Composite probiotic inulin gel biomass preparation as well as preparation method and application thereof

Abstract

The invention discloses a composite probiotic inulin gel biomass preparation as well as a preparation method and application thereof. Relates to the technical field of food. The raw materials comprise composite probiotics and inulin gel. The invention further provides a preparation method and application. The oral compound probiotic inulin gel preparation provided by the invention is good in safety, and is used for preventing ionizing radiation hematopoietic system injury and radiation enteritis; wherein the inulin gel has a certain protection effect on gastric acid and intestinal juice, and probiotics are protected from being damaged. The inulin gel can prolong the retention of the probiotics in the intestinal tract and enhance the colonization effect of the probiotics in the intestinal tract. The compound probiotics have a certain synergistic effect.

Description

Technical Field

[0001] The invention relates to the field of food technology, and more particularly to a composite probiotic inulin gel synbiotic preparation and a preparation method and application thereof. Background Art

[0002] Radiation can be divided into ionizing radiation and non-ionizing radiation. Ionizing radiation with high energy and strong penetrating power can cause harm to the body, such as medical X-rays, radioactive isotopes, nuclear energy equipment, etc. The damage caused by ionizing radiation to organisms is called ionizing radiation damage.

[0003] At the cellular level, ionizing radiation directly or indirectly interacts with the nucleic acids, proteins and cell membranes of cells, leading to DNA breakage, mutation and cell death. At the tissue level, ionizing radiation damage can affect the differentiation and proliferation ability of cells, leading to tissue damage, tumor lesions and organ dysfunction. Different types of tissues have different sensitivities to ionizing radiation. The radiation sensitivity of tissues is ranked from high to low in the order of bone marrow, intestine and brain.

[0004] Whole-body exposure to radiation can cause acute radiation syndrome, with the most obvious and severe damage to the hematopoietic system. When the body's radiation dose is greater than 3.5Gy, the blood system rapidly undergoes functional, morphological and quantitative changes, hindering cell division and proliferation, causing the hematopoietic system to fail to function normally, and even leading to infection and death. Patients receiving radiotherapy will inevitably suffer damage to the hematopoietic system. Radiation enteritis is a complication caused by radiation rays affecting the surrounding healthy intestinal tissue during radiotherapy for patients with abdominal and pelvic tumors. With the increasing incidence of abdominal tumors and the widespread application of radiation technology, the incidence of radiation enteritis has also increased. About 70% of patients will experience early acute radiation intestinal damage within a few days after receiving radiotherapy. The main clinical symptoms are severe diarrhea, bloody stools, abdominal pain and weight loss, which seriously interfere with the patient's subsequent treatment and affect the patient's survival and prognosis.

[0005] Ionizing radiation causes intestinal flora imbalance, resulting in a significant increase in the number of pathogenic bacteria, dysfunction of symbiotic bacteria, release of inflammatory factors, and a decrease in short-chain fatty acids.

[0006] Studies have shown that there are significant differences in microbial communities before and after irradiation, and regulating intestinal flora can help restore the body's health.

[0007] At present, there is a lack of safe and effective drugs for the prevention and treatment of hematopoietic system damage and radiation enteritis caused by ionizing radiation in clinical practice. As a small molecule cytoprotectant that needs to be injected, amifostine has a short half-life, strict administration time, and side effects such as vomiting, nausea, and hypotension, which limits its clinical application.

[0008] Therefore, developing an oral, safe and effective radiation protection product to overcome the above technical deficiencies is an urgent problem that technicians in this field need to solve. Summary of the invention

[0009] In view of this, the present invention provides a composite probiotic inulin gel synbiotic preparation and its preparation method and application. Among them, probiotics and prebiotics, as important components of dietary supplements, have excellent regulatory effects on intestinal microbiota. Probiotics interact with intestinal microorganisms, improve the intestinal barrier, regulate immune function, and produce organic acids and antimicrobial peptides. Prebiotics can be selectively utilized by host microorganisms to produce short-chain fatty acids, promote the growth of probiotics, inhibit the proliferation of pathogens, and lower intestinal pH. Synbiotic preparations are a combination of probiotics and prebiotics, which have a protective effect on ionizing radiation damage and are considered to be more effective in promoting body health.

[0010] In order to achieve the above object, the present invention adopts the following technical solution:

[0011] A composite probiotic inulin gel preparation, the raw materials of which include composite probiotics and inulin gel.

[0012] Preferably, the inulin gel is prepared from an inulin solution, and the mass concentration of the inulin solution is 45% to 55%.

[0013] Preferred: Compound probiotics including concentration of 5×10 8 CFU / mL of Clostridium butyricum, Bifidobacterium adolescentis, and Akkermansia muciniphila, with a volume ratio of 1:1:1; each 1mL of inulin gel contains 1.5×10 9 CFU complex probiotics.

[0014] The present invention also provides a method for preparing any of the above-mentioned composite probiotic inulin gel preparations, comprising the following steps:

[0015] (1) Preparation of inulin gel: adding inulin to water, heating to obtain an inulin solution, stopping heating, and allowing to stand to form a non-flowing inulin gel;

[0016] (2) Preparation of inulin gel: Three probiotics, Clostridium butyricum, Bifidobacterium adolescentis, and Akkermansia muciniphila, were respectively cultured to the logarithmic phase under an anaerobic environment, centrifuged, and the bacterial precipitate was collected and the supernatant was removed; the bacterial solution was resuspended with physiological saline, centrifuged again, and the centrifugation operation was repeated; the bacteria were resuspended with physiological saline, and then Clostridium butyricum and Bifidobacterium adolescentis were centrifuged again, and Akkermansia muciniphila was centrifuged to obtain a precipitate, which was resuspended with physiological saline; then the suspensions of the three probiotics were mixed to obtain a composite probiotic suspension, which was centrifuged to obtain the composite probiotic, and the supernatant was aspirated and discarded;

[0017] (3) The composite probiotics (precipitate) are suspended with the prepared inulin gel to obtain a composite probiotics inulin gel preparation.

[0018] Preferably, step (1) is specifically as follows: adding inulin to water, heating at 80° C. for 5 minutes to obtain an inulin solution, stopping heating, and allowing to stand at 4° C. to form a non-flowing inulin gel.

[0019] Preferably, step (2) is specifically as follows: three probiotics, Clostridium butyricum, Bifidobacterium adolescentis, and Akkermansia muciniphila, are taken respectively, and cultured to the logarithmic phase under an anaerobic environment, and centrifuged at 4°C: Clostridium butyricum and Bifidobacterium adolescentis centrifuged at 5000 rpm for 5 min, and Akkermansia muciniphila centrifuged at 5000 rpm for 10 min; the bacterial precipitate is collected and the supernatant is removed; the bacterial solution is resuspended with physiological saline, and centrifuged again: Clostridium butyricum and Bifidobacterium adolescentis centrifuged at 5000 rpm for 5 min, and Akkermansia muciniphila centrifuged at 5000 rpm for 10 min, and the process is repeated 3 times; the bacteria are resuspended with physiological saline to a concentration of 1×10 8 CFU / mL; then, Clostridium butyricum and Bifidobacterium adolescentis were centrifuged at 5000 rpm for 5 min, and Akkermansia muciniphila was centrifuged at 5000 rpm for 10 min to obtain a precipitate; the precipitate was resuspended in physiological saline to a concentration of 5×10 8 CFU / mL; then the suspensions of the three probiotics were mixed in a volume ratio of 1:1:1 to obtain a composite probiotic suspension, and then centrifuged at 5000 rpm for 10 min to obtain the composite probiotics, and the supernatant was aspirated and discarded.

[0020] Preferably: the expanded culture in step (2) includes: culturing Clostridium butyricum: taking a frozen Clostridium butyricum strain in the laboratory and dissolving it in a 37°C water bath, taking 100 μL of the bacterial solution and adding it to a shaking tube containing 9 mL of MRS broth liquid culture medium, transferring it to an anaerobic culture bag, and placing it in a 37°C constant temperature and humidity chamber, and standing for anaerobic culture for 24 to 48 hours; culturing Bifidobacterium adolescentis: dissolving the lyophilized powder of Bifidobacterium adolescentis into a suspension with 200 μL of sterile water, adding it to a shaking tube containing 9 mL of MRS broth liquid culture medium, transferring it to an anaerobic culture bag, and placing it in a 37°C constant temperature and humidity chamber, and standing for anaerobic culture for 24 to 48 hours;

[0021] Cultivation of Akkermansia muciniphila: Take 1 mL of Akkermansia muciniphila solution and add it to 9 mL of modified brain heart infusion medium, transfer it to an anaerobic culture bag, and place it in a constant temperature and humidity chamber at 37°C for anaerobic culture for 72 hours.

[0022] The present invention also provides the use of any of the above-mentioned composite probiotic inulin gel synbiotic preparations in the preparation of medicines and health products.

[0023] Preferred: Medicines: Medicines that have a protective effect against ionizing radiation damage.

[0024] Preferred: Ionizing radiation injury: radiation enteritis and hematopoietic system damage.

[0025] Furthermore, ionizing radiation damage refers to radiation enteritis and hematopoietic system damage caused by ionizing radiation.

[0026] Furthermore, the timing of administration of the composite probiotic inulin gel preparation is:

[0027] Selected from before the whole body or abdomen is irradiated, after the whole body or abdomen is irradiated, a combination of before and after the whole body or abdomen is irradiated, preferably before the whole body or abdomen is irradiated, more preferably a combination of before and after the whole body or abdomen is irradiated.

[0028] When the composite probiotic inulin gel preparation is taken orally before the whole body or abdomen is irradiated, it is taken 1 to 3 times a day, preferably once a day. The time is selected from 14 days before the irradiation to the day of the irradiation, preferably from 7 days before the irradiation to the day of the irradiation, and more preferably from 2 days before the irradiation to the day of the irradiation.

[0029] When the composite probiotic inulin gel preparation is taken orally after the whole body or abdomen is irradiated, it is taken 1 to 3 times a day, preferably once a day. The time is selected from the day of irradiation to 14 days after irradiation, preferably from the day of irradiation to 7 days after irradiation, and more preferably from the day of irradiation to 2 days after irradiation.

[0030] When the combined oral probiotic inulin gel preparation is selected before and after the whole body or abdomen is irradiated, it is taken 1 to 3 times a day, preferably once a day. The time is selected from 14 days before irradiation to 14 days after irradiation, preferably from 7 days before irradiation to 7 days after irradiation, and more preferably from 2 days before irradiation to 3 days after irradiation.

[0031] It can be seen from the above technical scheme that, compared with the prior art, the present invention discloses a composite probiotic inulin gel synbiotic preparation and a preparation method and application thereof, and the technical effects achieved are:

[0032] The oral composite probiotic inulin gel preparation has good safety, and there is no strict limit on the oral dosage for protecting the hematopoietic system from ionizing radiation damage and radiation enteritis (it can be selected from 2mL to 100mL each time, preferably from 5mL to 50mL, and more preferably from 10mL to 25mL).

[0033] Inulin gel has a certain protective effect on gastric acid and intestinal fluid, protecting probiotics from being destroyed. Inulin gel can prolong the retention of probiotics in the intestine and enhance the colonization effect of probiotics in the intestine. Compound probiotics have a certain synergistic effect. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on the provided drawings without paying creative work.

[0035] Figure 1 The accompanying drawings are diagrams showing the relationship between the state of inulin and the concentration of inulin provided by the present invention, wherein A: the state after adding different amounts of inulin to water and ultrasonication; B: the state of the inulin solution after heating; C: the state after being placed at 4°C overnight; and D: the state of inverted observation.

[0036] Figure 2 The accompanying drawings are SEM images of inulin gels of different concentrations provided by the present invention, wherein 40% inulin gel: inulin gel-40); 50% inulin gel: inulin gel-50; 60% inulin gel: inulin gel-60, and the scale is 50 μm.

[0037] Figure 3 The accompanying drawings are diagrams showing the injectability and viscosity of inulin gels with different percentage contents provided by the present invention, wherein A: 50% inulin gel can be smoothly squeezed out from a 1 mL syringe needle; B: viscosity changes of 40% and 50% inulin gels.

[0038] Figure 4 The accompanying figure is a diagram of the gastrointestinal environmental tolerance of the composite probiotic inulin gel synbiotic preparation provided by the present invention, wherein A: plate colonies of the positive control group, bacteria in gastric juice / intestinal juice group, and synbiotic preparation in gastric juice / intestinal juice group; B: optical density value OD 600 ; C: SEM image of the freeze-dried composite probiotic inulin gel preparation, arrows indicate probiotics; *P<0.05, **P<0.01.

[0039] Figure 5 The accompanying drawings are diagrams showing the retention of 50% inulin gel and 50% inulin solution provided by the present invention in the intestinal tract of mice at different times after intragastric administration.

[0040] Figure 6 The accompanying drawing shows that the oral composite probiotic inulin gel synbiotic preparation provided by the present invention promotes the recovery of body weight and physical activity of mice after irradiation, wherein A: body weight; B: total distance; C: immobility time; D: movement trajectory; n=4, in Figures B and C, *P<0.05, ***P<0.001; in Figure A, **P<0.01, the synbiotic preparation is compared with inulin gel; ##P<0.01, ###P<0.001, the synbiotic preparation is compared with the composite probiotic; ns: no statistical difference.

[0041] Figure 7 The accompanying drawings show the oral composite probiotic inulin gel preparation provided by the present invention for preventing and treating radiation-induced intestinal tissue damage, wherein A: colon appearance, B: length statistics, n=3; C: pathological section of the small intestine of mice 3.5 days after radiotherapy: H&E, Ki67 and AB / PAS.

[0042] Figure 8 The accompanying drawings show the effect of the oral composite probiotic inulin gel preparation provided by the present invention on preventing and treating radiation-induced intestinal tissue inflammation, wherein A: IL-6 in intestinal tissue; B: TNF-α; C: IL-10 content; n=4; *P<0.05, **P<0.01, ***P<0.001, ns: no statistical difference.

[0043] Fig. 9 The accompanying figure shows that the oral composite probiotic inulin gel preparation provided by the present invention reduces the intestinal permeability of irradiated mice, *P<0.05, **P<0.01, ***P<0.001, ns: no statistical difference.

[0044] Fig.10 The accompanying drawing is a diagram showing that the oral composite probiotic inulin gel syngeneic preparation provided by the present invention promotes the expression of tight junction proteins in the intestinal tissue of irradiated mice.

[0045] Fig.11 The accompanying drawing is a diagram showing the changes in the diversity of intestinal flora of irradiated mice after the oral composite probiotic inulin gel synbiotic preparation provided by the present invention, wherein A: Chao1, B: Shannon microbial community index; n=4; *P<0.05, **P<0.01, ***P<0.001, ns: no statistical difference.

[0046] Fig.12 The accompanying drawing is a diagram showing the changes in the richness of intestinal flora and the relative abundance of specific bacterial species in irradiated mice after the oral composite probiotic inulin gel synbiotic preparation provided by the present invention, wherein A: Blautia; B: f_Lachnospiraceae; C: Erysipelatoclostridium; D: Parasutterella; E: relative abundance heat map at the family level; n=4, *P<0.05, **P<0.01, ***P<0.001, ns: no statistical difference.

[0047] Fig.13 The accompanying drawing shows the effect of the oral composite probiotic inulin gel provided by the present invention on the content of short-chain fatty acids in the intestine of irradiated mice, n=3, *P<0.05, **P<0.01, ***P<0.001, ns: no statistical difference.

[0048] Fig.14The accompanying drawing shows the effect of the oral composite probiotic inulin gel synbiotic preparation provided by the present invention on the body weight of mice with whole-body radiation damage, n=3, **P<0.01, comparison between the synbiotic preparation group and the inulin gel group; #P<0.05, ##P<0.01, ###P<0.001, comparison between the synbiotic preparation group and the composite probiotic group.

[0049] Fig.15 The accompanying drawing is a diagram showing that the oral composite probiotic inulin gel synbiotic preparation provided by the present invention promotes blood cell recovery in mice subjected to whole body irradiation, wherein A: red blood cells, B: platelets, C: white blood cells, n=4, *P<0.05, **P<0.01, ***P<0.001, comparison between the synbiotic preparation group and the inulin gel group; # P<0.05, ## P<0.01, ### P<0.001, compared between the synbiotic preparation group and the compound probiotic group.

[0050] Fig.16 The accompanying drawing shows the increase of IL-3 and IL-10 content in irradiated mice by the oral composite probiotic inulin gel preparation provided by the present invention, wherein A: IL-3 content in blood, B: IL-10 content, n=4, *P<0.05, **P<0.01, ***P<0.001, ns: no statistical difference.

[0051] Fig.17 The accompanying drawing is a graph showing the effect of the oral composite probiotic inulin gel preparation provided by the present invention on the femoral tissue of whole-body irradiated mice.

[0052] Fig.18 The accompanying drawing is a graph showing the effect of the oral composite probiotic inulin gel preparation provided by the present invention on the spleen tissue of whole-body irradiated mice.

[0053] Fig.19 The attached figure shows the effect of the oral composite probiotic inulin gel preparation provided by the present invention on increasing the organ index of mice subjected to whole body irradiation, n=3, **P<0.01, ***P<0.001. ns: no statistical difference. DETAILED DESCRIPTION

[0054] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0055] The embodiment of the invention discloses a composite probiotic inulin gel synbiotic preparation and a preparation method and application thereof.

[0056] The microorganisms, reagents, and raw materials not mentioned in the examples are all existing or commercially available, and the processes not mentioned are conventional processes and will not be described in detail here.

[0057] For example, microorganisms: Clostridium butyricum (BNCC186498), Bifidobacterium adolescentis (BNCC 185974), and Akkermansia muciniphila (BNCC341917).

[0058] Example 1

[0059] A composite probiotic inulin gel preparation comprises the following steps:

[0060] (1) Preparation of inulin gel: inulin was added to water and heated at 80°C for 5 minutes to obtain a 50% inulin solution. The water was then heated and allowed to stand at 4°C to form a non-flowing inulin gel.

[0061] (2) Preparation of inulin gel: Three probiotics, Clostridium butyricum, Bifidobacterium adolescentis, and Akkermansia muciniphila, were respectively cultured in an anaerobic environment (culture of Clostridium butyricum: Clostridium butyricum strain frozen in the laboratory was placed in a 37°C water bath to dissolve, 100 μL of bacterial solution was taken in a clean bench and added to a shaking tube containing 9 mL of MRS broth liquid culture medium, transferred to an anaerobic culture bag, and placed in a 37°C constant temperature and humidity chamber for anaerobic culture for 24 to 48 h; culture of Bifidobacterium adolescentis: Bifidobacterium adolescentis freeze-dried powder was dissolved in 200 μL of sterile water to form a suspension, and added to 9 mL of The tube containing MRS broth liquid culture medium was transferred to an anaerobic culture bag and placed in a constant temperature and humidity chamber at 37°C for anaerobic culture for 24 to 48 hours; Culture of Akkermansia muciniphila: 1 mL of Akkermansia muciniphila liquid was added to 9 mL of modified brain heart infusion medium (on the basis of brain heart infusion medium, L-threonine (2230821001) and N-acetyl-D-glucosamine (KYEA610) were added, see: Pang Xiaoyang, Lü Jiaping, Meng Xiangyu, Zhang Shuwen, Wang Yunna, Li Xu, Xie Ning, Culture medium for Akkermansia muciniphila and its preparation method. p.20; Wang Min, Shi Changping, A culture medium for Akkermansia muciniphila and its preparation method. p.20 Anaerobic culture method of Akkermansia albicans.p.12.), transfer to an anaerobic culture bag, and place in a 37°C constant temperature and humidity chamber, and culture anaerobically for 72 hours) to the logarithmic phase, centrifuge at 4°C (5000 rpm for 5 min for Clostridium butyricum and Bifidobacterium adolescentis, 5000 rpm for 10 min for Akkermansia muciniphila), collect the bacterial precipitate, and remove the supernatant; resuspend the bacterial solution with saline, centrifuge again (5000 rpm for 5 min for Clostridium butyricum and Bifidobacterium adolescentis, 5000 rpm for Akkermansia muciniphila), repeat 3 times; resuspend the bacteria with saline to a concentration of 1×10 8 CFU / mL; then, Clostridium butyricum and Bifidobacterium adolescentis were centrifuged at 5000 rpm for 5 min, and Akkermansia muciniphila was centrifuged at 5000 rpm for 10 min to obtain a precipitate; the precipitate was resuspended in physiological saline to a concentration of 5×10 8 CFU / mL; then the suspensions of the three probiotics were mixed at a volume ratio of 1:1:1 to obtain a composite probiotic suspension, and then centrifuged at 5000 rpm for 10 min to obtain a composite probiotic (precipitate), and the supernatant was aspirated and discarded;

[0062] (3) The composite probiotics (precipitate) were suspended with the prepared inulin gel to obtain a composite probiotic inulin gel preparation (1.5×10 9 CFU complex probiotics).

[0063] Comparative experiment 1

[0064] Relationship between inulin gel formation and concentration

[0065] Method: Weigh 300 mg, 400 mg, 500 mg and 600 mg of inulin respectively, add them into 1 mL of water, mix them by ultrasonic, transfer them to a water bath preheated to 80°C, heat them for 5 minutes to obtain 30%, 40%, 50% and 60% inulin solutions, place them in a 4°C refrigerator and let them stand for 24 hours to observe whether inulin gel is formed.

[0066] Results: Ultrasound was observed to fully mix inulin in water, and a solution was formed after heating. After standing at 4°C overnight, 30% and 40% inulin solutions showed stratification, and flowed after inversion, but could not form a gel. However, 50% and 60% inulin solutions formed a non-flowing inulin gel ( Figure 1 ). Whether inulin gel is formed depends on the concentration. The higher the inulin content, the easier it is to form a gel. After heating, inulin dissolves faster and can be transformed from a solution to a gel after cooling, forming an interlaced three-dimensional network structure inside. There are many gaps in the middle of the network structure, and a gel state is formed after absorbing water molecules. Figure 1 Shows the state of inulin solutions of different concentrations after standing.

[0067] Comparative experiment 2

[0068] Investigation on the properties of inulin gel

[0069] Materials: inulin solution or inulin gel, fluorescein isothiocyanate (FITC)-inulin conjugate, scanning electron microscope (SEM), rheometer, small animal in vivo imaging instrument. C57BL / 6J mice, male, SPF grade, weight 20±2g.

[0070] Methods: Inulin solution or inulin gel was placed on the sample plate of the rheometer. The test temperature was 37℃. The fixed stress frequency test was carried out with a scanning frequency of 0.1-100Hz. The storage modulus and loss modulus were recorded respectively, and the viscosity of the inulin solution or inulin gel was measured. The inulin solution or inulin gel was added to liquid nitrogen for brittle fracture. After freeze-drying, it was cut into ultra-thin slices with a blade and pasted on the sample stage with conductive glue. The internal porous structure and microscopic morphology were observed by SEM. The morphology of the inulin gel-encapsulated composite probiotics was observed by SEM. FITC-labeled inulin gel and inulin solution were gavaged into mice, respectively, with a gavage dose of 200μL, and their intestinal retention was observed.

[0071] Bifidobacterium adolescentis (Ba) and the synbiotic preparation of Bifidobacterium adolescentis encapsulated in inulin gel (BIG) were incubated in artificial gastric fluid (SGF) for 1.5 hours, and then transferred to artificial intestinal fluid (SIF) for further incubation for 3 hours (the bacteria in gastric juice / intestinal juice group and the synbiotic preparation in gastric juice / intestinal juice group, respectively). Ba incubated in sterile PBS was used as the positive control group to investigate their growth.

[0072] Results: The internal structure of inulin solution or inulin gel was observed by SEM. The interior of inulin gel showed a cross-linked porous structure, and the pore size became relatively denser as the inulin concentration increased. The 40% inulin solution (marked as Inulin Gel-40 in the figure) could not form a gel after inversion, and the internal cross-linked structure of the gel after freeze-drying was mostly broken, the pore size was large and irregular, and it was easy to break after drying. The 50% inulin gel (marked as Inulin Gel-50 in the figure) and the 60% inulin gel (marked as Inulin Gel-60 in the figure) could form a gel after inversion, and the internal structure was cross-linked, the pore structure was relatively complete, and the pore size was small ( Figure 2 ).

[0073] The higher the inulin concentration, the stronger the internal intermolecular forces, the tighter the internal cross-linking, and the denser the pore size. 50% inulin gel has good needle permeability and is easy to absorb ( Figure 3 Middle A); 60% inulin gel is difficult to absorb.

[0074] The maximum shear viscosity of 40% inulin solution and 50% inulin gel were 0.136 and 35.04 Pa·s, respectively. As the concentration of inulin gel increased, the shear viscosity also increased ( Figure 3 (middle B).

[0075] SEM shows the morphology of inulin gel-encapsulated composite probiotics ( Figure 4 In middle C), arrows indicate probiotics, including rod-shaped, slender rod-shaped, and various types of probiotics growing in pairs. Different probiotics are evenly distributed in the inulin gel, and their surface is covered with an inulin gel layer.

[0076] After incubation with SGF and SIF (bacteria in gastric juice / intestinal juice group), Ba without inulin gel did not grow colonies after 24 hours of culture on the culture plate, indicating that Ba may be killed by gastric acid, the activity of the strain decreased, and it was difficult to grow and reproduce ( Figure 4 Middle A). After the composite probiotics encapsulated by inulin gel were continuously incubated in SGF and SIF (combined preparation in gastric juice / intestinal juice group), a large number of live bacteria were still able to grow and reproduce after 24 hours of culture, indicating that inulin gel can effectively protect the composite probiotics from gastrointestinal damage. Figure 4 In B, the OD value of bacteria was measured at 600nm by an ELISA instrument over time, indicating that more probiotics survived after continuous incubation in SGF and SIF in the composite probiotics encapsulated in inulin gel. The OD value of BIG increased to about 0.8 over time, which was not significantly different from the parallel control group. However, the OD value of unencapsulated Ba (bacteria in gastric juice / intestinal juice group) did not increase over time, which was significantly different from the combined preparation in gastric juice / intestinal juice group, indicating that there was no reproduction of live bacteria, which was consistent with the plate results.

[0077] 50% inulin gel has a certain viscosity and can stay in the intestine for a long time, targeting the colon. After encapsulating probiotics, it can help probiotics to play a role in the intestine. Figure 5 As shown, the 50% inulin solution has a short action time in the intestine, quickly passes through the stomach in 0.5 hours, and is mostly concentrated in the ileum and cecum in 1 hour; after 8 hours of observation, the fluorescence intensity is greatly reduced, and the inulin solution quickly passes through the colorectum. After 12 hours, the intestinal fluorescence intensity is close to the minimum, indicating that most of the inulin solution is excreted. In contrast, inulin gel has a longer retention time in the small intestine and colon. After 12 hours of gavage, strong fluorescence can still be seen in the cecum and colon. This shows that inulin gel can be used as an effective bioadhesive to prolong its retention in the intestine ( Figure 5 ).

[0078] Comparative test 3

[0079] Effect of oral inulin gel combination preparation on the prevention and treatment of radiation enteritis

[0080] Materials: C57BL / 6J mice, male, SPF grade, weighing 20±2g, 70 mice in total. Synbiotic prepared according to Example 1, 50% inulin gel (IG) prepared according to Example 1, and composite probiotics (MSP) prepared according to Example 1.

[0081] Methods: C57BL / 6J male mice were randomly divided into 5 groups, 14 mice in each group, and then divided into healthy group, model group, compound probiotics group, inulin gel group and synbiotic preparation group. Two days before irradiation and 1 hour before irradiation on the same day, the model group (0.2 mL / mouse, normal saline), the compound probiotics group (0.2 mL / mouse, 6×10 8 CFU / mL), inulin gel group (0.2mL / mouse), and synbiotic preparation group (0.2mL / mouse). All groups were given drugs by gavage 2 days before irradiation, and the drugs were given once a day until the 3rd day after irradiation.

[0082] Results: The irradiated mice had severe intestinal damage, which hindered the acquisition of nutrients, led to weight loss and decreased activity of the mice ( Figure 6 ). For the healthy group, all mice lost weight from irradiation to 5 days after irradiation. Compared with the radiation enteritis model group, the weight of each dosing group decreased less in the early stage and increased faster in the later stage. The difference in weight between the synbiotic groups and the model group was statistically significant (P<0.01). Most importantly, the mice in the synbiotic group showed the greatest weight recovery, even without significant difference compared with the healthy group ( Figure 6 ).

[0083] The open field test (OFT) was used to evaluate the activity rate of irradiated mice and reflect the recovery of radiation damage. The small intestine of mice was severely damaged after irradiation, resulting in a decrease in their motility. Compared with the radiation enteritis model group, the overall movement path of mice in the synbiotic preparation group increased and the immobility time was significantly reduced ( Figure 6 ), indicating that the synbiotic preparation significantly promoted the physical recovery of irradiated mice.

[0084] Comparative experiment 4

[0085] Oral inulin gel synbiotics alleviate radiation-induced intestinal damage and inflammation

[0086] Methods: The mice were anesthetized and killed on the 1st, 3.5th and 7th day after irradiation. The small intestinal tissue 5 cm below the lower end of the stomach was obtained and fixed in 4% paraformaldehyde solution for 24 hours. The tissue pathological morphology was observed under a microscope after staining with hematoxylin-eosin (H&E).

[0087] Materials: Inverted fluorescence microscope, 4% paraformaldehyde fixative.

[0088] Results: Radiation enteritis can shorten the colorectum of mice. Compared with the normal group, the colorectum of mice in the model group was significantly shortened, and the colorectal length of mice in the syngeneic preparation group was significantly restored (P<0.001, Figure 7 A and B in the figure). Pathological examination further showed that the combined preparation had excellent preventive and therapeutic effects on radiation enteritis ( Figure 7 (C). The radiation enteritis model group showed a large number of missing small intestinal crypts, severe structural damage, and significantly shortened intestinal villi. After treatment, the small intestinal morphology of the composite probiotic group was slightly better than that of the model group, but the crypts were accompanied by a large number of vacuoles and varying degrees of villus shortening. The inulin gel group showed a significant recovery of the number of crypts and villus length. The number of crypts in the synbiotic group increased significantly, and the small intestinal villi were well-shaped and uniform in length, with no significant difference from the healthy group ( Figure 7 C, H&E). To further explore the proliferation capacity of small intestinal crypt cells, the expression of Ki 67 in the small intestinal crypts was detected on day 3.5 ( Figure 7 C, Ki 67). Therefore, the synbiotics enhanced the expression of Ki 67 in the small intestinal crypts, indicating that the crypt cells are highly proliferative. Compared with the model group, the crypt proliferation of the compound probiotics group and the inulin gel group was improved, but their effects were not as good as those of the synbiotics group. The amount of mucin stained by AB-PAS in the synbiotics group was similar to that in the healthy group, indicating that the mucus barrier in the small intestinal tissue was restored ( Figure 7 In conclusion, oral administration of synbiotic preparations can repair the intestinal mucosal barrier, protect the integrity of the hindgut epithelium, promote crypt proliferation, alleviate intestinal tissue pathological damage, and contribute to the recovery of intestinal function.

[0089] Comparative experiment 5

[0090] Effects of oral administration of inulin gel syngeneic preparation on immune factors in irradiated mice

[0091] Materials: Mouse tumor necrosis factor (TNF-α), mouse interleukin-6 (IL-6), and mouse interleukin-10 (IL-10) detection kits.

[0092] Methods: On the 7th day after irradiation, mice were euthanized and 1 cm of small intestinal tissue was dissected (5 cm below the stomach). 0.1 g of small intestinal tissue was weighed and placed in a tissue homogenizer tube. 0.9 mL of 4°C pre-cooled physiological saline was added. Grinding steel balls (one large and two small) were placed in a pre-cooled high-speed tissue grinder centrifuge tube rack. The grinding frequency was 60 Hz and the grinding time was 180 seconds. The tissue was fully ground. High-speed centrifugation (4°C, 5000×g) was performed for 10 minutes. The supernatant was carefully aspirated and quickly aliquoted for storage.

[0093] Take the ELISA kits of IL-6, TNF-α and IL-10 and equilibrate them at room temperature. Perform the analysis according to the detailed steps of the ELISA kits.

[0094] The contents of IL-6 and TNF-α in small intestinal tissue in the radiation enteritis disease model were analyzed.

[0095] The results showed that compared with the normal group, the levels of IL-6 and TNF-α in the model group were significantly increased, with significant statistical differences (P<0.001). After treatment with inulin gel, compound probiotics and synbiotic preparations, the three reduced the levels of IL-6 and TNF-α, and the synbiotic preparations significantly reduced the levels of IL-6 (P<0.001) and TNF-α (P<0.01) compared with the model group. Figure 8 A, B). IL-10 is mainly produced by macrophages and is the most studied anti-inflammatory factor. It can inhibit the expression of inflammatory cytokines and plays an important role in the intestinal immune system. Compared with the normal group, the expression of IL-10 in the model group mice after irradiation was significantly decreased (P<0.001); while compared with the model group, the expression of IL-10 in the synbiotic preparation group was significantly upregulated ( Figure 8 C, P<0.001). Irradiation can damage the intestinal immune system of mice, leading to the release of a large number of inflammatory factors. Treatment with synbiotic preparations can regulate intestinal immunity, increase the expression of immune factors, regulate intestinal immune balance, and improve the intestinal microenvironment.

[0096] Comparative test 6

[0097] Oral administration of inulin gel syngeneic preparation reduces intestinal permeability in irradiated mice

[0098] Materials: Fluorescein isothiocyanate-dextran (FITC-Dextran)

[0099] Methods: On the 14th day after irradiation, mice were fasted for 12 hours and gavaged with 600 mg / kg FITC-dextran (4kDa) at a volume of 0.2 mL / mouse. Orbital blood was collected 4 hours later, and serum was collected after centrifugation at 1500×g for 15 minutes at room temperature. Serum (0.1 mL per sample) was transferred to a 96-well plate with a black bottom, and the fluorescence intensity was measured using a multifunctional microplate reader (excitation wavelength 485 nm; emission wavelength 520 nm).

[0100] Results: Radiation enteritis caused intestinal barrier damage and intestinal dysfunction. The overall intestinal permeability was evaluated by detecting the degree of intestinal absorption of fluorescently labeled macromolecular compounds into the blood. Compared with the normal group, the FITC-dextran content in the blood of mice in the model group was high, indicating that the intestinal barrier of mice was severely damaged, with a significant statistical difference (P<0.001). Compared with the model group, the FITC-dextran content in the peripheral blood of mice in the synbiotic preparation group was low, and the intestinal permeability was low, indicating that the synbiotic preparation effectively repaired the intestinal barrier, with a significant statistical difference ( Fig. 9 , P<0.001).

[0101] Comparative experiment 7

[0102] Oral administration of inulin gel synbiotics promotes the expression of tight junction proteins in intestinal tissue of irradiated mice

[0103] Methods: Tight junction protein ZO-1 and occludin protein in small intestinal tissue were detected by fluorescence staining.

[0104] Results: The expression levels of ZO-1 and occludin proteins in the small intestine tissue of mice in the model group and the compound probiotic group were significantly decreased. After intervention with the synbiotic preparation and inulin gel, the expression levels of ZO-1 and occludin proteins were upregulated. Compared with the inulin gel group, the expression of occludin in the synbiotic preparation group was higher and its effect was more obvious ( Fig.10 , Immunofluorescence images of A ZO-1 and B occludin staining in small intestinal tissues on day 3.5). Therefore, oral administration of the synbiotic preparation can increase the expression of tight junction proteins, improve the intestinal epithelial barrier, and promote the recovery of intestinal function in irradiated mice.

[0105] Comparative experiment 8

[0106] Oral administration of inulin gel synbiotics improves the recovery of intestinal microecology in irradiated mice

[0107] Methods: On the 7th day after mouse irradiation, fresh feces of mice were collected in sterilized 1.5 mL centrifuge tubes, frozen in liquid nitrogen immediately after collection, and stored at -80°C for analysis. After being taken out, 16srRNA sequencing of intestinal flora was performed.

[0108] Results: Chao1 and Shannon indices are important indicators for measuring α diversity, such as Fig.11 The results showed that compared with the normal group, Chao1 and Shannon indexes in the model group decreased significantly (P<0.01), reducing the intestinal species richness. There was no statistical difference between the synbiotic preparation group and the normal group, indicating that the synbiotic preparation can increase the diversity of intestinal flora after irradiation, improve the intestinal flora structure, and restore intestinal homeostasis.

[0109] Statistical analysis was performed on several bacteria with significant differences at the genus level ( Fig.12 ), after the mice were irradiated, the relative abundance of f_Lachnospiraceae and Blautia in the intestine decreased compared with the healthy group (P<0.001 and P<0.05). The proportion of conditional pathogens and pathogenic bacteria in the model group increased significantly. For example, the relative abundance of harmful genera such as Parasutterelal and Erysipelatoclostridium increased significantly (P<0.001, P<0.001), which led to intestinal flora disorder and seriously affected the repair of intestinal mucosal barrier. The relative abundance of probiotics such as f_Lachnospiraceae and Blautia in the intestine of mice in the synbiotic preparation group was higher than that in the model group, with statistical differences (P<0.001). The species distribution stacking diagram analysis was performed on the 11 families with the highest abundance in the samples on the 7th day after irradiation, in which the length of each column represents the proportion of each bacteria. Fig.12 In the middle E, the distribution of the microbiota in the model group was significantly different from that in the normal group, indicating that radiation caused changes in the composition of the intestinal microbiota. The distribution of the microbiota in the synbiotic preparation group was slightly different from that in the normal group, and the number of microbiota species increased significantly.

[0110] In summary, the combined preparation can increase the richness and diversity of beneficial bacteria in the intestines of irradiated mice, inhibit the growth of harmful bacteria, regulate the balance of intestinal flora in mice, play a positive role in the recovery of intestinal mucosal barrier function, inhibit intestinal inflammation, target the intestine, and improve radiation enteritis damage.

[0111] Comparative experiment 9

[0112] Oral administration of inulin gel syngeneic preparation increases the content of short-chain fatty acids in feces of irradiated mice

[0113] Methods: On the 7th day after mouse irradiation, fresh feces of mice were collected in sterilized 1.5mL centrifuge tubes, frozen in liquid nitrogen immediately after collection, and stored at -80℃ for analysis of short-chain fatty acids (SCFAs). The method used GC-MS, and the main steps included standard sample preparation, standard sample pretreatment, sample pretreatment, and on-machine detection.

[0114] Short-chain fatty acids (SCFAs) are produced by intestinal bacteria fermenting dietary fiber and absorbed by intestinal epithelial cells, providing ATP and energy to cells and maintaining normal intestinal physiological functions. Among them, acetate, propionate and butyrate are the most produced SCFAs. Whether the protective effect of synbiotic preparations is related to changes in the content of short-chain fatty acids in intestinal contents was analyzed.

[0115] The results show that Fig.13 ), compared with the normal group, the content of propionic acid (P<0.01) and butyric acid (P<0.001) in the model group decreased significantly. Compared with the model group, the content of acetic acid in the inulin gel group increased significantly (P<0.01). The content of butyric acid in the compound probiotic group increased (P<0.05), while there was no significant change in acetic acid and propionic acid. The content of acetic acid (P<0.001), propionic acid (P<0.01) and butyric acid in the synbiotic group increased significantly (P<0.001). Therefore, the synbiotic may improve the intestinal barrier by increasing the content of short-chain fatty acids in the intestine of irradiated mice.

[0116] Comparative experiment 10

[0117] Effects of oral administration of inulin gel composites on body weight of mice with whole-body radiation injury

[0118] Materials: C57BL / 6J mice, male, SPF grade, weighing 22±2g, 50 mice in total.

[0119] Body weight can reflect the health status of mice to a certain extent.

[0120] like Fig.14 As shown in the figure, from the first to the fourth day after 6.5Gy whole-body irradiation, the weight of the irradiated mice decreased, the mice were less active, and curled up together. From the fifth to the ninth day, the weight of the irradiated mice recovered, the mice moved freely, and their condition was improved compared with the previous one. Compared with the model group, the weight of the mice in the synbiotic preparation group recovered faster, and the weight returned to the pre-irradiation level on the 11th day after irradiation. The weight continued to rise in the later period, and there was no statistical difference with the normal group. The weight of the inulin gel group and the compound probiotic group showed a slow growth trend, and the weight returned to the pre-irradiation level on the 17th day after irradiation. On the 10th day after irradiation, the model lost weight again and died on the 10th day. The results showed that mice with whole-body irradiation had anorexia, malaise, and significantly worse weight in the early stage. Intervention can improve the weight loss of mice. Compared with the intervention of inulin gel and compound probiotics, the synbiotic preparation can better help mice recover, and their weight can also return to normal levels faster.

[0121] Comparative experiment 11

[0122] Oral administration of inulin gel syngeneic preparation restored peripheral blood count in irradiated mice

[0123] Methods: 20 μL of blood was collected from the tail vein of mice on day 0 before whole-body irradiation and on days 1, 7, 10, 14, 18, 22, and 28 after irradiation, and mixed with 0.48 mL of cell diluent. The number of white blood cells, red blood cells, platelets, and hemoglobin was determined using a hematology analyzer.

[0124] Results: After irradiation, the number of red blood cells in mice was severely reduced, with the model group decreasing the most and the synbiotic group decreasing the least. It was also observed that the number of red blood cells in the synbiotic group began to increase on the 14th day after irradiation, while the number of red blood cells in the inulin gel group and the compound probiotic group still showed a downward trend, and only on the 18th day did the number of red blood cells show an upward trend ( Fig.15 Middle A). The leukocyte count of the irradiated mice decreased sharply. After 14 days after irradiation, the leukocyte count of the drug-treated groups showed an upward trend, with the growth rate of the synbiotic preparation group being faster, but it has not yet returned to normal levels ( Fig.15 Platelets have a coagulation effect. After irradiation, the platelet content of mice in the model group dropped sharply until the 14th day after irradiation. During the blood collection process, it was found that the blood collection wounds of mice in the model group were difficult to heal, and the blood fluidity was relatively large. Compared with the model group, the platelet count in the drug intervention group did not decrease on the 7th day after irradiation, and remained stable, and began to increase after the 10th day. On the 18th day, there was no statistical difference in the platelet content between the Hesheng preparation group and the normal group ( Fig.15 Middle B). The platelet content of mice in the inulin gel group and the compound probiotic group was still at a low level. Therefore, the combined preparation can improve the hematopoietic system damage caused by whole-body irradiation, restore the reduction of white blood cells, red blood cells and platelets after irradiation, and effectively reduce complications such as infection caused by hematopoietic system damage.

[0125] Comparative experiment 12

[0126] Inulin gel syngeneic preparation increases IL-3 and IL-10 levels in irradiated mice

[0127] IL-3, also known as multi-colony stimulating factor, is a hematopoietic growth factor that mainly acts on early hematopoiesis. It has a positive regulatory effect on bone marrow hematopoiesis and can promote the reconstruction of the hematopoietic microenvironment.

[0128] like Fig.16 As shown in A, the IL-3 content of mice decreased significantly after irradiation (P<0.001), indicating that whole-body radiation inhibited the proliferation of hematopoietic factors and reduced the expression of IL-3. The IL-3 content in the serum of mice in the inulin gel group and the compound probiotic group was no different from that in the model group. An increase in the IL-3 content was observed in a few mice, and there were certain individual differences. The IL-3 content in the serum of mice in the combined preparation group increased significantly (P<0.001), and there was no difference from the normal group, which promoted the recovery of the hematopoietic function of irradiated mice.

[0129] The IL-10 content in the serum of mice in the model group was significantly decreased (P<0.001, Fig.16 B), indicating that the immune system function of irradiated mice was reduced, which easily led to infection in mice. The IL-10 content in the compound probiotic group was no different from that in the model group, and the mice still had inflammation and had not recovered. The IL-10 content in the synbiotic group was significantly increased compared with the model group (P<0.001), and there was no difference with the normal group, indicating that oral synbiotics can regulate the expression of immune factors and enhance the anti-infection ability of mice after irradiation.

[0130] Comparative experiment 13

[0131] Inulin gel syngeneic preparation reduces femoral damage in irradiated mice

[0132] Methods: On the 7th day after whole-body irradiation, the femurs of the hind limbs were dissected out and the muscles attached to the femurs were wiped clean with gauze. The dissected femurs were immediately immersed in a pre-prepared 4% paraformaldehyde solution for fixation, which can denature and coagulate the proteins in the tissues, thereby maintaining the original morphological structure of the cells, and preparing bone marrow hematoxylin-eosin (H&E) pathological sections.

[0133] Bone marrow is the main hematopoietic organ in the human body, contains hematopoietic stem cells, and is very sensitive to radiation.

[0134] like Fig.17 As shown in the figure, the nucleated cells in the femur of the model group decreased, the bone parenchyma was severely lost, and there were many vacuolar structures. Compared with the model group, the nucleated cells in the inulin gel group did not increase significantly, but there were fewer vacuolar structures. The number of nucleated cells in the composite probiotic group increased compared with the model group, but the vacuolar structure was more serious. The number of nucleated cells in the synbiotic group increased significantly, and the vacuolar structure was less. Therefore, the synbiotic can improve the hematopoietic function of the bone marrow to a certain extent and effectively protect the hematopoietic system from damage caused by whole-body irradiation.

[0135] Comparative experiment 14

[0136] Oral administration of inulin gel reduces spleen damage in mice subjected to whole-body irradiation

[0137] Methods: On the 7th day after whole-body irradiation, the spleens of the irradiated mice were removed by dissection and fixed in 4% paraformaldehyde solution. Bone marrow hematoxylin-eosin (H&E) and Ki67 pathological sections were prepared and observed under an inverted fluorescence microscope.

[0138] Results: The spleen plays an important role in the human immune system and hematopoietic system. It contains a small amount of hematopoietic stem cells that can play a hematopoietic function. The spleen is also one of the organs that are highly sensitive to radiation.

[0139] Fig.18(H&E and Ki67 stained spleen tissue). The mice were dissected on the 7th day after irradiation. After HE staining, the red pulp and white pulp (blue-purple) of the spleen sections in the normal group were clearly demarcated, while the boundary between the red and white pulp in the spleen tissue of the mice in the model group was blurred, and there was almost no white pulp after staining. The HE section results of the inulin gel group and the compound probiotics were similar to those of the model group. The sections of the synbiotic group showed that the boundary between the red and white pulp was more obvious and clear, and the area of ​​the white pulp was also larger. The Ki 67 results were consistent with the HE results. There were almost no positive cells in the model group, and the Ki 67 positive cells in the spleen tissue of the synbiotic preparation were significantly more than those in the model group.

[0140] Comparative test 15

[0141] Oral administration of inulin gel increases organ indexes in mice exposed to whole-body irradiation

[0142] Methods: On the 7th day after whole-body irradiation, mice were weighed and dissected. Four mice were selected from each group. The skin was cut open along the longitudinal axis of the body to completely expose the abdominal cavity. The small intestine and stomach were carefully opened to expose the spleen. The tissues and fat connected to the spleen were cut off to separate the spleen. The separated spleen was placed in 4°C precooled saline for weighing. The spleen tissue immersed in saline was dried with absorbent paper to remove the surface saline, placed on a balance for weighing, and weighed separately. The organ index = organ (mg) / body weight (g) was calculated.

[0143] The spleen is an important immune organ in the human body and is sensitive to ionizing radiation. Therefore, the spleen index can be used as an indicator of the function of the immune system.

[0144] The spleen index of irradiated mice Fig.19 As shown. The spleen of mice in the whole-body irradiation model group was atrophied (P<0.001), indicating that the whole-body radiation injury model was successfully established. However, there was no significant difference in the spleen index between the inulin gel group and the compound probiotic group and the model group. The spleen index of the synbiotic preparation group was significantly higher than that of the model group, inulin gel group and compound probiotic group (P<0.01), and there was no difference with the normal group, indicating that the synbiotic preparation can promote the recovery of immune organs and reduce radiation damage to the spleen, which is consistent with the results of spleen sections.

[0145] The various embodiments in this specification are described in a progressive manner, and each embodiment focuses on the differences from other embodiments. The same or similar parts between the various embodiments can be referenced to each other.

[0146] The above description of the disclosed embodiments enables one skilled in the art to implement or use the present invention. Various modifications to these embodiments will be apparent to one skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention will not be limited to the embodiments shown herein, but rather to the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A composite probiotic inulin gel preparation, characterized in that: Ingredients include: Complex probiotics and inulin gel.

2. The composite probiotic inulin gel preparation according to claim 1, characterized in that: The inulin gel is prepared from an inulin solution, and the mass concentration of the inulin solution is 45% to 55%.

3. The composite probiotic inulin gel preparation according to claim 1, characterized in that: The composite probiotics include a concentration of 5×10 8 CFU / mL of Clostridium butyricum, Bifidobacterium adolescentis, and Akkermansia muciniphila, with a volume ratio of 1:1:1; each 1 mL of the inulin gel contains 1.5×10 9 CFU complex probiotics.

4. The method for preparing the composite probiotic inulin gel preparation according to any one of claims 1 to 3, characterized in that: The following steps are involved: (1) Preparation of inulin gel: adding inulin to water, heating to obtain an inulin solution, stopping heating, and allowing to stand to form a non-flowing inulin gel; (2) Preparation of inulin gel: Three probiotics, Clostridium butyricum, Bifidobacterium adolescentis, and Akkermansia muciniphila, were respectively cultured to the logarithmic phase under an anaerobic environment, centrifuged, and the bacterial precipitate was collected and the supernatant was removed; the bacterial solution was resuspended with physiological saline, centrifuged again, and the centrifugation operation was repeated; the bacteria were resuspended with physiological saline, and then Clostridium butyricum and Bifidobacterium adolescentis were centrifuged again, and Akkermansia muciniphila was centrifuged to obtain a precipitate, which was resuspended with physiological saline; then the suspensions of the three probiotics were mixed to obtain a composite probiotic suspension, which was centrifuged to obtain the composite probiotic, and the supernatant was aspirated and discarded; (3) The composite probiotics (precipitate) are suspended with the prepared inulin gel to obtain a composite probiotics inulin gel preparation.

5. The method for preparing the composite probiotic inulin gel preparation according to claim 4, characterized in that: Step (1) is specifically as follows: adding inulin to water, heating at 80° C. for 5 minutes to obtain an inulin solution, stopping heating, and allowing to stand at 4° C. to form a non-flowing inulin gel.

6. The preparation method according to claim 5, characterized in that: Step (2) is specifically as follows: three probiotics, Clostridium butyricum, Bifidobacterium adolescentis, and Akkermansia muciniphila, are respectively taken and cultured to the logarithmic phase under an anaerobic environment, and centrifuged at 4°C: Clostridium butyricum and Bifidobacterium adolescentis are centrifuged at 5000 rpm for 5 min, and Akkermansia muciniphila is centrifuged at 5000 rpm for 10 min; Collect the bacterial precipitate and remove the supernatant; resuspend the bacterial solution with saline and centrifuge again: centrifuge Clostridium butyricum and Bifidobacterium adolescentis at 5000 rpm for 5 min, and Akkermansia muciniphila at 5000 rpm for 10 min, repeat 3 times; resuspend the bacteria with saline to a concentration of 1×10 8 CFU / mL; then, Clostridium butyricum and Bifidobacterium adolescentis were centrifuged at 5000 rpm for 5 min, and Akkermansia muciniphila was centrifuged at 5000 rpm for 10 min to obtain a precipitate; the precipitate was resuspended in physiological saline to a concentration of 5×10 8 CFU / mL; then the suspensions of the three probiotics were mixed in a volume ratio of 1:1:1 to obtain a composite probiotic suspension, and then centrifuged at 5000 rpm for 10 min to obtain the composite probiotics, and the supernatant was aspirated and discarded.

7. The preparation method according to claim 6, characterized in that: The expanded culture in step (2) includes: culturing Clostridium butyricum: taking a frozen Clostridium butyricum strain from the laboratory and dissolving it in a 37°C water bath, taking 100 μL of the bacterial solution and adding it to a shaking tube containing 9 mL of MRS broth liquid culture medium, transferring it to an anaerobic culture bag, and placing it in a 37°C constant temperature and humidity chamber, and anaerobic culture for 24 to 48 hours; culturing Bifidobacterium adolescentis: dissolving the lyophilized powder of Bifidobacterium adolescentis into a suspension with 200 μL of sterile water, adding it to a shaking tube containing 9 mL of MRS broth liquid culture medium, transferring it to an anaerobic culture bag, and placing it in a 37°C constant temperature and humidity chamber, and anaerobic culture for 24 to 48 hours; Cultivation of Akkermansia muciniphila: Take 1 mL of Akkermansia muciniphila solution and add it to 9 mL of modified brain heart infusion medium, transfer it to an anaerobic culture bag, and place it in a constant temperature and humidity chamber at 37°C for anaerobic culture for 72 hours.

8. Use of the composite probiotic inulin gel preparation according to any one of claims 1 to 3 in the preparation of medicines and health products.

9. The use according to claim 8, characterized in that Said medicine: a medicine that has a protective effect against ionizing radiation damage.

10. The use according to claim 9, characterized in that The ionizing radiation damage: radiation enteritis and hematopoietic system damage.

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