Application of ligusticum wallichii polysaccharide in preparation of medicine for regulating intestinal flora of inflammatory bowel disease

Abstract

The invention discloses application of ligusticum chuanxiong hort polysaccharide in preparation of medicines for regulating intestinal flora of inflammatory bowel diseases, belongs to the technical field of polysaccharide extraction process optimization and intestinal flora regulation, and particularly provides an economical and efficient extraction process and new application of the ligusticum chuanxiong hort polysaccharide, and the ligusticum chuanxiong hort polysaccharide is used for preparing a preparation or a composition. The intestinal flora disorder symptom in the inflammatory bowel disease state is relieved and treated by improving the relative abundance of probiotic flora, reducing the relative abundance of harmful bacteria, adjusting the intestinal flora structure and the like. Ligusticum wallichii polysaccharide up-regulates the relative abundance of probiotics such as bacteroides, clostridium praeparatum and the like and down-regulates the relative abundance of pathogenic bacteria such as proteobacteria, gamma proteobacteria, enterobacteriaceae, escherichia and the like so as to improve ulcerative colitis of mice, reverse abnormally proliferating bacteria in mice with inflammatory bowel diseases and improve the intestinal flora disorder state of the mice with inflammatory bowel diseases.

Description

Technical Field

[0001] The present invention relates to the field of traditional Chinese medicine, and particularly to the application of ligusticum sinense olivcv. chaxiong polysaccharide in the preparation of drugs for regulating the intestinal flora of inflammatory bowel disease. Background Art

[0002] Inflammatory bowel disease (IBD) is a chronic non-specific intestinal inflammatory disease that is prone to recurrence, including Crohn's disease (CD) and ulcerative colitis (UC). Research shows that important links in the pathogenesis of IBD include: intestinal oxidative stress injury, immune abnormalities, increased intestinal permeability, and microbial dysbiosis.

[0003] More and more studies have shown that the intestinal flora plays an important role in regulating UC. A large number of microorganisms in the intestine are in a balanced state under normal circumstances and play an important role in maintaining intestinal health. However, when the intestinal flora is imbalanced, the number of harmful bacteria will increase, leading to damage to the intestinal mucosa and thus triggering intestinal inflammation. In addition, the intestinal flora structure of patients with intestinal inflammation will also change, manifested as a decrease in the number of certain bacteria and an increase in the number of other bacteria. These changes may further aggravate the symptoms of intestinal inflammation, forming a vicious cycle. Therefore, intestinal flora imbalance is considered to be one of the important factors in the occurrence and development of intestinal inflammation.

[0004] Traditional Chinese medicine polysaccharides have been proven to be an important means of improving inflammatory bowel disease. A large number of studies have shown that traditional Chinese medicine polysaccharides are closely related to the intestinal flora in the process of improving inflammatory bowel disease. Ligusticum sinense Olivcv. Chaxiong, as a traditional Chinese medicinal material, has unique advantages in the treatment of inflammatory diseases. "Compendium of Materia Medica" states that ligusticum sinense olivcv. chaxiong "treats intestinal stasis, abdominal pain and diarrhea, and those with a slow pulse", and classifies it as "qi-promoting and blood-activating medicine". In addition, there are also records in "Synopsis of the Golden Chamber" about ligusticum sinense olivcv. chaxiong treating intestinal diseases. The medicinal value of polysaccharides, as the main component of ligusticum sinense olivcv. chaxiong, in regulating intestinal inflammation is worthy of further research, especially the effect of ligusticum sinense olivcv. chaxiong polysaccharide (LSOP) on the homeostasis of the intestinal flora in the state of intestinal inflammation has not been reported. Summary of the Invention

[0005] The purpose of the present invention is to solve at least one of the technical problems existing in the prior art, and to provide the application of ligusticum sinense olivcv. chaxiong polysaccharide in the preparation of drugs for regulating the intestinal flora of inflammatory bowel disease.

[0006] The technical solution of the present invention is as follows: The application of a ligusticum sinense olivcv. chaxiong polysaccharide in the preparation of drugs for regulating the intestinal flora of inflammatory bowel disease, wherein the ligusticum sinense olivcv. chaxiong polysaccharide comprises mannose, rhamnose, galacturonic acid, glucose, galactose, and arabinose in a mass ratio of 0.5 - 0.6:0.6 - 0.7:6 - 7:82 - 83:4 - 5:5 - 6; The tea lovage polysaccharide can increase the relative abundance of Bacteroidetes and Faecalibacterium prausnitzii, and downregulate the relative abundance of Proteobacteria, Gammaproteobacteria, Enterobacteriaceae and Escherichia, so as to regulate the intestinal flora structure.

[0007] Furthermore, the medicine uses the polysaccharide of Lophatherum chuanxiong as the active ingredient and contains a pharmaceutically acceptable carrier, wherein the weight percentage of the polysaccharide of Lophatherum chuanxiong in the preparation is 0.01-99.99%, and the rest is the pharmaceutically acceptable carrier.

[0008] Furthermore, the carrier is selected from at least one of sorbitol, mannitol, thioglycolic acid, methionine, EDTA sodium salt, sodium chloride, potassium chloride, glycine, sucrose, lactose, cyclodextrin, talc, calcium sulfate, sodium stearate, cellulose, gelatin, and glycerol.

[0009] Furthermore, the dosage form of the drug is capsule, tablet, granule, injection, sustained-release agent, oral solution or pill.

[0010] The invention also discloses a method for extracting cha xiong polysaccharide, comprising the following steps: S1: crushing the Chaxiong herbal slices, sieving, and obtaining Chaxiong herbal powder; S2: Take the tea lovage powder, add water to extract, obtain the extract, filter the pulp extract while hot, combine the tea lovage filtrate, concentrate under reduced pressure, precipitate with alcohol, let stand and then filter, add water to redissolve the precipitate, and obtain the tea lovage crude polysaccharide solution; S3: After removing protein with chloroform and n-butanol, the product was purified with a dialysis bag and freeze-dried to obtain the polysaccharide of Rhizoma Chuanxiong.

[0011] Furthermore, in the step S1, the product is sieved through a 100-200 mesh sieve.

[0012] Further, in step S2, the extraction temperature is 85-95°C, the liquid-to-solid ratio after adding water is 1:25-35, the number of extractions is 3-5 times, and the extraction time for each time is 2-3h; In step S2, the mixture is concentrated under reduced pressure to a crude drug content of 0.2-0.3 g / mL, and the alcohol precipitation end point is 80% alcohol content by volume; Furthermore, in step S3, the volume ratio of chloroform to n-butanol is 4:1.

[0013] In step S3, the cut-off relative molecular weight of the dialysis bag is 8000-14000.

[0014] The beneficial effects of the present invention are as follows: The present invention provides an economical and efficient extraction process and new uses of chaxiong polysaccharide for preparing a preparation or composition. By increasing the relative abundance of probiotic flora, reducing the relative abundance of harmful bacteria, and regulating the intestinal flora structure, etc., it alleviates and treats the symptoms of intestinal flora disorder in the state of inflammatory bowel disease. Chaxiong polysaccharide up-regulates the relative abundance of probiotics such as Bacteroidetes and Faecalibacterium prausnitzii, and down-regulates the relative abundance of pathogenic bacteria such as Proteobacteria, Gammaproteobacteria, Enterobacteriaceae, and Escherichia to improve ulcerative colitis in mice, reverse the abnormally proliferating bacteria in mice with inflammatory bowel disease, and improve the disordered state of intestinal flora in mice with inflammatory bowel disease. Description of the Drawings

[0015] Figure 1 It is the result of single-factor experiment on the preparation process of chaxiong polysaccharide.

[0016] Figure 2 It is the response surface analysis diagram and contour map; Figure 3 It is the result of the effect of polysaccharide on the body weight change rate, DAI score and colon length of mice; among them, compared with the M group, indicates P < 0.05, indicates P < 0.001, indicates P < 0.0001; compared with the C group, # indicates P < 0.0001.

[0017] Figure 4 It is the dilution curve; Figure 5 It is the result of PcoA analysis; Figure 6 It is the result of NMDS analysis; Figure 7 It is the species composition analysis - phylum level; Figure 8 It is the Venn diagram; Figure 9 It is the species composition heat map - genus level, where M1 - M6 are mice No. 1 - 6 in the M group, SA1 - SA6 are mice No. 1 - 6 in the SASP group, and C1 - C6 are mice No. 1 - 6 in the C group; Figure 10 It is the abundance value of Amulumruptor, where, compared with the M group, indicates P < 0.0001, and ns indicates no significant difference; Figure 11 It is the relative abundance of Duncaniella of the genus Dunnella, where, compared with the M group, indicates P < 0.0001, and ns indicates no significant difference; Figure 12is the relative abundance of Prevotella. Among them, compared with the M group, indicates P < 0.0001, and ns indicates no significant difference; Figure 13 is the relative abundance of Escherichia. Among them, compared with the M group, indicates P < 0.0001; Figure 14 is the relative abundance of Helicobacter-D. Among them, compared with the M group, indicates P < 0.0001; Figure 15 is the cladogram of LEfSe analysis; Figure 16 is the C-M LDA bar chart; Figure 17 is the M-CX LDA bar chart; Figure 18 is the HPLC chromatogram of the monosaccharide composition analysis of Cha-xiong polysaccharide. Among them, 1 - mannose, 2 - rhamnose, 3 - galacturonic acid, 4 - glucose, 5 - galactose, 6 - xylose, 7 - arabinose. Detailed implementation mode

[0018] The embodiments of the present invention are described in detail below. The following described embodiments are exemplary and are only used to explain the present invention, and should not be construed as a limitation to the present invention. For those without specific technologies or conditions noted in the embodiments, they shall be carried out according to the technologies or conditions described in the literature in the art or according to the product specifications. For the reagents or instruments without the manufacturer noted, they are all conventional products that can be obtained through commercial purchase.

[0019] The reagents used in the following embodiments: Cha-xiong; Sulfasalazine; Chloroform; n-Butanol; D-Anhydrous glucose (batch number 110833~201908) was purchased from the National Institutes for Food and Drug Control; Concentrated sulfuric acid; Phenol; Coomassie brilliant blue G-250; Absolute ethanol; Petroleum ether (60℃~90℃); 4% Paraformaldehyde general tissue fixative; Xylene; n-Butanol; HE staining kit; Interleukin (IL)-1β, IL-6, and Tumor necrosis factor α (TNF-α) ELISA kits.

[0020] The instruments used in the following embodiments: Chinese herbal medicine grinder; Electric heating mantle; Vacuum concentration instrument; Freeze dryer; Multifunctional microplate reader; BAS124S electronic balance (Sartorius Scientific Instruments Beijing Co., Ltd.); Ultraviolet spectrophotometer; RM2125 paraffin slicer (Leica Company, Germany); Automatic scanning microscope - VS200.

[0021] Animals used in the following examples: 60 male C57BL / 6J mice, 6 - 8 weeks old, weighing 18 - 21 g, were purchased from Liaoning Changsheng Biotechnology Co., Ltd., and the source institution license number is SCXK(Liao)2020 - 0001. The animals were housed in the Institute of Traditional Chinese Medicine Health Industry, China Academy of Chinese Medical Sciences, and the experimental animal license number is SYXK(Gan)2023 - 0008. All animal experiments were approved by the Animal Welfare and Ethics Committee of the Institute of Traditional Chinese Medicine Health Industry, China Academy of Chinese Medical Sciences (approval number: 2023001).

[0022] Example 1 Polysaccharides from Ligusticum wallichii Franch were prepared by hot water extraction. Through single - factor experiments and response - surface experiments, the extraction process parameters of polysaccharides from Ligusticum wallichii Franch were optimized, the factors and range intervals that significantly affected the extraction yield of polysaccharides from Ligusticum wallichii Franch were screened out, and an economical, efficient and complete extraction process route of polysaccharides from Ligusticum wallichii Franch was obtained.

[0023] (1) Preparation of polysaccharides from Ligusticum wallichii Franch a. Single - factor experiments The Ligusticum wallichii Franch medicinal materials were crushed and passed through a 100 - mesh sieve. Accurately weigh 10 g of Ligusticum wallichii Franch powder for each portion, and use extraction temperature (50, 60, 70, 80, 90, 100 °C), liquid - to - solid ratio (10, 15, 20, 25, 30, 35, 40 times), extraction time (1, 1.5, 2, 2.5, 3, 3.5, 4 h), and extraction times (1, 2, 3, 4, 5, 6 times) as influencing factors to design the extraction experiment plan.

[0024] b. Response - surface experiments Based on the experimental results of single - factor analysis, according to the principle of response - surface analysis method, Design Expert (version 6.0) software was used to establish an experimental mathematical model by Box - Benhnken central composite experimental design (see Table 1), and a total of 27 groups of extraction experiments were obtained to optimize the above four extraction parameters.

[0025] Table 1 Experimental design parameters for response - surface analysis

[0026] c. Preparation of polysaccharides from Ligusticum wallichii Franch Refer to the optimal extraction process to prepare the water extract of Ligusticum wallichii Franch, concentrate it under reduced pressure to a concentration of 0.25 g / mL of crude drug content, use 80% alcohol content as the end point of alcohol precipitation, stand overnight at 4 °C and then filter, redissolve the precipitate in water to obtain the crude polysaccharide solution of Ligusticum wallichii Franch. After removing proteins by the Sevag method (chloroform - n - butanol 4﹕1), it was refined by dialysis bag (cut - off relative molecular mass 8000 - 14000) and freeze - dried to obtain the de - proteinized polysaccharides from Ligusticum wallichii Franch.

[0027] d. Analysis of monosaccharide composition of chaxiong polysaccharide Take 5 mg of Camellia chuanxiong polysaccharide, put it in a stoppered test tube, add 2 mL of 2 mol / L trifluoroacetic acid, and hydrolyze it at 110 °C in the dark for 6 h. After cooling to room temperature, add 1 mL of methanol solution (to promote the volatilization of trifluoroacetic acid), evaporate the methanol with nitrogen, repeat the operation 3 times to remove trifluoroacetic acid, and finally add purified water to make up to 1 mL to obtain polysaccharide hydrolyzate. 500 μL of the mixed monosaccharide standard solution (mannose, rhamnose, galacturonic acid, glucose, galactose, xylose, arabinose) and polysaccharide hydrolyzate were respectively pipetted into 5 mL stoppered test tubes, and 1 mL of 0.3 mol / L NaOH solution and 1 mL of 0.5 mol / L PMP methanol solution were added in sequence, vortexed and mixed, reacted in a water bath at 70 ℃ for 70 min, removed, left to cool to room temperature, neutralized with 1 mL of 0.3 mol / L HCL, and then an equal volume of chloroform (to remove the derivatization reagent PMP) was added, vortexed and mixed, left to stand, the chloroform layer was discarded, and the extraction was repeated 3 times, and the supernatant (aqueous phase) was collected. The supernatant was filtered with a 0.22 μm filter membrane and stored, and then injected into the chromatographic analysis.

[0028] Chromatographic conditions: Chromatographic column: Shim-packGIS (4.6×250 mm, 5μm), mobile phase: A0.1 M potassium dihydrogen phosphate (pH=6.85): B acetonitrile = 83%:17%, column temperature: 30℃, flow rate: 1.0 mL / min, injection volume: 20 μL, detector wavelength: 245 nm.

[0029] (2) Modeling and drug administration After 7 days of adaptive feeding, except for the normal group, the experimental groups were fed with 3.0% DSS solution (dextran sulfate sodium solution) instead of drinking water. The mice were divided into 6 groups (n=10 / group) and randomly divided into normal (C, drinking water) group, model (M, 3.0% DSS solution) group, sulfasalazine (SASP, 100 mg / kg) group, low-dose chaxiong polysaccharide (CX-L, 50 mg / kg) group, medium-dose chaxiong polysaccharide (CX-M, 100 mg / kg) group, and high-dose chaxiong polysaccharide (CX-H, 150 mg / kg) group. Each group of animals received treatment from day 1 to day 7. On day 8, after fasting for 12 hours but not water, the mice were killed by cervical dislocation, and the feces of mice with high-dose chaxiong polysaccharide were collected to study the regulatory effect of chaxiong polysaccharide on the intestinal flora of mice, that is, the CX group in the experiment used high-dose chaxiong polysaccharide.

[0030] (3) Evaluation of mouse weight, colon length and DAI score During the entire experimental period, the body weight, colon length, stool characteristics and blood in stool of each group of mice were observed and recorded, and the DAI score was performed according to the scoring criteria in Table 2.

[0031] DAI = weight loss rate + stool characteristics + blood in stool Table 2 DAI scoring criteria

[0032] (4) 16S rRNA sequencing The intestinal contents of mice were collected using centrifuge tubes and stored at -80°C. The 16S rRNA sequencing of the mouse intestinal flora was completed in cooperation with Shanxi Xinzhida Technology Development Co., Ltd.

[0033] The results of the single factor experiment are as follows: Figure 1 The research ranges were determined to be extraction temperature 85-95 ℃, liquid-to-solid ratio 25-35 times, extraction time 2.5-3.5 h, and extraction times 3-5 times.

[0034] The results of the response surface experiment are as follows: The experimental design of each group and the yield of tea lovage polysaccharide are shown in Table 3, and then the optimal process parameters of each influencing factor were obtained, and an economical, efficient and complete extraction process route of tea lovage polysaccharide was obtained.

[0035] Table 3 Response surface analysis results for optimizing the extraction yield of polysaccharides from Rhizoma Chuanxiong

[0036] SAS 8.0 software was used to analyze the data in Table 3 to obtain the response surface analysis diagram and contour map ( Figure 2 ), the figure shows the optimal interval of each parameter, and the binary regression equation is established as follows:

[0037] By analyzing the above binary regression equation, the optimal extraction process of tea lovage polysaccharide was finally determined as follows: extraction temperature 93.7℃, liquid-to-solid ratio 1:28, extraction times 4 times, extraction time 2.6 h, and the optimal polysaccharide extraction rate was 9.4% (as shown in Table 4).

[0038] Table 4 Optimal extraction process of polysaccharides from Rhizoma Chuanxiong

[0039] The results of the evaluation of the effect of Chaxiong polysaccharide on the body weight and DAI score of mice are as follows ( Figure 3 ): During the experiment, mice in group C were stable, with shiny hair and steady weight gain. Mice in group M were listless and had severe blood in their stools. After sulfasalazine and chaxiong polysaccharide were administered, the mice's condition improved. Figure 3As shown, compared with group C, the body weight of mice in group M decreased significantly, and the DAI score increased significantly (P < 0.0001); after administration of ligusticum wallichii polysaccharide, compared with group M, the body weight change rate and DAI score in each dose group of ligusticum wallichii polysaccharide also showed a similar trend, the body weight of mice increased significantly, and the DAI score decreased significantly. Compared with group C, the colon length of mice treated with DSS was significantly shortened (P < 0.0001); after administration of ligusticum wallichii polysaccharide, the colon length was significantly restored (P < 0.0001).

[0040] The results of the regulation of the intestinal flora of mice by ligusticum wallichii polysaccharide are as follows: The rarefaction curve ( Figure 4 ) The curve is flat, indicating that the sequencing results are sufficient to reflect the microbial diversity contained in the current samples. Principal coordinate analysis (PCoA analysis) and non-metric multidimensional scaling (NMDS) analysis were performed. The farther the distance, the greater the difference in the microbial communities in the two samples. From the PCoA results Figure 5 and the NMDS results Figure 6 It can be seen that the microbial abundances of each group are significantly separated, indicating that the intestinal microbial compositions of each experimental group are different. The stress coefficient results show that the stress value is 0.0000971, indicating a good fitting effect.

[0041] Analyzing the intestinal microbial composition of mice at the phylum level, as Figure 7 shown, group C (normal group) is mainly composed of Bacteroidota, Firmicutes A, D, etc. Compared with group C, the relative abundance of Bacteroidota in group M decreased from 35.51% to 28.27%, and the pathogenic Proteobacteria appeared in the group, with a relative abundance of 17.53%. Compared with group M, the relative abundance of Bacteroidota in group CX increased to 51.70%, and the relative abundance of Proteobacteria decreased to 1.17%. The results of the species composition analysis show that ligusticum wallichii polysaccharide can intervene in UC by upregulating the relative abundance of Bacteroidota and decreasing the relative abundance of pathogenic Proteobacteria.

[0042] As Figure 8 shown, 157 overlapping ASV / OTUs were found among group C, group M, group SASP and group CX through the petal diagram. In addition, group C had 4440 unique ASV / OTUs, group M had 2820 unique ASV / OTUs, group SASP had 3088 unique ASV / OTUs, and group CX had 3110 unique ASV / OTUs. The results show that DSS induction reduced the diversity of the intestinal flora, while ligusticum wallichii polysaccharide can reverse the diversity of the intestinal flora and maintain the balance of the intestinal flora.

[0043] From Figures 9 - 14The relative abundances of the microbial compositions at the genus level were shown, and the species compositions of the intestinal flora of mice in different groups were different. In group C, the relative abundances of Amulumruptor, Mailhella, CAG-510, and Lactobacillus were higher. Escherichia, Helicobacter_D, and Bacteroides_H were more abundant in DSS-induced UC mice. Bifidobacterium and Phocaeicola_A were highly expressed in the SASP group. Prevotella and Duncaniella were more abundant in the mice with UC treated with chaxiong polysaccharide. By comparing the compositional differences in the intestinal microbiome among groups, compared with group M, the relative abundances of Amulumruptor, Duncaniella, and Prevotella in groups C and CX were significantly increased (P < 0.0001), indicating that chaxiong polysaccharide might improve intestinal inflammation in UC mice by upregulating Amulumruptor, Duncaniella, and Prevotella and downregulating Escherichia and Helicobacter.

[0044] The species that were significantly different among groups at all levels were analyzed by the LEfSe (LDA Effect Size) cladogram ( Figure 15 ). The LDA threshold was set at 4.0. Compared with group C ( Figure 16 ), 50 intestinal microbiota in group M changed (22 were upregulated and 28 were downregulated). The abundances of p_Proteobacteria, Gamnaproteobacteria, Enterobacteriaceae_A, Enterobacterales_A, Escherichia, etc. were upregulated; the abundances of Lachnospiraceae, Lachnospirales, f_Muribaculaceae, Lactobacillaceae, etc. were downregulated. Compared with group M ( Figure 17),(38 gut microbiota flora in the CX group changed (20 up-regulated and 18 down-regulated). Those with increased abundance included Bacteroidota, Bacteroidia, Bacteroidales, Muribaculaceae, Faecalibaculum, etc. Studies have shown that Faecalibaculum can produce short-chain fatty acids (SCFAs), which can regulate the immune system, promote the growth of beneficial flora, and inhibit the reproduction of harmful bacteria; Proteobacteria, Gammaproteobacteria, Enterobacterales_A, Enterobacteriaceae_A, Escherichia, etc. had decreased abundance, reversing the changes in the flora structure caused by intestinal inflammation and promoting the restoration of the normal structure level of the intestinal flora in UC mice. It is inferred that chaxiong polysaccharide may improve ulcerative colitis in mice by up-regulating the relative abundance of probiotics such as Bacteroidetes and Faecalibaculum, and down-regulating the relative abundance of pathogenic bacteria such as Proteobacteria, Gammaproteobacteria, Enterobacteriaceae, and Escherichia.

[0045] At the same time, the components of the chaxiong deproteinized polysaccharide prepared by the optimal process of Example 1 were analyzed, and the analysis results are shown in Figure 18 . It can be seen from the figure that the chaxiong polysaccharide in this example is mainly composed of 6 monosaccharides: mannose, rhamnose, galacturonic acid, glucose, galactose, and arabinose. According to the peak area ratio calculation, the proportions of each monosaccharide are 0.518%, 0.694%, 6.396%, 82.160%, 4.347%, and 5.886% respectively. The results show that chaxiong polysaccharide is a natural polysaccharide mainly composed of glucose.

[0046] Different from other natural polysaccharides, the prior art improves enteritis by regulating the relative abundance of flora such as Bacteroidota and Firmicutes. The chaxiong polysaccharide of this application mainly improves ulcerative colitis by regulating the relative abundance of intestinal microbiota flora such as Bacteroidetes, Faecalibaculum, Proteobacteria, Gammaproteobacteria, Enterobacteriaceae, and Escherichia.

[0047] The above embodiments only represent the preferred embodiments of the present invention, and the description is relatively specific and detailed, but it should not be construed as a limitation on the scope of the invention patent. It should be noted that for those of ordinary skill in the art, according to the technical solutions and concepts described above, various other corresponding changes and deformations can also be made, and all these changes and deformations should fall within the protection scope of the claims of the present invention.

Claims

1. Use of chaxiong polysaccharide in the preparation of a drug for regulating the intestinal flora of inflammatory bowel disease, characterized in that, The tea xiong polysaccharide includes mannose, rhamnose, galacturonic acid, glucose, galactose, and arabinose with a mass ratio of 0.5 - 0.6:0.6 - 0.7:6 - 7:82 - 83:4 - 5:5 - 6; The tea xiong polysaccharide can increase the relative abundances of Bacteroidetes and Faecalibacterium prausnitzii, and down - regulate the relative abundances of Proteobacteria, Gammaproteobacteria, Enterobacteriaceae, and Escherichia, so as to regulate the intestinal flora structure.

2. Use of the chaxiong polysaccharide according to claim 1 in the preparation of a drug for regulating the intestinal flora of inflammatory bowel disease, characterized in that, The drug uses the tea xiong polysaccharide as the active ingredient of the drug and also contains a pharmaceutically acceptable carrier, wherein the weight percentage of the tea xiong polysaccharide in the preparation is 0.01 - 99.99%, and the rest is the pharmaceutically acceptable carrier.

3. Use of a chaxiong polysaccharide according to claim 2 in the preparation of a drug for regulating the intestinal flora of inflammatory bowel disease, characterized in that, The carrier is selected from at least one of sorbitol, mannitol, mercaptoacetic acid, methionine, sodium EDTA, sodium chloride, potassium chloride, glycine, sucrose, lactose, cyclodextrin, talcum powder, calcium sulfate, sodium stearate, cellulose, gelatin, and glycerol.

4. Use of a chaxiong polysaccharide according to claim 1 in the preparation of a drug for regulating the intestinal flora of inflammatory bowel disease, characterized in that, The dosage form of the drug is capsule, tablet, granule, injection, sustained - release preparation, oral liquid, or dripping pill.

5. A method for extracting tea rhizome polysaccharide used in the application as described in claim 1, characterized in that, It includes the following steps: S1: Crush the tea xiong slices and sieve them to obtain tea xiong powder; S2: Take the tea xiong powder, extract it with water to obtain an extract, filter the extract while it is hot, combine the tea xiong filtrates, concentrate under reduced pressure, precipitate with alcohol, filter by suction after standing, and redissolve the precipitate in water to obtain a crude tea xiong polysaccharide solution; S3: Remove proteins with chloroform and n - butanol, then refine it with a dialysis bag and freeze - dry it to obtain tea xiong polysaccharide.

6. The extraction method according to claim 5, wherein in step S1, it is sieved through a 100 - 200 mesh sieve.

7. The extraction method according to claim 5, wherein in step S2, the extraction temperature is 85 - 95 °C, the liquid - to - solid ratio after adding water is 1:25 - 35, the extraction times are 3 - 5 times, and the extraction time for each time is 2 - 3 h; in step S2, it is concentrated under reduced pressure to a drug content of 0.2 - 0.3 g / mL, and the alcohol precipitation end point is an alcohol content of 80% by volume.

8. The extraction method according to claim 5, wherein in step S3, the volume ratio of chloroform to n - butanol is 4:1; in step S3, the cut - off relative molecular weight of the dialysis bag is 8000 - 14000.

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