Application of chaxiong polysaccharide in the preparation of drugs for regulating intestinal flora in inflammatory bowel disease

Abstract

The present invention discloses an application of tea lovage polysaccharide in the preparation of a drug for regulating intestinal flora in inflammatory bowel disease, and belongs to the technical field of polysaccharide extraction process optimization and intestinal flora regulation. Specifically, the present invention provides an economical and efficient extraction process and new use of tea lovage polysaccharide for the preparation of a preparation or composition. By increasing the relative abundance of probiotic flora, reducing the relative abundance of harmful bacteria, regulating the structure of intestinal flora, etc., the symptoms of intestinal flora disorder in inflammatory bowel disease are alleviated and treated. Tea lovage polysaccharide improves ulcerative colitis in mice by upregulating the relative abundance of probiotics such as Bacteroides and Faecalibacterium prausnitzii, and downregulating the relative abundance of pathogenic bacteria such as Proteobacteria, Gammaproteobacteria, Enterobacteriaceae and Escherichia, thereby reversing the abnormal proliferation of bacteria in mice with inflammatory bowel disease and improving the disordered intestinal flora in mice with inflammatory bowel disease.

Description

Technical Field

[0001] The present invention relates to the field of traditional Chinese medicine, and in particular to an application of a chaxiong polysaccharide in the preparation of a medicine for regulating intestinal flora in inflammatory bowel disease. Background Art

[0002] Inflammatory bowel disease (IBD) is a chronic, nonspecific intestinal inflammatory disease that is prone to relapse and includes Crohn's disease (CD) and ulcerative colitis (UC). Research has shown that key components of IBD's pathogenesis include intestinal oxidative stress, 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 the UC process. Under normal circumstances, a large number of microorganisms in the intestine are in a balanced state and play an important role in maintaining intestinal health. However, when the intestinal flora is unbalanced, harmful bacteria will increase, causing damage to the intestinal mucosa, thereby 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 shown to be an important tool for improving inflammatory bowel disease. Numerous studies have demonstrated that these polysaccharides are closely linked to the intestinal microbiota in improving inflammatory bowel disease. Chaxiong (Ligusticum sinense Olivcv. Chaxiong), a traditional Chinese medicinal herb, possesses unique advantages in treating inflammatory diseases. The Compendium of Materia Medica states that Chaxiong "treats small intestinal stagnation, abdominal pain, diarrhea, and slow pulse," and categorizes it as a "Qi-activating and blood-activating medicine." Furthermore, the Synopsis of the Golden Chamber also mentions Chaxiong as a treatment for intestinal diseases. The medicinal value of polysaccharides, the main component of Chaxiong, in regulating intestinal inflammation warrants further investigation. In particular, the effects of Chaxiong polysaccharides (LSOP) on intestinal microbiota homeostasis during intestinal inflammation have 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 a use of angelica sinensis polysaccharide in the preparation of a drug for regulating intestinal flora in inflammatory bowel disease.

[0006] The technical solutions of the present invention are as follows:

[0007] A use of a tea lovage polysaccharide in preparing a drug for regulating intestinal flora in inflammatory bowel disease, wherein the tea lovage 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;

[0008] 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.

[0009] Furthermore, the medicine uses the tea lovage polysaccharide as the active ingredient and contains a pharmaceutically acceptable carrier, wherein the weight percentage of the tea lovage polysaccharide in the preparation is 0.01-99.99%, and the rest is the pharmaceutically acceptable carrier.

[0010] 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.

[0011] Furthermore, the dosage form of the drug is capsule, tablet, granule, injection, sustained-release agent, oral solution or pill.

[0012] The present invention also discloses a method for extracting tea lovage polysaccharide, comprising the following steps:

[0013] S1: Grinding the Chaxiong slices and sieving to obtain Chaxiong powder;

[0014] 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 it stand and then filter, add water to redissolve the precipitate to obtain the tea lovage crude polysaccharide solution;

[0015] S3: After removing protein with chloroform and n-butanol, the product was purified with a dialysis bag and freeze-dried to obtain the tea lovage polysaccharide.

[0016] Furthermore, in step S1, the product is sieved through a 100-200 mesh sieve.

[0017] Furthermore, 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 2-3 hours;

[0018] 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% by volume of alcohol content;

[0019] Furthermore, in step S3, the volume ratio of chloroform to n-butanol is 4:1.

[0020] In step S3, the cut-off relative molecular weight of the dialysis bag is 8000-14000.

[0021] The beneficial effects of the present invention are as follows: the present invention provides an economical and efficient extraction process and new use of tea lovage polysaccharide for preparing a preparation or composition. By increasing the relative abundance of probiotic bacteria, reducing the relative abundance of harmful bacteria, and regulating the structure of intestinal flora, the symptoms of intestinal flora disorder in inflammatory bowel disease are alleviated and treated. Tea lovage polysaccharide improves ulcerative colitis in mice by upregulating the relative abundance of probiotics such as Bacteroides and Faecalibacterium prausnitzii, and downregulating the relative abundance of pathogenic bacteria such as Proteobacteria, Gammaproteobacteria, Enterobacteriaceae, and Escherichia, thereby reversing the abnormal proliferation of bacteria in mice with inflammatory bowel disease and improving the disordered intestinal flora in mice with inflammatory bowel disease. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 These are the results of a single-factor experiment on the preparation process of Rhizoma Chuanxiong polysaccharides.

[0023] Figure 2 Response surface analysis diagram and contour map;

[0024] Figure 3 The results show the effects of polysaccharides on the body weight change rate, DAI score and colon length of mice; compared with the M group, Indicates P < 0.05, Indicates P < 0.001, Indicates P < 0.0001; compared with group M, #### indicates P < 0.0001.

[0025] Figure 4 is the dilution curve;

[0026] Figure 5 The results of the PcoA analysis were obtained;

[0027] Figure 6 This is the NMDS analysis result;

[0028] Figure 7 For species composition analysis - phylum level;

[0029] Figure 8 For Venn diagram;

[0030] Figure 9 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;

[0031] Figure 10 is the abundance value of Amulumruptor, among which, compared with the M group, indicates P < 0.0001, ns indicates no significant difference;

[0032] Figure 11is the relative abundance of Duncanilla, among which, compared with group M, indicates P < 0.0001, ns indicates no significant difference;

[0033] Figure 12 is the relative abundance of Prevotella, among which, compared with group M, indicates P < 0.0001, ns indicates no significant difference;

[0034] Figure 13 is the relative abundance of Escherichia, among which, compared with the M group, indicates P < 0.0001;

[0035] Figure 14 is the relative abundance of Helicobacter-D, among which, compared with the M group, indicates P < 0.0001;

[0036] Figure 15 Analyze the cladogram for LEfSe;

[0037] Figure 16 is the CM LDA histogram;

[0038] Figure 17 It is the M-CX LDA histogram;

[0039] Figure 18 This is the HPLC chromatogram of the monosaccharide composition analysis of the tea lovage polysaccharide, including 1-mannose, 2-rhamnose, 3-galacturonic acid, 4-glucose, 5-galactose, 6-xylose, and 7-arabinose. DETAILED DESCRIPTION

[0040] The following embodiments of the present invention are described in detail. The embodiments described below are exemplary and are only used to explain the present invention, and are not to be construed as limiting the present invention. Where specific techniques or conditions are not specified in the embodiments, the techniques or conditions described in the literature in this field or the product specifications are used. Where the manufacturer of the reagents or instruments is not specified, all are conventional products that can be obtained commercially.

[0041] Reagents used in the following examples:

[0042] Rhizoma Chuanxiong, sulfasalazine, chloroform, n-butanol, and D-anhydrous glucose (batch numbers 110833–201908) were purchased from the China Food and Drug Administration. Concentrated sulfuric acid, phenol, Coomassie Brilliant Blue G-250, anhydrous ethanol, petroleum ether (60°C–90°C), 4% paraformaldehyde universal tissue fixative, xylene, n-butanol, a HE staining kit, and interleukin (IL)-1β, IL-6, and tumor necrosis factor α (TNF-α) ELISA kits were also purchased.

[0043] The instruments used in the following examples are:

[0044] Chinese herbal medicine grinder; electric heating mantle; vacuum concentrator; freeze dryer; multifunctional microplate reader; BAS124S electronic balance (Sartorius Scientific Instruments Beijing Co., Ltd.); ultraviolet spectrophotometer; RM2125 paraffin sectioner (Leica, Germany); fully automatic slide scanner - VS200.

[0045] Animals used in the following examples:

[0046] Sixty C57BL / 6J male mice, 6-8 weeks old, weighing 18-21 g, were purchased from Liaoning Changsheng Biotechnology Co., Ltd. under institutional license number SCXK (Liao) 2020-0001. The animals were maintained at the Institute of Traditional Chinese Medicine and Health Industry, China Academy of Chinese Medical Sciences under license number SYXK (Gan) 2023-0008. All animal experiments were approved by the Animal Welfare and Ethics Committee of the Institute of Traditional Chinese Medicine and Health Industry, China Academy of Chinese Medical Sciences (approval number 2023001).

[0047] Example 1

[0048] Tea lovage polysaccharide was prepared by hot water extraction. Through single factor experiments and response surface experiments, the extraction process parameters of tea lovage polysaccharide were optimized, and the factors and ranges that significantly affected the extraction yield of tea lovage polysaccharide were screened out, resulting in an economical, efficient, and complete extraction process for tea lovage polysaccharide.

[0049] (1) Preparation of Tea Loquat Polysaccharide

[0050] a. Single-factor experiment

[0051] Chaxiong herb was ground and passed through a 100-mesh sieve. Accurately weigh 10 g of Chaxiong powder per portion. An extraction protocol was designed using 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 number of extractions (1, 2, 3, 4, 5, 6) as influencing factors.

[0052] b. Response surface experiment

[0053] Based on the experimental results of univariate analysis and the principle of response surface analysis, Design Expert (version 6.0) software was used to establish an experimental mathematical model using the Box-Benhnken central composite experimental design (see Table 1). A total of 27 groups of extraction experiments were obtained to optimize the above four extraction parameters.

[0054] Table 1 Experimental design parameters for response surface analysis

[0055]

[0056] c. Preparation of Tea-Qiong Polysaccharide

[0057] An aqueous extract of Chaxiong rhizome was prepared according to the optimal extraction process and concentrated under reduced pressure to a crude drug content of 0.25 g / mL. Alcohol precipitation was performed at 80% alcohol content, and the extract was allowed to stand overnight at 4°C before filtration. The precipitate was then reconstituted with water to obtain a crude Chaxiong rhizome polysaccharide solution. Protein was removed using the Sevag method (chloroform-n-butanol 4:1), followed by purification using a dialysis bag (molecular weight cut-off range 8,000-14,000), and freeze-dried to obtain the deproteinized Chaxiong rhizome polysaccharide.

[0058] d. Analysis of monosaccharide composition of chaxiong polysaccharide

[0059] Take 5 mg of tea xiong polysaccharide, place 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 the volume 1 mL to obtain the polysaccharide hydrolyzate. 500 μL each of a mixed monosaccharide standard solution (mannose, rhamnose, galacturonic acid, glucose, galactose, xylose, and arabinose) and a polysaccharide hydrolyzate were transferred to a 5 mL stoppered test tube. 1 mL of 0.3 mol / L NaOH solution and 1 mL of 0.5 mol / L PMP methanol solution were added, sequentially. The mixture was vortexed and incubated in a 70°C water bath for 70 min. The mixture was removed, allowed to stand, and cooled to room temperature. 1 mL of 0.3 mol / L HCl was added for neutralization. An equal volume of chloroform (to remove the derivatization reagent, PMP) was added, vortexed, allowed to stand, and the chloroform layer was discarded. The extraction was repeated three times, and the supernatant (aqueous phase) was collected. The supernatant was filtered through a 0.22 μm filter and stored for chromatographic analysis.

[0060] Chromatographic conditions:

[0061] Chromatographic column: Shim-pack GIS (4.6×250 mm, 5 μm), mobile phase: A 0.1 M potassium dihydrogen phosphate (pH = 6.85): B acetonitrile = 83%:17%, column temperature: 30°C, flow rate: 1.0 mL / min, injection volume: 20 μL, detector wavelength: 245 nm.

[0062] (2) Modeling and drug administration

[0063] After 7 days of adaptive feeding, mice in all experimental groups, except the control group, were fed 3.0% DSS solution (dextran sulfate sodium solution) instead of drinking water. Mice were randomly divided into six groups (n = 10 / group) and randomly assigned to a control (C, drinking water) group, a model (M, 3.0% DSS solution) group, a sulfasalazine (SASP, 100 mg / kg) group, a low-dose (CX-L, 50 mg / kg) group, a medium-dose (CX-M, 100 mg / kg) group, and a high-dose (CX-H, 150 mg / kg) group. Each group received treatment from day 1 to day 7. On day 8, after a 12-hour fast without water deprivation, the mice were sacrificed by cervical dislocation. Feces from mice receiving the high-dose (CX-H) treatment were collected to investigate the regulatory effects of CX-H on the intestinal microbiota. In this experiment, the CX group received the high-dose (CX-H) treatment.

[0064] (3) Evaluation of mouse weight, colon length and DAI score

[0065] The body weight, colon length, stool characteristics, and blood in stool of each group of mice were observed and recorded throughout the experiment, and the DAI score was calculated according to the scoring criteria in Table 2.

[0066] DAI = weight loss rate + stool characteristics + blood in stool

[0067] Table 2 DAI scoring criteria

[0068]

[0069] (4) 16S rRNA sequencing

[0070] Mouse intestinal contents were collected using centrifuge tubes and stored frozen at −80°C. 16S rRNA sequencing of the mouse intestinal microbiota was performed in collaboration with Shanxi Xinzhida Technology Development Co., Ltd.

[0071] 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.

[0072] The results of the response surface experiment are as follows: The experimental group designs and the yield results of tea lovage polysaccharides 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 for tea lovage polysaccharides was obtained.

[0073] Table 3 Results of response surface analysis to optimize the extraction yield of polysaccharides from Rhizoma Chuanxiong

[0074]

[0075] SAS 8.0 software was used to analyze the data in Table 3 and 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:

[0076]

[0077] By analyzing the above binary regression equation, the optimal extraction process of tea xiong polysaccharide was finally determined to be: 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).

[0078] Table 4 Optimal extraction process of angelica sinensis polysaccharide

[0079]

[0080] The results of the evaluation of Chaxiong polysaccharide on the body weight and DAI score of mice are as follows ( Figure 3 During the experiment, the mice in group C were in a stable state, with shiny fur and steady weight gain. The mice in group M were listless and had severe bloody stools. After the administration of sulfasalazine and chaxiong polysaccharide, the mice's condition improved. Figure 3 As shown in the results, compared with group C, the body weight of mice in group M decreased significantly, and the DAI score increased significantly (P < 0.0001). After the administration of tea xiong polysaccharide, the body weight change rate and DAI score of each tea xiong polysaccharide dose group also showed similar trends compared with group M, with the body weight of mice increasing significantly and the DAI score decreasing significantly. Compared with group C, the colon length of mice treated with DSS was significantly shortened (P < 0.0001). After the administration of tea xiong polysaccharide, the colon length was significantly restored (P < 0.0001).

[0081] The results of the regulation of Chaxiong polysaccharide on the intestinal flora of mice are as follows: Rarefaction curve ( Figure 4 ) The curve is flat, indicating that the sequencing results are sufficient to reflect the diversity of the bacterial communities contained in the current sample. Principal coordinate analysis (PCoA analysis) and non-metric multidimensional scaling (NMDS) analysis were performed. The greater the distance, the greater the difference in the microbial communities between the two samples. From the PCoA results Figure 5 and NMDS results Figure 6It can be seen that the microbial abundance of each group is clearly separated, indicating that the intestinal microbial composition of each experimental group is different. The stress coefficient results show that its stress value is 0.0000971, indicating a good fitting effect.

[0082] The composition of mouse intestinal microorganisms was analyzed at the phylum level, such as Figure 7 As shown, group C (normal group) was primarily composed of Bacteroidetes, Firmicutes A and D, and other phyla. Compared to group C, the relative abundance of Bacteroidetes in group M decreased from 35.51% to 28.27%, and pathogenic Proteobacteria emerged in the group with a relative abundance of 17.53%. Compared to group M, the relative abundance of Bacteroidetes in group CX increased to 51.70%, while the relative abundance of Proteobacteria decreased to 1.17%. Species composition analysis suggests that chaxiong polysaccharides can intervene in UC by increasing the relative abundance of Bacteroidetes and reducing the relative abundance of pathogenic Proteobacteria.

[0083] like Figure 8 As shown in the figure, the petal plot revealed 157 overlapping ASV / OTUs between groups C, M, SASP, and CX. Furthermore, group C had 4,440 unique ASV / OTUs, group M had 2,820 unique ASV / OTUs, group SASP had 3,088 unique ASV / OTUs, and group CX had 3,110 unique ASV / OTUs. These results suggest that DSS induces a decrease in the diversity of the intestinal microbiota, while tea lovage polysaccharides can reverse this decrease and maintain a balanced intestinal microbiota.

[0084] from Figures 9-14The relative abundance of microbial composition at the genus level revealed different species composition in the gut microbiota between mice in different groups. In group C, Amulumruptor, Mailhella, CAG-510, and Lactobacillus were more abundant. Escherichia, Helicobacter D, and Bacteroides H were more abundant in mice with DSS-induced UC. Bifidobacterium and Phocaeicola A were more highly expressed in the SASP group. Prevotella and Duncaniella were more abundant in mice with UC treated with Chaxiong polysaccharide. Comparing the differences in the composition of the intestinal microbiome among the groups, the relative abundance of Amulumruptor, Duncanella, and Prevotella in groups C and CX were significantly increased compared with group M (P < 0.0001), indicating that tea lovage polysaccharide may improve intestinal inflammation in UC mice by upregulating Amulumruptor, Duncanella, and Prevotella, and downregulating Escherichia and Helicobacter.

[0085] Through LEfSe (LDA Effect Size) branch diagram ( Figure 15 ) and LDA histogram analysis were used to identify species with significant differences between groups at all levels. The LDA threshold was set at 4.0. Compared with group C ( Figure 16 ), 50 groups of intestinal microbial flora in group M changed (22 groups up-regulated, 28 groups down-regulated), the abundance of Proteobacteria (p_Proteobacteria), Gammaproteobacteria (Gamnaproteobacteria), Enterobacteriaceae (Enterobacteriaceae_A), Enterobacterales (Enterobacterales_A), Escherichia (Escherichia) increased; Lachnospiraceae (Lachnospiraceae), Lachnospirales (Lachnospirales), f_Muribaculaceae, Lactobacillaceae (Lactobacillaceae) decreased. Compared with group M ( Figure 17), 38 groups of intestinal microbial flora in the CX group changed (20 groups upregulated and 18 groups downregulated), including Bacteroidetes (p_Bacteroidota), Bacteroidetes (c_Bacteroidia), Bacteroidetes (o_Bacteroidales), f_Muribaculaceae, Faecalibaculum (g_Faecalibaculum), etc. The abundance of those increased included Bacteroidetes (p_Bacteroidota), Bacteroidetes (c_Bacteroidia), Bacteroidetes (o_Bacteroidales), f_Muribaculaceae, Faecalibaculum (g_Faecalibaculum), etc. Studies have shown that Faecalibaculum can produce short-chain fatty acids (SCFA). These short-chain fatty acids can regulate the immune system, promote the growth of beneficial bacteria, and inhibit the reproduction of harmful bacteria; the abundance of Proteobacteria (p_Proteobacteria), Gammaproteobacteria (c_Gamnaproteobacteria), Enterobacteriales (o_Enterobacterales_A), Enterobacteriaceae (f_Enterobacteriaceae_A), Escherichia (g_Escherichia), etc. was downregulated, reversing the changes in the microbial structure caused by intestinal inflammation and promoting the restoration of the normal structural level of the intestinal microbial flora in UC mice. It can be inferred that tea xiong polysaccharide may improve ulcerative colitis in mice by upregulating the relative abundance of probiotics such as Bacteroidetes and Faecalibacterium prausnitzii, and downregulating the relative abundance of pathogenic bacteria such as Proteobacteria, Gammaproteobacteria, Enterobacteriaceae and Escherichia.

[0086] At the same time, the components of the tea lovage proteoglycan prepared by the optimal process in Example 1 were analyzed, and the analysis results are shown in Figure 18 As can be seen from the figure, the tea lovage polysaccharide of this embodiment is mainly composed of six monosaccharides: mannose, rhamnose, galacturonic acid, glucose, galactose, and arabinose. According to the peak area ratio, the proportion of each monosaccharide is 0.518%, 0.694%, 6.396%, 82.160%, 4.347%, and 5.886%, respectively. The results show that the tea lovage polysaccharide is a natural polysaccharide mainly composed of glucose.

[0087] Unlike other natural polysaccharides, the existing technology improves enteritis by regulating the relative abundance of bacteria such as Bacteroidetes and Firmicutes. The tea lovage polysaccharide of this application mainly improves ulcerative colitis by regulating the relative abundance of intestinal flora such as Bacteroidetes, Faecalibacterium prausnitzii, Proteobacteria, Gammaproteobacteria, Enterobacteriaceae and Escherichia.

[0088] The above-described embodiments merely represent preferred implementations of the present invention. While the descriptions thereof are relatively specific and detailed, they should not be construed as limiting the scope of the present invention. It should be noted that a person skilled in the art would be able to make various other corresponding changes and modifications based on the technical solutions and concepts described above, and all such changes and modifications should fall within the scope of protection of the claims of the present invention.

Claims

1. A use of tea xiong polysaccharide in the preparation of a drug for regulating intestinal flora in inflammatory bowel disease, characterized in that: The tea lovage polysaccharide is composed of 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; The extraction method of the tea lovage polysaccharide comprises the following steps: S1: crushing the Chaxiong slices and sieving to obtain Chaxiong powder; S2: extracting the tea lovage powder with water to obtain an extract, filtering the extract while hot, combining the tea lovage filtrates, concentrating under reduced pressure, precipitating with alcohol, letting it stand and then filtering, re-dissolving the precipitate with water to obtain a 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 tea lovage polysaccharide; 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 2-3 hours; In step S3, the cut-off relative molecular weight of the dialysis bag is 8000-14000.

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

3. The use of a tea lovage polysaccharide according to claim 2 in the preparation of a drug for regulating intestinal flora in inflammatory bowel disease, characterized in that: 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.

4. The use of a tea lovage polysaccharide according to claim 1 in preparing a drug for regulating intestinal flora in inflammatory bowel disease, characterized in that: The dosage form of the medicine is capsule, tablet, granule, injection, sustained-release preparation, oral solution or pill.

5. A method for extracting tea lovage polysaccharide used in the application of claim 1, characterized in that: The following steps are involved: S1: crushing the Chaxiong slices and sieving to obtain Chaxiong powder; S2: extracting the tea lovage powder with water to obtain an extract, filtering the extract while hot, combining the tea lovage filtrates, concentrating under reduced pressure, precipitating with alcohol, letting it stand and then filtering, re-dissolving the precipitate with water to obtain a tea lovage crude polysaccharide solution; S3: After removing protein with chloroform and n-butanol, the product was purified by dialysis bag and freeze-dried to obtain the tea lovage polysaccharide; 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 2-3 hours; In step S3, the cut-off relative molecular weight of the dialysis bag is 8000-14000.

6. The extraction method according to claim 5, characterized in that In the step S1, the product is passed through a 100-200 mesh sieve.

7. The extraction method according to claim 5, characterized in that 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% by volume of alcohol content.

8. The extraction method according to claim 5, characterized in that In step S3, the volume ratio of chloroform to n-butanol is 4:1.

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