Composition combining probiotics and prebiotics as well as preparation method and application of composition

Abstract

The invention provides a composition combining probiotics and prebiotics as well as a preparation method and application of the composition, and relates to the technical field of biological medicines. The composition disclosed by the invention comprises the following components: compound probiotics and compound prebiotics, the compound probiotics are lactobacillus crispatus and plant lactobacillus in a mass ratio of (0.5-2): (3-8); the preservation number of the lactobacillus crispatus is CGMCC (China General Microbiological Culture Collection Center) No.26680, and the preservation number of the plant lactobacillus is CGMCC No.26679; the composite prebiotics are galactooligosaccharide, isomaltooligosaccharide, raffinose, spirulina and inulin, and the mass ratio of the galactooligosaccharide to the isomaltooligosaccharide to the raffinose to the spirulina to the inulin is (2.5-5): (2-4): (2-4): (0.5-3): (0.5-1.5). The prebiotics with specific components and proportions are compounded, so that the activity of the compound probiotics is protected, and the effects of treating inflammation and inhibiting harmful bacteria of vaginitis are improved.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedicine, and in particular relates to a composition of combined probiotics and prebiotics, and a preparation method and application thereof. Background Art

[0002] Vaginitis is one of the common gynecological diseases. Common types of vaginitis include Trichomonas vaginitis, vulvovaginal candidiasis, and bacterial vaginosis. The pathogenic bacteria that cause vaginitis mainly include Candida albicans, Staphylococcus aureus, Shigella flexneri, Shigella 301, Salmonella enteritidis, Escherichia coli O157, Salmonella typhimurium, beta-hemolytic Streptococcus, and Pseudomonas aeruginosa. Therefore, the first choice for drugs to treat vaginitis is antibacterial drugs with the purpose of sterilization and anti-inflammatory.

[0003] Probiotics are a class of active microorganisms that are beneficial to the host. They are the general name for active beneficial microorganisms that can produce definite health effects to improve the host microecological balance and play a beneficial role. There are many probiotics that are beneficial to the human body. Chinese invention patent CN115386519A discloses a lactobacillus composition, a probiotic preparation, and its preparation method and application. In the lactobacillus composition, by the synergistic cooperation of Lactobacillus crispatus DH200 and Lactobacillus plantarum DH230, vaginitis and urinary tract infection are both better improved. However, the activity of probiotics is often affected by the environment, and the survival rate is low, and the effect is unstable. There is a problem that the activity is reduced during storage and use, and the number of probiotics is significantly reduced, thereby affecting the probiotics to play a role. Therefore, it is necessary to protect the activity of probiotics.

[0004] Prebiotics are a kind of food ingredients that are not easily digested. They have a beneficial effect on the host and improve the host's health by selectively stimulating the growth and activity of one or several bacteria. Chinese invention patent CN107115364A discloses a composite probiotic and its preparation method; the composite probiotic is composed of more than ten probiotics and more than two prebiotics, the probiotics include bifidobacterium flora, Lactobacillus casei flora, Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus fermentum, Lactobacillus rhamnosus and Streptococcus thermophilus, and the prebiotics include soybean oligosaccharides and oligofructose; however, the invention adds more than ten kinds of probiotics, which not only increases the production cost, but also greatly increases the strict control of the production environment; even if prebiotics are added, due to the differences in the activity, living environment, etc. of different probiotics, prebiotics cannot protect the activity and effective bacterial count of multiple probiotics.

[0005] Chinese invention patent CN115531425A discloses a vaginal preparation containing active probiotics and its preparation method. The invention promotes the recovery of vaginal microecology by using exogenous prebiotics and probiotics. The preparation method of the preparation adopts multi-layer packaging technology to store prebiotics and probiotics in separate areas, preventing the problem of prebiotics being consumed due to post-fermentation during product storage, and allowing probiotics to be stored at room temperature without losing activity. However, the vaginal preparation prepared by the invention is only a suppository, and the preparation method is complicated, which is not conducive to industrial production.

[0006] In view of this, it is necessary to find a composition combining probiotics and prebiotics, compounding prebiotics of specific types and amounts, protecting the antibacterial activity of probiotics, improving the anti-inflammatory effect, and thus improving the therapeutic effect on vaginitis. Summary of the invention

[0007] In view of the problems existing in the prior art, the present invention provides a composition of combined probiotics and prebiotics, and a preparation method and application thereof. Prebiotics with specific ingredients and proportions are compounded to protect the activity of the composite probiotics, improve the treatment of inflammation and the inhibitory effect on harmful bacteria of vaginitis, and bring about a significant antibacterial effect.

[0008] To achieve the above purpose, the technical solution adopted by the present invention is as follows: First, the present invention provides a composition of combined probiotics and prebiotics, comprising components: composite probiotics and composite prebiotics; The composite probiotic is a mixture of Lactobacillus crispatus and Lactobacillus plantarum in a mass ratio of 0.5-2:3-8; the Lactobacillus crispatus is deposited as CGMCC No. 26680, and the Lactobacillus plantarum is deposited as CGMCC No.26679; The composite prebiotics are galacto-oligosaccharide, isomaltooligosaccharide, raffinose, spirulina and inulin, and the mass ratio of galacto-oligosaccharide, isomaltooligosaccharide, raffinose, spirulina and inulin is 2.5-5:2-4:2-4:0.5-3:0.5-1.5.

[0009] Preferably, the mass ratio of galacto-oligosaccharide, isomaltooligosaccharide, raffinose, spirulina and inulin is 3-4:2.5-3.5:2.5-3.5:1-2:0.8-1.2.

[0010] Further preferably, the mass ratio of galacto-oligosaccharide, isomaltooligosaccharide, raffinose, spirulina and inulin is 3.5:3:3:1.5:1.

[0011] Preferably, the composition comprises the following components in parts by weight: 0.5-2 parts of Lactobacillus crispatus, 3-8 parts of Lactobacillus plantarum, 2.5-5 parts of galacto-oligosaccharides, 2-4 parts of isomaltooligosaccharides, 2-4 parts of raffinose, 0.5-3 parts of spirulina and 0.5-1.5 parts of inulin.

[0012] Further preferably, the composition comprises the following components, in parts by weight: 0.8-1.2 parts of Lactobacillus crispatus, 4-6 parts of Lactobacillus plantarum, 3-4 parts of galacto-oligosaccharides, 2.5-3.5 parts of isomaltooligosaccharides, 2.5-3.5 parts of raffinose, 1-2 parts of spirulina and 0.8-1.2 parts of inulin.

[0013] More preferably, the composition comprises the following components, in parts by weight: 1 part of Lactobacillus crispatus, 5 parts of Lactobacillus plantarum, 3.5 parts of galacto-oligosaccharides, 3 parts of isomaltooligosaccharides, 3 parts of raffinose, 1.5 parts of spirulina and 1 part of inulin.

[0014] Then, the present invention provides a method for preparing the above composition, comprising the steps of: mixing the composite probiotics and the composite prebiotics to obtain the composition.

[0015] Preferably, the mixing process is carried out at a temperature controlled at 5-30°C.

[0016] Furthermore, the present invention provides a probiotic preparation, comprising the above composition.

[0017] Preferably, the probiotic preparation further comprises a component: a drug carrier.

[0018] The pharmaceutical carrier used can be solid, liquid or gas. Examples of solid carriers include lactose, kaolin, sucrose, talc, gelatin, agar, pectin, gum arabic, magnesium stearate and stearic acid. Examples of liquid carriers include syrup, peanut oil, olive oil and water. Examples of gaseous carriers include carbon dioxide and nitrogen.

[0019] When preparing the composition for oral dosage form, any convenient pharmaceutical medium can be used. For example, water, ethanol, oil, alcohol, flavoring agent, preservative, coloring agent, etc. can be used to form oral liquid preparations, such as suspensions and solutions; while carriers, such as starch, sugars, microcrystalline cellulose, diluents, granulating agents, emulsifiers, lubricants, binders, disintegrants can be used to form oral solid preparations, such as powders, capsules and tablets. Tablets and capsules are the preferred oral dosage units using solid pharmaceutical carriers due to their ease of administration. Tablets can be coated using standard aqueous or non-aqueous techniques.

[0020] Tablets containing the Chinese medicine composition or extract of the present invention can be prepared by tableting or molding, and one or more auxiliary ingredients or adjuvants can be used. The active ingredient can be tableted in a free-flowing form (such as powder or granules) in a suitable machine, and can be mixed with an adhesive, a lubricant, an inert diluent, a surfactant or a dispersant to prepare a tablet. Molded tablets can be molded in a suitable machine, i.e., a powdered compound mixture moistened with an inert liquid diluent. Each tablet preferably contains about 0.05 mg to about 5 g of active ingredient, and each sachet or capsule preferably contains about 0.05 mg to about 5 g of active ingredient. For example, a preparation intended for oral administration to the human body may contain about 0.5 mg to about 5 g of active drug, mixed with an appropriate and convenient carrier material, which may account for about 5% to 95% of the total composition. The unit dosage form usually contains about 1 mg to about 2 g of active ingredient, usually 25 mg, 50 mg, 100 mg, 200 mg, 300 mg, 400 mg, 500 mg, 600 mg, 800 mg or 1000 mg.

[0021] The pharmaceutical composition suitable for parenteral administration of the present invention can be prepared as an aqueous solution or suspension of the active compound. Suitable surfactants such as hydroxypropylcellulose can be included. Dispersions can also be prepared in glycerol, liquid polyethylene glycol and oil mixtures thereof. In addition, preservatives can be added to prevent the harmful growth of microorganisms.

[0022] Medicine of the present invention can be in the form suitable for topical use, for example aerosol, cream, ointment, lotion, powder or the like. In addition, composition can be in the form suitable for transdermal administration device. Can use Chinese medicine composition of the present invention, prepare these prescriptions by conventional processing method. For example, by mixing hydrophilic material and water, and about 5wt% to about 10wt% of compound, prepare cream or ointment with required consistency.

[0023] The medicine of the present invention can be in a form suitable for rectal administration, wherein the carrier is a solid. Preferably, the mixture is made into a unit dose suppository. Suitable carriers include cocoa butter and other materials commonly used in the art. Suppositories can be prepared by first forming a composition containing a softened or melted carrier, followed by cooling and shaping in a mold.

[0024] In addition to the above-mentioned carrier components, the above-mentioned pharmaceutical preparations may include (if applicable) one or more additional carrier components, such as diluents, buffers, flavoring agents, adhesives, surfactants, thickeners, lubricants, preservatives (including antioxidants), etc. In addition, other excipients, such as lactose, starch, cellulose derivatives, magnesium stearate, stearic acid, etc., colorants and flavoring agents, etc., may be added. The preparation is made isotonic with the blood of the intended recipient. The components containing the Chinese medicine composition of the present invention can also be prepared in the form of powder or concentrate.

[0025] Preferably, the dosage form of the probiotic preparation is not limited to pills, capsules, granules, oral liquids, powders, tablets, lozenges, pastes and the like, and suitable drug carriers in the art may be selected for different dosage forms.

[0026] Finally, the present invention provides the use of the above composition in preparing a drug for treating inflammation of the female reproductive system.

[0027] Preferably, the application is application in the preparation of a drug for treating female vaginitis.

[0028] Further preferably, the vaginal pathogens of vaginitis are not limited to: Candida albicans, Staphylococcus aureus, Shigella flexneri, Shigella 301, Salmonella enteritidis, Escherichia coli, Salmonella typhimurium, beta-hemolytic Streptococcus and Listeria monocytogenes.

[0029] Compared with the prior art, the present invention has the following beneficial effects: 1. In the present invention, galacto-oligosaccharides, isomaltooligosaccharides, raffinose, spirulina and inulin are selected as composite prebiotics. The components of the composite prebiotics interact with each other, which is conducive to being decomposed and utilized by probiotics, promoting Lactobacillus crispatus and Lactobacillus plantarum to exert antibacterial and anti-inflammatory effects, inhibiting the growth of harmful bacteria, and simplifying the formula to the greatest extent while improving the improvement effect on vaginitis.

[0030] 2. The composite prebiotic of the present invention is obtained by matching five specific ingredients with each other. The reasonable proportion of each component can protect the probiotics to still have high activity under high acid and high bile salt conditions, thereby promoting the growth and activity of probiotics and improving the antibacterial and anti-inflammatory effects.

[0031] 3. The present invention can achieve the protective effect on probiotics by using a small amount of prebiotics, the formula of the composition and probiotic preparation is simple, and the production conditions are easy to control.

[0032] Collection Instructions Deposit number: CGMCC No. 26680; Classification name: Lactobacillus crispatus Lactobacillus crispatus ; Storage time: February 27, 2023; Depository: General Microbiology Center, China Microbiological Culture Collection Administration; Abbreviation of depository unit: CGMCC; Collection address: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0033] Collection Instructions Deposit number: CGMCC No. 26679; Classification name: Lactobacillus plantarum Lactiplantibacillus plantarum ; Storage time: February 27, 2023; Depository: General Microbiology Center, China Microbiological Culture Collection Administration; Abbreviation of depository unit: CGMCC; Collection address: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. DETAILED DESCRIPTION

[0034] The following non-limiting examples can enable those skilled in the art to more fully understand the present invention, but do not limit the present invention in any way. The following content is merely an exemplary description of the scope of the present invention, and those skilled in the art can make various changes and modifications to the present invention based on the disclosed content, which should also fall within the scope of the present invention.

[0035] When the embodiments give numerical ranges, it should be understood that, unless otherwise specified in the present invention, both endpoints of each numerical range and any numerical value between the two endpoints can be selected. Unless otherwise defined, all technical and scientific terms used in the present invention have the same meanings as those generally understood by those of ordinary skill in the art to which the present invention belongs.

[0036] The present invention is further described below by way of specific examples. Unless otherwise specified, the various chemical reagents used in the examples of the present invention are obtained through conventional commercial channels.

[0037] In the following examples, the polymerization degree of the polygalactose is 2-6, and the polymerization degree of the oligogalactose is 4; the polymerization degree of the oligomaltosan is 2-10, and the polymerization degree of the oligomaltosan is 8. For the above components, products from different manufacturers have no significant effect on the effects.

[0038] In the present invention, the preservation number of the Lactobacillus crispatus is CGMCC No. 26680, and the preservation number of Lactobacillus plantarum is CGMCC No. 26679. The viable cell count of the Lactobacillus crispatus is 1×10 7 CFU / -10×10 7 CFU / g, the number of viable bacteria of Lactobacillus plantarum is 1×10 8 CFU / g-10×10 8 CFU / g.

[0039] Example 1 A composition of combined probiotics and prebiotics, comprising the following components by weight: 1 part of Lactobacillus crispatus, 5 parts of Lactobacillus plantarum, 3.5 parts of galacto-oligosaccharide, 3 parts of isomaltooligosaccharide, 3 parts of raffinose, 1.5 parts of spirulina and 1 part of inulin. The above components are mixed uniformly at room temperature to obtain the composition.

[0040] Example 2 A composition of combined probiotics and prebiotics, comprising the following components by weight: 0.8 parts of Lactobacillus crispatus, 4 parts of Lactobacillus plantarum, 3 parts of galacto-oligosaccharide, 2.5 parts of isomaltooligosaccharide, 2.5 parts of raffinose, 1 part of spirulina and 0.8 parts of inulin. The above components are mixed uniformly at room temperature to obtain the composition.

[0041] Example 3 A composition of combined probiotics and prebiotics, comprising the following components by weight: 1.2 parts of Lactobacillus crispatus, 6 parts of Lactobacillus plantarum, 4 parts of galacto-oligosaccharide, 3.5 parts of isomaltooligosaccharide, 3.5 parts of raffinose, 2 parts of spirulina and 1.2 parts of inulin. The above components are mixed uniformly at room temperature to obtain the composition.

[0042] Example 4 A composition of combined probiotics and prebiotics, comprising the following components by weight: 0.5 parts of Lactobacillus crispatus, 3 parts of Lactobacillus plantarum, 2.5 parts of galacto-oligosaccharide, 2 parts of isomaltooligosaccharide, 2 parts of raffinose, 0.5 parts of spirulina and 0.5 parts of inulin. The above components are mixed uniformly at room temperature to obtain the composition.

[0043] Example 5 A composition of combined probiotics and prebiotics, comprising the following components by weight: 2 parts of Lactobacillus crispatus, 8 parts of Lactobacillus plantarum, 5 parts of galacto-oligosaccharide, 4 parts of isomaltooligosaccharide, 4 parts of raffinose, 3 parts of spirulina and 3 parts of inulin. The above components are mixed uniformly at room temperature to obtain the composition.

[0044] Comparative Example 1 The difference from Example 1 is that the ratio of Lactobacillus crispatus and Lactobacillus plantarum is different. Specifically: A composition of combined probiotics and prebiotics, comprising the following components by weight: 0.3 parts of Lactobacillus crispatus, 5.7 parts of Lactobacillus plantarum, 3.5 parts of galacto-oligosaccharide, 3 parts of isomaltooligosaccharide, 3 parts of raffinose, 1.5 parts of spirulina and 1 part of inulin. The above components are mixed uniformly at room temperature to obtain the composition.

[0045] Comparative Example 2 The difference from Example 1 is that the components of the composite prebiotics are different, and isomaltooligosaccharide is replaced by galactose.

[0046] Comparative Example 3 The difference from Example 1 is that the components of the composite prebiotics are different, and raffinose is replaced by stachyose.

[0047] Comparative Example 4 Different from Example 1, the components of the composite prebiotics are galacto-oligosaccharide, isomaltooligosaccharide, spirulina and inulin. Specifically: A composition of combined probiotics and prebiotics, comprising, by weight, 1 part of Lactobacillus crispatus, 5 parts of Lactobacillus plantarum, 3.5 parts of galacto-oligosaccharides, 3 parts of isomaltooligosaccharides, 1.5 parts of spirulina and 1 part of inulin. The above components are mixed uniformly at room temperature to obtain the composition.

[0048] Comparative Example 5 The difference from Example 1 is that the component of the composite prebiotic is raffinose. Specifically: A composition of combined probiotics and prebiotics, comprising the following components by weight: 1 part of Lactobacillus crispatus, 5 parts of Lactobacillus plantarum, and 3 parts of raffinose. The above components are mixed uniformly at room temperature to obtain the composition.

[0049] Comparative Example 6 Different from Example 1, the ratio of each component in the composite prebiotics is different, specifically: A composition of combined probiotics and prebiotics, comprising the following components by weight: 1 part of Lactobacillus crispatus, 5 parts of Lactobacillus plantarum, 1 part of galacto-oligosaccharide, 1 part of isomaltooligosaccharide, 1 part of raffinose, 4 parts of spirulina and 5 parts of inulin. The above components are mixed uniformly at room temperature to obtain the composition.

[0050] Test 1 Acid resistance and bile salt resistance test 1. Acid resistance test (1) The composition of each example and comparative example was cultured in an MRS liquid culture medium in a constant temperature incubator at 37° C. for 24-36 hours to prepare a bacterial solution of the composition; no prebiotics were added to the control group.

[0051] (2) Prepare PBS buffer at pH = 2, take 1 mL of bacterial culture solution that has been cultured for 24-36 hours, centrifuge at 3000 rpm for 10 minutes, discard the supernatant, wash twice with PBS, add 1 mL of PBS at pH = 2 to resuspend, place in a constant temperature incubator at 37°C for 4 hours, then dilute and plate, and count the viable bacteria (Lactobacillus plantarum). Repeat three times. The viable bacteria count results of the acid resistance experiment are shown in Table 1.

[0052] Table 1

[0053] In Table 1, # P <0.05, ## P <0.01, compared with the group in Example 1.

[0054] As can be seen from Table 1, the composite prebiotics of the present invention can protect probiotics, have high tolerance under strong acidic conditions, and can still grow in a strong acidic environment of pH = 2. The results of Comparative Examples 4 and 5 show that in the composition of the present invention, the prebiotic components interact with each other to bring about a synergistic effect of protecting the activity of probiotics; the results of Comparative Examples 2, 3 and 6 show that the specific prebiotic ingredients and ratios not described in the present invention have a low protective effect on probiotics.

[0055] 2. Bile salt tolerance test (1) The composition of each example and comparative example was cultured in MRS liquid culture medium in a 37°C constant temperature incubator for 24-36 hours to prepare a bacterial solution of the composition; (2) Take 10 mL of the culture solution that has been cultured for 24 hours, centrifuge at 3000 rpm for 10 minutes, discard the supernatant, wash twice with PBS, and resuspend the bacteria in 5 mL of MRS medium. Add bile salts to prepare a 0.2% MRS culture medium. Take 200 μL of the resuspended bacterial solution and put it into the prepared 10 mL culture medium and culture it at 37°C for 24-36 hours, then dilute and plate for counting. Repeat 3 times. The results of the viable bacterial count (Lactobacillus crispatus) of the bile salt tolerance test are shown in Table 2. No prebiotics were added to the control group.

[0056] Table 2

[0057] In Table 2, # P <0.05, ## P <0.01, compared with the group in Example 1.

[0058] As can be seen from Table 2, the composite prebiotics of the present invention can protect probiotics, have a higher tolerance at a higher bile salt concentration, can still grow in a 0.2% bile salt environment, and have a significantly increased number of viable bacteria ( P <0.01); the results of comparative examples 4 and 5 show that the composite prebiotic composition of the present invention has the effect of synergistically protecting the activity of probiotics by the interaction of the prebiotic components; the results of comparative examples 2, 3 and 6 show that the specific prebiotic ingredients and proportions not described in the present invention have a low protective effect on probiotics.

[0059] Test 2 Antibacterial performance (1) inoculating each composition into MRS liquid culture medium, culturing in a carbon dioxide incubator at 37° C. for 24-36 hours, and centrifuging at 3000 rpm for 10 minutes to obtain a treated composition; (2) 1×10 9CFU / mL 30μL of Candida albicans, Staphylococcus aureus, Listeria monocytogenes were coated in LB solid culture medium. The Oxford cup was gently placed on the plate, 250μL of the treated composition was pipetted into the Oxford cup, and cultured at 37°C. The diameter of the inhibition zone was measured after 8 hours.

[0060] The results of the inhibition zone diameter (mm) of each group are shown in Table 3.

[0061] Table 3

[0062] In Table 3, # P <0.05, ## P <0.01, compared with the group in Example 1.

[0063] It can be seen from the results of the antibacterial zone diameter in Table 3 that the antibacterial performance test results show that the composition of the combined probiotics and prebiotics of the present invention significantly inhibits the growth of Candida albicans, Staphylococcus aureus, and Listeria monocytogenes, and the antibacterial effect is significantly better than that of the comparative example. The composition of comparative example 1 changes the ratio of Lactobacillus crispatus and Lactobacillus plantarum, so that the synergistic effect between the two probiotics is destroyed, and the antibacterial effect is significantly reduced ( P <0.01, P <0.05); Comparative Examples 2-5 changed the composition of prebiotics, the protective effect on probiotics decreased, the probiotics played a reduced role, and thus significantly affected the size of the inhibition zone; Comparative Example 6 changed the ratio of prebiotic components, and the antibacterial effect was significantly reduced ( P <0.01, P <0.05). Therefore, only by using the combined probiotics and prebiotics composition of the present invention can a significantly improved antibacterial effect be achieved ( P <0.01, P <0.05).

[0064] Experiment 3 Animal experiment 1. Purchase and living environment of experimental animals Animals were purchased from Hunan Slake Jingda Experimental Animal Co., Ltd. 104 BALB / c female mice (SPF level, 6-8 weeks old, weighing 20-22g). Mice had free access to water and food during the experiment. Animals were fed with standard pellet feed every day. Cleaned regularly every day. Mice were kept in sterile polypropylene cages under specific pathogen-free conditions. They were maintained in a room with controlled temperature (22℃±2℃) and humidity (50%±15%) with a light-dark cycle (12h:12h), and provided with standard diet and water. All mice used in this study were caged one week before the start of the study to adapt to housing conditions.

[0065] 2. Establishment of animal model BALB / c female mice were subcutaneously injected with 0.05 mL of 2 mg / mL estradiol phthalate injection for 3 consecutive times, once every 2 days. On the 6th day, the vagina of the mice was first flushed with sterile PBS solution (pH = 8.5) 3 times (5 minutes between each flushing), and beta-hemolytic streptococci, Staphylococcus aureus, and Escherichia coli O157 were prepared at a volume ratio of 1:1:2 to a concentration of 1×10 9 CFU / mL of mixed bacterial solution; cut the medical absorbable gelatin sponge into 0.5cm×0.5cm size, inject 20μL of infected bacterial solution into the sponge, and plug the sponge injected with bacteria into the mouse vagina (inserted into the mouse vagina about 1cm-1.5cm, the injection amount of each bacteria is 0.025mL / 100g, 1 time / d); when the mouse vagina is obviously congested, red and swollen with a large amount of purulent secretions, it indicates that the mouse BV model (bacterial vaginitis model) is successfully prepared; estradiol benzoate injection is given in the same way, and on the 6th day, a surgical absorbable hemostatic sponge injected with 20μL of normal saline is plugged into the mouse vagina as a blank control group.

[0066] 3. Animal model grouping and drug treatment (1) Animal model groups: The mice were grouped according to their weight using a randomized block design analysis of variance method, with a total of 13 groups, each with 8 mice, including: a blank control group, a model control group, Example 1-Example 5 groups, and Comparative Example 1-Comparative Example 6 groups.

[0067] (2) Drug treatment: Blank control group (C): flush the vagina with 100 μL normal saline once a day for 14 consecutive days; Model control group (M): flush the vagina with 100 μL normal saline once a day for 14 consecutive days; Example group: Take the composition of each example (containing 1g of probiotics) to wash the vagina once a day for 14 consecutive days; Comparative Example Group: The composition of each comparative example (containing 1 g of probiotics) was taken to flush the vagina once a day for 14 consecutive days.

[0068] 4. qPCR detection The reverse transcription and qPCR reaction system kit (purchased from TaKaRa, catalog number RR047A) was used to detect the levels of IL-1β, IL-10, and IL-17 in vaginal tissue by qPCR according to the instructions. The test results are shown in Table 4.

[0069] Table 4

[0070] In Table 4, # P <0.05, ## P <0.01, ### P <0.001, compared with Example 1; ★ P <0.05, ★★ P <0.01, ★★★ P <0.001, compared with the model control group; △ P <0.05, △△ P <0.01, △△△ P <0.001, compared with the normal control group.

[0071] As can be seen from Table 4, compared with the blank control group, the expression levels of IL-1β and IL-17 in the model control group were significantly increased, and the expression of these pro-inflammatory factors in the Example 1-Example 5 groups was significantly reduced ( ★★★ P <0.001); Although the expression of pro-inflammatory factors can be reduced in the comparative examples 1 to 6, the inhibitory effect is significantly reduced compared with the example group ( # P <0.05, ## P <0.01). The expression of anti-inflammatory factor IL-10 showed an opposite trend.

[0072] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, rather than to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions of the technical solution of the present invention by ordinary technicians in this field do not deviate from the essence and scope of the technical solution of the present invention.

Claims

1. A composition combining probiotics and prebiotics, characterized in that: Including components: compound probiotics and compound prebiotics; The composite probiotic is a mixture of Lactobacillus crispatus and Lactobacillus plantarum in a mass ratio of 0.5-2:3-8; the Lactobacillus crispatus is deposited as CGMCC No. 26680, and the Lactobacillus plantarum is deposited as CGMCC No. 26679; The composite prebiotics are galacto-oligosaccharide, isomaltooligosaccharide, raffinose, spirulina and inulin, and the mass ratio of galacto-oligosaccharide, isomaltooligosaccharide, raffinose, spirulina and inulin is 2.5-5:2-4:2-4:0.5-3:0.5-1.

5.

2. The composition according to claim 1, characterized in that The mass ratio of the galacto-oligosaccharide, isomaltooligosaccharide, raffinose, spirulina and inulin is 3-4:2.5-3.5:2.5-3.5:1-2:0.8-1.

2.

3. The composition according to claim 2, characterized in that The mass ratio of the galacto-oligosaccharide, isomaltooligosaccharide, raffinose, spirulina and inulin is 3.5:3:3:1.5:

1.

4. The composition according to claim 1, characterized in that The composition comprises the following components in parts by weight: 0.5-2 parts of Lactobacillus crispatus, 3-8 parts of Lactobacillus plantarum, 2.5-5 parts of galacto-oligosaccharide, 2-4 parts of isomaltooligosaccharide, 2-4 parts of raffinose, 0.5-3 parts of spirulina and 0.5-1.5 parts of inulin.

5. The composition according to claim 4, characterized in that The composition comprises the following components in parts by weight: 0.8-1.2 parts of Lactobacillus crispatus, 4-6 parts of Lactobacillus plantarum, 3-4 parts of galacto-oligosaccharide, 2.5-3.5 parts of isomaltooligosaccharide, 2.5-3.5 parts of raffinose, 1-2 parts of spirulina and 0.8-1.2 parts of inulin.

6. The composition according to claim 5, characterized in that The composition comprises the following components in parts by weight: 1 part of Lactobacillus crispatus, 5 parts of Lactobacillus plantarum, 3.5 parts of galacto-oligosaccharides, 3 parts of isomaltooligosaccharides, 3 parts of raffinose, 1.5 parts of spirulina and 1 part of inulin.

7. A method for preparing the composition according to any one of claims 1 to 6, characterized in that: The method comprises the steps of: mixing the compound probiotics and the compound prebiotics to obtain a composition.

8. The preparation method according to claim 7, characterized in that: The temperature of the mixing process is controlled at 5-30°C.

9. A probiotic preparation, characterized in that: The invention comprises the composition according to any one of claims 1 to 6.

10. Use of the composition in the preparation of a drug for treating inflammation of the female reproductive system, characterized in that: The composition is the composition according to any one of claims 1 to 6 or the composition prepared by the preparation method according to any one of claims 7 to 8.

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