide provided by the present invention (groups C, M, and P are the control group, model group, and test group, respectively);

[0033] Figure 6Microscopic examination staining diagrams of the effect of the Tupistra chinensis Bak. polysaccharide provided by the present invention on the intestinal epithelial tissue morphology of mice with intestinal inflammation caused by pathogenic Escherichia coli (the CK, MK, and PK groups are the control group, model group, and experimental group, respectively);

[0034] Figure 7 Diagrams of the effect of the Tupistra chinensis Bak. polysaccharide provided by the present invention on interleukin and antioxidant enzymes in the serum of mice with intestinal inflammation caused by pathogenic Escherichia coli (the CK, MK, and PK groups are the control group, model group, and experimental group, respectively);

[0035] Figure 8 Diagrams of the effect of the Tupistra chinensis Bak. polysaccharide provided by the present invention on the intestinal flora structure of mice with intestinal inflammation caused by pathogenic Escherichia coli (the CK, MK, and PK groups are the control group, model group, and experimental group, respectively);

[0036] Figure 9 Diagrams of the effect of the Tupistra chinensis Bak. polysaccharide provided by the present invention on the possible metabolic pathways of the intestinal flora of mice with intestinal inflammation caused by pathogenic Escherichia coli (the CK, MK, and PK groups are the control group, model group, and experimental group, respectively). Detailed implementation manners

[0037] Next, the technical solutions in the embodiments of the present invention will be clearly and completely described in conjunction with the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present invention.

[0038] The experimental methods used in the following embodiments are all conventional methods unless otherwise specified.

[0039] The materials, reagents, etc. used in the following embodiments can be obtained from commercial channels unless otherwise specified.

[0040] The present invention provides a preparation method of a Tupistra chinensis Bak. polysaccharide with antibacterial and anti-inflammatory effects, including the following steps:

[0041] (1) Defatting and impurity removal of Tupistra chinensis Bak.: Select an appropriate amount of the rhizomes of Tupistra chinensis Bak., sieve to remove impurities, dry the screened rhizomes and crush them into fine powder. Wrap the crushed rhizome fine powder with gauze and put it into a Soxhlet extractor. Use an ethanol solution with a concentration of 95% as the solvent for continuous cyclic extraction (the mass-to-volume ratio of the rhizome fine powder of Tupistra chinensis Bak. to the ethanol solution with a concentration of 95% is 1 g:(2 - 5) mL)), until the extract becomes colorless. Collect the residue after defatting through the above process; then transfer the defatted residue to a flask, reflux twice with an ethanol solution with a concentration of 95% at 60 °C (the mass-to-volume ratio of the defatted residue to the ethanol solution with a concentration of 95% is 1 g:(2 - 5) mL), 1 h each time). After the reflux is completed, take out the residue and dry it.

[0042] (2) Extraction of Tupistra chinensis Bak. polysaccharide: Put the residue dried in step (1) into a flask. According to the mass-to-volume ratio of the dried residue to distilled water of 1 g:(3 - 10) mL, add distilled water, reflux and extract 3 times in a constant temperature water bath at 90 °C, 2 h each time. Filter after each extraction, collect the filtrate, and combine all the filtrates; use a vacuum concentrator to concentrate the combined filtrate to 1 / 10 of the original volume to form a concentrated filtrate.

[0043] (3) Preparation of Tupistra chinensis Bak. polysaccharide: Slowly add an ethanol solution with a concentration of 30 - 95% (the volume ratio of the concentrated filtrate to the ethanol solution is 1:(0.8 - 1.5)) to the concentrated filtrate in step (2), and continuously stir at the same time for alcohol precipitation treatment. Use a suction filtration device to separate the precipitate and the supernatant; wash the precipitate with absolute ethanol and place it in a drying oven at 50 °C to dry to constant weight. Crush the dried precipitate to obtain light brown particles, that is, obtain Tupistra chinensis Bak. polysaccharide.

[0044] In the above step (1), the most preferred mass-to-volume ratio of the rhizome fine powder of Tupistra chinensis Bak. to the ethanol solution with a concentration of 95% during the defatting process is 1 g:4 mL; the most preferred mass-to-volume ratio of the defatted residue to the ethanol solution with a concentration of 95% during the reflux process is 1 g:3 mL.

[0045] In the above step (2), the most preferred mass-to-volume ratio of the dried residue to distilled water is 1 g:6 mL.

[0046] In the above step (3), it is most preferred to perform precipitation treatment on the concentrated filtrate successively with ethanol solutions with concentrations of 40%, 60%, and 80%, and dry the three precipitates (40%, 60%, and 80% ethanol precipitates) to constant weight in a drying oven at 50 °C and then mix them in a ratio of 1:1:1. It includes the following steps:

[0047] (i) 40% ethanol precipitation: Slowly add an ethanol solution with a concentration of 40% to the concentrated filtrate while continuously stirring. After standing for 12 - 24 h, a precipitate is obtained. Use a suction filtration device to separate the 40% ethanol precipitate and the supernatant, and wash with absolute ethanol;

[0048] (ii) 60% ethanol precipitation: Retain the supernatant from step (i), slowly add an ethanol solution with a concentration of 60% to it while continuously stirring. After standing for 12 - 24 h, a precipitate is obtained. Use a suction filtration device to separate the 60% ethanol precipitate and the supernatant, and wash with absolute ethanol;

[0049] (iii) 80% ethanol precipitation: Retain the supernatant from step (ii), slowly add an ethanol solution with a concentration of 80% to it while continuously stirring and standing for 12 - 24 h. Then a precipitate is obtained. Use a suction filtration device to separate the 60% ethanol precipitate and the supernatant, and wash with absolute ethanol;

[0050] (iv) Place the precipitates of 40% ethanol, 60% ethanol, and 80% ethanol obtained above in a drying oven at 50 °C and dry to a constant weight. Then mix them evenly according to a mass ratio of 1:1:1. Next, crush the evenly mixed precipitate, and finally obtain the Tupistra chinensis Bak. polysaccharide.

[0051] The present invention also discloses the Tupistra chinensis Bak. polysaccharide prepared by the above preparation method.

[0052] The present invention also discloses the application of the above Tupistra chinensis Bak. polysaccharide in pharmaceutical products with antibacterial and anti - inflammatory effects.

[0053] Among them, the above - mentioned pharmaceutical products with antibacterial and anti - inflammatory effects can be pharmaceutical products for preventing or treating diarrhea or inflammatory bowel disease.

[0054] The following further illustrates the technical solution of the present invention in conjunction with the accompanying drawings and specific embodiments (where: Figure 1 is the flow chart of the preparation and efficacy verification of the Tupistra chinensis Bak. polysaccharide provided by the present invention):

[0055] Example 1

[0056] The preparation method of the Tupistra chinensis Bak. polysaccharide with antibacterial and anti - inflammatory effects in this example is as follows:

[0057] (1) Defatting and impurity removal of Tupistra chinensis Bak.: Select an appropriate amount of the rhizomes of Tupistra chinensis Bak., sieve to remove impurities, dry the screened rhizomes and crush them into fine powder. Wrap the crushed rhizome fine powder with gauze and put it into a Soxhlet extractor. Use 95% ethanol solution as the solvent for continuous cyclic extraction according to the mass-to-volume ratio of the rhizome fine powder of Tupistra chinensis Bak. to 95% ethanol solution of 1 g:4 mL until the extract becomes colorless. Collect the medicinal residues defatted through the above process. Then transfer the defatted medicinal residues to a flask, and use 95% ethanol solution for reflux extraction 2 times at 60 °C according to the mass-to-volume ratio of the defatted medicinal residues to 95% ethanol solution of 1 g:3 mL, 1 h each time. After the reflux is completed, take out the medicinal residues and dry them.

[0058] (2) Extraction of polysaccharides from Tupistra chinensis Bak.: Put the dried medicinal residues in step (1) into a flask, add distilled water according to the mass-to-volume ratio of the dried medicinal residues to distilled water of 1 g:6 mL, reflux and extract in a constant temperature water bath at 90 °C for 3 times, 2 h each time. Filter after each extraction, collect the filtrates, and combine all the filtrates. Use a vacuum concentrator to concentrate the combined filtrates to 1 / 10 of the original volume to form a concentrated filtrate.

[0059] (3) Preparation of polysaccharides from Tupistra chinensis Bak.: (i) 40% ethanol precipitation: Slowly add a 40% ethanol solution to the concentrated filtrate in step (2) while constantly stirring. Let it stand for 12 - 24 h to obtain a precipitate. Use a suction filtration device to separate the 40% ethanol precipitate and the supernatant, and wash with absolute ethanol; (ii) 60% ethanol precipitation: Retain the supernatant in step (i), slowly add a 60% ethanol solution to it while constantly stirring. Let it stand for 12 - 24 h to obtain a precipitate. Use a suction filtration device to separate the 60% ethanol precipitate and the supernatant, and wash with absolute ethanol; (iii) 80% ethanol precipitation: Retain the supernatant in step (ii), slowly add an 80% ethanol solution to it while constantly stirring and let it stand for 12 - 24 h to obtain a precipitate. Use a suction filtration device to separate the 60% ethanol precipitate and the supernatant, and wash with absolute ethanol; (iv) Place the precipitates of 40% ethanol, 60% ethanol, and 80% ethanol above in a drying oven at 50 °C and dry to constant weight, then mix them evenly according to the mass ratio of 1:1:1, and then crush the evenly mixed precipitate, that is, obtain the polysaccharides of Tupistra chinensis Bak.

[0060] Example 2

[0061] The preparation method of polysaccharides from Tupistra chinensis Bak. with antibacterial and anti-inflammatory effects in this example is as follows:

[0062] (1) Defatting and impurity removal of Tupistra chinensis Bak.: Select an appropriate amount of the rhizomes of Tupistra chinensis Bak., sieve and remove impurities. Dry the screened rhizomes and crush them into fine powder. Wrap the crushed rhizome fine powder with gauze and put it into a Soxhlet extractor. According to the mass - volume ratio of the rhizome fine powder of Tupistra chinensis Bak. to the 95% ethanol solution of 1 g:2 mL, use the 95% ethanol solution as the solvent for continuous cyclic extraction until the extract becomes colorless. Collect the medicinal residues defatted through the above process. Then transfer the defatted medicinal residues to a flask. According to the mass - volume ratio of the defatted medicinal residues to the 95% ethanol solution of 1 g:2 mL, under the condition of 60 °C, use 95% ethanol for reflux 2 times, 1 h each time. After the reflux is completed, take out the medicinal residues and dry them.

[0063] (2) Extraction of Tupistra chinensis Bak. polysaccharide: Put the dried medicinal residues in step (1) into a flask. According to the mass - volume ratio of the dried medicinal residues to distilled water of 1 g:3 mL, add distilled water, and reflux and extract in a 90 °C constant - temperature water bath 3 times, 2 h each time. Filter after each extraction, collect the filtrate, and combine all the filtrates. Use a vacuum concentrator to concentrate the combined filtrate to 1 / 10 of the original volume to form a concentrated filtrate.

[0064] (3) Preparation of Tupistra chinensis Bak. polysaccharide: Slowly add the 95% ethanol solution to the concentrated filtrate in step (2), and the volume ratio of the concentrated filtrate to the 95% ethanol solution is 1:1. Stir continuously at the same time for alcohol precipitation treatment. Use a suction filtration device to separate the precipitate and the supernatant. Wash the precipitate with absolute ethanol and place it in a drying oven at 50 °C to dry to constant weight. Crush the dried precipitate to obtain light - brown particles, that is, obtain Tupistra chinensis Bak. polysaccharide.

[0065] Example 3

[0066] The preparation method of Tupistra chinensis Bak. polysaccharide with antibacterial and anti - inflammatory effects in this example is as follows:

[0067] (1) Defatting and impurity removal of Tupistra chinensis Bak.: Select an appropriate amount of the rhizomes of Tupistra chinensis Bak., sieve and remove impurities. Dry the screened rhizomes and crush them into fine powder. Wrap the crushed rhizome fine powder with gauze and put it into a Soxhlet extractor. According to the mass - volume ratio of the rhizome fine powder of Tupistra chinensis Bak. to the 95% ethanol solution of 1 g:5 mL, use the 95% ethanol solution as the solvent for continuous cyclic extraction until the extract becomes colorless. Collect the medicinal residues defatted through the above process. Then transfer the defatted medicinal residues to a flask. According to the mass - volume ratio of the defatted medicinal residues to the 95% ethanol solution of 1 g:5 mL, under the condition of 60 °C, use 95% ethanol for reflux 2 times, 1 h each time. After the reflux is completed, take out the medicinal residues and dry them.

[0068] (2) Extraction of Tupistra chinensis Bak. polysaccharide: Put the dried medicinal residues in step (1) into a flask, add distilled water according to the mass-volume ratio of the dried medicinal residues to distilled water of 1 g: 10 mL, reflux and extract 3 times in a constant temperature water bath at 90 °C for 2 h each time. After each extraction, filter, collect the filtrate, and combine all the filtrates; use a vacuum concentrator to concentrate the combined filtrate to 1 / 10 of the original volume to form a concentrated filtrate.

[0069] (3) Preparation of Tupistra chinensis Bak. polysaccharide: Slowly add a 95% ethanol solution to the concentrated filtrate in step (2), and the volume ratio of the concentrated filtrate to the 95% ethanol solution is 1: 1.5. At the same time, continuously stir for alcohol precipitation treatment. Use a suction filtration device to separate the precipitate and the supernatant; wash the precipitate with absolute ethanol and place it in a drying oven at 50 °C to dry to constant weight. Crush the dried precipitate to obtain light brown particles, that is, obtain Tupistra chinensis Bak. polysaccharide.

[0070] Example 4

[0071] This example is a protection test of the Tupistra chinensis Bak. polysaccharide prepared in Example 1 above against intestinal inflammation.

[0072] In this example, mice were used as experimental animals and the experiment was carried out under standard hygienic conditions. Thirty mice (body weight 15 - 18 g) were randomly divided into 3 groups of 10 each, namely the control group (C), the model group (M), and the experimental group (P). The mice in the experimental group (P) were intragastrically administered the Tupistra chinensis Bak. polysaccharide prepared in Example 1 every day, with a dose of 200 mg / kg·bw; the mice in the control group (C) and the model group (M) were intragastrically administered an equal volume of normal saline; on the 14th day after the start of the experiment, the mice in the above model group (M) and experimental group (P) were intraperitoneally injected with a 20 mg / kg·bw lipopolysaccharide (LPS) solution, and the mice in the control group (C) were intragastrically administered an equal volume of normal saline. All the mice were sacrificed 12 h after modeling and samples were collected for subsequent analysis, as follows.

[0073] (1) Effects of Tupistra chinensis Bak. polysaccharide on the tissue morphology of the jejunum, ileum, and spleen of LPS (lipopolysaccharide)-mediated intestinal inflammation mice.

[0074] Take about 1 cm of the jejunum, ileum, and spleen from the mouse samples of each group above 3Tissue samples of appropriate size were fixed using 4% paraformaldehyde solution. These tissues were subjected to a conventional dehydration process, embedded in paraffin, and cut into sections approximately 3 μm thick. The sections were then dewaxed with xylene and dehydrated step by step with ethanol, followed by H&E staining (hematoxylin-eosin staining). After staining, dehydration was carried out step by step with ethanol again, and then cleared with xylene. Finally, the sections were sealed with neutral balsam. The prepared sections were observed under an optical microscope to record the pathological changes in the jejunum, ileum, and spleen tissues.

[0075] As Figure 2 shown, it can be seen from the figure that: the control group (C) showed normal intestinal tissue structure, with tightly and regularly arranged intestinal villi, intact epithelial cells, and no obvious inflammatory cell infiltration; and the spleen structure was normal, with a clear white pulp area and normal red blood cell content in the red pulp. In the model group (M), obvious pathological changes occurred after LPS treatment: intestinal epithelial cells showed exfoliation, intestinal villi ruptured and shortened, and there was infiltration of inflammatory cells; at the same time, in the spleen, it was observed that the white pulp area shrank and the red blood cell content in the red pulp also decreased significantly; this indicates that LPS not only caused damage to the intestinal tissue structure and intestinal barrier function, but also significantly reduced the lymphocytes and red blood cells in the spleen, resulting in impaired immune function. In the experimental group (P), after treatment with the tupistra root polysaccharide of Example 1, obvious improvement effects could be seen: the intestinal morphology became regular and orderly, and the intestinal villi were closely packed, indicating an effective alleviation of intestinal inflammation; at the same time, the spleen structure also returned to normal, which shows that the tupistra root polysaccharide not only reduced the intestinal inflammatory response induced by LPS, but also reversed the abnormal changes in the spleen caused by LPS. This indicates that the tupistra root polysaccharide prepared by the present invention has the ability to protect the intestinal barrier, reduce the inflammatory response, and help restore the immune function damaged by LPS stimulation.

[0076] (2) Effects of tupistra root polysaccharide on serum interleukin and antioxidant enzymes in LPS-mediated intestinal inflammation mice.

[0077] 1 mL of blood was collected from each mouse in the above groups by eye socket blood collection. After standing at 4°C for 2 h, the supernatant was aspirated after centrifugation at 3000 rpm for 10 min. According to the method described in the ELISA kit instructions, the contents of interleukin 6, interleukin 10, interleukin 1β, tumor necrosis factor α (TNF-α), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), malondialdehyde (MDA), and total antioxidant capacity (T-AOC) in the serum were detected.

[0078] As Figure 3As shown in the figure, it can be seen that: in the model group (M), after treatment with LPS, the antioxidant enzymes (SOD, GSH-Px) and total antioxidant capacity (T-AOC) in the serum of mice were significantly decreased, while the level of malondialdehyde (MDA) increased, indicating that the antioxidant capacity of the body decreased after LPS treatment; the levels of inflammatory factors (IL-6, IL-10, IL-1β, TNF-α) increased, indicating that LPS induced an inflammatory response in the mouse body. After treatment with the tupistra root polysaccharide of the present invention in the experimental group (P), the antioxidant enzymes (SOD, GSH-Px) and total antioxidant capacity (T-AOC) in the serum of mice increased significantly, and the level of malondialdehyde (MDA) decreased, indicating that it can enhance the antioxidant capacity of the body; at the same time, the levels of inflammatory factors (IL-6, IL-10, IL-1β, TNF-α) decreased, indicating that the tupistra root polysaccharide has a certain protective effect on the intestinal inflammation induced by LPS. This shows that the tupistra root polysaccharide of the present invention shows significant effects in enhancing the antioxidant capacity of the body and reducing the inflammatory response, and can effectively counteract the damage caused by LPS.

[0079] (3) Effect of tupistra root polysaccharide on the intestinal flora structure of mice with LPS-mediated intestinal inflammation.

[0080] Take 0.2 grams of intestinal contents from the above-mentioned groups of mice as samples. After extracting total DNA, use specific primers to perform PCR amplification on the V3-V4 region of the bacterial 16S rRNA gene. The obtained products are used to construct a library with a kit for sequencing, and the relative abundances and changes of dominant strains at different taxonomic levels are analyzed to evaluate the effect of tupistra root polysaccharide on the intestinal microbial community structure.

[0081] As Figure 4 shown in the figure, it can be seen that: (i) at the phylum level, the abundance of Bacteroidota was the highest in the control group (C), and it decreased significantly after treatment with LPS in the model group (M), indicating that the balance state in the intestine may be disrupted after LPS treatment, resulting in a decrease in beneficial bacteria; while after treatment with the tupistra root polysaccharide of the present invention in the experimental group (P), the abundance of Bacteroidota increased slightly, indicating that the tupistra root polysaccharide helps to restore the normal level of this phylum. At the same time, the abundances of two subgroups of Firmicutes_D and Firmicutes-A were relatively high in the control group (C), and the abundances of these two subgroups decreased significantly in the model group (M) due to LPS treatment; after treatment with tupistra root polysaccharide in the experimental group (P), the abundances of Firmicutes_D and Firmicutes_A recovered; further illustrating the role of the tupistra root polysaccharide of the present invention in restoring the intestinal microbial community structure.

[0082] (ⅱ) At the genus level: After the model group (M) was treated with LPS, the abundances of the harmful genera Paramuribaculum, Duncaniella, UBA7173, CAG-269, Tidjanibacter, Eubacterium_F, and UBA9715 increased significantly. Among them, the increase in the abundance of Duncaniella was related to insulin resistance and colonic inflammatory responses. Eubacterium might promote metabolic disorders by regulating adipose tissue macrophage polarization. The increase in the abundance of Paramuribaculum was significantly correlated with the increase in pro-inflammatory cytokine levels, indicating that after the model group (M) was treated with LPS, it led to intestinal microbiota imbalance, and the increase in harmful genera exacerbated the inflammatory response. Moreover, after the model group (M) was treated with LPS, it also led to a significant decrease in the abundances of the beneficial genera Butyricimonas and Parabacteroides_B_86206. Among them, Butyricimonas is a butyrate-producing bacterium that is crucial for maintaining intestinal health, and the genus Parabacteroides can regulate bile acid metabolism, protect the intestinal barrier, and inhibit the growth of pathogenic bacteria. After the test group (P) was treated with Tupistra chinensis Bak. polysaccharide, the abundances of the harmful genera Paramuribaculum, Duncaniella, UBA7173, CAG-269, Tidjanibacter, Eubacterium_F, and UBA9715 decreased significantly and even approached the normal level; the abundances of the beneficial genera Butyricimonas and Parabacteroides_B_86206 recovered somewhat, indicating that Tupistra chinensis Bak. polysaccharide helped to restore the intestinal microbiota structure and reduce the inflammatory response. It can also be seen from the figure that after the model group (M) was treated with LPS, the abundance of Limosilactobacillus increased significantly, which might be a response mechanism of the body to the inflammatory response induced by LPS; after the test group (P) was treated with Tupistra chinensis Bak. polysaccharide, the abundance of Limosilactobacillus decreased and approached the normal level, so that Limosilactobacillus no longer needed to exist at a high abundance to combat inflammation or repair damage.

[0083] This shows that treatment with LPS led to severe imbalance of the intestinal microbiota in mice, increased the abundance of potentially harmful bacteria, and reduced the number of beneficial bacteria, thus exacerbating intestinal inflammation; in contrast, treatment with Tupistra chinensis Bak. polysaccharide of the present invention could partially reverse these negative effects. By regulating the butyrate-producing bacteria, bile acid-FXR signaling pathway, etc. in the intestine, it promoted the growth of beneficial bacteria and inhibited the proliferation of harmful bacteria, thereby restoring the normal structure of the intestinal microbiota and showing the effect of reducing intestinal inflammation.

[0084] (4) Effects of Tupistra chinensis Bak. polysaccharide on the metabolic pathways of intestinal flora in LPS-mediated intestinal inflammation mice.

[0085] The possible effects of changes in intestinal flora on metabolism were compared and analyzed through the MetaCyc and KEGG databases. From Figure 5It can be seen that in the control group (C), significant differences were shown in metabolic pathways such as 5-aminoimidazole ribonucleotide biosynthesis II (PWY-6122), 5-aminopyrimidine ribonucleotide biosynthesis (PWY-6277), L-lysine biosynthesis VI (PWY-5097), UDP-N-acetyl-D-glucosamine biosynthesis I (UDPNAGSYN-PWY), D-galactose degradation I (Leloir pathway) (PWY-6317), etc. These pathways are mainly involved in the biosynthesis of nucleotides and amino acids and the metabolism of carbohydrates. At the same time, significant differences were also shown in specific metabolic pathways such as secondary bile acid biosynthesis, lysine biosynthesis, primary bile acid biosynthesis, Staphylococcus aureus infection, etc., indicating that under normal conditions, these metabolic pathways are crucial for maintaining the balance of the gut microbial community and host health. In the model group (M), significant differences were found in metabolic pathways such as uroporphyrinogen III II metabolism (from glycine) (PWY5189), L-arginine degradation II (AST pathway) (AST-PWY), bacterial hexitol degradation pathway (HEXITOLDEGSUPER-PWY), etc. These pathways are mainly involved in amino acid metabolism and carbohydrate degradation. At the same time, the metabolic pathway of lysine degradation showed a relatively high LDA score in the model group (M), indicating that lysine degradation was significantly activated after LPS treatment. This phenomenon may be due to the impact of the inflammatory response induced by LPS on the metabolic activities of gut microbes, leading to abnormal activation of certain amino acid metabolic pathways and thus affecting the overall metabolic balance. After treatment with Tupistra chinensis Bak. polysaccharide in the experimental group (P), a similar trend to that in the control group (C) was shown in metabolic pathways such as histidine metabolism and peroxisome, indicating that Tupistra chinensis Bak. polysaccharide may restore the gut microbial community structure and its function damaged by LPS by regulating specific metabolic pathways. Tupistra chinensis Bak. polysaccharide can help restore the gut microbiota imbalance caused by LPS and then promote gut health and reduce the inflammatory response by regulating multiple metabolic pathways, including nucleotide synthesis, amino acid synthesis, carbohydrate metabolism, and bile acid metabolism.

[0086] Example 5

[0087] This example is a protection test of the tupistra chinensis bak. polysaccharide prepared in Example 1 above against pathogenic Escherichia coli. In this example, mice were used as experimental animals, treated with the product prepared in Example 1 above, and then an intestinal inflammation injury model was prepared by orally administering pathogenic Escherichia coli to verify the protective effect of the present invention on intestinal inflammation caused by pathogenic Escherichia coli.

[0088] In this example, mice were used as experimental animals and the experiment was carried out under standard sanitary conditions. Thirty mice (body weight 15 - 18 g) were selected and randomly divided into 3 groups of 10 each, namely the control group (CK), the model group (MK), and the test group (PK). During the experiment, the mice in the test group (PK) were intragastrically administered the tupistra chinensis bak. polysaccharide solution prepared in Example 1 every day, with a dose of 200 mg / kg·bw; the mice in the control group (CK) and the model group (MK) were intragastrically administered an equal volume of normal saline; starting from the 14th day after the start of the experiment, the mice in the above-mentioned model group (MK) and test group (PK) were intragastrically administered 8×10 9 CFU of pathogenic Escherichia coli (dissolved in 0.2 mL of PBS solution), and the mice in the control group (CK) were intragastrically administered 0.2 mL of PBS (phosphate buffer solution). All mice were sacrificed 24 h after modeling and samples were collected for subsequent analysis, as follows.

[0089] (1) Effects of tupistra chinensis bak. polysaccharide on the morphology of intestinal epithelial tissue in mice with Escherichia coli-mediated intestinal inflammation.

[0090] Approximately 1 cm 3 tissue samples of the same size were taken from the jejunum of each group of mice in this example and fixed with 4% paraformaldehyde solution. The samples were dehydrated according to the standard procedure and embedded in paraffin. The processed samples were cut into sections approximately 3 μm thick, dewaxed with xylene and dehydrated step by step with ethanol. Next, H&E staining (i.e., hematoxylin and eosin staining) was performed. After staining, dehydration was carried out step by step with ethanol again, and after transparency treatment with xylene, mounting was performed with neutral gum. The pathological changes of intestinal epithelial tissue were observed and recorded under an electron microscope.

[0091] As Figure 6 shown, it can be seen from the figure that: after treatment with pathogenic Escherichia coli, the intestinal wall of the model group (MK) was significantly thinned, and obvious atrophy, deformation and even rupture of intestinal villi occurred, accompanied by infiltration of inflammatory cells, indicating that pathogenic Escherichia coli caused obvious damage to the intestinal wall structure. In the mice pretreated with the tupistra chinensis bak. polysaccharide of the present invention in the test group (PK), it can be seen that the length of intestinal villi was restored, the arrangement of villi was more orderly, and at the same time, the infiltration of inflammatory cells was significantly reduced, indicating that tupistra chinensis bak. polysaccharide helps to reduce the intestinal wall inflammatory response and tissue damage caused by pathogenic Escherichia coli, thereby protecting intestinal health.

[0092] (2) Effects of Tupistra chinensis Bak. polysaccharide on serum interleukin and antioxidant enzymes in mice with Escherichia coli-mediated intestinal inflammation.

[0093] Collect 1 mL of blood from each mouse in the above groups by eye socket blood collection. After standing at 4 °C for 2 h, centrifuge at 3000 rpm for 10 min and then aspirate the supernatant. According to the method in the ELISA kit instruction manual, detect the contents of interleukin 6 (IL-6), interleukin 10 (IL-10), interleukin 1β (IL-1β), tumor necrosis factor α (TNF-α), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), malondialdehyde (MDA), and total antioxidant capacity (T-AOC) in the serum.

[0094] As Figure 7 shown, it can be seen from the figure that: in the model group (MK), treatment with Escherichia coli significantly reduced the antioxidant levels in the serum of mice, including superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and total antioxidant capacity (T-AOC); at the same time, the levels of malondialdehyde (MDA) and inflammatory factors such as interleukin 6, interleukin 10, interleukin 1β, and tumor necrosis factor α (TNF-α) increased; indicating that pathogenic Escherichia coli caused an increase in oxidative stress and exacerbation of the inflammatory response in vivo. In the mice pretreated with Tupistra chinensis Bak. polysaccharide of the present invention in the experimental group (PK), it was observed that the antioxidant indexes (SOD, GSH-Px, T-AOC) in the serum of mice tended to return to normal levels, indicating that its antioxidant capacity was enhanced; at the same time, the level of malondialdehyde (MDA) showed a downward trend, further confirming the effectiveness of Tupistra chinensis Bak. polysaccharide in reducing oxidative damage; and, the levels of factors related to inflammation (including IL-6, IL-10, IL-1β, TNF-α) also showed a downward trend after treatment with Tupistra chinensis Bak. polysaccharide; this shows that Tupistra chinensis Bak. polysaccharide helps to alleviate the inflammatory response caused by pathogenic Escherichia coli. Thus, it shows that Tupistra chinensis Bak. polysaccharide can not only improve the antioxidant status in mice, but also effectively inhibit the inflammatory response, thereby protecting the body from damage caused by pathogenic Escherichia coli.

[0095] (3) Effects of Tupistra chinensis Bak. polysaccharide on the intestinal flora structure of mice with Escherichia coli-mediated intestinal inflammation.

[0096] Take 0.2 g of intestinal contents from the above groups of mice as samples. After extracting total DNA, use specific primers to perform PCR amplification on the V3-V4 region of the bacterial 16S rRNA gene. The obtained products are used to construct a library with a kit and then used for sequencing, and analyze the relative abundances and changes of dominant strains at different taxonomic levels to evaluate the effects of Tupistra chinensis Bak. polysaccharide on the intestinal microbial community structure.

[0097] As Figure 8 shown, compared with the control group (CK), the abundances of genera such as Alloprevotella (p < 0.05), UBA7173 (p < 0.05), UBA3263 (p < 0.05), Muribaculum (p < 0.05), and Eubacterium_R (p < 0.05) in the model group (MK) were significantly increased after treatment with pathogenic Escherichia coli. In the experimental group (PK), after pretreatment with the Tupistra chinensis Bak. polysaccharide of the present invention, the abundances of these genera were decreased compared with the model group (MK), indicating that the Tupistra chinensis Bak. polysaccharide helps to restore the normal intestinal microbial community structure. In addition, pathogenic Escherichia coli in the model group (MK) also led to a significant decrease in the abundances of multiple beneficial genera, including UBA3282 (p < 0.01), Adlercreutzia_404257, Desulfovibrio_R_446353 (p < 0.05), Lawsonibacter (p < 0.01), UBA9715 (p < 0.05), Eubacterium_J (p < 0.01), 14-2 (p < 0.05), Dysosmobacter (p < 0.05), Borkfalkia (p < 0.05), Angelakisella (p < 0.05), Acutalibacter (p < 0.05), Merdibacter, Coprocola (p < 0.05), Massilioclostridium (p < 0.01), Lachnoclostridium_B (p < 0.01), Emergencia (p < 0.05), Cupidesulfovibrio (p < 0.05), and Staphylococcus (p < 0.05). After pretreatment with the Tupistra chinensis Bak. polysaccharide of the present invention in the experimental group (PK), the abundances of each genus except Merdibacter were restored, and the abundances of other genera except Adlercreutzia-404257, Borkfalkia, and Massilioclostridium had been restored to a level close to that of the control group.

[0098] Existing studies have shown that an increase in the abundance of Alloprevotella is closely related to gastrointestinal diseases, a decrease in the abundance of Adlercreutzia is associated with insulin resistance, and an increase in the abundance of Muribaculum is related to diseases such as diabetes. This indicates that these genera of bacteria may have a negative impact on health when present in high abundance and are regarded as harmful bacteria. In contrast, the abundance of Desulfovibrio increases during the treatment of alcoholic fatty liver in mice, while a decrease in the abundance of Lawsonibacter is associated with regulating intestinal health, preventing inflammation, and reducing the risk of cancer. An increase in the abundance of Eubacterium_J helps alleviate colitis in mice. These are all important examples of beneficial bacteria. In particular, Lawsonibacter, Eubacterium_J, Dysosmobacter, Angelakisella, and Lachnoclostridium_B are all butyrate-producing bacteria, and their abundances increase after treatment with tupistra polysaccharide. Butyric acid, as the preferred energy source for intestinal epithelial cells, can promote the proliferation of intestinal villi and enhance the absorption and utilization of nutrients by the intestine. Therefore, tupistra polysaccharide can not only promote the growth of beneficial bacteria but also inhibit the growth of harmful bacteria by adjusting the structure of the intestinal microbial community, especially by increasing the abundance of butyrate-producing bacteria, thereby restoring the intestinal flora disorder caused by Escherichia coli infection.

[0099] In summary, tupistra polysaccharide not only helps inhibit the growth of harmful bacteria but also promotes the growth of beneficial bacteria by adjusting the structure of the intestinal microbial community, especially by increasing the abundance of butyrate-producing bacteria, thereby restoring the intestinal flora disorder caused by Escherichia coli infection and maintaining intestinal health.

[0100] (4) Effects of tupistra polysaccharide on potential metabolic pathways of the intestinal flora in mice with Escherichia coli-mediated intestinal inflammation.

[0101] The possible effects of changes in the intestinal flora on metabolism were analyzed by comparison using the MetaCyc and KEGG databases. Figure 9It can be seen that metabolic pathways such as the biosynthesis of L-glutamic acid and L-glutamine (PWY-5505), the recovery of adenosylcobalamin from cobinamide II (PWY-6269), the biosynthesis of L-arginine I (ARGSYN-PWY), the biosynthesis of L-arginine IV (PWY-7400), the fermentation of pyruvate to acetone (PWY-6588), the fermentation of acetyl-CoA to butyric acid (PWY-5676), purine nucleotide degradation II (PWY-6353), guanosine nucleotide degradation III (PWY-6608), and adenosine nucleotide degradation II (SALVADEHYPOX-PWY) in the model group (MK) showed significantly higher LDA scores than those in other groups. However, the biotin biosynthesis I pathway (BIOTIN BIOSYNTHESIS) in the experimental group (PK) showed a significantly increased LDA score. This indicates that tupistra root polysaccharide may restore the function of the gut microbiota damaged by Escherichia coli by adjusting specific metabolic pathways. The significant increase in the biotin biosynthesis I pathway in the experimental group (PK) suggests that tupistra root polysaccharide may promote the growth or activity of beneficial microorganisms in the intestine, thus contributing to overall metabolic health. At the same time, the activation of various amino acid and nucleotide metabolic pathways and the presence of the butyric acid synthesis pathway observed in the control group (CK) further illustrate the important role of gut microbiota in maintaining host metabolic balance. Tupistra root polysaccharide can promote the production of certain beneficial metabolites (such as butyric acid) and enhance the biosynthesis of important nutrients such as biotin, for example, improving energy metabolism and antioxidant status. Thus, it can be seen that tupistra root polysaccharide may restore the function of the gut microbiota damaged by Escherichia coli by adjusting butyric acid synthesis, nucleotide degradation, biotin synthesis, etc.

[0102] The above has introduced in detail a tupistra root polysaccharide with antibacterial and anti-inflammatory effects, its preparation method and application. Specific examples are used in this article to elaborate on the principle and implementation mode of the present invention. The description of the above embodiments is only used to help understand the method and its core idea of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and modifications can still be made to the present invention, and these improvements and modifications also fall within the protection scope of the claims of the present invention.

Claims

1. A method for preparing a polysaccharide having antibacterial and anti-inflammatory effects, characterized in that: The following steps are involved: (1) Degreasing and removing impurities from the open-mouthed arrow: Select an appropriate amount of rhizomes of the open-mouthed arrow and screen them to remove impurities. The screened rhizomes are dried and crushed into fine powder. The crushed rhizome powder is wrapped with gauze and placed in a Soxhlet extractor. A 95% ethanol solution is used as a solvent for continuous cyclic extraction until the extract becomes colorless. The degreased residue is collected after the above process. The degreased residue is then transferred to a flask and refluxed twice at 60° C. with a 95% ethanol solution for 1 hour each time. After the reflux is completed, the residue is taken out and dried. (2) Extraction of C. cerana polysaccharide: Place the dried residue in step (1) into a flask, add distilled water, and reflux extract in a 90°C constant temperature water bath for 3 times, each time for 2 h. Filter after each extraction, collect the filtrate, and combine all the filtrates; The combined filtrate was concentrated to 1 / 10 of the original volume using a vacuum concentrator to form a concentrated filtrate; (3) Preparation of open-mouthed arrow polysaccharide: slowly add ethanol solution to the concentrated filtrate in step (2) while constantly stirring, perform alcohol precipitation treatment, and use a suction filtration device to separate the precipitate and the supernatant; wash the precipitate with anhydrous ethanol, and place it in a drying oven at 50°C to constant weight, and crush the dried precipitate to obtain light brown particles, that is, obtain open-mouthed arrow polysaccharide.

2. The method for preparing the open-mouthed arrow polysaccharide with antibacterial and anti-inflammatory effects according to claim 1, characterized in that: In step (1), the mass volume ratio of the fine powder of the open arrowroot stem to the ethanol solution with a concentration of 95% during the defatting process is 1g: (2-5)mL; the mass volume ratio of the defatted medicinal residue to the ethanol solution with a concentration of 95% during the reflux process is 1g: (2-5)mL.

3. The method for preparing the open-mouthed arrow polysaccharide with antibacterial and anti-inflammatory effects according to claim 2, characterized in that: In the degreasing process, the mass volume ratio of the fine powder of the open arrowroot stem to the ethanol solution with a concentration of 95% is 1g:4mL; in the reflux process, the mass volume ratio of the degreased medicinal residue to the ethanol solution with a concentration of 95% is 1g:3mL.

4. The method for preparing the open-mouthed arrow polysaccharide with antibacterial and anti-inflammatory effects according to claim 1, characterized in that: In step (2), the mass volume ratio of the dried medicinal residue to distilled water is 1g: (3~10)mL.

5. The method for preparing the open-mouthed arrow polysaccharide with antibacterial and anti-inflammatory effects according to claim 4, characterized in that: The mass volume ratio of the dried medicinal residue to distilled water is 1 g:6 mL.

6. The method for preparing the open-mouthed arrow polysaccharide with antibacterial and anti-inflammatory effects according to claim 1, characterized in that: In step (3), the concentration of the ethanol solution is 30-95%; the volume ratio of the concentrated filtrate to the ethanol solution is 1:(0.8-1.5).

7. The method for preparing the open-mouthed arrow polysaccharide with antibacterial and anti-inflammatory effects according to claim 6, characterized in that: The step (3) comprises the following steps: (i) 40% ethanol precipitation: Slowly add 40% ethanol solution to the concentrated filtrate while stirring continuously. After standing for 12 to 24 hours, a precipitate is obtained. The 40% ethanol precipitate and the supernatant are separated by a suction filtration device and washed with anhydrous ethanol. (ii) 60% ethanol precipitation: retain the supernatant from step (i), slowly add a 60% ethanol solution thereto, while stirring continuously, and let stand for 12 to 24 hours to obtain a precipitate. Use a suction filtration device to separate the 60% ethanol precipitate and the supernatant, and wash with anhydrous ethanol; (iii) 80% ethanol precipitation: retain the supernatant from step (ii), slowly add 80% ethanol solution thereto, and stir continuously while standing for 12 to 24 hours to obtain a precipitate, separate the 60% ethanol precipitate and the supernatant using a suction filtration device, and wash with anhydrous ethanol; (iv) The 40% ethanol precipitate, the 60% ethanol precipitate, and the 80% ethanol precipitate were placed in a drying oven at 50° C. and dried to constant weight, and then mixed evenly at a mass ratio of 1:1:1, and then the mixed precipitates were crushed to finally obtain the open arrow polysaccharide.

8. The open-mouthed arrow polysaccharide obtained by the preparation method according to any one of claims 1 to 7.

9. Use of the open-mouthed arrow polysaccharide described in claim 8 in a pharmaceutical product with antibacterial and anti-inflammatory effects.

10. The use according to claim 9, characterized in that: The pharmaceutical product with antibacterial and anti-inflammatory effects is a pharmaceutical product used for preventing or treating diarrhea or inflammatory bowel disease.

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