Application of spinosad neutral polysaccharide in preparing medicine for preventing and / or treating ulcerative colitis

Abstract

The present invention provides an application of a thorn sugar neutral polysaccharide in preparing a drug for preventing and / or treating ulcerative colitis, and belongs to the field of medical technology. Animal experimental results show that after the thorn sugar neutral polysaccharide is used in a mouse model of ulcerative colitis, the phenomenon of colon shortening can be alleviated, the production of harmful bacteria can be inhibited, the abundance of Helicobacter genus can be reduced, the number of beneficial bacteria can be increased, and the abundance of Lactobacillus genus, Bacteroides genus and Oscillatory Spirochete genus can be increased. Therefore, the thorn sugar neutral polysaccharide of the present invention can be used as a drug for preparing a drug for preventing or treating ulcerative colitis.

Description

Technical Field

[0001] The invention belongs to the field of medical technology, and in particular relates to the application of spinosad neutral polysaccharide in preparing medicines for preventing and / or treating ulcerative colitis. Background Art

[0002] Ulcerative colitis (UC) is an inflammatory bowel disease characterized by long-term inflammation. It originates in the rectum and extends proximally to the colon, causing ulcers and bloody diarrhea. Clinical manifestations include diarrhea, abdominal pain, and even bloody stools. The course of the disease can last for decades or even a lifetime, which seriously reduces the quality of life of patients, shortens their lifespan, and increases the socioeconomic burden. Due to its high incidence, complexity, recurrence, difficulty in healing, and high risk of cancer in all age groups, the disease has become a global public health issue. The exact cause of ulcerative colitis is still unclear. The complex interaction of factors such as genetic susceptibility, impaired intestinal barrier, immune regulation disorders, and intestinal flora imbalance leads to the occurrence of the disease. Clinical first-line drugs such as aminosalicylic acids, hormones, and immunosuppressants have certain defects, limited efficacy, lack of specificity, and obvious toxic and side effects, leading to repeated attacks of ulcerative colitis and even gradual aggravation of the disease. Therefore, finding more effective and economical drugs for the treatment of ulcerative colitis is a hot topic and difficulty in research.

[0003] Studies in recent years have shown that Chinese herbal polysaccharides can effectively relieve and treat ulcerative colitis through a variety of processes, with the advantages of safety, clear effects, and few adverse reactions. Using polysaccharides as adjuvants for drugs or first-line drugs for the treatment of ulcerative colitis may be an effective treatment strategy that can overcome the shortcomings of current drugs for the treatment of intestinal diseases.

[0004] Alhagi Honey is extracted from Alhagi Honey, a plant of the Fabaceae family, Alhagi genus, and a semi-shrub. The leaves of Alhagi Honey are secreted and condensed into sugar particles. They are yellow-white in color and round or oval in shape. Alhagi Honey has a variety of biological activities, but domestic and foreign research on it focuses on enhancing immunity, antibacterial, and lowering blood sugar. There are no related reports on the treatment of ulcerative colitis. Summary of the invention

[0005] In view of this, the object of the present invention is to provide a use of a spinosad neutral polysaccharide in the preparation of a drug for preventing and / or treating ulcerative colitis.

[0006] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0007] Application of spinosad neutral polysaccharide in preparing medicine for preventing and / or treating ulcerative colitis.

[0008] Preferably, the extraction method of the neutral polysaccharide of the thorn sugar comprises: the thorn sugar is defatted, ethanol impurity removed, water extracted, dialyzed, alcohol precipitated, deproteinized, decolorized, and purified to obtain the neutral polysaccharide of the thorn sugar; the dialysis is carried out in a dialysis bag with a molecular weight cutoff of 500-2000Da, and the dialysis time is 12-36h.

[0009] Preferably, the alcohol precipitation comprises mixing the dialysate with anhydrous ethanol to obtain a mixed solution, and subjecting the mixed solution to alcohol precipitation at 2-5° C. for 20-30 hours; the volume percentage of ethanol in the mixed solution is 70-90%.

[0010] Preferably, the solid-liquid ratio of the water extraction is 1:5-10 g / mL, the temperature is 85-95° C., and the time is 0.5-1.5 h.

[0011] Preferably, the deproteinization adopts the Sevage method; and the decolorization adopts the shaking table static adsorption method.

[0012] Preferably, the purification comprises sequentially performing DEAE cellulose column elution separation and dextran gel column elution separation.

[0013] Preferably, the spinosad neutral polysaccharide can relieve the symptoms of shortened colon, loose stools and bloody stools in patients with ulcerative colitis.

[0014] Preferably, the neutral polysaccharide of Saccharum officinale can inhibit the production of harmful bacteria in the intestine and increase the number of beneficial bacteria.

[0015] Preferably, the spinose neutral polysaccharide can reduce the abundance of Helicobacter and increase the abundance of Lactobacillus, Bacteroides and Oscillatory Spirillum.

[0016] Compared with the prior art, the present invention has the following beneficial effects:

[0017] The present invention provides the use of a thorn sugar neutral polysaccharide in the preparation of a drug for preventing and / or treating ulcerative colitis. Animal experimental results show that after the thorn sugar neutral polysaccharide is used in a mouse model of ulcerative colitis, the phenomenon of colon shortening can be alleviated, the production of harmful bacteria can be inhibited, the abundance of Helicobacter can be reduced, and the abundance of Lactobacillus, Bacteroides and Oscillatory Spirochete can be increased. Therefore, the thorn sugar neutral polysaccharide of the present invention can be used as a drug for preventing or treating ulcerative colitis. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 is the glucose standard curve;

[0019] Figure 2 It is the UV spectrum of spinosa polysaccharide;

[0020] Figure 3 The following is the modeling flow chart;

[0021] Figure 4This is an analysis chart of the weight changes of mice;

[0022] Figure 5 is a mouse DAI analysis diagram; a is a representative visual diagram of mouse feces; b is a mouse DAI score diagram;

[0023] Figure 6 is the colon length of DSS-induced mice; a is a representative colon tissue picture; b is a colon length analysis chart;

[0024] Figure 7 is the colon pathological tissue analysis; a is the H&E staining of colon pathological tissue sections; b is the colon pathological histological scoring diagram;

[0025] Figure 8 This is the spleen index change diagram;

[0026] Fig. 9 Principal component analysis diagram of intestinal flora; graphs of different colors or shapes represent samples under different conditions; the scales of the horizontal and vertical axes are relative distances, representing the similarity distance of the samples under the influence of the principal component (PCo1 or PCo2);

[0027] Fig.10 It is a bar graph of the community composition of the intestinal flora at the genus level; each bar in the horizontal axis represents a sample, and the vertical axis represents the number of sequences at the classification level. The same color represents the same classification level;

[0028] Note: Figure 4-7 The data are expressed as mean ± SD. “###” compared with the control group, P < 0.001; “*” compared with the DSS-induced UC group, *P < 0.05, **P < 0.01, ***P < 0.001; DETAILED DESCRIPTION

[0029] The invention provides application of spinosad neutral polysaccharide in preparing medicine for preventing and / or treating ulcerative colitis.

[0030] In the present invention, the ulcerative colitis preferably includes ulcerative colitis.

[0031] In the present invention, the extraction method of the neutral polysaccharide of the thorn sugar preferably comprises: the thorn sugar is subjected to defatting, ethanol impurity removal, water extraction, dialysis, alcohol precipitation, deproteinization, decolorization, and purification to obtain the neutral polysaccharide of the thorn sugar; the dialysis is preferably carried out in a dialysis bag with a molecular weight cutoff of preferably 500-2000Da, more preferably 1000Da; the dialysis time is preferably 12-36h, more preferably 24h; the alcohol precipitation preferably comprises mixing the dialysate with anhydrous ethanol to obtain a mixed solution, preferably alcohol precipitation at 2-5°C for 20-30h, more preferably alcohol precipitation at 4°C The mixture is allowed to settle for 24 hours; the volume percentage of ethanol in the mixed solution is preferably 70-90%, more preferably 80%; the solid-liquid ratio of the water extraction is preferably 1:5-10 g / mL, more preferably 1:9 g / mL, the temperature is preferably 85-95°C, more preferably 90°C; the time is preferably 0.5-1.5h, more preferably 1h; the deproteinization is preferably carried out by the Sevage method; the decolorization is preferably carried out by a shaking table static adsorption method; the purification preferably includes sequentially performing DEAE cellulose column elution separation and dextran gel column elution separation. The present invention uses a 1000Da dialysis bag, which can just distinguish the neutral polysaccharide of thorn sugar from oligosaccharides, monosaccharides and proteins, and is convenient for removing sugars and proteins with smaller molecular weights, with less polysaccharide loss and higher yield; the dialysis time of 24 hours is the preferred time, less crude polysaccharides are obtained if the alcohol precipitation is insufficient for less than 24 hours, and more pigment tannin impurities are precipitated for more than 24 hours, which affects subsequent separation; the 80% ethanol precipitation of the present invention can precipitate most of the polysaccharides and improve the yield; the extraction pretreatment (petroleum ether defatting and ethanol impurity removal) of the present invention reduces the introduction of impurities and has a higher total sugar content; the present invention adopts shaking table static adsorption during decolorization, and the dialysis speed is fast and the amount is large, which improves efficiency and is more suitable for large-scale industrial applications.

[0032] In the present invention, the spiny sugar neutral polysaccharide can alleviate the symptoms of shortened colon, loose stools, and bloody stools in patients with ulcerative colitis; the spiny sugar neutral polysaccharide can inhibit the production of harmful bacteria in the intestine and increase the number of beneficial bacteria; the spiny sugar neutral polysaccharide can reduce the abundance of Helicobacter and increase the abundance of Lactobacillus, Bacteroides, and Oscillatory Spirochete. After the spiny sugar neutral polysaccharide is used in a mouse model of ulcerative colitis, the present invention can alleviate the phenomenon of shortened colon, inhibit the production of harmful bacteria, reduce the abundance of Helicobacter, increase the number of beneficial bacteria, and increase the abundance of Lactobacillus, Bacteroides, and Oscillatory Spirochete. The experimental results show that the spiny sugar neutral polysaccharide of the present invention has potential medicinal value in the treatment of ulcerative colitis.

[0033] The raw materials involved in the present invention have no special limitation on their sources, and commercially available products in the art may be used.

[0034] The technical solutions provided by the present invention are described in detail below in conjunction with the embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0035] Example 1

[0036] Extraction of Neutral Polysaccharides from Saccharum officinale

[0037] 1. Extraction of crude polysaccharides from Saccharum officinale

[0038] Take 100g of the medicinal material of the thorn sugar, dry and crush it, and degrease it by refluxing in a water bath at 60°C with petroleum ether in a ratio of 1:3. After filtering, the precipitate is refluxed in a water bath with 95% ethanol in a ratio of 1:3 to remove alcohol-soluble substances. After filtering, the precipitate is mixed with water in a ratio of 1:9 and refluxed in a constant temperature water bath at 90°C for 1h. After filtering, the filtrate is concentrated to 1 / 5 of the original volume by a rotary vacuum evaporator, dialyzed for 24h using a 1000Da molecular weight dialysis bag, and the dialysate in the bag is concentrated again, and anhydrous ethanol is added to 80% (v / v) alcohol precipitation concentration, and alcohol precipitation is carried out at 4°C for 24h. The precipitate is collected by filtration and centrifugation, and washed with ethanol, acetone and ether in turn, and finally a light yellow dry powder is obtained, which is the thorn sugar crude polysaccharide powder ASP.

[0039] The phenol-sulfuric acid method was used to determine the total sugar content in the polysaccharide samples.

[0040] (1) Draw a glucose standard curve: Prepare 40 μg / mL of glucose mother solution in a 500 mL volumetric flask, and pipette 0.4 mL, 0.6 mL, 0.8 mL, 1.0 mL, 1.2 mL, 1.4 mL, 1.6 mL, and 1.8 mL into 8 test tubes, respectively, and add water to 2.0 mL. Then add 1.0 mL of phenol solution (6%) and 5.0 mL of concentrated sulfuric acid, mix thoroughly, let stand at room temperature, and react for 20 minutes. Measure the absorbance A at 490 nm, and use water as a blank. Draw a glucose standard curve with the standard glucose concentration as the horizontal axis and the absorbance A as the vertical axis ( Figure 1 ), the regression equation is y = 0.5473x + 0.1365 (R 2 =0.9862). This indicates that within the range of 40-200 μg / mL, the standard glucose concentration has a good linear relationship with the absorbance value at 490 nm.

[0041] 4.6 mg of crude thorny sugar polysaccharide powder ASP was prepared into 1 mg / mL crude thorny sugar polysaccharide solution. The absorbance A at 490 nm was measured to be 2.324. The total sugar content in the sample was 3.9969 mg after being substituted into the standard curve. After calculation (extraction rate / %=total sugar content / sample mass×100%), the total sugar extraction rate in the crude thorny sugar polysaccharide was 81.57%.

[0042] 2. Purification of crude polysaccharides from Saccharum officinale

[0043] (1) Sevage method for protein removal

[0044] Kjeldahl nitrogen analyzer detection step 1 The protein content in the crude polysaccharide powder of the thorn sugar is 1.78%. Take the crude polysaccharide ASP obtained in step 1 and dissolve it in water. Add 1 / 5 volume of n-butanol-chloroform mixed solution to it, centrifuge at 4000r / min for 20min, remove the precipitate, repeat the operation for more than 3 times until the protein component is completely removed. The supernatant after removing the protein is concentrated under reduced pressure, add anhydrous ethanol to precipitate at 4°C for 24h, and vacuum freeze-dry.

[0045] Using UV spectroscopy to scan in the wavelength range of 185nm to 800nm, no absorption peaks were shown at 260nm and 280nm ( Figure 2 ), indicating that the protein was completely removed and there were no protein and nucleic acid impurities.

[0046] (2) Decolorization by static adsorption

[0047] The deproteinized crude polysaccharide of the spinosa truncatula was decolorized by using a macroporous resin (AB-8) using a static adsorption method. That is, 600 mL of the spinosa truncatula polysaccharide solution (25 mg / mL) was mixed with 250 g of the macroporous resin AB-8 (wet weight) in a conical flask, placed on a shaker, and shaken at room temperature at 200 rpm for 12 hours. The filtrate was collected by filtration and dialyzed in deionized water for 72 hours until the charge in the deionized water no longer changed. Finally, the dialyzate was concentrated and collected, and freeze-dried.

[0048] (3) DEAE cellulose column elution

[0049] A glass chromatography column with a specification of 2.6cm×60cm was used. 5g of crude polysaccharide powder was weighed and dissolved in 50mL of ultrapure water. The supernatant was centrifuged and added to the sample. The sample was eluted with ultrapure water and 0.1-0.9mol / L gradient NaCl solution, respectively. The elution rate was 1.0mL / min, and 15mL of eluent was used in each tube. The polysaccharide content in each eluent was detected at 490nm by the phenol-sulfuric acid method, and the elution curve was drawn with the absorbance value versus the tube number. The eluents were combined according to the elution curve, and the components eluted with the aqueous solution were collected.

[0050] (4) Sepharose column elution

[0051] Weigh 5g of the thorn sugar polysaccharide component after elution from the DEAE cellulose column, completely dissolve it with 20mL ultrapure water, centrifuge, take the supernatant, and slowly drip it along the tube wall with a rubber-tipped dropper. Elute with ultrapure water at a flow rate of 0.3mL / min, 10mL per tube, and collect 30 tubes. Follow up detection at 490nm using the phenol-sulfuric acid method, combine the eluates, concentrate under reduced pressure, and freeze-dry. Refined thorn sugar neutral polysaccharide is obtained.

[0052] Example 2

[0053] Animal experiments

[0054] 1. Animal grouping and drug administration

[0055] The experimental process is as follows Figure 3 As shown, 56 Kunming mice, all male, weighing 18-22g, 8 mice in each group, were randomly divided into:

[0056] Model group (Control): 4% DSS (dextran sulfate sodium) was continuously administered by gavage until the end of administration, and water was freely available during the period;

[0057] Blank control group (DSS): free drinking water every day, and normal saline was given by gavage on the 9th day of modeling;

[0058] Positive control group (5-ASA): 0.7 mL of 4% DSS solution was intragastrically administered 4 times a day, and 10 mg / mL 5-aminosalicylic acid (120 mg / mL) was intragastrically administered on the 9th day of modeling;

[0059] Three groups were treated with T. spinulosa: low-dose (200 mg / kg, AP-L), medium-dose (400 mg / kg, AP-M), and high-dose (800 mg / kg, AP-H) T. spinulosa polysaccharide. Each group was intragastrically administered with 0.7 mL of 4% DSS solution 4 times a day. After the 9th day of modeling, different doses of T. spinulosa polysaccharide aqueous solution were intragastrically administered, and 4% DSS was given daily.

[0060] There were 8 mice in each of the above groups;

[0061] Experimental group of co-housing: 4 mice with ulcerative colitis induced by 4% DSS but not treated (Cohouse+DSS), and 4 mice with ulcerative colitis induced by 4% DSS but treated with high-dose of Acanthopanax polysaccharide (Cohouse+AP-H), marked them and placed them in the same cage to observe the changes in the mice without treatment and the mice treated with treatment.

[0062] 2. Evaluate the disease changes in mice

[0063] The weight, fecal characteristics and blood in the stool of mice were recorded every day to evaluate the disease activity index (DAI). The DAI value is the average of the scores of weight loss percentage, stool shape and occult blood in stool.

[0064] The percentage of weight loss of experimental mice (%): no change in weight was recorded as 0 points; 1-5 points were recorded as 1 point; 6-10 points were recorded as 2 points; 11-15 points were recorded as 3 points; more than 15 points were recorded as 4 points;

[0065] Normal mouse stool was scored as 0 points; loose stool was scored as 2 points; and watery stool was scored as 4 points;

[0066] Normal mouse stool and no anal bleeding were scored as 0 points; positive occult blood was scored as 2 points; and overt bleeding was scored as 4 points, as shown in the following table.

[0067] Table 1 Disease activity index scoring criteria

[0068]

[0069] 3. Sample collection

[0070] On the 22nd day, the mice were fasted for 12 hours and blood was collected from their eyeballs. The supernatant was centrifuged at 3000r / min for 10 minutes at 4°C and stored at -80°C for biochemical analysis. Fresh feces of the mice were collected and stored at -80°C for intestinal flora detection. After the administration, the mice were killed by cervical dislocation, and the colon from the cecum to 1 cm above the anus was removed. The colon tissue of the mice was collected, the colon length was measured, and it was fixed in 4% paraformaldehyde solution for HE staining and tissue section observation.

[0071] Spleen index measurement

[0072] The spleen was dissected, washed and weighed, and the mouse spleen index was calculated. The spleen index was the ratio of the spleen to the total weight of the mouse.

[0073] 4. Histopathological analysis

[0074] At the end of the drug administration, it was observed that mice in the same cage group voluntarily ate the feces of their own kind.

[0075] After the administration, mice were killed by cervical dislocation, and the colon was removed from the cecum to 1 cm above the anus, and its length was measured. After the colon tissue was fixed in formalin, it was embedded according to the conventional method, cut into slices with a thickness of about 4-5 μm, and fixed on an adhesive slide. Hematoxylin-eosin (H&E) staining was used. First, the slides were dewaxed in xylene for 5 minutes; then placed in different concentrations of ethanol for 5 minutes each, washed with water for 2 minutes; then blued with hematoxylin for 2 minutes, rinsed with tap water; then placed in hydrochloric acid alcohol for a few seconds, rinsed with tap water; then stained with eosin stain for 2 minutes, rinsed with tap water twice; finally dehydrated, transparent, and sealed with neutral gum, and histological observation and analysis were performed under a microscope. Inflammation scores were scored according to body weight, fecal characteristics, and blood in stool. The data were compared using the one-way analysis of variance (anova) method. P<0.05 was considered statistically significant. All statistical analyses were performed using SPSS software and Graph-pad prism7 software, and the results are expressed as mean ± SEM (standard error of the mean). Figures 4 to 8 .

[0076] Depend on Figures 4 to 8It can be seen that compared with the blank control group, the DAI score of the model group was significantly increased, the colon length was significantly shortened, and the mice had blood in their stools, listlessness, and messy hair, indicating that the colon of the mice was inflamed. After treatment with three different concentrations of thorny sugar neutral polysaccharides, the DAI scores of the mice decreased to varying degrees, the phenomenon of shortened colon length was alleviated, and the symptoms of inflammation were alleviated.

[0077] In this field, colon length and spleen index are important indicators of colitis inflammation phenotype. Ulcerative colitis is often manifested by shortened colon, bloody stools, and intestinal flora imbalance. Restoring shortened colon and adjusting the intestinal flora environment can directly explain the relationship with ulcerative colitis. Shortened colon and adjusted intestinal flora environment can prove that the effect of prevention and / or treatment can be achieved; during the same cage, animals feed on feces (coprophagy), or ingest feces through self-cleaning, which can indicate that the animal microbial community is homogenized, and the effect of treating colitis inflammation can be achieved.

[0078] like Figure 4 As shown, the weight of mice in the control group gradually increased, while the weight of mice in the DSS group continued to decrease from day 8. In contrast, the AP-H group significantly improved the weight loss of mice in a dose-dependent manner (P<0.001).

[0079] Representative photos of feces are shown in Figure 5 a. Spinach neutral polysaccharide can alleviate the bloody and loose stools of DSS-induced ulcerative colitis mice, and the DAI score was significantly reduced after treatment with Spinach neutral polysaccharide (P < 0.001). The AP-H group and the Cohouse-AP800 group showed the most obvious disease recovery, and the effect was similar to that of the positive drug 5-aminosalicylic acid ( Figure 5 b).

[0080] Representative photographs of the colon are shown in Figure 6 a, indicating that thorn sugar neutral polysaccharide can alleviate the colon shortening of DSS-induced ulcerative colitis mice. Figure 6 As shown in b, compared with the control group mice, the colon length was significantly shortened after DSS treatment, 5.72±0.21 cm in the DSS group and 9.53±0.44 cm in the control group (P<0.001), while the 5-ASA group (7.48±0.55 cm) and AP-H group (7.95±0.74 cm) could significantly alleviate DSS-induced colon shortening (P<0.001).

[0081] Figure 7H&E staining showed that the colon tissue structure of the control group mice was basically normal, with intact colon mucosa and regular arrangement of goblet cells in the crypts. Compared with the control group, the mice in the DSS group had severe mucosal ulceration, inflammatory cells infiltrated into the mucosal tissue, goblet cells disappeared, and crypts were destroyed. These changes were reflected in significantly higher pathological scores (P<0.001). After treatment with spinosad neutral polysaccharide, the colonic lesions were improved and the pathological scores decreased (P<0.001). However, the AP-L group still showed more severe mucosal ulceration, inflammatory cell infiltration, and crypt cell loss, and the pathological score did not decrease significantly. In contrast, the AP-H group, Cohouse+DSS group, and Cohouse+AP-H group showed more obvious improvements, showing significantly reduced mucosal ulceration, reduced inflammatory cell infiltration, and crypt recovery.

[0082] like Figure 8 As shown, compared with the control group (2.71±0.23 mg / g), the spleen of mice in the DSS group was significantly enlarged (8.63±1.52 mg / g), while the 5-ASA group (4.92±1.17 mg / g), AP-H group (4.83±0.93 mg / g), and Cohouse-AP800 group (5.25±0.91 mg / g) could significantly reduce spleen enlargement (P<0.001).

[0083] In short, schisandra neutral polysaccharide can treat DSS-induced ulcerative colitis in a dose-dependent manner, and 400mg / kg and 800mg / kg doses of schisandra neutral polysaccharide can significantly alleviate the symptoms of DSS mice. And co-housing with mice treated with 800mg / kg schisandra neutral polysaccharide can increase the recovery of DSS-induced ulcerative colitis symptoms in mice, indicating that schisandra neutral polysaccharide may achieve therapeutic effects by changing intestinal flora.

[0084] 5. 16S rRNA gene sequencing and microbiome analysis

[0085] The V3-V4 region of the bacterial 16S ribosomal RNA (rRNA) gene was amplified using FastPfu primers (338F and 806R) polymerase PCR. The amplicon was then purified by gel extraction and quantified using quantitative fluorescence-st. The purified amplicons were pooled at equimolar concentrations and sequenced end-to-end using an Illumina MiSeq instrument.

[0086] Microbiome analysis results Figures 9-10 shown.

[0087] Immune regulation disorder is the key direct pathogenesis of ulcerative colitis, and intestinal flora is an important triggering factor in this immune damage process. Fig. 9The test results show that there are significant differences in the composition structure between the control group and the DSS group. DSS destroyed the intestinal microbial community structure, while the intestinal microbial community of each treatment group moved along the PCo1 and PCo2 directions. As the dose of strychnine neutral polysaccharides increased, the clustering of the microbial community moved closer to the control group, indicating that strychnine neutral polysaccharides protected the ecological disorder caused by DSS. The microbial community structure of the Cohouse+DSS group and the Cohouse+AP-H group changed, and was closer to the control group than the AP-H group. Fig.10 The intestinal flora of mice was analyzed at the genus level. Lactobacillus, Bacteroides, Prevotella and Helicobacter were the most abundant genera. The intestinal flora of mice in the DSS group was seriously unbalanced. Compared with the Control group, the composition and structure of the intestinal flora of mice in the DSS group changed greatly. The abundance of Lactobacillus, Bacteroides and Oscillatory Spirochete was significantly reduced, and the abundance of Helicobacter was significantly increased. Compared with the DSS group, the abundance of Helicobacter in each group decreased after treatment with spiny sugar neutral polysaccharide, while the abundance of Lactobacillus, Bacteroides and Oscillatory Spirochete increased. Among them, the effect of AP-H group increased, and the flora level was reversed, indicating that spiny sugar neutral polysaccharide can inhibit the production of harmful bacteria and regulate the intestinal flora by increasing beneficial bacteria.

[0088] In summary, spiny saccharide polysaccharide has a certain protective and preventive effect on DSS-induced ulcerative colitis, improves the changes and deterioration of the number and structure of intestinal flora in mice caused by ulcerative colitis, increases the beneficial bacteria in the intestine, and thus significantly improves the pathological symptoms of ulcerative colitis. At the same time, the same cage-reared group of mice voluntarily eats the same kind of feces, which also verifies that the microorganisms in the mouse feces can also influence each other and regulate the structure and number of microorganisms in the intestine.

[0089] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. The use of spinosad neutral polysaccharide in the preparation of a drug for treating ulcerative colitis, characterized in that: The extraction method of the neutral polysaccharide of the thorn sugar comprises: defatting, removing impurities with ethanol, extracting with water, dialysis, alcohol precipitation, deproteinizing, decolorizing and purifying the thorn sugar to obtain the neutral polysaccharide of the thorn sugar; The dialysis was performed in a dialysis bag with a molecular weight cut-off of 1000 Da, and the dialysis time was 24 h; The neutral polysaccharide of thorn sugar can inhibit the production of harmful bacteria in the intestine and increase the number of beneficial bacteria; the neutral polysaccharide of thorn sugar can reduce the abundance of Helicobacter and increase the abundance of Lactobacillus, Bacteroides and Oscillatory Spirillum; The alcohol precipitation comprises mixing the dialysate with anhydrous ethanol to obtain a mixed solution, and alcohol precipitation at 2-5° C. for 20-30 hours; the volume percentage of ethanol in the mixed solution is 70-90%; The water extraction has a solid-liquid ratio of 1:5-10 g / mL, a temperature of 85-95° C., and a time of 0.5-1.5 h; The deproteinization adopts the Sevage method; the decolorization adopts the shaking table static adsorption method; The purification comprises sequentially performing elution separation using a DEAE cellulose column and elution separation using a dextran gel column.

2. The use according to claim 1, characterized in that: The spinosad neutral polysaccharide can relieve the symptoms of shortened colon, loose stools and bloody stools in patients with ulcerative colitis.

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