Lactobacillus fermentum as well as compound preparation and application thereof

Abstract

The invention provides lactobacillus fermentum as well as a compound preparation and application thereof, and relates to the technical field of microorganisms. Compared with a strain in the prior art, the lactobacillus fermentum HXJS4-1 disclosed by the invention has a better effect in the aspect of nucleoside degradation and can realize rapid nucleoside degradation. In addition, the compound has a stronger inhibition effect on xanthine oxidase, and the activity of the xanthine oxidase is effectively reduced. In mouse experiments, the strain also has an obvious uric acid reducing effect, also has a certain anti-inflammatory effect, can be used in the prevention and treatment process of hyperuricemia and gout, and provides a new probiotic or drug material for the treatment of hyperuricemia and gout. In addition, eating of the strain has no obvious influence on indexes of a sample body, and it is proved that the strain has the characteristic of high safety and can be widely applied to the fields of medicine and food.

Description

Technical Field

[0001] The present application relates to the field of microbial technology, and in particular to a strain of Lactobacillus fermentum and its composite preparation and application. Background Art

[0002] Hyperuricemia (HUA) is a chronic clinical syndrome in which the blood uric acid content significantly exceeds the normal range due to purine metabolism disorder or abnormal uric acid excretion. It generally refers to the fasting blood uric acid level of two different days higher than 420μmol / L for men and higher than 360μmol / L for women under normal purine diet. Gout (Gout) is a crystalline arthritis caused by monosodium urate deposition, which is directly related to hyperuricemia caused by purine metabolism disorder. In recent years, hyperuricemia in my country has shown a significant upward and younger trend. Hyperuricemia is not only the early stage of gout, but also an independent risk factor for hypertension, diabetes, coronary heart disease, and chronic kidney disease. Gout has become the "fourth high" after diabetes, hypertension, and hyperlipidemia, causing serious harm to people's health.

[0003] Clinically, hyperuricemia is generally prevented and treated in three ways. The first way is to reduce the production of uric acid, using drugs such as xanthine oxidase (XOD) inhibitors, such as allopurinol and febuxostat; the second way is to increase the excretion of uric acid, using drugs such as URAT1 inhibitors, such as benzbromarone and lesinade; the third way is to degrade uric acid, using drugs such as rasburicase and purikaxib. However, xanthine oxidase inhibitors and URAT1 inhibitors are chemical drugs, which have certain damage to liver and kidney function and have strong toxic side effects. Although urate oxidase drugs have better efficacy, they have strong immunogenicity and are generally used as second-line drugs.

[0004] Under normal circumstances, there are about 1200 mg of uric acid in the human body, about 600 mg is newly generated every day, and 600 mg is excreted at the same time, which is in a balanced state. About 70% of the uric acid excreted by the human body is excreted through the kidneys, and the rest is excreted with feces or further decomposed and metabolized by intestinal flora. A small part is excreted in the human intestine or decomposed by intestinal flora. In recent years, improving traditional metabolic diseases such as hyperuricemia based on intestinal microecology has become a new development direction. Many studies have shown that regulating intestinal microecology through probiotics, prebiotics, etc. plays an important role in regulating serum uric acid and uric acid distribution in the body and helps in the treatment of hyperuricemia and gout.

[0005] It can be seen that the current drugs used for hyperuricemia have many problems, high side effects, and are not suitable for long-term use. While more and more probiotics have been shown to have good effects in preventing or relieving hyperuricemia, their efficacy in the actual auxiliary treatment of HUA and its related diseases needs to be improved urgently. Summary of the invention

[0006] In view of the above problems, the present invention aims to screen out probiotics and related preparations that can better reduce blood uric acid levels, relieve hyperuricemia and gout, and tolerate carbon and / or nitrogen source substrates.

[0007] On the one hand, the present application provides a strain of Lactobacillus fermentum HXJS4-1, which is deposited in the General Microbiological Center of China Microbiological Culture Collection Administration with a deposit number of CGMCC NO: 29677.

[0008] On the other hand, the present application also provides a strain of Lactobacillus paracasei HXXJ-008, which is deposited in the General Microbiological Center of China Microbiological Culture Collection Administration with a deposit number of CGMCC NO: 30483.

[0009] On the other hand, the present application also provides a preparation, which comprises any one or more of Lactobacillus fermentum, Lactobacillus paracasei and / or Lactobacillus plantarum.

[0010] Preferably, the fermented Lactobacillus is the fermented Lactobacillus (Lactobacillus fermentum) HXJS4-1,

[0011] More preferably, the Lactobacillus paracasei is the Lactobacillus paracasei HXXJ-008.

[0012] Most preferably, the plant lactobacillus is plant lactobacillus (Lactobacillus plantarum) HXJS3-1, which is deposited in the General Microbiological Center of China Microbiological Culture Collection Administration with a deposit number of CGMCC NO: 29676.

[0013] In a preferred embodiment, the fermented Lactobacillus is Lactobacillus fermented HXJS4-1, which is deposited in the General Microbiological Center of China National Microbiological Culture Collection Administration with a deposit number of CGMCC NO: 29677.

[0014] Preferably, the fermented lactobacillus may include fermented lactobacillus HXJS4-1, fermented lactobacillus HXJS4-1 bacterial agent, fermented lactobacillus HXJS4-1 dead bacteria suspension, fermented lactobacillus HXJS4-1 metabolites, and fermented lactobacillus HXJS4-1 extracts.

[0015] In a preferred embodiment, the Lactobacillus paracasei is Lactobacillus paracasei HXXJ-008, which is deposited in the General Microbiological Center of China National Microbiological Culture Collection Committee with a deposit number of CGMCC NO: 30483.

[0016] Preferably, the Lactobacillus paracasei may include Lactobacillus paracasei HXXJ-008, a Lactobacillus paracasei HXXJ-008 bacterial agent, a dead bacteria suspension of Lactobacillus paracasei HXXJ-008, a metabolite of Lactobacillus paracasei HXXJ-008, or an extract of Lactobacillus paracasei HXXJ-008.

[0017] Preferably, the Lactobacillus plantarum may include Lactobacillus plantarum HXJS3-1, a Lactobacillus plantarum HXJS3-1 bacterial agent, a dead bacteria suspension of Lactobacillus plantarum HXJS3-1, a metabolite of Lactobacillus plantarum HXJS3-1, or an extract of Lactobacillus plantarum HXJS3-1.

[0018] It will be understood by those skilled in the art that known excipients and solvents may be added to the preparation without affecting the activity and efficacy of the strain.

[0019] The excipients may be appropriate solvents, propellants, solubilizers, cosolvents, emulsifiers, colorants, adhesives, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, glidants, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-adhesives, integrators, penetration enhancers, pH regulators, buffers, plasticizers, surfactants, foaming agents, defoamers, thickeners, inclusion agents, humectants, absorbents, diluents, flocculants and deflocculating agents, filter aids, release retardants, etc.

[0020] Furthermore, the preparation can be prepared by a common method, such as directly mixing the strains. One or more diluents or pharmaceutically acceptable carriers can also be added. In a preferred embodiment, the preparation can be prepared as an oral preparation.

[0021] Furthermore, the total number of viable bacteria of the fermented Lactobacillus, the paracasei Lactobacillus and / or the plantarum Lactobacillus is greater than 1×10 8 CFU.

[0022] Preferably, 1×10 8 CFU-1×10 12 CFU.

[0023] The total number of viable bacteria of the fermented Lactobacillus, paracasei Lactobacillus and / or plantarum Lactobacillus may be 1×10 8 CFU, 1×10 9 CFU, 1×10 10 CFU, 1×10 11 CFU, 1×10 12 CFU and above.

[0024] In a preferred embodiment, the sum of the viable counts of Lactobacillus fermentum, Lactobacillus paracasei and / or Lactobacillus plantarum and their composite strains is greater than 1×10 8 CFU.

[0025] It is understood by those skilled in the art that the amount of bacteria can be adjusted according to the actual situation, as long as the sum of the total viable counts of the three strains mixed randomly is greater than 1×10 8 CFU is sufficient. This application only gives an example and does not impose any specific upper limit on it.

[0026] In this application, for the first time, a combination of fermented Lactobacillus HXJS4-1, Lactobacillus paracasei HXXJ-008, and Lactobacillus plantarum HXJS3-1 was used to achieve better effects of lowering uric acid content in hyperuricemia and reducing inflammatory factors, proving that there is a synergistic effect between the three strains, and the effect can reach the level of the positive drug group.

[0027] In a preferred embodiment, the preparation can be a medicine, probiotic yogurt, soy milk, compressed candy and / or solid beverage.

[0028] On the other hand, the present application also provides the use of the fermented Lactobacillus or the Lactobacillus paracasei or the preparation in degrading nucleosides and / or preparing degraded nucleoside products.

[0029] Preferably, the nucleoside comprises one or more of inosine, guanosine and adenosine; more preferably, the nucleoside comprises inosine and / or guanosine.

[0030] Preferably, Lactobacillus fermentum HXJS4-1.

[0031] Preferably, the degradation is rapid degradation.

[0032] In this application, Lactobacillus fermentum HXJS4-1 is used for the first time to degrade nucleosides. Among them, Lactobacillus fermentum HXJS4-1 can achieve rapid degradation of nucleosides. In a specific embodiment, Lactobacillus fermentum HXJS4-1OD 600 When the value is 4.0, the inosine-guanosine degradation solution with a final concentration of about 1.26 mM can be completely degraded within 1 hour.

[0033] Preferably, the process of degrading nucleosides is not affected by carbon sources and / or nitrogen sources, that is, when sugar substances or proteins are contained, the fermented Lactobacillus or the Lactobacillus paracasei described in the present application can still maintain efficient nucleoside degradation capabilities.

[0034] Preferably, the carbon source can be selected from glucose, sucrose, starch, glycerol, etc., and the nitrogen source can be selected from yeast extract powder, peptone, yeast extract, beef extract, corn steep liquor, etc.

[0035] More preferably, the concentration of the carbon source may be 1-5 g / L, and the concentration of the nitrogen source may be 1-5 g / L.

[0036] The concentration of the carbon source can be any value among 1g / L, 2g / L, 3g / L, 4g / L, and 5g / L, and the concentration of the nitrogen source can be any value among 1g / L, 2g / L, 3g / L, 4g / L, and 5g / L.

[0037] In a preferred embodiment, the carbon source is sucrose and / or glucose, and the nitrogen source is yeast and its extract.

[0038] On the other hand, the present application also provides the use of the fermented Lactobacillus or the Lactobacillus paracasei or the preparation in inhibiting xanthine oxidase (XOD) activity and / or in preparing a xanthine oxidase (XOD) inhibition product.

[0039] In this application, for the first time, fermented lactobacillus HXJS4-1 and / or Lactobacillus paracasei HXXJ-008 are used to verify the xanthine oxidase inhibition ability. Among them, the inhibition rate of xanthine oxidase of fermented lactobacillus HXJS4-1 can reach 71.5%. That is, this application provides a new microbial material for xanthine oxidase inhibitors and related products.

[0040] On the other hand, the present application also provides the use of the fermented Lactobacillus or the Lactobacillus paracasei or the preparation for inhibiting inflammation for non-disease diagnosis and treatment purposes and / or preparing an inflammation-inhibiting product.

[0041] Among them, inhibiting inflammation is achieved by reducing the content of inflammatory factors or increasing the content of anti-inflammatory factors. The inflammation-related factors include but are not limited to interleukin-1β (IL-1β), IL-6, TNF-α, IL-10, and IL-11.

[0042] This application uses for the first time Lactobacillus fermentum HXJS4-1, Lactobacillus plantarum HXJS3-1 and / or Lactobacillus paracasei HXXJ-008 to inhibit inflammatory factors and reduce the number of inflammatory-related factors in the sample.

[0043] In a preferred embodiment, the present application specifically verifies the content of inflammatory-related factors in the liver and kidney of the sample, and proves that Lactobacillus fermentum HXJS4-1, Lactobacillus plantarum HXJS3-1 and Lactobacillus paracasei HXXJ-008 and their combination can effectively reduce the content of inflammatory factors in the liver and kidney and increase the content of anti-inflammatory factors.

[0044] The highest inhibition rates of IL-1β were 77.5% and 81.93%, respectively;

[0045] The highest inhibition rates of IL-6 were 84.7% and 71.78%, respectively;

[0046] The highest inhibition rates of TNF-α were 77.51% and 79.45% respectively;

[0047] The highest recovery rates for IL-10 were 70.55% and 73.1%, respectively;

[0048] The highest inhibition rates of IL-11 could reach 77.77% and 83.19% respectively.

[0049] Preferably, the sample is a hyperuricemia sample.

[0050] On the other hand, the present application also provides the use of the fermented Lactobacillus or the Lactobacillus paracasei or the preparation in the preparation of a product for reducing uric acid levels.

[0051] Preferably, the uric acid level may be the uric acid content in urine or the uric acid content in serum.

[0052] This application is the first to use Lactobacillus fermentum HXJS4-1, Lactobacillus plantarum HXJS3-1 and / or Lactobacillus paracasei HXXJ-008 to prepare products for reducing uric acid levels and for use in reducing uric acid levels.

[0053] On the other hand, the present application also provides the use of the fermented Lactobacillus or the Lactobacillus paracasei or the preparation in the preparation of a product for preventing and / or treating hyperuricemia.

[0054] Among them, prevention and / or treatment of hyperuricemia can be achieved by inhibiting xanthine oxidase (XOD) activity, reducing the content of inflammatory factors while increasing the content of anti-inflammatory factors and reducing the content of uric acid.

[0055] On the other hand, the present application also provides the use of the fermented Lactobacillus or the Lactobacillus paracasei or the preparation in the preparation of a product for preventing and / or treating gout.

[0056] The fermented Lactobacillus HXJS4-1, Lactobacillus plantarum HXJS3-1 and / or Lactobacillus paracasei HXXJ-008 and preparations thereof described in the present application can effectively reduce the probability of further conversion of hyperuricemia into gout while reducing the risk of hyperuricemia, thereby playing a role in preventing and / or treating gout.

[0057] Specifically, since an increase in serum uric acid content can cause an individual to suffer from hyperuricemia, and hyperuricemia can cause an increase in the concentration of inflammatory factors in the body, thereby increasing the risk of gout in an individual. In addition, studies have shown that an increase in the concentration of inflammatory factors may lead to an increase in XOD activity, and XOD can convert xanthine into uric acid and increase the production of uric acid. Therefore, in this application, the production of uric acid is reduced by inhibiting the activity of XOD, and at the same time, the risk of gout in an individual is reduced by reducing the level of inflammatory factors.

[0058] On the other hand, the present application also provides the use of the fermented Lactobacillus and / or the Lactobacillus paracasei and / or the preparation in improving immunity.

[0059] On the other hand, the present application also provides the use of the Lactobacillus fermentum and / or the Lactobacillus paracasei and / or the preparation in the preparation of an IL-11 inhibitor.

[0060] Furthermore, the IL-11 inhibitor can be used in oral preparations and / or external preparations for anti-aging and / or life extension.

[0061] Preferably, the IL-11 inhibitor can be used to prevent or treat various age-related diseases selected from cardiovascular fibrosis, pulmonary fibrosis, cancer, diabetic complications, mitochondrial dysfunction, inflammation and cellular senescence, thrombocytopenia, muscle weakness and weakness, coronary artery disease, metabolic decline, coronary artery disease, hypertension, pulmonary hypertension, cerebrovascular disease, aortic disease, and other vascular diseases.

[0062] In addition, the fermented Lactobacillus HXJS4-1, the Lactobacillus plantarum HXJS3-1 and / or the Lactobacillus paracasei HXXJ-008 described in the present application are highly safe and have no significant effect on other indicators of the body after taking, which proves that they are safe and non-toxic.

[0063] The present invention has the following beneficial effects:

[0064] The strain fermented lactobacillus HXJS4-1 disclosed in the present invention has a better effect in nucleoside degradation than the prior art strains, can achieve rapid degradation of nucleosides, and the process of degrading nucleosides is not affected by carbon sources and / or nitrogen sources, that is, when containing carbohydrates or proteins, the fermented lactobacillus described in the present application can still maintain efficient nucleoside degradation ability; and, it has a stronger inhibitory effect on xanthine oxidase, effectively reducing the activity of xanthine oxidase. And the strain also has a significant uric acid-lowering effect on hyperuricemia mice, and has a certain anti-inflammatory effect.

[0065] The present invention also provides a composite preparation of fermented Lactobacillus HXJS4-1, plantarum Lactobacillus HXJS3-1 and Lactobacillus paracasei HXXJ-008, which proves that the three strains have a synergistic effect and can be used in the prevention and treatment of hyperuricemia and gout, providing a new probiotic or drug material for the treatment of hyperuricemia and gout.

[0066] The present invention also provides the use of the fermented lactobacillus and / or the Lactobacillus paracasei and / or the preparation in the preparation of IL-11 inhibitors. The oral preparations and / or external preparations can be used for anti-aging and / or life extension, and can also be used to prevent or treat various age-related diseases selected from cardiovascular fibrosis, pulmonary fibrosis, cancer, diabetic complications, mitochondrial dysfunction, inflammation and cell senescence, thrombocytopenia, muscle weakness and weakness, coronary artery disease, metabolic decline, coronary artery disease, hypertension, pulmonary hypertension, cerebrovascular disease, aortic disease, and other vascular diseases.

[0067] In addition, consumption of the above-mentioned strains has no obvious effect on the body indicators of the samples, proving that they are highly safe and can be widely used in the fields of medicine and food. BRIEF DESCRIPTION OF THE DRAWINGS

[0068] The drawings described herein are used to provide a further understanding of the present application and constitute a part of the present application. The illustrative embodiments of the present application and their descriptions are used to explain the present application and do not constitute an improper limitation on the present application. In the drawings:

[0069] Figure 1 is a schematic diagram of the morphology of strain HXJS4-1;

[0070] Figure 2 is a schematic diagram of the morphology of strain HXXJ-008;

[0071] Figure 3 is a statistical graph of IL-1β content in the liver and kidney of mice in Example 8;

[0072] Figure 4 is a statistical graph of IL-6 content in the liver and kidney of mice in Example 8;

[0073] Figure 5 is a statistical graph of XOD activity in mouse liver and serum in Example 9;

[0074] Figure 6 is a statistical graph of serum uric acid levels of mice on days 0, 7, 14, and 21 in Example 9;

[0075] Figure 7 is a statistical graph of uric acid content in mouse urine in Example 9;

[0076] Figure 8 is a statistical graph of IL-1β content in the liver and kidney of mice in Example 9;

[0077] Fig. 9 is a statistical graph of TNF-α content in the liver and kidney of mice in Example 9;

[0078] Fig.10 is a statistical graph of IL-6 content in the liver and kidney of mice in Example 9;

[0079] Fig.11 is a statistical graph of IL-11 content in the liver and kidney of mice in Example 9;

[0080] Fig.12 is a statistical graph of IL-10 content in the liver and kidney of mice in Example 9;

[0081] Deposit of biological materials:

[0082] A strain of Lactobacillus fermentum HXJS4-1 was deposited in the General Microbiology Center of China Microorganism Culture Collection Administration on January 19, 2024, with the deposit number CGMCC NO: 29677, and the deposit address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, China, Institute of Microbiology, Chinese Academy of Sciences.

[0083] A strain of Lactobacillus plantarum HXJS3-1 was deposited in the General Microbiology Center of China Microorganism Culture Collection Administration on January 19, 2024, with the deposit number CGMCC NO: 29676, and the deposit address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, China, Institute of Microbiology, Chinese Academy of Sciences.

[0084] A strain of Lactobacillus paracasei HXXJ-008 was deposited in the General Microbiology Center of China Microorganism Culture Collection Administration on April 30, 2024, with the deposit number CGMCC NO: 30483, and the deposit address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, China, Institute of Microbiology, Chinese Academy of Sciences. DETAILED DESCRIPTION

[0085] In order to more clearly explain the overall concept of the present application, the following is described in detail in conjunction with the accompanying drawings of the specification by way of embodiment. In the following description, a large number of specific details are provided to provide a more thorough understanding of the present invention. However, it is obvious to those skilled in the art that the present invention can be implemented without one or more of these details. In other examples, in order to avoid confusion with the present invention, some technical features well known in the art are not described.

[0086] If no specific conditions are specified in the examples, the experiments were carried out according to conventional conditions or conditions recommended by the manufacturer.

[0087] Unless otherwise specified, in the following embodiments, the reagents or instruments used without indicating the manufacturer are all conventional products that can be purchased commercially.

[0088] Unless otherwise stated, the experimental methods, detection methods, and preparation methods disclosed in the present invention all adopt conventional techniques in microbiology, biochemistry, analytical chemistry, cell culture, and related fields conventional in the technical field.

[0089] The culture medium and reagent components involved in the following examples are as follows:

[0090] MRS solid medium: 10g peptone, 5g beef powder, 20g glucose, 4g yeast powder, 5g sodium acetate, 2g potassium dihydrogen phosphate, 0.2g magnesium sulfate, 2g ammonium citrate, 0.05g manganese sulfate, 1mL Tween 80, 15g agar powder, add to 1000mL distilled water, stir evenly to dissolve, divide into portions, and sterilize at 121℃ high temperature and high pressure for 15min. This medium is mainly used for the separation and cultivation of lactic acid bacteria.

[0091] MRS broth medium: 10g peptone, 5g beef powder, 20g glucose, 4g yeast powder, 5g sodium acetate, 2g potassium dihydrogen phosphate, 0.2g magnesium sulfate, 2g triammonium citrate, 0.05g manganese sulfate, 1mL Tween 80, add to 1000mL distilled water, heat and boil to dissolve, divide into portions, and sterilize at 121℃ for 15min. This medium is mainly used for the culture of lactic acid bacteria.

[0092] PCA medium: 5g tryptone, 2g yeast extract powder, 1g glucose, 15g agar, dissolved in 1000mL distilled water, sterilized at 121℃ for 20min. This medium is mainly used for lactic acid bacteria colony counting.

[0093] Nucleoside degradation solution: Dissolve 33.7 mg of inosine and 35.7 mg of guanosine in 100 mM K3PO4 solution to prepare inosine-guanosine degradation solution with a final concentration of about 1.26 mM.

[0094] HPLC mobile phase: Weigh 3.4 g KH2PO4 and dissolve it in 950 mL ultrapure water, then add 50 mL methanol to prepare 1 L of mobile phase.

[0095] Xanthine oxidase solution: Dissolve xanthine oxidase (100 U) in 0.2 mol / L phosphate buffer (pH 7.5) to 0.5 U / mL and store at 4°C.

[0096] 2 mM xanthine substrate: Accurately weigh 30.422 mg of xanthine standard, first dissolve it with 0.1 mol / L NaOH solution by ultrasonication, and then dilute to 100 mL with 0.2 mol / L phosphate buffer (pH 7.5) to prepare a 2 mM xanthine substrate solution.

[0097] 100mM K3PO4 buffer: weigh 21.23g K3PO4, add 995mL ultrapure water, add about 5mL phosphoric acid after dissolving, adjust pH to 7.0, and place at room temperature for use.

[0098] In addition, the "water" mentioned in the present invention includes any feasible water that can be used in the art, such as deionized water, distilled water, ion exchange water, double distilled water, high-purity water, and purified water.

[0099] In the following examples, unless otherwise specified, % means wt %, i.e. weight percentage.

[0100] Example 1 Preliminary screening of Lactobacillus fermentum strains

[0101] The commercially available slurry solution was diluted 10 times, 100 times, and 1000 times, respectively, and applied to MRS solid medium containing 1% CaCO3, three parallels per group, and cultured at 37°C. After three days of culture, 25 colonies of different morphologies were picked from the single colonies with obvious transparent circles on the plate, inoculated into MRS broth medium, and cultured at 37°C and 220rpm for 24h. Then transferred to new MRS broth medium, cultured for 16h, and the nucleoside degradation ability was tested after the bacterial solution became turbid.

[0102] Example 2 Determination of Nucleoside Degradation Ability

[0103] In this example, the nucleoside degradation ability of the strain obtained in Example 1 was determined. At the same time, the fermented Lactobacillus BNCC138617 purchased from Beina Chuanglian Biotechnology Co., Ltd. was used as a control strain for detection.

[0104] The method for detecting the nucleoside degradation ability comprises: using MRS broth medium to measure the OD of the fermentation liquid of Example 1 or Lactobacillus plantarum BNCC336943; 600 Adjust to 4.0. Take 1mL of fermentation broth and centrifuge at 4℃, 8000rpm for 2min, discard the supernatant and collect the bacteria. Add 750μL of saline to the bacteria to wash the bacteria, repeat the washing twice, and collect the washed bacterial cells. Add 750μL of nucleoside degradation solution to the above-collected bacterial cells, and use equal volumes of saline and MRS broth culture medium as blank controls. Set the bacterial cells inactivated at 100℃ for 10min as the heat inactivated control. Incubate the prepared nucleoside degradation system at 37℃ and 120rpm for 30min, 60min, 90min, and 120min respectively. After the reaction is completed, inactivate the system at 100℃ for 10min and centrifuge at 12000rpm for 2min. Use a 0.22μm filter membrane to filter the supernatant into a liquid phase vial for detection.

[0105] The nucleoside content was determined by high performance liquid chromatography, with the mobile phase being 25 mM KH2PO4 containing 5% methanol. An ODS-3 (4.6×250 mm, 5 μm) chromatographic column was used. Detection conditions: PDA detector, detection wavelength 254 nm, column temperature 35°C, flow rate 1 mL / min, injection volume 20 μL, retention time 20 min.

[0106] The calculation formula of nucleoside degradation rate is: (initial nucleoside concentration - residual nucleoside concentration after reaction) / initial nucleoside concentration × 100%

[0107] Finally, a lactic acid bacterium HXJS4-1 with high nucleoside degradation ability was screened out, and its nucleoside degradation ability is shown in Table 1.

[0108] Table 1 Nucleoside degradation ability of HXJS4-1

[0109]

[0110] From the results in Table 1, it can be seen that compared with the control strain BNCC138617, the strain HXJS4-1 can quickly degrade nucleosides and can almost completely degrade nucleosides within 1 hour.

[0111] Example 3 Identification of strains

[0112] (1) Colony morphology characteristics

[0113] The strain HXJS4-1 obtained in Example 2 is a Gram-positive bacterium that grows well in MRS medium, with an optimal growth temperature of 37°C and an optimal growth pH of 6.5. It does not form spores and has no flagella. It enters the stable phase after 16 hours of shaking culture. The plaques on the MRS solid medium (plate) are milky white, round, with neat edges, opaque, and smooth surface. Figure 1 shown.

[0114] (2) Molecular Biological Identification

[0115] In this embodiment, 16S rDNA sequencing was performed on strain HXJS4-1: a few colonies on the MRS solid plate were picked and added to 20 μL of the prepared PCR reaction solution. The composition of the reaction solution is shown in Table 2. PCR amplification was performed according to the pre-set program (as shown in Table 3), and then 4 μL was taken for DNA gel electrophoresis verification, and a band was found at about 1400 bp. The primer sequences involved in the amplification were: 27F: 5'-AGAGTTTGATCCTGGCTCAG-3'; 1492R: 5'-TACGG YTACCTTGTTACGACTT-3'.

[0116] The remaining PCR reaction solution was sent for sequencing, and the sequencing work was completed by Qingke Biotechnology Co., Ltd. The obtained 16SrDNA sequence (as shown in SEQ ID No. 1) was BLAST-matched on NCBI and homology-matched with the sequence information of known strains to identify the classification of strain HXJS4-1. The strain was finally identified as Lactobacillus fermentum. The fermented Lactobacillus fermentum HXJS4-1 was deposited in the General Microbiology Center of the China Microbiological Culture Collection Administration on January 19, 2024, with a preservation address of No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, and a preservation number of CGMCC NO: 29677.

[0117] Table 2 PCR reaction system

[0118]

[0119] Table 3 PCR reaction program

[0120]

[0121] 16S rDNA sequence of Lactobacillus fermentum HXJS4-1 (SEQ ID No. 1):

[0122]

[0123] Considering that the food consumed by humans contains not only nucleosides but also many carbohydrates and proteins, the degradation rate of nucleosides by fermented lactobacillus HXJS4-1 in the presence of different carbohydrate substrates was tested in this embodiment. The specific substrates and detection methods are as follows:

[0124] Substrate 1: 1.26 mM inosine-1.26 mM guanosine-2 g / L sucrose-2 g / L yeast extract;

[0125] Substrate 2: 1.26 mM inosine-1.26 mM guanosine-2 g / L glucose-2 g / L yeast extract;

[0126] Substrate 3: 1.26 mM inosine-1.26 mM guanosine.

[0127] Take OD 600 The fermented Lactobacillus HXJS4-1 bacterial solution with a value of 4.0 was washed with physiological saline and then added with equal volumes of substrate 1, 2, and 3 solutions for co-incubation for 2 hours, with physiological saline and nucleoside degradation solution as blank controls. The determination method is the same as in Example 2, and the nucleoside degradation ability of fermented Lactobacillus HXJS4-1 under different conditions is shown in Table 4 below.

[0128] Table 4 Nucleoside degradation ability of strains in the presence of carbon and nitrogen sources

[0129]

[0130] The experimental results in Table 4 show that the presence of carbon and nitrogen sources does not affect the degradation of nucleosides by the strain.

[0131] Example 5 XOD inhibition ability determination

[0132] Xanthine oxidase can catalyze xanthine to produce uric acid, and the product uric acid has a characteristic absorption peak at 293nm. The change in the absorbance value of the reaction system within a certain period of time is measured by an ELISA instrument, which characterizes the amount of uric acid generated and can measure the change in xanthine oxidase activity.

[0133] In the present embodiment, the xanthine oxidase (XOD) inhibition ability was determined for Lactobacillus fermentum HXJS4-1, and the purchased Lactobacillus fermentum BNCC138617 was used as a control during the determination. The specific determination method is as follows:

[0134] Take 0.75mL of xanthine substrate solution and mix with an equal volume of fermentation broth (OD 600The bacterial solution of Lactobacillus fermentum HXJS4-1 or Lactobacillus fermentum BNCC138617 with a value of 4.0) was mixed, and finally 0.5 mL of xanthine oxidase solution (0.5 U / mL) pre-warmed at 25°C for 20 min was added. The reaction system was placed at 25°C for 30 min and the absorbance was detected at a wavelength of 293 nm. Three parallels were used for each group.

[0135] Among them, the reaction groups and reaction systems are as follows:

[0136] Control sample ①: 0.75 mL xanthine + 0.5 mL xanthine oxidase + 0.75 mL PBS;

[0137] Control sample ②: 0.75 mL xanthine + 0.5 mL PBS + 0.75 mL fermentation liquid;

[0138] Test sample ③: 0.75 mL xanthine + 0.5 mL xanthine oxidase + 0.75 mL fermentation liquid;

[0139] Control sample ④: 0.75 mL xanthine + 1.25 mL PBS.

[0140] The xanthine oxidase activity was detected and the XOD inhibition rate was calculated as follows. The XOD inhibition rate results are shown in Table 5 below.

[0141] XOD inhibition rate calculation formula: ((absorbance value of control sample ① - absorbance value of control sample ④) - (absorbance value of test sample ③ - absorbance value of control sample ②)) / (control sample ① - control sample ④) × 100%

[0142] Table 5 XOD inhibition rate

[0143] Strain name XOD inhibition rate HXJS4-1 71.5％ BNCC138617 11.3％

[0144] From the results in Table 5, it can be seen that the inhibition rate of xanthine oxidase by the fermented Lactobacillus HXJS4-1 screened and obtained in the present application can reach 71.5%, while the inhibition rate of xanthine oxidase by the control bacteria is only 11.3%.

[0145] Example 6 Screening of Lactobacillus paracasei HXXJ-008

[0146] The sample collectors selected cow dung with intact morphology in the Narat Grassland in Xinjiang. Wearing disposable sterile gloves, use 75% alcohol to disinfect the sampling shovel and sampling spoon, shovel the cow dung to dig out the core part of 5-10g, seal it in a sterile sampling bag, record the sample information, and store it in a sample refrigerated insulation box (with built-in ice packs). On the same day, the low-temperature cold chain express was mailed to the Beijing laboratory for experiments. Weigh 1 g of cow dung sample, place it in a sterile homogenizer cup, add 10 mL of sterile saline, use a vortex shaker to mix for 1-2 min to make a suspension, and filter the suspension with 4 layers of sterile gauze. Dilute the filtered suspension 10 times, 100 times, and 1000 times, respectively, and apply it on MRS solid culture medium containing 1% CaCO3. After three days of cultivation, pick 25 colonies of different morphologies from single colonies with obvious transparent circles on the plate, and screen a strain of Lactobacillus paracasei by sequencing 16SrDNA, named HXXJ-008.

[0147] Example 7 Identification of Lactobacillus paracasei HXXJ-008

[0148] (1) Colony morphology characteristics

[0149] The strain HXXJ-008 obtained in Example 6 is a Gram-positive bacterium that grows well in MRS medium, with an optimal growth temperature of 37°C and an optimal growth pH of 6.5. It does not form spores and has no flagella. It enters the stable phase after 16 hours of shaking culture. The plaques on the MRS solid plate are milky white, round, with neat edges, opaque, and smooth surface. Figure 2 shown.

[0150] (2) Molecular Biological Identification

[0151] 16S rDNA sequencing: Pick a few colonies on the MRS solid plate and add them to 20 μL of the prepared PCR reaction solution. The composition of the reaction solution is shown in Table 6. Perform PCR amplification according to the pre-set program (as shown in Table 7). Then take 4 μL to run DNA gel electrophoresis for verification. A band was found at around 1400 bp. The primer sequences involved in the amplification are: 27F: 5'-AGAGTTTGATCCTGGCTCAG-3'; 1492R: 5'-TACGGYTACCTTGTTACGACTT-3'.

[0152] The remaining PCR reaction solution was sent for sequencing, and the sequencing work was completed by Qingke Biotechnology Co., Ltd. The obtained 16SrDNA sequence (as shown in SEQ ID No. 2) was compared by BLAST on NCBI, and the homology was compared with the sequence information of the known strain to identify the strain, and the final strain was identified as Lactobacillus paracasei. And the Lactobacillus fermentum HXXJ-008 was deposited in the General Microbiology Center of the China Microbiological Culture Collection Administration on April 30, 2024, and the deposit address is No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing, China, Institute of Microbiology, Chinese Academy of Sciences, and the deposit number is CGMCC NO: 30483.

[0153] Table 6 PCR reaction system

[0154]

[0155] Table 7 PCR reaction program

[0156]

[0157]

[0158] 16S rDNA sequence of Lactobacillus paracasei HXXJ-008 (SEQ ID No. 2):

[0159] TTAGACGGCTCCGCTCCCTAAAAGGGTTACGCCACCGGCTTCGGGTGTTACAAACTCTC

[0160] ATGGTGTGACGGGCGGTGTGTACAAGGCCCGGGAACGTATTCACCGCGGCGTGCTGAT

[0161] CCGCGATTACTAGCGATTCCGACTTCGTGTAGGCGAGTTGCAGCCTACAGTCCGAACT

[0162] GAGAATGGCTTTAAGAGATTAGCTTGACCTCGCGGTCTCGCAACTCGTTGTACCATCCA

[0163] TTGTAGCACGTGTGTAGCCCAGGTCATAAGGGGCATGATGATTTGACGTCATCCCCACC

[0164] TTCCTCCGGTTTGTCACCGGCAGTCTTACTAGAGGTGCCCAACTAAATGCTGGCAACTA

[0165] GTCATAAGGGTTGCGCTCGTTGCGGGACTTAACCCAACATCTCACGACACGAGCTGAC

[0166] GACAACCATGCACCACCTGTCATTTTGCCCCCGAAGGGGAAACCTGATCTCTCAGGTG

[0167] ATCAAAAAGATGTCAAGACCTGGTAAGGTTCTTCGCGTTGCTTCGAATTAAACCACATG

[0168] CTCCACCGCTTGTGCGGGCCCCCGTCAATTCCTTTTGAGTTTCAACCTTGCGGTCGTACT

[0169] CCCCAGGCGGAATGCTTAATGCGTTAGCTGCGGCACTGAAGGGCGGAAACCCCTCCAA

[0170] CACCTAGCATTCATCGTTTACGGCATGGACTACCAGGGTATCTAATCCTGTTCGCTACCC

[0171] ATGCTTTCGAGCTCAGCGTCAGTTACAGACCAGACAGCCGCCTTCGCCACTGGTGTT

[0172] CTTCCATATATCTACGCATTTCACCGCTACACATGGAGTTCCACTGTCCTCTTCTGCACT

[0173] CAAGTTTCCCAGTTTCCGATGCGCTTCCTCGGTTAAGCCGAGGGCTTTCACATCAGACT

[0174] TAAAAAACCGCCTGCGCTCGCTTTACGCCCAATAAATCCGGATAACGCTTGCCACCTAC

[0175] GTATTACCGCGGCTGCTGGCACGTAGTTAGCCGTGGCTTTCTGGTTGGATACCGTCACG

[0176] CCGACAACAGTTACTCTGCCGACCATTCTTCTCCAACAACAGAGTTTTACGACCCGAA

[0177] AGCCTTCTTCACTCACGGCGGCGTTGCTCCATCAGACTTGCGTCCATTGTGGAAGATTCC

[0178] CTACTGCTGCCTCCCGTAGGAGTTTGGGCCGTGTCTCAGTCCCAATGTGGCCGATCAA

[0179] CCTCTCAGTTCGGCTACGTATCATCGCCTTGGTGAGCCATTACCTCACCAACTAGCTAAT

[0180] ACGCCGCGGGTCCATCCAAAAGCGATAGCTTACGCCATCTTTCAGCCAAGAACCATGC

[0181] GGTTCTTGGATCTATGCGGTATTAGCATCTGTTTCCAAATGTTATCCCCACTTAAGGGC

[0182] AGGTTACCCACGTGTTACTCACCCGTCCGCCACTCGTTCCATGTTGAATCTCGGTGCAA

[0183] GCACCGATCATCAACAAGAACTCGTTCGACTTGCA

[0184] Example 8 Lactobacillus paracasei HXXJ-008 mouse experiment

[0185] 1. Modeling and feeding of hyperuricemia mice

[0186] Eighteen male mice, 4 weeks old and weighing 16-20 g, were purchased and allowed to acclimate for one week before the experiment. The mice were housed under standard laboratory conditions of 22°C ± 1°C and 60% relative humidity, 12 h light / 12 h dark cycle (7:00-19:00 light), and fed with standard commercial mouse food. The probiotic (Lactobacillus paracasei HXXJ-008) was fed at a rate of 1×10 9 CFU / pc. Divided into the following groups on average:

[0187] (1) Blank group: The mice were fed with basal feed and gavaged with 0.5 mL of sterile skim milk every day.

[0188] (2) Model group: mice were fed a basal diet and gavaged with 300 mg / kg / d of potassium oxonate (OP) and 10 g / kg / d of yeast extract powder in sterile skim milk.

[0189] (3) HUA+HXXJ-008: Feeding a basic diet containing 300 mg / kg / d oxonate potassium (OP) and 10 g / kg yeast extract powder, and the number of viable probiotics HXXJ-008 was 1×10 9 The probiotics were dissolved in an equal volume of skim milk with an equal concentration of CFU by intragastric administration.

[0190] 2. Detection of inflammatory factor indicators

[0191] On the 21st day of feeding, liver and kidney samples of mice were collected, washed with saline, collected in pre-weighed test tubes with 1 mL of PBS diluent, and stored at -20°C for use. Fresh liver and kidney were ground on ice with a handheld grinder, and then obtained under low-speed centrifugation (8000 rpm, 10 min) to obtain a sample suspension. Interleukin-1β (IL-1β) and IL-6 levels in liver and kidney were detected according to the instructions of the test kit provided by Beijing Biosci.

[0192] Test results see Figure 3-4 It can be seen that the Lactobacillus paracasei HXXJ-008 treatment group has a good inhibitory effect on the inflammatory factors interleukin-1β (IL-1β) and IL-6 in the liver and kidney of hyperuricemia mice. Figure 3 It can be seen that the inhibition rates of Lactobacillus paracasei HXXJ-008 on the inflammatory factor IL-1β in the liver and kidney of hyperuricemia mice were 42.82% and 64.09%, respectively. Figure 4 It can be seen that the inhibition rates of Lactobacillus paracasei HXXJ-008 on the inflammatory factor IL-6 in the liver and kidney of hyperuricemia mice were 44.67% and 58.99%, respectively, and it has a good effect in inhibiting inflammatory factors.

[0193] Example 9 Compound strain mouse experiment

[0194] 1. Modeling and feeding of hyperuricemia mice

[0195] 60 male mice, 4 weeks old, weighing 16-20 g, were purchased and allowed to adapt for one week before the experiment. The mice were housed under standard laboratory conditions of 22°C ± 1°C and 60% relative humidity, 12 hours of light / 12 hours of dark cycle (7:00-19:00 light), fed with standard commercial mouse food, and divided equally into the following groups:

[0196] (1) Blank group: The mice were fed with basal feed and gavaged with 0.5 mL of sterile skim milk every day.

[0197] (2) Model group: mice were fed a basal diet and gavaged with 300 mg / kg / d of potassium oxonate (OP) and 10 g / kg / d of yeast extract powder in sterile skim milk.

[0198] (3) HUA+ treatment group 1: The animals were fed a basal diet containing 300 mg / kg / d oxolane potassium (OP) and 10 g / kg yeast extract powder, and the probiotic HXJS3-1 with a viable count of 1 × 10 9 The probiotics were dissolved in an equal volume of skim milk with an equal concentration of CFU by intragastric administration.

[0199] (4) HUA+ treatment group 2: The animals were fed a basal diet containing 300 mg / kg / d oxolane potassium (OP) and 10 g / kg yeast extract powder, and the probiotic HXJS4-1 with a viable count of 1 × 10 9 CFU were gavaged with sterile skim milk of equal volume and concentration.

[0200] (5) HUA+ treatment group 3: The animals were fed a basal diet containing 300 mg / kg / d oxolane potassium (OP) and 10 g / kg yeast extract powder, and the probiotic HXXJ-008 with a viable count of 1 × 10 9 CFU were gavaged with sterile skim milk of equal volume and concentration.

[0201] (6) HUA+ treatment group 4: The rats were fed a basal diet containing 300 mg / kg / d oxytetracycline potassium (OP) and 10 g / kg yeast extract powder, as well as equal amounts of probiotics HXJS3-1, HXJS4-1, and HXXJ-008. The total number of live bacteria in the diet was 1 × 10 7 CFU were gavaged with sterile skim milk of equal volume and concentration.

[0202] (7) HUA+ treatment group 5: The rats were fed a basal diet containing 300 mg / kg / d oxytetracycline potassium (OP) and 10 g / kg yeast extract powder, as well as equal amounts of probiotics HXJS3-1, HXJS4-1, and HXXJ-008. The total number of live bacteria in the diet was 1 × 10 8 CFU were gavaged with sterile skim milk of equal volume and concentration.

[0203] (8) HUA+ treatment group 6: The rats were fed a basal diet containing 300 mg / kg / d oxytetracycline potassium (OP) and 10 g / kg yeast extract powder, as well as equal amounts of probiotics HXJS3-1, HXJS4-1, and HXXJ-008. The total number of live bacteria in the diet was 1 × 10 9CFU were gavaged with sterile skim milk of equal volume and concentration.

[0204] (9) HUA+ treatment group 7: The rats were fed a basal diet containing 300 mg / kg / d oxytetracycline potassium (OP) and 10 g / kg yeast extract powder, as well as equal amounts of probiotics HXJS3-1, HXJS4-1, and HXXJ-008. The total number of live bacteria in the diet was 1 × 10 11 CFU were gavaged with sterile skim milk of equal volume and concentration.

[0205] (10) HUA+ positive drug group: The mice were fed a basal diet containing 300 mg / kg / d of potassium oxyphosphate (OP) and 10 g / kg / d of yeast extract powder, and an equal volume and concentration of sterile skim milk containing 5 mg / kg / d of allopurinol was administered by gavage.

[0206] Feed according to the above standards for 21 days.

[0207] 2. Test indicators and results

[0208] (1) Uric acid content detection

[0209] A Urine

[0210] During the 21-day feeding period, mice in each group were moved into clean empty cages every three days for 1 h to collect urine samples. Fresh urine from mice was diluted 10 times with distilled water and placed in a 60°C water bath to ensure that there was no urate deposition in the urine samples. Then, uric acid content was tested for all samples according to the instructions provided by the UA detection kit (Jiancheng, Nanjing, China), and the test was performed immediately after the sample was collected.

[0211] The effects of each treatment group on the uric acid content in the urine of hyperuricemia mice are shown in Figure 7 It can be seen that oral administration of probiotics to mice can significantly reduce the uric acid level in the urine of hyperuricemia mice. Among them, HUA+ treatment group 6 and HUA+ treatment group 7 have the most significant effects, which are equivalent to the effect of purinol in the positive drug group. The effect of HUA+ treatment group 4 is equivalent to the effect of single bacteria, and the effect of HUA+ treatment group 5 is significantly better than that of single bacteria. This shows that when the three bacteria are used together, the uric acid-lowering effect is enhanced with the increase of the number of live bacteria administered orally. The number of live bacteria administered orally is greater than 1×10 8 CFU / each can significantly reduce uric acid in urine.

[0212] B Blood

[0213] Blood was collected from each group of mice on days 0, 7, 14, and 21 and allowed to stand for 30 min. The collected blood samples were centrifuged at 4000 rpm for 20 min to obtain serum, and the uric acid content was detected according to the instructions provided by the UA detection kit (Jiancheng, Nanjing, China).

[0214] Effects of each treatment group on serum uric acid levels in hyperuricemia mice Figure 6 It can be seen that all probiotic treatment groups can effectively inhibit the uric acid content in the serum of hyperuricemia mice. Among them, HUA+ treatment group 6 and HUA+ treatment group 7 had the highest inhibition rates, with the inhibition rates of uric acid in serum being 56.81% and 59.85%, respectively, which were comparable to the positive drug group. With the increase of the number of viable bacteria administered orally, the uric acid-lowering effect was also enhanced. The number of viable bacteria administered orally with the three strains of viable bacteria was greater than 1×10 8 CFU / each can significantly reduce uric acid in urine.

[0215] (2) XOD activity index detection

[0216] On the 21st day of feeding, the mouse liver and blood samples were collected, and then the collected liver samples were rinsed with physiological saline, collected in a pre-weighed test tube with 1 mL of PBS diluent, and stored at -20°C for use. The fresh liver was ground on ice with a handheld grinder, and then a liver sample suspension was obtained under low-speed centrifugation (8000 rpm, 10 min). XOD was detected in the liver sample (according to the instructions provided by Nanjing Jiancheng XOD detection kit). The collected blood samples were centrifuged at 4000 rpm for 20 min to obtain serum, and the XOD activity in the serum was detected according to the instructions provided by Nanjing Jiancheng XOD detection kit.

[0217] The inhibition of XOD activity in the liver and serum of hyperuricemia mice by each treatment group is shown in Figure 5 It can be seen that each probiotic treatment group has a certain inhibitory effect on the XOD activity in the liver and serum of hyperuricemia mice, among which HUA+ treatment group 6 and HUA+ treatment group 7 have the highest inhibition rates, with the inhibition rates of HUA+ treatment group 6 being 85.71% and 78.21, respectively, and the inhibition rates of HUA+ treatment group 7 being 86.93% and 78.69%, respectively, which are equivalent to the positive drug group and can effectively inhibit the production of uric acid from purine. With the increase of the number of viable bacteria after intragastric administration, the uric acid-lowering effect is also enhanced. The number of viable bacteria after intragastric administration of the three strains of viable bacteria is greater than 1×10 8 CFU / can significantly and effectively inhibit the production of uric acid from purine.

[0218] (3) Detection of inflammatory factor indicators

[0219] On the 21st day of feeding, liver and kidney samples of mice were collected, washed with saline, collected in pre-weighed test tubes with 1 mL of PBS diluent, and stored at -20 °C for use. Fresh liver and kidney were ground on ice with a handheld grinder, and then obtained under low-speed centrifugation (8000 rpm, 10 min) The levels of interleukin-1β (IL-1β), IL-6, tumor necrosis factor TNF-α, and IL-11 in the liver and kidney were detected according to the instructions of the detection kit provided by Beijing Biosci.

[0220] Test results see Figure 8-11 It can be seen that each probiotic treatment group has a good inhibitory effect on the inflammatory factors interleukin-1β (IL-1β), IL-6, tumor necrosis factor TNF-α, and IL-11 in the liver and kidney of hyperuricemia mice. With the increase of the number of live bacteria perfusion, the uric acid-lowering effect is also enhanced. The number of live bacteria perfusion of the three live bacteria is greater than 1×10 8 CFU / each can significantly inhibit inflammatory factors such as interleukin-1β (IL-1β), IL-6, tumor necrosis factor TNF-α, and IL-11. Figure 8 It can be seen that HUA+ treatment group 6 and HUA+ treatment group 7 had the best inhibitory effect on the inflammatory factor IL-1β in the liver and kidney of hyperuricemia mice. The inhibition rates of HUA+ treatment group 6 were 76.61% and 80.83%, respectively, and the inhibition rates of HUA+ treatment group 7 were 77.5% and 81.95%, respectively, which were comparable to the positive drug group. Fig. 9 It can be seen that HUA+ treatment group 6 and HUA+ treatment group 7 have the best inhibitory effect on the inflammatory factor TNF-α in the liver and kidney of hyperuricemia mice. The inhibition rates of HUA+ treatment group 6 were 76.92% and 77.43%, respectively, and the inhibition rates of HUA+ treatment group 7 were 77.51% and 79.45%, respectively, second only to the positive drug group. Fig.10 It can be seen that HUA+ treatment group 6 and HUA+ treatment group 7 had the best inhibitory effect on the inflammatory factor IL-6 in the liver of hyperuricemia mice, among which the inhibition rates of HUA+ treatment group 6 were 83.1% and 71.35%, and the inhibition rates of HUA+ treatment group 7 were 84.7% and 71.78%, respectively, second only to the positive drug group. Fig.11 It can be seen that HUA+ treatment group 6 and HUA+ treatment group 7 had the best inhibitory effect on the inflammatory factor IL-11 in the liver and kidneys of hyperuricemia mice, among which the inhibition rates of HUA+ treatment group 6 were 76.96% and 81.67%, respectively, and the inhibition rates of HUA+ treatment group 7 were 77.77% and 83.19%, respectively, which were second only to the positive drug group.

[0221] (4) Detection of anti-inflammatory factor IL-10

[0222] The anti-inflammatory factor IL-10 in the liver and kidney of hyperuricemia mice was detected using the inflammatory factor index detection method.

[0223] Test results see Fig.12 It can be seen that HUA+ treatment groups 1-7 have good recovery effects on the anti-inflammatory factor IL-10 in the liver and kidneys of hyperuricemia mice, among which HUA+ treatment groups 5, HUA+ treatment groups 6 and HUA+ treatment groups 7 have more significant recovery effects on the anti-inflammatory factor IL-10 in the liver and kidneys of hyperuricemia mice. The inhibition rates of HUA+ treatment group 6 were 66.7% and 71.56%, respectively, and the inhibition rates of HUA+ treatment group 7 were 70.55% and 73.1%, respectively, which are better than those of the positive drug group.

[0224] In summary, strains HXJS3-1, HXJS4-1, and HXXJ-008 can significantly reduce the blood uric acid, urine uric acid, XOD activity in liver and serum, and the levels of inflammatory factors (IL-1β, IL-6, TNF-α, IL-11) in liver and kidney of uricemia mice, and increase the level of anti-inflammatory factor IL-10; however, the combination of the three bacteria has a better effect. When the three bacteria are used in combination, when the number of viable bacteria is 1×10 7 CFU / each, the effect is equivalent to that of a single bacterium. When the number of viable bacteria per gavage is greater than 1×10 8 CFU / unit, it can better prevent and treat hyperuricemia, and its effect is close to or equivalent to that of the positive drug allopurinol.

[0225] Example 10 Application Example

[0226] (1) Preparation of probiotic yogurt containing fermented Lactobacillus HXJS4-1

[0227] The fermented Lactobacillus HXJS4-1 was activated in MRS culture medium and the OD 600 When the pH value is 0.8-1.2, take 1 mL of bacterial solution and centrifuge it at 8000 rpm in a low-temperature centrifuge for 5 minutes, wash it three times with sterile saline, add the cleaned strain into 100 mL of sterile cow's milk and ferment it at 42°C for 8-10 hours, then inoculate thermophilic Streptococcus and Bulgarian Lactobacillus for secondary fermentation to form probiotic yogurt.

[0228] (2) Preparation of soy milk containing fermented Lactobacillus HXJS4-1

[0229] Select high-quality soybeans, wash them three times with clean water, soak them until the soybean hulls can be removed, add boiling water to grind them, then keep them warm at a temperature above 80°C for 10 minutes, filter and centrifuge to obtain the supernatant to obtain crude soy milk, add fermented Lactobacillus HXJS4-1 when the temperature drops to 37°C, and add the fermented Lactobacillus HXJS4-1 in an amount corresponding to the amount in Example 10(1). Store the mixture under refrigeration at 4°C to obtain soy milk containing fermented Lactobacillus HXJS4-1.

[0230] (3) Preparation of Tablet Candy Containing Lactobacillus fermentum HXJS4-1

[0231] According to weight proportions, 50 parts of sorbitol, 15 parts of microcrystalline cellulose, 10 parts of fermented Lactobacillus HXJS4-1 and 5 parts of magnesium stearate are mixed, and the mixture is subjected to mixed dry granulation and tableting to obtain a tablet having a specification of 0.5 g / tablet and a viable bacterial count of 10 billion CFU.

[0232] (4) Preparation of solid beverage containing fermented Lactobacillus HXJS4-1

[0233] The solid beverage comprises 50 parts of glucose, 15 parts of maltodextrin, 10 parts of fermented lactobacillus HXJS4-1, 10 parts of soybean peptide powder, 10 parts of resistant dextrin and 8 parts of isomaltooligosaccharide in parts by weight, which are fully mixed and packaged.

[0234] (5) Preparation of solid beverage containing Lactobacillus fermentum HXJS4-1, Lactobacillus plantarum HXJS3-1 and Lactobacillus paracasei HXXJ-008

[0235] The solid beverage comprises 50 parts of glucose, 15 parts of maltodextrin, 10 parts of fermented lactobacillus HXJS4-1, 10 parts of plant lactobacillus HXJS3-1, 10 parts of lactobacillus paracasei HXXJ-008, 10 parts of soybean peptide powder, 10 parts of resistant dextrin and 8 parts of isomaltooligosaccharide in parts by weight, which are fully mixed and packaged.

[0236] The above is only an embodiment of the present application and is not intended to limit the present application. For those skilled in the art, the present application may have various changes and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application should be included in the scope of the claims of the present application.

Claims

1. A strain of Lactobacillus fermentum HXJS4-1, characterized in that: It is deposited in the General Microbiology Center of China Microbiological Culture Collection Administration with the deposit number CGMCCNO: 29677.

2. A strain of Lactobacillus paracasei HXXJ-008, characterized in that: It is deposited in the General Microbiology Center of China Microorganism Culture Collection Administration with the deposit number CGMCC NO: 30483.

3. A preparation, characterized in that The preparation comprises any one or more of Lactobacillus fermentum, Lactobacillus paracasei and / or Lactobacillus plantarum, Preferably, the fermented Lactobacillus is the fermented Lactobacillus (Lactobacillusfermentum) HXJS4-1 according to claim 1, More preferably, the Lactobacillus paracasei is the Lactobacillus paracasei HXXJ-008 according to claim 2, Most preferably, the plant lactobacillus is plant lactobacillus (Lactobacillus plantarum) HXJS3-1, which is deposited in the General Microbiological Center of China Microbiological Culture Collection Administration with a deposit number of CGMCC NO: 29676.

4. The preparation according to claim 3, characterized in that The total number of viable bacteria of the fermented Lactobacillus, Lactobacillus paracasei and / or Lactobacillus plantarum is greater than 1×10 8 CFU.

5. the application of lactobacillus fermentum as claimed in claim 1 or Lactobacillus paracasei as claimed in claim 2 or the arbitrary described preparation of claim 3-4 in degraded nucleosides and / or preparation degraded nucleosides product.

6. the application of lactobacillus fermentum as claimed in claim 1 or Lactobacillus paracasei as claimed in claim 2 or the arbitrary described preparation of claim 3-4 in suppressing xanthine oxidase activity and / or in preparing xanthine oxidase inhibition product.

7. the application of the fermented lactobacillus as claimed in claim 1 or the Lactobacillus paracasei as claimed in claim 2 or the arbitrary described preparation of claim 3-4 in the inhibition of inflammation of non-disease diagnosis and treatment purpose and / or the preparation inhibition of inflammation product.

8. Application of the fermented lactobacillus as claimed in claim 1 or the Lactobacillus paracasei as claimed in claim 2 or the arbitrary preparation as claimed in claim 3-4 in preparing a product that reduces uric acid levels.

9. the application of Lactobacillus fermentum as claimed in claim 1 or Lactobacillus paracasei as claimed in claim 2 or the arbitrary described preparation of claim 3-4 in the preparation prevention and / or treatment of hyperuricemia and / or gout product.

10. Use of the fermented Lactobacillus according to claim 1 or the Lactobacillus paracasei according to claim 2 or the preparation according to any one of claims 3-4 in the preparation of an IL-11 inhibitor; preferably, the IL-11 inhibitor can be used in oral preparations and / or external preparations for anti-aging and / or life extension.

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