Synbiotics composition based on mixed probiotics and medicinal and edible polysaccharides and application

Abstract

The invention belongs to the technical field of functional food, and particularly relates to a synbiotics composition based on mixed probiotics and medicinal and edible polysaccharides and application. The synbiotic composition comprises a probiotic component and a plant polysaccharide component according to a specific ratio, wherein the probiotic component comprises plant lactobacillus CGMCC (China General Microbiological Culture Collection Center) No.34512 and lactobacillus paracasei CGMCC 1.12731; the plant polysaccharide is selected from polygonatum kingianum polysaccharide and dendrobium devonianum polysaccharide. According to the technical scheme, through a probiotic-polysaccharide synergistic effect mechanism, the intestinal flora structure and metabolism can be effectively adjusted, and the intestinal microecological balance of IBD mice is effectively rebuilt. Compared with the prior art, the synbiotics composition overcomes the defects that a traditional IBD treatment medicine is remarkable in liver and kidney toxicity, the effect of single use of probiotics or plant polysaccharide is limited, and an existing synbiotics composition is difficult to adjust intestinal flora to completely recover to a normal state, and a safe and effective nutrition intervention strategy is provided.

Description

Technical Field

[0001] The present invention belongs to the technical field of functional foods, and in particular relates to a synbiotic composition based on mixed probiotics and medicinal and edible polysaccharides and its application. Background Art

[0002] Inflammatory bowel disease (IBD) is an idiopathic disease affecting the ileum, rectum, and colon. Its pathological progression begins with damage to the colonic mucosa and submucosa, manifesting clinically with core symptoms such as abdominal pain, diarrhea, and bloody stools. As the disease progresses, it can lead to penetrating intestinal lesions and the risk of cancer. Current clinical treatments primarily rely on 5-aminosalicylic acid compounds, corticosteroids, and immunosuppressants. However, long-term use can lead to serious side effects such as liver and kidney damage and gastrointestinal reactions, necessitating the development of safer and more effective alternative therapies.

[0003] In recent years, probiotic therapy has shown promising promise in the treatment of IBD. Studies have shown that probiotics such as Lactobacillus can improve IBD symptoms by regulating intestinal flora balance and enhancing intestinal barrier function. However, single probiotic preparations suffer from low colonization efficiency and limited functionality, significantly limiting their therapeutic effectiveness.

[0004] Synbiotics, as a combination of probiotics and prebiotics, can theoretically overcome these shortcomings through synergistic effects. Complementary approaches often use oligosaccharides in combination with Lactobacillus / Bifidobacterium, while synergistic approaches require the design of bacterial-element combinations based on metabolic associations. Existing technology systems generally suffer from two major bottlenecks: First, the development of prebiotics derived from plant polysaccharides that are both medicinal and edible is severely insufficient, failing to fully utilize the synergistic effects of their structural properties and microbial metabolism; second, while existing synbiotic preparations can regulate microbial structure (reducing harmful bacteria / increasing the abundance of beneficial bacteria), it is difficult to fully restore the microbial community to a normal state.

[0005] Polysaccharides from plants with both medicinal and edible properties hold great potential for the treatment of IBD due to their unique structural characteristics and bioactivity. Natural polysaccharides such as Polygonatum yunnanensis polysaccharide (PKP) and Dendrobium candidum polysaccharide (DCP) are particularly noteworthy for their well-defined molecular structure, excellent water solubility, robust gastrointestinal digestibility, and significant prebiotic activity. Summary of the Invention

[0006] This invention addresses the significant side effects of existing inflammatory bowel disease (IBD) medications, the insufficient efficacy of probiotics or plant polysaccharides alone, and the difficulty of fully restoring normal microbial structures with existing synbiotic combinations. The invention provides a synbiotic composition based on a mixture of probiotics and edible polysaccharides and its application. This synbiotic composition significantly regulates intestinal microbial flora and metabolism through synergistic effects, effectively restoring the intestinal microecological balance of IBD mice.

[0007] The technical solution adopted by the present invention to achieve the above-mentioned purpose is:

[0008] A synbiotic composition based on mixed probiotics and edible and medicinal polysaccharides, comprising:

[0009] (a) Probiotics, consisting of Lactobacillus plantarum CGMCC No. 34512 and Lactobacillus paracasei CGMCC 1.12731;

[0010] (b) plant polysaccharide selected from Polygonatum yunnanensis polysaccharide and / or Dendrobium officinale polysaccharide.

[0011] Furthermore, the total viable count of Lactobacillus plantarum and Lactobacillus paracasei was 5.88×10 9 -5×10 11 CFU / g.

[0012] Furthermore, the ratio of the number of viable Lactobacillus plantarum bacteria to the number of viable Lactobacillus paracasei bacteria is 1:3-3:1.

[0013] Furthermore, when the purity of the Polygonatum yunnanensis polysaccharide is 70.45%, the added amount is 58.8-200.0 mg / g; when the purity of the Dendrobium officinale polysaccharide is 74.34%, the added amount is 58.8-200.0 mg / g.

[0014] Furthermore, the polygonatum yunnanensis polysaccharide contains β-D-Fruf-(2→,→1,2)-β-D-Fruf-(6→,→1)-β-D-Fruf-(2→ and →1)-α-D-Glcp-(6→) as sugar chain residues, and has a molecular weight of 3-8 kDa.

[0015] Furthermore, the mannose content of the purple dendrobium polysaccharide is ≥70%, and the molecular weight is 25-30kDa.

[0016] Furthermore, when used on humans, the recommended daily intake for adults is: 27.5-379.3 mg / kg.

[0017] Application of the above-mentioned synbiotic composition in regulating intestinal flora.

[0018] Use of the above-mentioned synbiotic composition in preparing a preparation for preventing and treating inflammatory bowel disease.

[0019] Compared with the prior art, the present invention has the following beneficial effects:

[0020] 1. Synergistic effect of polysaccharides and compound probiotics

[0021] The experimental studies presented in this paper have yielded important conclusions: when polysaccharides or compound probiotics were used alone, diarrhea and bloody stool symptoms in IBD mice showed significant dose-dependent improvement; however, the combined synbiotic group showed significantly better intervention effects than the single-ingredient groups (p<0.05). This was demonstrated in three key ways: first, the remission rate for diarrhea and bloody stool symptoms increased by 30%-45%; second, the rate of weight loss in mice was significantly slowed; and finally, the disease activity index decreased by approximately 40%. These data fully demonstrate the significant synergistic effect of synbiotics in the treatment of IBD.

[0022] 2. The role of remodeling intestinal flora

[0023] The results showed that the Beta diversity F statistic of the synbiotic intervention group was significantly lower than that of the single intervention group (p<0.05). This result indicates that the intestinal flora structure of the synbiotic group is closer to that of the normal group. Although existing studies have confirmed that probiotics or polysaccharides used alone can regulate the flora structure, including reducing the abundance of harmful bacteria and increasing the proportion of beneficial bacteria, these single intervention measures are often difficult to completely restore the intestinal flora to a normal state. The innovative discovery of this study is that it has been confirmed for the first time through experimental data that a synbiotic combination can more effectively restore the intestinal microecological balance of IBD model mice. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 This is the separation and extraction process of polysaccharides in the present invention.

[0025] Figure 2 DCP monosaccharide composition

[0026] Figure 3 PKP monosaccharide composition

[0027] Figure 4 Gas chromatography-mass spectrometry analysis of PKP

[0028] Figure 5 Effects of different synbiotic combinations on the abundance of intestinal flora phyla under in vitro culture conditions

[0029] Figure 6 Content of short-chain fatty acids (propionic acid, butyric acid) in in vitro fermentation broth of intestinal flora

[0030] Figure 7 Effects of different synbiotic combinations on the changes in disease index (DAI) in mice

[0031] Figure 8 Effects of different synbiotic combinations on colon length changes in mice

[0032] Figure 9 Effects of PKP, probiotics and their synbiotics on changes in DAI in colitis mice

[0033] Figure 10 Effects of PKP, probiotics and their synbiotics on colon length changes in colitis mice

[0034] Figure 11 Colon H&E staining

[0035] Figure 12 Effects of PKP, probiotics and their synbiotics on intestinal flora at the phylum level

[0036] Figure 13 Effects of PKP, probiotics and their synbiotics on intestinal flora at the genus level

[0037] Figure 14 Beta diversity of intestinal flora and similarity of flora between groups DETAILED DESCRIPTION

[0038] The present invention will be described in further detail below with reference to the examples and accompanying drawings, but the embodiments of the present invention are not limited thereto. Where specific conditions are not specified in the examples, the experimental methods used are conventional methods, and the reagents and instruments used are all commercially available conventional products.

[0039] Unless otherwise specified, all extraction reagents used in the present invention are analytically pure reagents.

[0040] 1. Preparation of Synbiotic Composition Raw Materials

[0041] The compound probiotics used mainly include Lactobacillus paracasei and Lactobacillus plantarum.

[0042] Lactobacillus paracasei (LA-PA) was purchased from China General Microorganism Culture Collection Center with the strain number CGMCC 1.12731.

[0043] Lactobacillus plantarum DL0405 (LA-PL) was isolated from homemade yogurt in Ordos and deposited in the General Microbiology Center of China Culture Collection Administration with the deposit number CGMCC No. 34512 and the deposit date 2025-05-12.

[0044] In this embodiment, each strain in the composite probiotics can be prepared using any method in the prior art for preparing a live bacteria liquid or live bacteria powder.

[0045] For example, the bacterial solution containing live bacteria is prepared by a method comprising the following steps:

[0046] Step 1: Prepare MRS liquid culture medium, sterilize at 115°C for 20 min, and set aside.

[0047] Step 2: Inoculate the bacterial strain into the culture medium at an inoculum size of 1%, and culture at 37° C. for 18 hours until the logarithmic growth phase to obtain a live bacterial liquid.

[0048] For another example, the bacterial powder containing live bacteria is prepared by a method comprising the following steps:

[0049] Step 1: Prepare MRS liquid culture medium, sterilize at 115°C for 20 min, and set aside.

[0050] Step 2: Inoculate the culture medium with a 1% inoculum of the bacteria and culture at 37°C for 18 hours until the logarithmic growth phase.

[0051] Step 3: Centrifuge at 6000 rpm for 10 minutes to obtain bacterial cells, prefreeze at -80°C for 12 hours, and freeze-dry in a vacuum freeze dryer for 24 hours to obtain bacterial powder. During the freeze-drying process, an appropriate amount of protective agent, such as skim milk powder, sucrose, or ascorbic acid, should be added to the probiotics.

[0052] In this embodiment, the PKP or DCP polysaccharides used were extracted from Polygonatum yunnanensis and Dendrobium officinale by hot water extraction and purified by steps such as degreasing, decolorization, and protein removal.

[0053] The specific steps are as follows: First, the raw materials must be pretreated. Take the rhizome of Polygonatum dahliae or the branch of Dendrobium officinale, wash it, cut it into sections, dry it at 60°C to constant weight, crush it through a 20-mesh sieve, and seal it for later use. Weigh the powder separately, add 85% ethanol at a solid-liquid ratio of 1:8, stir it at room temperature for 24 hours to remove lipids and fat-soluble pigments, filter it, and dry it at 50°C to constant weight. Weigh the defatted powder, add purified water at a solid-liquid ratio of 1:20, and extract it in a 65°C waterbath with stirring for 2 hours. After filtering, repeat the extraction twice (1.5 hours and 1 hour), and combine the filtrates to obtain an aqueous extract containing polysaccharides. The aqueous extract is concentrated by rotary evaporation at 55°C, then added with 4 volumes of anhydrous ethanol, allowed to stand at 4°C overnight, and centrifuged and freeze-dried to obtain crude polysaccharides. Once the crude polysaccharides have dissolved, add Sevage reagent (chloroform:n-butanol = 4:1) at a ratio of 4:1, shake it for 20 minutes, centrifuge it, and remove the supernatant. Repeat this process until no white flocs remain (protein is completely removed). Add 4% activated carbon, decolorize in a 35°C water bath for 1 hour, filter and dialyze (3500Da molecular weight cutoff) for 48 hours, add 4 times anhydrous ethanol to the dialysate, let it stand at 4°C overnight, centrifuge and evaporate to remove the solvent, and freeze-dry to obtain Yunnan Polygonatum polysaccharide (PKP) or Dendrobium candidum polysaccharide (DCP).

[0054] The specific separation and purification process is as follows Figure 1 shown.

[0055] 2. Plant Polysaccharide Analysis

[0056] (1) Purity determination method of plant polysaccharides: The purity of polysaccharides was determined by the phenol-sulfuric acid method.

[0057] Preparation of color developer: Mix 5% phenol solution and concentrated sulfuric acid at a ratio of 1:5 (v:v) and cool in an ice-water bath.

[0058] Standard Curve Determination: Prepare a 1 mg / mL fructose standard solution. Accurately measure 0.0 mL, 0.2 mL, 0.4 mL, 0.6 mL, 0.8 mL, and 1.0 mL of the reference solution into 1.5 mL sample tubes. Add purified water to 1.0 mL and mix thoroughly. Transfer 200 μL of each standard solution to a centrifuge tube, add 1200 μL of the color developer, mix thoroughly, and place in boiling water at 100°C for 25 minutes. Measure the absorbance of the solution at a detection wavelength of 490 nm to construct a concentration-absorbance standard curve.

[0059] Determination of sample purity: The operation process is consistent with the determination of the standard solution. After obtaining the absorbance, the sample concentration is calculated using the standard curve and then converted into sample purity.

[0060] Calculation formula:

[0061] y=5.748x-0.1059

[0062] R 2 =0.9939

[0063] The polysaccharide content in the sample was calculated as mass fraction ω according to the standard curve and repeated three times.

[0064] (2) Monosaccharide composition determination method: 1-phenyl-3-methyl-5-pyrazolone pre-column derivatization high performance liquid chromatography was used for determination using the PMP-HPLC-PDA method.

[0065] Polysaccharide samples need to be pretreated with acid hydrolysis and derivatization:

[0066] Acid Hydrolysis: Accurately weigh 10 mg of Polygonatum odoratum polysaccharide into a stoppered test tube, add 2 mL of 2 M trifluoroacetic acid (TFA) solution, mix thoroughly, seal, and acid hydrolyze at 120°C for 4 h. After the reaction, dry with nitrogen, add methanol for washing, and dry three times to remove excess TFA. Dissolve in 2 mL of purified water to obtain the polysaccharide acid hydrolysis solution.

[0067] Derivatization: Add 1 mL of 0.3 M sodium hydroxide (NaOH) solution and 1.2 mL of 0.5 M PMP methanol solution to the polysaccharide acid hydrolysis solution, mix thoroughly, seal the tube, and derivatize at 70°C for 1 hour. After the reaction, cool to room temperature. Add 1 mL of 0.3 M hydrochloric acid (HCl) solution to adjust the pH. Extract three times with equal volumes of chloroform, and the aqueous phase is used as the test solution.

[0068] Derivatization of Monosaccharide Standards: Prepare 1 mg / mL monosaccharide standards and perform the same derivatization as for polysaccharide samples for HPLC analysis. Monosaccharide standards include: D-mannose (Man), L-rhamnose (Rha), D-glucuronic acid (GlcA), D-galacturonic acid (GalA), D-glucose (Glc), G-galactose (Gal), D-xylose (Xyl), and L-arabinose (Ara).

[0069] Chromatographic conditions: A YMC-Pack Pro C18 RS capillary column (4.6 mm × 250 mm × 5 mm) equipped with a PDA detector was used; the mobile phase was acetonitrile: 0.02 mol / L ammonium acetate solution = 20:80, the flow rate was 1 mL / min, and the injection volume was 20 mL. (3) PKP methylation reaction method:

[0070] 10 mg of PKP and 50 mg of sodium hydride (NaH) were dissolved in 6 mL of dimethyl sulfoxide (DMSO) and stirred under nitrogen for 12 h. 2 mL of pre-cooled methyl iodide was slowly added dropwise on ice and reacted in the dark under nitrogen for 2 h. The reaction was repeated three times and then terminated with 3 mL of distilled water. The reaction solution was extracted three times with chloroform and the chloroform layer was washed with water to remove DMSO. The nitrogen was blown dry and the extract was examined by infrared spectroscopy to find no hydroxyl bands (3200-3700 cm -1 ) indicates complete methylation. The completely methylated product was reacted with 2mL 1M trifluoroacetic acid (TFA) at 90°C for 150min. Add 2mL 2M ammonia water and vortex to remove excess acid. The hydrolyzed product was reduced with excess sodium borodeuteride (NaBD4) overnight, neutralized with 50% acetic acid to a pH of 6-7, blown dry with nitrogen, washed with methanol and blown dry, and repeated three times. Add 2mL acetic anhydride and 2mL pyridine and acetylate at 90°C for 2h, and add 2mL purified water to terminate the reaction. Extract with chloroform, remove the lower organic phase, dry it with anhydrous sodium sulfate (Na2SO4), and perform gas chromatography-mass spectrometry after passing through a 0.22μm oil filter membrane.

[0071] Chromatographic Conditions: Analyses were performed using an Orbitrap high-resolution gas chromatography-mass spectrometer equipped with an electron impact ionization (EI) source and an HP-5 capillary column (30 m × 0.25 mm, 0.25 μm). High-purity nitrogen was used as the carrier gas, with an injection volume of 1 μL and a split ratio of 10:1. The ion source and quadrupole temperatures were both 230°C. Analytes were detected in full-scan mode with a mass scan range (m / z) of 30–600. The temperature program was as follows: an initial temperature of 140°C for 2 min, then a temperature increase of 3°C / min to 230°C, where it was maintained for 3 min.

[0072] 3. Preparation method of synbiotic composition

[0073] The preparation of synbiotic composition mainly includes two key steps: probiotic mixing and prebiotic addition. First, the probiotic LA-PL and LA-PA powders are fully mixed in a ratio of 1:3 to 3:1 to ensure that the total viable count of the final mixed powder is controlled at 1×10 9 to 1×10 11 CFU / g range. Subsequently, polygonatum odoratum polysaccharide and / or purple dendrobium polysaccharide are selected as prebiotic ingredients. In this embodiment, polygonatum odoratum polysaccharide with a purity of 70.45% or purple dendrobium polysaccharide with a purity of 74.34% are selected, and are uniformly mixed with the prepared mixed probiotic powder according to an addition amount of 58.8-200.0 mg / g. Through the organic combination of these two steps, a synbiotic composition with a specific number of viable bacteria and prebiotic content is finally obtained. It should be noted that in actual production, polygonatum odoratum polysaccharide and purple dendrobium polysaccharide of different purities can be selected according to demand, and the balance of the formula can be maintained by adjusting the addition amount in proportion.

[0074] 4. Evaluation method of the effect of probiotics on antibacterial effect

[0075] The pathogenic bacteria to be tested were inoculated into liquid LB medium at a 1% inoculum volume and diluted to 1.0×10 8 CFU / mL. The above strains were also inoculated into MRS medium at a 1% inoculation rate, cultured at 37°C for 24 hours, and then the fermentation liquid was centrifuged in a centrifuge at 10,000 rpm / min for 5 minutes, and the supernatant fermentation liquid was taken. The antibacterial experiment was carried out by the punching method. Solid LB culture medium was prepared and poured into plates. There was about 20 mL of culture medium in each plate. After solidification and cooling, 100 μL of pathogenic bacteria was inoculated into each plate, and the plate was evenly spread. Then three holes with a diameter of 0.6 cm were evenly punched on the plate, and 50 μL of strain fermentation liquid was added to the wells. Three replicates were set for each strain fermentation liquid. The plates were turned upside down and cultured in a 37°C incubator for 12 hours. The diameter of the inhibition zone was measured.

[0076] The MRS medium formulation (g / L) is: peptone 10.0, beef extract powder 8.0, yeast extract powder 4.0, glucose 20.0, dipotassium hydrogen phosphate 2.0, triammonium citrate 2.0, sodium acetate 5.0, magnesium sulfate 0.2, manganese sulfate 0.04, Tween 80 1.0. pH: 5.7 ± 0.2.

[0077] 5. Animal Experimental Research Subjects

[0078] Female Balb / c mice weighing 18–20 g were purchased from the SPF Laboratory Animal Center of Liaoning Changsheng Biotechnology Co., Ltd.

[0079] 6. In vitro culture methods of intestinal flora

[0080] (1) Culture medium configuration

[0081] Weigh 59 g of GAM broth and add 1000 mL of purified water. After cooling, add 500 μL of 1% hemin solution, 500 μL of 0.2% vitamin K solution, and 1 mL of 1% resazurin solution. Heat and boil to dissolve. Add 1 g of L-cysteine hydrochloride and adjust the pH to 7.2 ± 0.2. Dispense 20 mL of the broth into vials. Replace the broth with nitrogen for 3 minutes to remove dissolved oxygen. Seal the vials and sterilize them by high-pressure steam at 115°C for 20 minutes.

[0082] (2) Preparation of intestinal flora

[0083] Feces of normal Balb / c mice (n=3) were collected and treated with phosphate buffered saline (PBS) containing 20% glycerol in an anaerobic incubator. Fecal microorganisms were resuspended at a ratio of 0.1 g feces: 1 mL PBS, allowed to stand, and the supernatant was collected and stored in a -80°C refrigerator until use.

[0084] 7. Evaluation Methods for In Vitro Intervention Effects of Synbiotics on Intestinal Microbiota

[0085] (1) Experimental methods

[0086] Four treatment groups were set up for comparative study: a blank control group (NC) received no treatment; an E. coli infection group (EC) received 200 μL of E. coli culture medium with an OD600 value of 1 added to the intestinal flora culture medium; and the Polygonatum dahliae polysaccharide synbiotic intervention group (ELP) and the Dendrobium officinale polysaccharide synbiotic intervention group (ELD) received the same 200 μL of E. coli culture medium and 20 mg of the corresponding polysaccharide synbiotic. All treatment groups used in vitro intestinal flora culture medium to adjust the culture system to a final volume of 20 ml to ensure consistency in experimental conditions.

[0087] Each group was incubated in a shaker at 37°C for 24 hours. After incubation, the cells were centrifuged at 12,000 rpm for 5 minutes, and the bacterial cells at the bottom of the centrifuge tube were collected and stored at -20°C for subsequent analysis. The fermentation broth was used for the determination of short-chain fatty acids.

[0088] (2) Method for determining intestinal flora abundance

[0089] The bacteria collected in the above experiment were sent to Hangzhou Guhe Biotechnology Co., Ltd. for sequencing, and a triangular phase diagram of the relative abundance of each bacteria was drawn to represent the abundance ratio of specific microorganisms in different groups.

[0090] (3) Determination of short-chain fatty acids in fermentation broth

[0091] To prepare the standard curve, weigh an appropriate amount of each short-chain fatty acid standard and dilute each to a final concentration of 3 mg / mL acetic acid and 0.4 mg / mL butyric acid using 50% (v / v) ethanol. Perform a serial dilution to create a series of mixed standard solutions with varying concentrations. Thoroughly mix 1 mL of each supernatant and the standard solutions (acetic acid and butyric acid) at each concentration with excess ammonium persulfate. Add 1 mL of anhydrous ethanol, vortex for 10 seconds, and centrifuge for 5 minutes. The supernatant is then used as the test solution.

[0092] Chromatographic conditions: Analyses were performed using a gas chromatograph equipped with an FID detector and an FFAP capillary column (30 m × 0.25 mm × 0.25 μm). High-purity nitrogen was used as the carrier gas, with an injection volume of 1, a split ratio of 1:10, a flow rate of 2.96 mL / min, and injector and detector temperatures of 250°C.

[0093] Heating program: initial temperature was 80 °C for 2 min, then the temperature was increased to 180 °C at a rate of 10 °C / min and maintained at this temperature for 5 min.

[0094] 8. Evaluation Methods of Biobiotics for Preventing and Improving Inflammatory Bowel Disease in Vivo

[0095] Balb / c mice were used as experimental mice. After a week of acclimation, they were divided into groups and administered intragastrically for 21 days. On the 15th day, 2.5% dextran sulfate sodium (DSS) was added to the drinking water of all mice except the control group to induce an inflammatory bowel disease model. DSS induction lasted for seven days, and the model was established on the 21st day. The mice were then sacrificed and samples were collected.

[0096] 8.1 Evaluation of Polygonatum sibiricum Polysaccharide Synbiotics and Dendrobium officinale Synbiotics for Preventing Inflammatory Bowel Disease in Vivo (1) Experimental Methods

[0097] The experiment was divided into four groups, with 8 mice in each group: the control group (NC), the model group (DSS), the Yunnan Polygonatum polysaccharide synbiotic group (ELP), and the purple dendrobium polysaccharide synbiotic group (ELD). During the IBD modeling period, each mouse was weighed at a fixed time every day, and the mouse feces and blood in the stool were monitored at the same time. After the end of the experiment, the disease activity index (DAI) was plotted. DAI is the sum of weight loss (no weight loss = 0, 1%-5% weight loss = 1, 6%-10% weight loss = 2, 11%-20% weight loss = 3, and >20% weight loss = 4), stool consistency (normal = 0, loose = 2, and diarrhea = 4), and the degree of blood in the stool (no blood = 0, a small amount of blood in the stool = 2, and a large amount of blood in the stool = 4), reflecting the health status of IBD mice.

[0098] 8.2 Evaluation of Different Doses of Polygonatum sibiricum Polysaccharide Synbiotics for Preventing Inflammatory Bowel Disease in Vivo

[0099] (1) Experimental methods

[0100] The experiment was divided into eight groups, with 8 mice in each group, namely the control group (NC), model group (DSS), positive drug group (5-aminosalicylic acid (5-ASA) group), probiotics alone group (LA), low-dose Polygonatum sibiricum polysaccharide group (PKP-L), high-dose Polygonatum sibiricum polysaccharide group (PKP-H), low-dose Polygonatum sibiricum polysaccharide synbiotic group (Syn-L), and high-dose Polygonatum sibiricum polysaccharide synbiotic group (Syn-H). The animal experimental method was the same as 8.1. During the IBD modeling period, each mouse was weighed at a fixed time every day, and the feces and blood in the stool of the mice were monitored at the same time, and DAI was drawn. In addition to the determination of DAI, the following indicators were also evaluated: colon length and lesion status, and the regulatory effect on intestinal flora.

[0101] (2) Methods for detecting colon length and lesion status

[0102] The mouse colon segments were taken and their lengths were measured. The colons were immediately placed in 4% paraformaldehyde fixative, embedded, and stained with HE and AB. The cross-sectional sections were used to count the colon morphology and the number of colonic goblet cells.

[0103] (3) Detection methods for regulatory effects on intestinal flora

[0104] Feces were collected from each group of mice and sent to Hangzhou Guhe Biotechnology Co., Ltd. for sequencing. The abundance of intestinal microbes in each group was analyzed, and short-chain fatty acids were measured. The F statistic was used to analyze their effects on gut microbial structure. The F statistic is an important statistic used to measure the relative magnitude of differences between groups and within groups. It plays a key role in analysis of variance, evaluating the significance of differences by comparing the sum of squares between groups to the sum of squares within groups.

[0105] The calculation formula for the F statistic is:

[0106] F = (between-group sum of squares / between-group degrees of freedom) / (within-group sum of squares / within-group degrees of freedom)

[0107] 9. Analysis results of Polygonatum sibiricum polysaccharides and Dendrobium officinale polysaccharides

[0108] The phenol-sulfuric acid method showed that the polysaccharide content in Dendrobium officinale was about 22.25%, and the polysaccharide content in Polygonatum yunnanensis was 29.3%. The results of PMP-HPLC showed that Man was the main monosaccharide component in Dendrobium officinale polysaccharide ( Figure 2 ), where Man:Glc=72:7.5; Polygonatum dahliae polysaccharide may contain Fru, Man, Glc, Gal ( Figure 3Fru has been shown to readily convert to Man and Glc under high temperature and strong acid conditions. Due to its structural characteristics, it cannot be detected after PMP derivatization. Therefore, the abundant Man and Glc may be derived from Fru, indicating that Fru and Glc are the primary components of Polygonatum yunnanensis.

[0109] Figure 4 The GC results of the polysaccharide of Polygonatum yunnanensis were analyzed by methylation method. Combined with the mass spectrometry data, the main mono / oligosaccharide fragments and glycosidic bond molar ratios are shown in 1. Combined with the GC-MS results, it was determined that Polygonatum yunnanensis mainly contains five sugar chain residues: β-D-Fruf-(2→, →1)-β-D-Fruf-(2→, →2)-β-D-Fruf-(6→, →1)-α-D-Glcp-(6→, →1,2)-β-D-Fruf-(6→

[0110] Table 1. PKP methylation mass spectrometry fragmentation analysis

[0111]

[0112] 10. Evaluation results of the effects of single bacteria and different proportions of compound probiotics on antibacterial effects

[0113] Example 1

[0114] The ratio of viable bacteria counts of LA-PL and LA-PA in the composite probiotics was 1:1;

[0115] Example 2

[0116] The ratio of viable bacteria counts of LA-PL and LA-PA in the composite probiotics was 2:1;

[0117] Example 3

[0118] The ratio of viable bacteria counts of LA-PL and LA-PA in the composite probiotics was 1:2;

[0119] Example 4

[0120] The ratio of viable bacteria counts of LA-PL and LA-PA in the composite probiotics was 1:3;

[0121] Example 5

[0122] The ratio of viable bacteria counts of LA-PL and LA-PA in the composite probiotics was 3:1;

[0123] According to the evaluation method of the effect of probiotics on antibacterial effect, the antibacterial ability of single bacteria and compound probiotics in different proportions was evaluated. The results are shown in Table 2:

[0124] Table 2. Evaluation table of antibacterial ability of probiotics

[0125]

[0126] As can be seen from the table: against E. coli and Salmonella pathogens, the compound probiotics showed better inhibitory effects than the single bacteria. When the ratio of LA-PL to LA-PA of the compound probiotics was 3:1, the inhibition diameter on the plate was the largest. 11. Evaluation results of the in vitro intervention effect of synbiotics on intestinal flora

[0127] Comparative Example 1

[0128] The blank group (NC) contained only culture medium without the addition of E. coli culture medium or other substances.

[0129] Comparative Example 2

[0130] Escherichia coli group (EC), add the OD 600 200 μL of 1% E. coli culture medium.

[0131] Example 6

[0132] In the intervention group of Polygonatum yunnanensis polysaccharide synbiotics (ELP), 200 μL was added to the culture medium and pre-cultured to OD 600 coli culture medium, and 20 mg of Polygonatum sibiricum polysaccharide synbiotics were added at the same time. The addition ratio of Polygonatum sibiricum polysaccharide in the synbiotics was 200 mg / g, and the total viable count of the composite probiotics was 5×10 9 CFU / g, among which the ratio of viable bacteria count of LA-PL and LA-PA in the compound probiotics is 3:1.

[0133] Example 7

[0134] In the Dendrobium officinale polysaccharide synbiotic intervention group (ELD), 200 μL was added to the culture medium and pre-cultured to OD 600 coli culture medium, and 20 mg of purple dendrobium polysaccharide synbiotics were added. The addition ratio of purple dendrobium polysaccharide in the synbiotics was 200 mg / g, and the total viable count of the composite probiotics was 5×10 10 CFU / g, among which the ratio of viable bacteria count of LA-PL and LA-PA in the compound probiotics was 3:1.

[0135] Studies have found that polysaccharides can maintain intestinal homeostasis and promote intestinal health by regulating the structure and function of intestinal flora. First, the effects of Dendrobium officinale synbiotics and Polygonatum dahliae synbiotics on improving intestinal flora were evaluated in vitro. Experiments were conducted based on the evaluation method of in vitro intervention effects of synbiotics on intestinal flora. The results are shown in Figure 2. Figure 5As shown. After the intestinal flora of mice were cultured in vitro, the flora of the NC group was mainly composed of Proteobacteria, Firmicutes, and Bacteroidota. Compared with NC, the abundance of Proteobacteria in the EC group increased significantly. After intervention by the ELP and ELD groups, the flora structure was adjusted. The abundance of Proteobacteria decreased in the ELP and ELD groups. Studies have shown that changes in the ratio of Firmicutes to Bacteroidota are closely related to intestinal health. Compared with the NC group, the F / B ratio of the EC group decreased. Firmicutes contains a large number of bacteria that can produce acetic acid and butyric acid. The decrease in the abundance of Firmicutes will lead to changes in the pH in the intestine and an increase in harmful bacteria. The F / B ratio in the ELP group has been restored.

[0136] Significant changes were also observed in the in vitro intestinal microbiota at the genus level. Compared with the NC group, the abundance of Escherichia coli was significantly increased in the EC group, while it decreased in the ELP group. Furthermore, the abundance of the probiotic Lactobacillus was significantly increased in the ELP group. Literature has reported that Alistipes can alleviate the severity of colitis. In this study, the abundance of Alistipes in the EC group decreased significantly compared with the NC group, from 0.26% in the NC group to 0.18% in the EC group. However, the abundance of Alistipes increased in the ELP group to 0.28%. A similar pattern was observed in the ELD group, but the improvement was more pronounced in the ELP group. This further suggests that Alistipes is a bacterium that can alleviate colitis. These results demonstrate that the synbiotic combination of Polygonatum sibiricum polysaccharide and Dendrobium officinale polysaccharide can significantly reduce and improve the abundance of Escherichia coli in the intestinal microbiota under in vitro culture conditions, suggesting its potential for the prevention and treatment of inflammatory bowel disease.

[0137] The effects of the above two synbiotics on short-chain fatty acids in intestinal flora were further analyzed. Figure 6 As shown, the production of various short-chain fatty acids in the EC group decreased to varying degrees. However, after intervention with the Polygonatum dahliae polysaccharide synbiotic, butyrate production increased significantly, reaching 1.7 times that of the EC group. However, the butyrate production in the Dendrobium officinale polysaccharide synbiotic group did not differ significantly from that in the EC group. Similarly, propionic acid production was also increased in the ELP group. This further confirms that Lactobacillus and polysaccharides can regulate the intestinal flora and thus affect the production of short-chain fatty acids, maintaining intestinal homeostasis by regulating the pH of the intestinal microenvironment.

[0138] It can be seen from this that the synbiotics composed of compound probiotics and plant polysaccharides have the effect of promoting the production of short-chain fatty acids by intestinal flora.

[0139] 12. Evaluation results of Biobiotics in preventing and improving inflammatory bowel disease in vivo

[0140] 12.1 Results of the In Vivo Evaluation of Polygonatum sibiricum Polysaccharide Synbiotics and Dendrobium officinale Synbiotics for the Prevention of Inflammatory Bowel Disease

[0141] Comparative Example 3

[0142] The control group (NC group) was gavaged with normal saline every day.

[0143] Comparative Example 4

[0144] The model group (DSS group) was gavaged with normal saline every day. On the 15th day after gavage, 2.5% DSS was added to the drinking water of the mice.

[0145] Example 8

[0146] The Polygonatum sibiricum polysaccharide synbiotic group (ELP group) was given 20 mg of Polygonatum sibiricum polysaccharide synbiotic to each mouse daily by gavage. The addition ratio of Polygonatum sibiricum polysaccharide in the Polygonatum sibiricum polysaccharide synbiotic was 200 mg / g, and the total viable count of the composite probiotics was 5×10 11 CFU / g, the ratio of viable bacteria counts of LA-PL and LA-PA in the compound probiotics was 1:3. After conversion, this treatment regimen was equivalent to giving mice 5×10 11 The dosage level of probiotics was 200 mg / kg (calculated based on the weight of mice of 20 g). On the 15th day after oral administration, 2.5% DSS was added to the drinking water of mice.

[0147] Example 9

[0148] The purple dendrobium polysaccharide synbiotic group (ELD group) was given 20 mg of purple dendrobium polysaccharide synbiotics to each mouse daily by gavage. The purple dendrobium polysaccharide synbiotics contained 200 mg / g of purple dendrobium polysaccharide, and the total viable count of the composite probiotics was 5×10 11 CFU / g, the ratio of viable bacteria counts of LA-PL and LA-PA in the compound probiotics was 1:3. After conversion, this treatment regimen was equivalent to giving mice 5×10 11 The dosage level of probiotics was 200 mg / kg and polysaccharide of Dendrobium officinale was 200 mg / kg (the weight of mice was calculated based on 20 g). On the 15th day after oral administration, 2.5% DSS was added to the drinking water of mice.

[0149] The experiment was conducted based on the evaluation method of Yunnan Polygonatum polysaccharide synbiotics and purple Dendrobium synbiotics for preventing inflammatory bowel disease in vivo. The results are as follows Figure 7As shown, both synbiotic combinations can reduce the disease index of mice caused by DSS and improve the colon lesions in mice. The synbiotic containing Polygonatum odoratum polysaccharide has a stronger effect.

[0150] 12.2 Evaluation of Different Doses of Polygonatum sibiricum Polysaccharide Synbiotics for Preventing Inflammatory Bowel Disease in Vivo

[0151] Comparative Example 5

[0152] The control group (NC group) was gavaged with normal saline every day.

[0153] Comparative Example 6

[0154] The model group (DSS group) was gavaged with normal saline every day. On the 15th day after gavage, 2.5% DSS was added to the drinking water of the mice.

[0155] Comparative Example 7

[0156] The positive drug group (5-ASA group) was orally administered 5-ASA at a dose of 100 mg / kg (based on a mouse body weight of 20 g) daily. On the 15th day after orally administering 5-ASA, 2.5% DSS was added to the drinking water of the mice.

[0157] Example 10

[0158] In the probiotics alone group (LA group), each mouse was given 20 mg of a specific ratio of compound probiotics by gavage daily, with a total viable count of 5×10 11 CFU / g, the ratio of viable bacteria count of LA-PL and LA-PA in the compound probiotics was 3:1. After conversion, this treatment regimen was equivalent to giving mice 5×10 11 CFU / kg probiotics (calculated based on 20 g mouse body weight) On the 15th day after oral administration, 2.5% DSS was added to the drinking water of the mice.

[0159] Example 11

[0160] The low-dose Polygonatum cyrtonema polysaccharide group (PKP-L group) was given 50 mg / kg Polygonatum cyrtonema polysaccharide to each mouse via oral gavage every day (calculated based on the weight of the mouse being 20 g). On the 15th day after oral gavage, 2.5% DSS was added to the drinking water of the mice.

[0161] Example 12

[0162] The high-dose Polygonatum cyrtonema polysaccharide group (PKP-H group) was given 200 mg / kg Polygonatum cyrtonema polysaccharide to each mouse by oral gavage every day (the mouse weight was calculated based on 20 g). On the 15th day after oral gavage, 2.5% DSS was added to the drinking water of the mice.

[0163] Example 13

[0164] The low-dose Polygonatum sibiricum polysaccharide synbiotic group (Syn-L group) was given 17 mg of Polygonatum sibiricum polysaccharide synbiotic to each mouse daily by gavage. The addition ratio of Polygonatum sibiricum polysaccharide in the Polygonatum sibiricum polysaccharide synbiotic was 58.8 mg / g, and the total viable count of the composite probiotics was 5.88×10 9 CFU / g, the ratio of viable bacteria count of LA-PL and LA-PA in the compound probiotics was 3:1. After conversion, this treatment regimen was equivalent to giving mice 5×10 9 The dosage level of probiotics was 50 mg / kg (calculated based on 20 g mouse body weight) and 2.5% DSS was added to the drinking water of mice on the 15th day after oral administration.

[0165] Example 14

[0166] The high-dose Polygonatum sibiricum polysaccharide synbiotic group (Syn-H group) was given 20 mg of Polygonatum sibiricum polysaccharide synbiotics to each mouse daily by gavage. The addition ratio of Polygonatum sibiricum polysaccharide in the Polygonatum sibiricum polysaccharide synbiotics was 200 mg / g, and the total viable count of the composite probiotics was 5×10 11 CFU / g, the ratio of viable bacteria count of LA-PL and LA-PA in the compound probiotics was 3:1. After conversion, this treatment regimen was equivalent to giving mice 5×10 11 The dosage levels of probiotics were 200 mg / kg and 200 mg / kg of Polygonatum odoratum polysaccharide (the mice were weighed 20 g). On the 15th day after oral administration, 2.5% DSS was added to the drinking water of the mice.

[0167] Experiments were conducted based on the evaluation method of different doses of Polygonatum sibiricum polysaccharide synbiotics in vivo for the prevention of inflammatory bowel disease. The results showed that the mice in the control group (NC group) had no abnormalities, and the mice in the model group (DSS group) did not show obvious abnormalities within the first 1-3 days of modeling. From the fourth day, they showed obvious symptoms of blood in the stool and diarrhea, and their weight dropped rapidly in the later stage of modeling, which are typical symptoms of inflammatory bowel disease; the mice in the positive pressure drug group (5-ASA group) had diarrhea, but no blood in the stool; the mice in the low-dose Polygonatum sibiricum polysaccharide group (PKP-L group) also had obvious diarrhea and blood in the stool. The diarrhea and blood in the mice in the probiotic group (LA group), high-dose Polygonatum sibiricum polysaccharide group (PKP-H group), low-dose Polygonatum sibiricum polysaccharide synbiotic group (Syn-L group), and high-dose Polygonatum sibiricum polysaccharide synbiotic group (Syn-H group) were improved, and the weight loss trend slowed down.

[0168] Judging from the results, Polygonatum sibiricum polysaccharide and compound probiotics had a certain improvement effect on IBD mice, and showed a certain dose dependence; while Polygonatum sibiricum polysaccharide synbiotics significantly improved the improvement effect, and the synbiotic group was more effective than Polygonatum sibiricum polysaccharide alone and compound probiotics alone.

[0169] Figure 9The figure shows the disease index of the mice at the end of the experiment. The DAI of the NC group remained at an extremely low level, while the disease index of the mice in the DSS group increased significantly, with the mice exhibiting significant diarrhea and bloody stools. The low-dose Polygonatum sibiricum group (PKP-L) alone also showed a significant increase in DAI, indicating that PKP-L had no effect on the DAI index of the mice. However, the disease index of the mice in the synbiotic group decreased significantly, indicating that the disease was significantly improved. This suggests that the synbiotic group significantly improved diarrhea, bloody stools, and weight loss in IBD mice compared to the polysaccharide and probiotic groups alone.

[0170] The colon length of mice in each group was further analyzed. Figure 10 As shown in the results, the colon length of mice in the DSS group was significantly shorter than that in the NC group, with an average length of only 5.4 cm. The colon length of mice in the 5-ASA, LA, PKP-L, and PKP-H groups increased to varying degrees. Furthermore, the Syn-H group had the best colon protection effect on mice, with an average colon length of 9.2 cm, close to that of normal mice. This indicates that both Polygonatum sibiricum polysaccharide and probiotics have a protective effect on the colon, and that Synbiotics has a better protective effect on the colon than either Polygonatum sibiricum polysaccharide or probiotics alone.

[0171] To further explore the improving effect of synbiotics on the mouse colon, the colon segments of mice in each group were immediately placed in 4% paraformaldehyde fixative, and the colon was embedded, stained with H&E, and AB; cross-sectional sections were used to count the colon morphology and the number of colon goblet cells.

[0172] After H&E staining, the colon of each experimental group was subjected to histological analysis. Figure 11 As shown, the results showed that the intact cells of the superstructure of the colon of the mice in the NC group were clearly visible, and there was no inflammatory infiltration; while the colon of the mice in the DSS group showed obvious lesions, the intestinal villi disappeared, the superstructure was destroyed, and obvious inflammatory infiltration appeared (indicated by arrows); compared with the PKP-L group, there was no obvious inflammatory infiltration in the colon tissue of the PKP-H group; the Syn-H group was significantly improved, the superstructure of the intestine was significantly restored, and there was no obvious inflammatory infiltration, indicating that the effect of synbiotics is better than the use of compound probiotics and Polygonatum sibiricum polysaccharide alone.

[0173] After AB staining, the colonic goblet cells were counted using Image J software.

[0174] Goblet cells are mucus-secreting cells distributed between the columnar epithelial cells of the mucosa. Their main function is to synthesize and secrete mucins, forming a mucosal barrier to protect epithelial cells. Next, the protective effect of Polygonatum sibiricum polysaccharide synbiotics on goblet cells was investigated.

[0175] Alcian blue (AB) staining results showed that the goblet cells in the colon of mice in the NC group were abundant; the goblet cells in the DSS group were almost lost; the number of goblet cells in the PKP-L group was also significantly reduced; and the number of goblet cells in the high-dose polysaccharide group was significantly restored; under the same area, the Syn-H group had the largest number of goblet cells and the strongest protective effect on colon tissue.

[0176] Further study the changes in the intestinal microbial structure of each group of mice to explore the connection between IBD and intestinal microorganisms.

[0177] At the phylum level, compared with the NC group, the abundance of Bacteroides increased and the abundance of Firmicutes decreased in the DSS group; this phenomenon was partially reversed after synbiotic intervention, which significantly increased the F / B ratio by increasing the abundance of Firmicutes and decreasing the abundance of Bacteroides ( Figure 12 Compared with the groups receiving Polygonatum sibiricum polysaccharide and probiotics alone, the Syn-H group showed more pronounced phylum-level regulation of the intestinal flora. Previous studies have shown that changes in the F / B ratio are closely associated with inflammation. Therefore, the increased F / B ratio in the Syn group may play a positive role in the metabolic regulation of inflammatory bowel disease.

[0178] At the genus level ( Figure 13 ), compared with the NC group, the abundance of Lachnospiraceae and Mucispirillum, which are recognized probiotics in inflammatory bowel disease, in the DSS group decreased significantly, to 10.7% and 1.3%, respectively. On the contrary, the abundance of Paraprevotella increased to 4.2%. After synbiotic intervention, the abundance of recognized probiotics increased significantly, such as the abundance of Lachnospiraceae increased to 20.9%, while the abundance of Paraprevotella decreased significantly to 0.7%. Figure 10 After synbiotic intervention, the bacterial flora of IBD mice became more similar to that of normal (NC) mice, indicating that synbiotics can effectively restore the changes in intestinal flora caused by DSS.

[0179] Beta diversity can reflect the similarity of intestinal flora between groups. The Beta diversity results of intestinal flora in each group showed that each drug-treated group could change the composition of intestinal flora of mice to varying degrees ( Figure 14The results showed that after DSS induction, the intestinal flora of the DSS group mice showed significant differences from those of the normal group. However, intervention with the Polygonatum sibiricum polysaccharide synbiotic significantly improved the intestinal flora of the mice. In Beta diversity, the F statistic represents the similarity coefficient between groups; a larger F statistic indicates a greater difference in intestinal flora from the NC group. The Syn-H group had the smallest F statistic compared to the NC group, indicating that the microbial composition of the two groups of mice was the most similar, suggesting that the Polygonatum sibiricum polysaccharide synbiotic has a good ability to regulate intestinal flora.

[0180] The dosage range of the plant polysaccharide in the edible and medicinal polysaccharide synbiotic provided by the present invention is 50-200 mg / kg. Based on a 1:9.1 ratio of human to mouse surface area for drug absorption, the recommended human dosage of the plant polysaccharide in the synbiotic is 5.5-22 mg / kg. Since the polysaccharide content in the synbiotic was measured to be 5.8%-20%, the recommended total human dosage range of the synbiotic is 27.5-379.3 mg / kg.

[0181] Probiotics are effective in treating inflammatory bowel disease, weight management, regulating blood lipids, and inhibiting the growth of harmful bacteria. The recommended dosage in public literature is 10 9 -10 12 CFU (Probiotics Branch of Chinese Institute of Food Science and Technology, Journal of Chinese Institute of Food Science and Technology, 2025, 5-15: 1-13.), and the experimental doses used in this study were all within this safety range.

[0182] Regarding the safety of plant polysaccharides, sufficient research has confirmed their safety. Studies have shown that 480 mg / kg of Yunnan Polygonatum polysaccharide has no significant negative impact on the kidneys of mice (Liu Zhijun, Journal of Traditional Chinese Medicine, 2021, 27(01):12-15.), and 0.2% Yunnan Polygonatum extract has no cytotoxicity to keratinocytes and fibroblasts (Shi Yang, Daily Chemical Industry, 2020, 50(11):788-798.). In addition, Purple Dendrobium has been approved as a local specialty food by Yunnan Province, and its safety has been officially recognized. Relevant literature points out that the content of purple Dendrobium polysaccharide is similar to that of Dendrobium officinale, and the dosage of Dendrobium officinale polysaccharide in animal experiments can be as high as 900 mg / kg, and it has no toxic side effects on the liver and kidneys (Can Jin, Carbohydrate Polymers, 2025, 123660.). These research data fully prove that the synbiotic product within the dosage range adopted by the present invention has reliable safety.

[0183] The present invention is described by way of example, and those skilled in the art will appreciate that various changes or equivalent substitutions may be made to these features and embodiments without departing from the spirit and scope of the present invention. Furthermore, under the teachings of the present invention, these features and embodiments may be modified to suit specific circumstances and materials without departing from the spirit and scope of the present invention. Therefore, the present invention is not limited to the specific embodiments disclosed herein, and all embodiments falling within the scope of the claims of this application are intended to be within the scope of the present invention.

Claims

1. A synbiotic composition based on mixed probiotics and edible polysaccharides, characterized in that: contain: (a) Probiotics, consisting of Lactobacillus plantarum CGMCC No. 34512 and Lactobacillus paracasei CGMCC 1.12731; (b) plant polysaccharide selected from Polygonatum yunnanensis polysaccharide and / or Dendrobium officinale polysaccharide.

2. The synbiotic composition according to claim 1, characterized in that The total viable count of Lactobacillus plantarum and Lactobacillus paracasei was 5.88×10 9 -5×10 11 CFU / g.

3. The synbiotic composition according to claim 2, characterized in that The ratio of the number of viable Lactobacillus plantarum bacteria to the number of viable Lactobacillus paracasei bacteria is 1:3-3:

1.

4. The synbiotic composition according to claim 1, characterized in that When the purity of the Polygonatum yunnanensis polysaccharide is 70.45%, the added amount is 58.8-200.0 mg / g; when the purity of the Dendrobium officinale polysaccharide is 74.34%, the added amount is 58.8-200.0 mg / g.

5. The synbiotic composition according to claim 1, characterized in that The polygonatum yunnanensis polysaccharide contains β-D-Fruf-(2→,→1,2)-β-D-Fruf-(6→,→1)-β-D-Fruf-(2→ and →1)-α-D-Glcp-(6→ as sugar chain residues, and has a molecular weight of 3-8 kDa.

6. The synbiotic composition according to claim 1, characterized in that The mannose content of the purple dendrobium polysaccharide is greater than or equal to 70%, and the molecular weight is 25-30 kDa.

7. The synbiotic composition according to claims 1-6, characterized in that When used on humans, the recommended daily intake for adults is: 27.5-379.3 mg / kg.

8. Use of the synbiotic composition according to claim 7 in regulating intestinal flora.

9. Use of the synbiotic composition according to claim 7 in preparing a preparation for preventing and treating inflammatory bowel disease.

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