Identification method of honeysuckle flower polysaccharide and application of honeysuckle flower polysaccharide in preparation of medicine for preventing or treating inflammatory bowel disease

Abstract

The invention belongs to the field of biological medicines, and particularly discloses an identification method of honeysuckle flower polysaccharide and application of the honeysuckle flower polysaccharide in preparation of medicines for preventing or treating inflammatory bowel diseases. A DSS solution is used for inducing a mouse inflammatory bowel disease model, honeysuckle polysaccharide FLP is used for gavage treatment, and mouse weight, disease activity indexes, colorectal tissue pathological structures, spleen pathological structures, HE staining, intestinal microbial structures, intestinal metabolite spectrums and the like are detected and analyzed. Results show that the honeysuckle flower polysaccharide can delay inflammatory bowel diseases of model animals in an oral administration manner, and has the characteristics of high targeting property, convenience in administration and the like. Good application prospects are realized in the aspect of treating the inflammatory bowel disease.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine, and specifically discloses a method for identifying Flos Lonicerae polysaccharide and its application in the preparation of drugs for preventing or treating inflammatory bowel disease. Background Art

[0002] Inflammatory bowel disease (IBD) is a chronic non-specific inflammatory disease, characterized by intestinal inflammation, tissue damage, abdominal pain, frequent and persistent diarrhea, weight loss, and rectal bleeding. According to the latest statistical data, in the past more than 20 years, the incidence and prevalence of IBD in China have shown an obvious increasing trend. It is estimated that by 2025, there will be 1.5 million IBD patients in the country.

[0003] Currently, the methods for treating inflammatory bowel disease clinically include traditional drugs, immunosuppressants, biological agents, antibiotics, and surgical treatment, etc. However, these methods still have their limitations, and there is an urgent need for new treatment methods to inhibit or delay the occurrence of inflammatory bowel disease clinically.

[0004] Traditional Chinese medicine believes that the occurrence of inflammatory bowel disease is closely related to etiologies such as qi stagnation, damp turbidity, and heat toxin. Therefore, the treatment should be mainly based on clearing heat and resolving dampness, cooling blood and promoting blood circulation to remove stasis, and astringing ulcers and promoting granulation. In recent years, natural polysaccharides have gradually become a new choice for treatment due to their good biological activities and low toxicity. As an important medicinal material in traditional Chinese medicine in China, Flos Lonicerae has been famous for its effects of clearing heat and detoxifying, cooling blood and stopping dysentery since ancient times. Modern scientific research reveals that Flos Lonicerae is rich in various bioactive components, among which polysaccharide compounds have attracted much attention due to their unique biological activities. Polysaccharides are high-molecular compounds formed by the connection of multiple monosaccharide molecules through glycosidic bonds, and have a wide range of biological functions, such as immune regulation, anti-inflammatory, antioxidant, etc. Flos Lonicerae Polysaccharide (FLP) is an effective component extracted from Flos Lonicerae, and has shown various pharmacological activities such as anti-inflammatory, antiviral, and antioxidant in diseases such as atopic dermatitis and allergic rhinitis.

[0005] For IBD, Flos Lonicerae Polysaccharide is also a promising treatment option. At present, there has been no in-depth analysis of its effect on inflammatory bowel disease, especially in terms of regulating the intestinal flora structure and metabolite profile. Summary of the Invention

[0006] In order to solve the above problems, the present invention discloses a method for identifying Flos Lonicerae polysaccharide and its application in the preparation of drugs for preventing or treating inflammatory bowel disease.

[0007] The technical solution of the present invention is as follows:

[0008] In the first aspect of the present invention, a method for identifying honeysuckle polysaccharide is provided. The method includes the following steps: subjecting the honeysuckle polysaccharide to infrared spectroscopy detection, monosaccharide composition detection, and transmission and scanning electron microscopy observation. The specific steps are as follows:

[0009] 1) Weigh the polysaccharide sample and potassium bromide according to a weight ratio of 1:50. After mixing evenly, press into a sheet with a thickness of 1 mm, and then use a Fourier transform infrared spectrometer for detection;

[0010] 2) Take a clean chromatographic vial, weigh the polysaccharide sample, add 2M trifluoroacetic acid (TFA) solution, and heat at 121 °C for 2 hours; pass nitrogen and dry; add 99.99% methanol for washing and then dry again. Repeat the methanol washing 2 - 3 times; add sterile water to dissolve and transfer it into the chromatographic vial for testing. The chromatographic system used is the Thermo ICS 5000+ ion chromatography system, and an electrochemical detector is used to analyze and detect the monosaccharide components; and compare the results with the standard products to qualitatively and quantitatively analyze the monosaccharide components in the polysaccharide. The volume - weight ratio of the polysaccharide sample to 2M trifluoroacetic acid (TFA) is 10 mg:1 ml;

[0011] 3) Dissolve a small amount of honeysuckle polysaccharide in 18.2 ultrapure water, then drop it onto a carbon - supported film. After natural drying, use a transmission electron microscope for observation to further understand the microstructure and properties of the honeysuckle polysaccharide FLP;

[0012] 4) Paste a small amount of honeysuckle polysaccharide powder on the sample stage, perform surface gold spraying treatment, and then use a scanning electron microscope for observation to further understand the microstructure and properties of the honeysuckle polysaccharide FLP.

[0013] Preferably, in step 1), the instrument resolution is 4.00 cm -1 , the scanning range is 4000 - 450 cm -1 , the number of scans is 32 times; the sampling gain is 8.0; the moving mirror speed is 0.4747; the aperture is 80.00; DTGS KBr detector; KBr beam splitter; infrared light source.

[0014] In the second aspect of the present invention, an application of honeysuckle polysaccharide in the preparation of drugs for preventing or treating inflammatory bowel disease is provided.

[0015] Preferably, in the above - mentioned application, every 1 mg of honeysuckle polysaccharide contains 4.6982 μg of Rha, 9.4605 μg of Gal, 10.6458 μg of Ara, 28.7586 μg of Gal - UA, and 78.8753 μg of Glu.

[0016] Preferably, in the above - mentioned application, it includes establishing a DSS - induced mouse IBD model to verify its effect.

[0017] Compared with the prior art, the present invention has the following beneficial effects:

[0018] The present invention discloses a method for identifying honeysuckle polysaccharide and its application in the preparation of drugs for preventing or treating inflammatory bowel disease. The honeysuckle polysaccharide is a polysaccharide component obtained from the traditional Chinese medicine honeysuckle. In the present invention, a mouse model of inflammatory bowel disease is induced by DSS solution, and the mice are treated by intragastric administration of honeysuckle polysaccharide FLP. The body weight, disease activity index, pathological structure of colorectal tissue, pathological structure of spleen, HE staining, intestinal microbial structure, intestinal metabolite profile, etc. of the mice are detected and analyzed. The results show that the honeysuckle polysaccharide can delay the inflammatory bowel disease of the model animals by oral administration, and has the characteristics of strong targeting and convenient administration. It has good application prospects in the treatment of inflammatory bowel disease. Brief Description of the Drawings

[0019] Figure 1 For the related identification of honeysuckle polysaccharide. Among them, A: Ion chromatogram analysis of FLP; B: Monosaccharide composition proportion diagram of FLP; C: Absolute content diagram of monosaccharides in FLP; D: Infrared spectrum identification diagram of FLP; E: Transmission electron microscope (TEM) morphology observation diagram of FLP; F: Scanning electron microscope (SEM) morphology observation diagram of FLP.

[0020] Figure 2 For the experimental results of verifying that honeysuckle polysaccharide can relieve DSS-induced colitis in mice. Among them: A: Line graph of mouse body weight; B: Evaluation of mouse disease activity index; C: Gross view of mouse colon; D: Gross view of mouse spleen; E: Western-blot detection of the expression of COX2, PCNA, Occludin, and Claudin-1 in colon tissue; F: qRT-PCR detection of the mRNA levels of inflammatory factors (IL-1β, IL-6, TNF-α, IL-10) in colon tissue; G: HE staining of spleen tissue (200×); H: HE staining of colon tissue (200×); I: IHC detection of the expression of PCNA in colon tissue. *P < 0.05, **P < 0.01, ***P < 0.001.

[0021] Figure 3Experimental results of honeysuckle polysaccharide in restoring the intestinal microbiota structure. Among them, A: Species rank abundance curve based on OTU; B: Dilution curve based on the total number of OTUs; C: Venn diagram of OTUs among the NC group, DSS group and FLP group; D: Relative abundance of intestinal microbiota species at the phylum level; E: ACE box plot of differences in α-diversity indices among groups; F: Analysis of β-diversity group differences based on bray-Curtis; G: PCoA analysis based on Weighted_Unifrac, UnweightedUnifrac and Bray Curtis distances; H: Cluster heat map of species abundances at the genus level within groups.

[0022] Figure 4 Experimental results of honeysuckle polysaccharide in alleviating intestinal metabolite disorders. Among them, A: PLS-DA score plot of metabolites in positive ion mode for three groups; B: PLS-DA score plot of metabolites in negative ion mode for three groups; C: PLS-DA model parameters in positive and negative ion modes; D: Proportion of the identified metabolites in each chemical classification; E: Venn diagram of differential metabolites among groups in positive ion mode; F: Venn diagram of differential metabolites among groups in negative ion mode; G: PCA analysis results of the overall samples in positive ion mode; H: PCA analysis results of the overall samples in positive ion mode; I: Volcano plot of differential metabolites between the NC group and DSS group in positive and negative ion modes; J: Volcano plot of differential metabolites between the FLP group and DSS group in positive and negative ion modes. Specific implementation manners

[0023] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

[0024] The reagents or instruments used in the embodiments of the present invention that are not marked with the manufacturer can all be conventional reagent products obtained through commercial purchase. Purchase source: Sichuan Vic Biotechnology Co., Ltd.; Product number: WKQ-0008285

[0025] Example 1

[0026] 1 Identification of honeysuckle polysaccharide

[0027] The honeysuckle polysaccharide was purchased from Sichuan Vic Biotechnology Co., Ltd. (WKQ-0008285). The identification method includes the following steps: infrared spectrum detection, monosaccharide composition detection, and transmission and scanning electron microscopy observation of the honeysuckle polysaccharide.

[0028] ①Weighed 4 mg of the polysaccharide sample and mixed it with 200 mg of potassium bromide (7758-02-3, Sigma, USA). After that, it was pressed into a sheet with a thickness of 1 mm, and then detected using a Fourier transform infrared spectrometer (Nicolet iZ-10, Thermo, USA). The instrument resolution was 4.00 cm-1, the scanning range was 4000 - 450 cm-1, and the number of scans was 32 times. The sampling gain was 8.0; the moving mirror speed was 0.4747; the aperture was 80.00; DTGS KBr detector; KBr beam splitter; infrared light source.

[0029] ②Took a clean chromatographic vial, weighed 10 mg of the polysaccharide sample, added 1 ml of 2M trifluoroacetic acid (TFA) solution (76-05-1, ANPEL, CHINA), and heated it at 121 °C for 2 hours. Passed nitrogen and dried it. Added 99.99% methanol (67-56-1, ANPEL, CHINA) for cleaning and then dried it again. Repeated the methanol cleaning 2 - 3 times. Added sterile water to dissolve it and transferred it into the chromatographic vial for measurement. The chromatographic system used was the Thermo ICS 5000+ ion chromatographic system (ICS 5000PLUS, Thermo, USA), and the electrochemical detector was used to analyze and detect the monosaccharide components. And the results were compared with the standards to qualitatively and quantitatively analyze the monosaccharide components in the polysaccharide.

[0030] ③Dissolved a small amount of honeysuckle polysaccharide in 18.2 ultrapure water, then dropped it onto the carbon support film (BZ1102, EMCN, CHINA). After natural drying, it was observed using a transmission electron microscope (JEOL 2100PLUS, JEOL, JAPAN) to further understand the microstructure and properties of FLP.

[0031] ④Pasted a small amount of polysaccharide powder on the sample stage, carried out surface gold spraying treatment, and then observed it using a scanning electron microscope (MIRA, TESCAN, Czech) to further understand the microstructure and properties of FLP.

[0032] 2 Identification results

[0033] FLP is a complex polysaccharide composed of glucose (Glu), galacturonic acid (Gal-UA), arabinose (Ara), galactose (Gal), and rhamnose (Rha) Figure 1 A). Among them, Glu accounted for the largest proportion among all monosaccharides, accounting for 59.56%, and the rest were Gal-UA accounting for 21.71%, Ara accounting for 8.04%, Gal accounting for 7.14%, and Rha accounting for 3.55% Figure 1B). Quantitative detection and analysis revealed that 1 mg of honeysuckle polysaccharide contained 4.6982 μg of Rha, 9.4605 μg of Gal, 10.6458 μg of Ara, 28.7586 μg of Gal-UA, and 78.8753 μg of Glu( Figure 1 C). In addition, the infrared spectrum identification map( Figure 1 D) The functional groups and structural characteristics in FLP were revealed by characteristic absorption peaks, providing strong support for further confirmation of its chemical structure. It can be seen from the infrared spectrum of honeysuckle polysaccharide that the absorption peak at 3260.96 cm-1 is the stretching vibration absorption peak of O-H, with a large intensity and a relatively wide peak. The absorption peaks in this region are characteristic peaks of carbohydrates. The absorption peak at 2920.84 cm-1 belongs to the stretching vibration of C-H; a benzene ring skeletal vibration absorption peak appears at 1592.22 cm-1, which is one of the most important peaks for determining the presence of a benzene ring; an absorption peak formed by the coupling of in-plane symmetric vibration of C-H, in-plane vibration of O-H, and stretching vibration of C=O appears at 1372.77 cm-1; an in-plane bending vibration absorption peak of O-H appears at 1260.04 cm-1; an absorption peak appears at 1043.03 cm-1, which belongs to the stretching vibration of C-O. To more intuitively understand the microstructure of FLP, transmission electron microscopy (TEM) and scanning electron microscopy (SEM) were also used for observation. It can be seen from the TEM image that the FLP powder was dissolved in deionized water for sample preparation, and it showed a relatively unique chain-like and network structure under the electron microscope( Figure 1 E). When observed by SEM at a magnification of 1500 times, FLP showed the common irregular flaky structure of polysaccharides, with some debris attached to the surface. When the magnification was 5000 and 10000 times, it could be seen that FLP was still relatively smooth, and the debris was attached to the surface. When the magnification was 20000 times, some fine cracks were observed on the surface of FLP. The morphology and structural details at the nanoscale, as well as the surface morphology and characteristics( Figure 1 F).

[0034] Example 2: Using the honeysuckle polysaccharide of Example 1 to treat the DSS-induced mouse inflammation model

[0035] An IBD model of mice was induced by DSS, and the intervention was carried out by intragastric administration of the aqueous solution of honeysuckle polysaccharide. The experiment was divided into a normal control group (NC group), a DSS-induced inflammation group (DSS group), and a honeysuckle polysaccharide intervention group (FLP group): 6-week-old male BALB / C mice were fed.

[0036] Except for the NC group, the other two groups started drinking 3% DSS solution from the first day. In the FLP group, 0.1 ml (40 mg / ml) was orally administered to each mouse every day, and the other two groups were given the same volume of sterile water. On the 10th day, all mice were sacrificed by cervical dislocation and tissue specimens such as the colorectum were isolated for further detection and analysis. According to the equivalent dose ratio table calculated by converting the body surface area of humans and animals, if it is applied to clinical in the future, it will probably require 28 - 38 ml of honeysuckle polysaccharide aqueous solution at 40 mg / ml.

[0037] 1. Observation indicators

[0038] The indicators were observed and recorded daily. The body weight change, stool characteristics, occult blood or blood in the stool of mice were observed daily, and the body weight change, change in stool characteristics, and presence or absence of blood in the stool were used as the disease activity index (DAI) to evaluate the symptoms of mice. The degree of damage to the colonic mucosa was observed under HE staining microscopy.

[0039] 2. Experimental results

[0040] By observing the body weight change of mice ( Figure 2 A), it was found that the body weight of mice in the DSS group continued to decline, while the downward trend of body weight in the FLP-treated group was significantly alleviated, preliminarily indicating that FLP has a relieving effect on IBD. The disease activity index (DAI) of mice further confirmed that FLP can reduce the IBD of mice ( Figure 2 B). The colon length of mice in the DSS group was shortened ( Figure 2 C), and the spleen was significantly enlarged ( Figure 2 D), while these pathological changes in the FLP-treated group of mice were significantly improved. In order to further explore the effect of FLP on the injury and repair of colon tissue, Western-blot analysis was performed. The results showed that FLP could inhibit the expression of the inflammation-related protein COX2, and at the same time promote the expression of the cell proliferation marker PCNA and the tight junction proteins Occludin and Claudin-1, indicating that FLP is helpful for the repair of colon tissue and the restoration of barrier function ( Figure 2 E). In addition, by detecting the mRNA levels of inflammatory factors in colon tissue by qRT-PCR, it was found that FLP could down-regulate the expression of pro-inflammatory factors IL-1β, IL-6, TNF-α, and up-regulate the expression of the anti-inflammatory factor IL-10, thereby regulating the inflammatory response of colon tissue ( Figure 2 F). At the histological level, the HE staining results of the spleen and colon tissue ( Figure 2 G,H) showed that the number and structure of lymphoid nodules in the spleen of mice in the FLP-treated group were restored, and the infiltration of inflammatory cells was reduced. The structure of the colon tissue was restored clearly, and the crypt structure was complete, further verifying the repair effect of FLP on tissue injury. Finally, the expression of PCNA in colon tissue was detected by IHCFigure 2 I), it was found that FLP could promote the proliferation of colon tissue cells, providing strong support for tissue repair and regeneration. In summary, these experimental results fully demonstrated that FLP had a significant alleviating effect on DSS-induced murine IBD.

[0041] Example 3

[0042] Regulating the intestinal flora in a DSS-induced murine inflammation model with the honeysuckle polysaccharide of Example 1

[0043] The 16S rDNA amplicon sequencing technology was adopted to analyze murine fecal samples. Microbial rank abundance curves were used to characterize the abundance and evenness of species. It could be seen that the flora abundance of the NC group was the highest, followed by the FLP group, while the DSS group had the lowest abundance ( Figure 3 A). The dilution curve tended to flatten with the increase in the number of sample sequences, indicating that the sequencing data volume of each group of samples was large enough and the results were reasonable. In addition, it could also be seen that under the condition of basically the same sequencing depth, the overall flora abundance of the FLP group was higher than that of the DSS group ( Figure 3 B). Through the Venn diagram analysis of OTUs, it was observed that there were 1639 OTUs in the NC group, while the number of OUTs in the DSS group decreased significantly, only 1371. The FLP group had 1744 OTUs, significantly restoring the flora diversity ( Figure 3 C). Previous studies have shown that the intestinal microbial community is mainly composed of Firmicutes, Bacteroidetes, Proteobacteria, and Actinobacteria, among which Firmicutes and Bacteroidetes are dominant. In IBD patients, the relative increase in the abundance of Proteobacteria is closely related to disease progression. The results of this study showed that after DSS treatment, the relative abundances of Bacteroidetes and Siderobacter were decreased, while the proportions of Proteobacteria, Desulfobacter, and Actinobacteria were increased. The addition of FLP effectively reversed these changes. In addition, FLP also significantly promoted the restoration of the abundances of microbial taxa such as Verrucomicrobia and unclassified bacteria ( Figure 3 D). The results of the clustering heatmap analysis further intuitively demonstrated the restoration effect of FLP on the microbial community structure. Multiple clusters in the FLP group were highly similar to the NC group, while the DSS group was significantly different in comparison ( Figure 3 H). In terms of α-diversity, the diversity index was significantly decreased after DSS treatment, while it was restored in the FLP group ( Figure 3 E). β-diversity analysis showed that there were significant differences between the DSS group and the NC group, while the differences between the FLP group and the NC group were not significant ( Figure 3F). PCoA analysis based on Weighted Unifrac, Unweighted Unifrac, and Bray Curtis distances also intuitively showed that FLP reversed the damage induced by DSS and approached the NC group. These findings together indicate that FLP can effectively alleviate colitis-induced microbial community imbalance, significantly promote the recovery of microbial diversity and structure, and play a positive role in regulating the intestinal microbial community structure and maintaining intestinal homeostasis.

[0044] Example 4

[0045] Regulating intestinal metabolites in a DSS-induced mouse inflammation model with the honeysuckle polysaccharide of Example 1

[0046] Using various methods such as PLS-DA (partial least squares discriminant analysis) and PCA (principal component analysis), the fecal metabolites of the DSS-induced IBD mouse model group, the colitis mouse model group treated with FLP (FLP group), and the normal control group (NC group) were deeply analyzed. The samples of the FLP group were significantly separated from those of the DSS group and were closer to the samples of the NC group, indicating that FLP treatment effectively regulated the metabolite profile of colitis mice and restored it to a normal state ( Figure 4 A, B). In addition, the Q2 values in the PLS-DA model parameters of this study model were all greater than 0.3, indicating that the model had high explanatory and predictive abilities, further confirming the reliability of the analysis results ( Figure 4 C). The identified metabolites covered 16 categories in total, including lipids and lipid-like molecules, organic acids and their derivatives, organic heterocyclic compounds, and oxygen-containing organic compounds. Lipids and lipid-like molecules constituted the largest metabolite group, accounting for 30.261%, which reflected the important role of intestinal microorganisms in fatty acid metabolism and lipid biosynthesis. Secondly, organic acids and their derivatives accounted for 20.577%, and these metabolites played a key role in energy metabolism, signal transduction, and intestinal environment regulation. The undefined category accounted for 12.896%, which might be due to the unclear or difficult-to-classify specific chemical classification of some metabolites. In addition, organic heterocyclic compounds accounted for 11.592%, and such compounds usually had various biological activities and might participate in various physiological and pathological processes ( Figure 4 D). Compared with the NC group, 77 metabolites were dysregulated in the positive ion mode and 131 metabolites were dysregulated in the negative ion mode in the DSS group. 59 metabolites were dysregulated in the positive ion mode and 99 metabolites were dysregulated in the negative ion mode in the FLP group. The number of dysregulated metabolites between the FLP group and the NC group was significantly less than that between the DSS group and the NC group, indicating that FLP treatment reduced the metabolite disorders caused by colitis ( Figure 4E, F). The results of PCA analysis of the overall samples further verified this conclusion, and in the positive ion mode, the samples of the FLP group were closer to those of the NC group in terms of metabolite profiles ( Figure 4 G, H). The volcano plot analysis showed that compared with the DSS and NC groups, the concentrations of significantly different metabolites between the FLP and NC groups decreased ( Figure 4 I, J). Therefore, FLP has a significant effect on regulating the metabolite profiles of colitis mice.

[0047] Based on the above embodiments, the following conclusions can be drawn: The present invention induces an inflammatory bowel disease model in mice with a DSS solution, and detects the body weight, disease activity index, pathological structure of colorectal tissue, pathological structure of spleen, HE staining, intestinal flora structure, and intestinal metabolite profiles of the mice. The results show that the polysaccharide derived from honeysuckle can delay inflammatory bowel disease in model animals by oral administration, with the characteristics of strong targeting and convenient administration, and has good application prospects in the treatment of inflammatory bowel disease.

[0048] The above are only a limited number of preferred embodiments of the present invention, and the descriptions thereof are relatively specific and detailed, but should not be construed as limiting the scope of the patent of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the concept of the present invention, several modifications and improvements can still be made, and these all belong to the protection scope of the present invention.

Claims

1. A method for identifying honeysuckle polysaccharide, characterized in that, The method described above includes the following steps: performing infrared spectrum detection, monosaccharide composition detection, and transmission and scanning electron microscopy observation on honeysuckle polysaccharide. The specific steps are as follows: 1) Weigh the polysaccharide sample and potassium bromide according to a weight ratio of 1:

50. After mixing evenly, press them into a sheet with a thickness of 1 mm, and then use a Fourier transform infrared spectrometer for detection; 2) Take a clean chromatographic vial, weigh the polysaccharide sample, add 2M trifluoroacetic acid (TFA) solution, and heat at 121 °C for 2 hours; pass nitrogen gas and dry it; add 99.99% methanol for cleaning and then dry it again. Repeat the methanol cleaning 2 - 3 times; add sterile water to dissolve and transfer it into the chromatographic vial for testing; the chromatographic system used is the Thermo ICS 5000+ ion chromatography system, and an electrochemical detector is used to analyze and detect the monosaccharide composition; and compare the results with the standard product to qualitatively and quantitatively analyze the monosaccharide composition in the polysaccharide; the volume - weight ratio of the polysaccharide sample to 2M trifluoroacetic acid (TFA) is 10 mg:1 ml; 3) Dissolve a small amount of honeysuckle polysaccharide in 18.2 ultrapure water, then drop it onto a carbon - supported film. After natural drying, use a transmission electron microscope for observation to further understand the microstructure and properties of honeysuckle polysaccharide FLP; 4) Paste a small amount of honeysuckle polysaccharide powder on the sample stage, perform surface gold spraying treatment, and then use a scanning electron microscope for observation to further understand the microstructure and properties of honeysuckle polysaccharide FLP.

2. The method according to claim 1, wherein In the said step 1), the instrument resolution is 4.00 cm -1 , the scanning range is 4000 - 450 cm -1 , the number of scans: 32 times; the sampling gain is 8.0; the moving mirror speed is 0.4747; the aperture is 80.00; DTGS KBr detector; KBr beam splitter; infrared light source.

3. Application of honeysuckle polysaccharide in the preparation of drugs for preventing or treating inflammatory bowel disease.

4. The application according to claim 3, characterized in that, Each 1 mg of honeysuckle polysaccharide contains 4.6982 μg of Rha, 9.4605 μg of Gal, 10.6458 μg of Ara, 28.7586 μg of Gal - UA, and 78.8753 μg of Glu.

5. The application according to claim 3, wherein In the application described above, it includes establishing a DSS - induced mouse IBD model to verify its effect.

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