Application of Bacillus coagulans BC99 in the preparation of a probiotic preparation for assisting in reducing uric acid levels

Abstract

The present invention relates to the application of Bacillus coagulans BC99 in the preparation of a probiotic preparation for assisting in reducing uric acid levels; the Bacillus coagulans BC99 is Bacillus coagulans with a preservation number of CGMCC No. 21801 Bacillus coagulans Strain BC99. The present invention discovers that Bacillus coagulans BC99 has very significant effects in assisting in reducing uric acid, specifically manifested in: reducing the serum uric acid, urea nitrogen, and creatinine levels of hyperuricemic rats; improving the xanthine oxidase activity of hyperuricemic rats; regulating the serum inflammatory factor levels of hyperuricemic rats and inhibiting the inflammatory reaction caused by elevated uric acid; effectively reducing the levels of glutamic oxaloacetic transaminase and glutamic pyruvic transaminase in hyperuricemic rats. At the same time, as a probiotic, Bacillus coagulans BC99 has high product safety and is not easily resistant

Description

Technical Field

[0001] The present invention belongs to the technical field of microbiology, and relates to a new use of Bacillus coagulans BC99, specifically to the application of Bacillus coagulans BC99 in the preparation of a probiotic preparation for assisting in reducing uric acid levels. Background Art

[0002] The cells in the human body undergo metabolism every day. The nucleic acids in the body will be broken down to generate purines, and the purines produced in the body and those ingested from food will be decomposed under the action of xanthine oxidase and adenosine deaminase in the body, and finally form uric acid. Uric acid is produced in the liver, muscle and adipose tissues, is the end product of purine metabolism, and is the main antioxidant in human plasma, which can protect cells or neurons from oxidative damage. Under normal circumstances, there is a dynamic balance among intestinal absorption of purines, uric acid production, and uric acid excretion through the kidneys to maintain the normal physiological state of the body. Due to changes in diet structure and lifestyle, the number of patients with hyperuricemia and gout has increased accordingly, which has become a common disease in modern society. However, problems such as drug dependence and difficult diet control brought about by traditional drug treatment and diet control also make the measures to relieve hyperuricemia have little effect. Therefore, it is urgent to develop new effective methods for preventing and relieving hyperuricemia symptoms.

[0003] At present, a large number of studies have shown that the increase in uric acid levels is closely related to liver injury. Uric acid is produced in the liver, and high uric acid will continuously produce pro-inflammatory factors and pro-oxidative mediators, further aggravating liver injury and forming a vicious cycle. Existing studies have shown that probiotics have shown great potential in preventing and treating liver injury and reducing inflammatory factors. As live microorganisms, probiotics can bring health benefits to the host in various ways when ingested in sufficient amounts. The mechanisms of action of probiotics in assisting in reducing uric acid mainly include improving enzyme activity, reducing the production of pro-inflammatory factors, and improving the metabolic function of the host.

[0004] The application of probiotics in assisting in reducing uric acid is still in its initial stage, and there are few strategies. Therefore, developing more probiotic agents for assisting in reducing uric acid can provide new ideas and methods for assisting in reducing uric acid and treating liver injury. Summary of the Invention

[0005] Aiming at the deficiencies of the prior art, the purpose of the present invention is to provide a new use of Bacillus coagulans BC99, specifically to provide the application of Bacillus coagulans BC99 in the preparation of a probiotic preparation for assisting in reducing uric acid levels.

[0006] To achieve the purpose of this invention, the present invention adopts the following technical solutions:

[0007] In a first aspect, the present invention provides the use of Bacillus coagulans BC99 in the preparation of a probiotic preparation for assisting in reducing uric acid levels;

[0008] The Bacillus coagulans BC99 is Bacillus coagulans with the preservation number of CGMCC No. 21801 Bacillus coagulans strain BC99.

[0009] The present invention has developed a new use of Bacillus coagulans BC99, providing a research basis for its wide application. At the same time, it has also developed a brand-new strategy for assisting in reducing uric acid levels. The present invention has found that Bacillus coagulans BC99 is very effective in assisting in reducing uric acid, specifically manifested in: (1) significantly reducing the serum uric acid, urea nitrogen, and creatinine levels of hyperuricemic rats; (2) significantly improving the activity of xanthine oxidase (XOD) in hyperuricemic rats; (3) regulating the levels of serum inflammatory factors such as TNF-α, IL-6, and IL-1β in hyperuricemic rats, and inhibiting the inflammatory response caused by the increase in uric acid; (4) effectively reducing the levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in hyperuricemic rats. At the same time, as a probiotic, Bacillus coagulans BC99 has high product safety and is not prone to generating resistance.

[0010] Preferably, the dosage form of the probiotic preparation includes solution, powder, tablet, granule, or capsule. The dosage form of the probiotic preparation involved in the present invention is not limited, including the most commonly used solution and powder, or further prepared capsule, tablet, or granule.

[0011] Preferably, the viable count in the probiotic preparation is not less than 1×10 9 CFU / mL or 1×10 9 CFU / g, such as 1×10 9 CFU / g (CFU / mL), 2×10 9 CFU / g (CFU / mL), 5×10 9 CFU / g (CFU / mL), 8×10 9 CFU / g (CFU / mL), 1×10 10 CFU / g (CFU / mL), 5×10 10 CFU / g (CFU / mL), 1×10 11 CFU / g (CFU / mL), 1×10 12 CFU / g (CFU / mL), 1×10 13 CFU / g (CFU / mL), etc. Other specific point values within this numerical range can be selected and will not be elaborated one by one here.

[0012] Preferably, the strains in the probiotic preparation further include Bifidobacterium bifidum with the preservation number of CGMCC No. 16923 Bifidobacterium bifidum strain BBi32.

[0013] The above-mentioned Bacillus coagulans BC99 can not only be used alone to reduce uric acid levels, but also can be used in combination with Bifidobacterium bifidum strain BBi32 to reduce uric acid levels.

[0014] The present invention also develops a brand-new strategy for reducing uric acid levels, that is, compounding and combining Bacillus coagulans BC99 strain and Bifidobacterium bifidum BBi32 strain, and it is found that the two can cooperate with each other and promote each other, and have synergistic effects in reducing serum uric acid, urea nitrogen, creatinine levels in hyperuricemic rats, improving xanthine oxidase activity in hyperuricemic rats, regulating serum inflammatory factor levels in hyperuricemic rats, and inhibiting inflammatory reactions caused by elevated uric acid. When the amount of bacteria used is the same, compared with the intervention methods of single BC99 strain or single BBi32 strain, the compounding of the two bacteria significantly improves the above effects. Therefore, using these two probiotics to prepare a probiotic preparation for assisting in reducing uric acid levels has good prospects. At the same time, the above two bacteria are both probiotics, the product has high safety, and it is not easy to produce resistance.

[0015] Preferably, the ratio of the viable count of BC99 strain to BBi32 strain is 1:10 - 10:1, such as 1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, etc. Other specific point values within this numerical range can be selected and will not be elaborated here one by one.

[0016] Based on the potential synergistic cooperation relationship between the above two strains, when they meet the above specific viable count ratio, their effects in reducing serum uric acid, urea nitrogen, creatinine levels in hyperuricemic rats, improving xanthine oxidase activity in hyperuricemic rats, regulating serum inflammatory factor levels in hyperuricemic rats, and inhibiting inflammatory reactions caused by elevated uric acid are more excellent.

[0017] Preferably, the dosage form of the probiotic preparation is a solution, which is prepared by the following method:

[0018] Inoculate the relevant strains into a culture medium for activation and fermentation culture to obtain a fermentation broth; after centrifuging the fermentation broth, resuspend it with a solvent to obtain a bacterial suspension or a mixed bacterial suspension.

[0019] Preferably, the dosage form of the probiotic agent is a freeze-dried powder, which is prepared by the following method:

[0020] Inoculate the relevant strains into a culture medium for activation and fermentation culture to obtain a fermentation broth; after centrifuging the fermentation broth, mix it with a cryoprotectant and then perform freeze-drying to obtain bacterial powder or mixed bacterial powder.

[0021] Preferably, the probiotic preparation further contains excipients; the excipients include any one or a combination of at least two of fillers, binders, wetting agents, disintegrants, emulsifiers, solubilizers, osmotic pressure regulators, colorants, pH regulators, antioxidants, bacteriostatic agents, or buffers.

[0022] In a second aspect, the present invention provides the use of Bacillus coagulans BC99 in the preparation of a probiotic preparation for improving hyperuricemia;

[0023] The Bacillus coagulans BC99 is the Bacillus coagulans with the preservation number of CGMCC No. 21801 Bacillus coagulans BC99 strain.

[0024] Based on the excellent efficacy of Bacillus coagulans BC99 in reducing uric acid levels, it can be used in the preparation of a probiotic preparation for improving hyperuricemia.

[0025] Preferably, the viable count of the probiotic preparation is not less than 1×10 9 CFU / mL or 1×10 9 CFU / g, such as 1×10 9 CFU / g (CFU / mL), 2×10 9 CFU / g (CFU / mL), 5×10 9 CFU / g (CFU / mL), 8×10 9 CFU / g (CFU / mL), 1×10 10 CFU / g (CFU / mL), 5×10 10 CFU / g (CFU / mL), 1×10 11 CFU / g (CFU / mL), 1×10 12 CFU / g (CFU / mL), 1×10 13 CFU / g (CFU / mL), etc. Other specific point values within this numerical range can be selected and will not be elaborated one by one here.

[0026] Preferably, the strains in the probiotic preparation further include the Bifidobacterium bifidum BBi32 strain with the preservation number of CGMCC No. 16923; the ratio of the viable count of the BC99 strain to the BBi32 strain is 1:10 - 10:1.

[0027] The present invention also develops a brand-new strategy for improving hyperuricemia, that is, combining Bacillus coagulans strain BC99 and Bifidobacterium bifidum strain BBi32. It is found that the two can cooperate and promote each other, showing synergistic effects in improving hyperuricemia. When the amount of bacteria used is the same, compared with the intervention methods using a single BC99 strain or a single BBi32 strain, the combination of the two bacteria significantly improves the above effects. Therefore, using these two probiotics to prepare probiotic preparations for improving hyperuricemia has good prospects.

[0028] Compared with the prior art, the present invention has the following beneficial effects:

[0029] The present invention develops a new use of Bacillus coagulans BC99, providing a research basis for its wide application. At the same time, it also develops a brand-new strategy for assisting in reducing uric acid levels. The present invention finds that Bacillus coagulans BC99 is very effective in assisting in reducing uric acid, specifically manifested as follows: (1) significantly reducing the serum uric acid, urea nitrogen, and creatinine levels of hyperuricemic rats; (2) significantly improving the activity of xanthine oxidase (XOD) in hyperuricemic rats; (3) regulating the levels of serum inflammatory factors such as TNF-α, IL-6, and IL-1β in hyperuricemic rats and inhibiting the inflammatory response caused by elevated uric acid; (4) effectively reducing the levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in hyperuricemic rats.

[0030] The present invention also develops a brand-new strategy of compound bacteria for reducing uric acid levels, that is, combining Bacillus coagulans strain BC99 and Bifidobacterium bifidum strain BBi32. It is found that the two can cooperate and promote each other, showing synergistic effects in reducing the serum uric acid, urea nitrogen, and creatinine levels of hyperuricemic rats, improving the activity of xanthine oxidase in hyperuricemic rats, regulating the levels of serum inflammatory factors in hyperuricemic rats, and inhibiting the inflammatory response caused by elevated uric acid. When the amount of bacteria used is the same, compared with the intervention methods using a single BC99 strain or a single BBi32 strain, the combination of the two bacteria significantly improves the above effects. Therefore, using these two probiotics to prepare probiotic preparations for assisting in reducing uric acid levels has good prospects. At the same time, both of the above two bacteria are probiotics, and the product has high safety and is not easy to produce resistance.

[0031] The taxonomic name of the BC99 strain involved in the present invention is Bacillus coagulans Bacillus coagulans The preservation unit is the General Microbiology Center of the China Committee for Culture Collection of Microorganisms. The preservation time is February 1, 2021, and the preservation number is CGMCC No. 21801. The address is: No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing.

[0032] The taxonomic name of the BBi32 strain involved in the present invention is Bifidobacterium bifidumBifidobacterium bifidum The preservation unit is the General Microbiology Center of the China Committee for Culture Collection of Microorganisms. The preservation date is December 10, 2018, and the preservation number is CGMCC No. 16923. The address is: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. Description of the Drawings

[0033] Figure 1 It is a statistical result graph of the body weight changes of rats in each group;

[0034] Figure 2 It is a statistical result graph of the serum uric acid content of rats in each group;

[0035] Figure 3 It is a statistical result graph of the activity of serum xanthine oxidase (XOD) of rats in each group;

[0036] Figure 4 It is a statistical result graph of the content of serum aspartate aminotransferase (AST) of rats in each group;

[0037] Figure 5 It is a statistical result graph of the amount of serum alanine aminotransferase (ALT) of rats in each group;

[0038] Figure 6 It is a statistical result graph of the level of serum inflammatory factor TNF-α of rats in each group;

[0039] Figure 7 It is a statistical result graph of the level of serum inflammatory factor IL-6 of rats in each group;

[0040] Figure 8 It is a statistical result graph of the level of serum inflammatory factor IL-1β of rats in each group;

[0041] Figure 9 It is a statistical result graph of the serum creatinine content of rats in each group;

[0042] Figure 10 It is a statistical result graph of the serum urea nitrogen content of rats in each group. Detailed Embodiments

[0043] The technical solution of the present invention will be further described below through specific embodiments. Those skilled in the art should understand that the embodiments are only for helping to understand the present invention and should not be regarded as specific limitations to the present invention.

[0044] The taxonomic name of the BC99 strain involved in the following content is Bacillus coagulans Bacillus coagulans The preservation date is February 1, 2021, and the preservation number is CGMCC No. 21801.

[0045] The taxonomic name of the BBi32 strain involved in the following content is Bifidobacterium bifidumBifidobacterium bifidum , with a preservation time of December 10, 2018 and a preservation number of CGMCC No. 16923.

[0046] The SPF-grade SD male rats involved in the following content were purchased from Shanghai Laboratory Animal Center; the ordinary feed was purchased from Shanghai Slack Co., Ltd.

[0047] Method for preparing BC99 or BBi32 bacterial suspension involved in the following test: Inoculate the required strain into MRS liquid medium, culture at 37 °C for 18 h for activation, and activate continuously for 2 times to obtain an activated solution; inoculate the activated solution into MRS liquid medium at an inoculation amount of 3% (v / v), culture at 37 °C to obtain a bacterial solution; centrifuge the bacterial solution at 4000 rpm at 4 °C for 5 min, filter to obtain bacterial cells, and resuspend the bacterial cells with PBS to obtain the bacterial suspension.

[0048] MRS medium: Peptone 10 g / L, Beef extract 10 g / L, Glucose 20 g / L, Sodium acetate 2 g / L, Yeast extract 5 g / L, Diammonium hydrogen citrate 2 g / L, K2PO4·3H2O 2.6 g / L, MgSO4·7H2O 0.1 g / L, MnSO4 0.05 g / L, Tween 80 1 mL / L, Cysteine hydrochloride 0.5 g / L. Example

[0049] This example explores the improvement effect of probiotic preparations on hyperuricemic rat models:

[0050] (1) Feeding and screening conditions:

[0051] Healthy male SPF-grade SD rats, 8 weeks old. These rats were raised in a controlled environment with the room temperature maintained at 23 - 27 °C, humidity at 45 - 55%, following a 12 h light / dark cycle. They could eat and drink freely. The bedding was changed 1 - 2 times a week. 0.4 mL of blood was collected from the tail vein on an empty stomach into a 1.5 mL EP tube, placed in a 4 °C refrigerator for 1 h, centrifuged at 4 °C and 3000 r / min for 15 min, and the serum was taken. The uric acid value was measured using a uric acid assay kit to exclude experimental rats with unstable uric acid. 70 rats with normal uric acid values were selected and adaptively raised for one week.

[0052] (2) Animal grouping:

[0053] After one week of adaptive feeding, the rats were randomly divided into 8 groups of 10 rats each: blank group, model group, BC99 group (intervened with BC99 bacterial solution), BBi32 group (intervened with BBi32 bacterial solution), ATCC31284 group (intervened with commercially available Bacillus coagulans ATCC31284 bacterial solution), combined group 1 (intervened with a combination of BC99 and BBi32 bacterial solutions at a viable bacteria count ratio of 10:1), combined group 2 (intervened with a combination of BC99 and BBi32 bacterial solutions at a viable bacteria count ratio of 1:10), and comparative combined group (intervened with a combination of commercially available Bacillus coagulans ATCC31284 bacterial solution and commercially available Bifidobacterium bifidum ATCC29521 bacterial solution at a viable bacteria count ratio of 10:1).

[0054] (3) Animal modeling method:

[0055] After one week of adaptive feeding, the rats were first fasted for 12 h and water-deprived for 4 h. Then, potassium oxonate (300 mg / kg) and hypoxanthine (300 mg / kg) were administered by gavage once a day for 7 consecutive days. After 7 days, blood was collected from the rat tail tip, and the serum uric acid content was detected using a uric acid assay kit. Rats with a serum uric acid content higher than 120 μmol / L were determined to have successfully modeled.

[0056] (4) Animal intervention method:

[0057] Blank group: Rats in the blank group (healthy rats) were gavaged with 10 mL of normal saline daily for 21 days;

[0058] Model group: Rats in the model group (model rats) were gavaged with 10 mL of normal saline daily for 21 days;

[0059] BC99 group: Rats in the BC99 group (model rats) were gavaged with 10 mL of BC99 bacterial suspension (concentration of 2×10 9 CFU / mL) daily for 21 days;

[0060] BBi32 group: Rats in the BBi32 group (model rats) were gavaged with 10 mL of BBi32 bacterial suspension (concentration of 2×10 9 CFU / mL) daily for 21 days;

[0061] ATCC31284 group: Rats in the ATCC31284 group (model rats) were gavaged with 10 mL of ATCC31284 bacterial suspension (concentration of 2×10 9 CFU / mL) daily for 21 days;

[0062] Combined group 1: Rats in the combined group 1 (model rats) were gavaged with 10 mL of a mixed bacterial suspension of BC99 and BBi32 (total concentration of 2×109 (CFU / mL), for 21 days;

[0063] Compound Group 2: Rats in Compound Group 2 (model rats) were gavaged with 10 mL of a mixed bacterial suspension of BC99 and BBi32 (total concentration of 2×10 9 CFU / mL) every day for 21 days;

[0064] Comparative Compound Group: Rats in the Comparative Compound Group (model rats) were gavaged with 10 mL of a mixed bacterial suspension of commercially available Bacillus coagulans ATCC31284 and commercially available Bifidobacterium bifidum ATCC29521 (total concentration of 2×10 9 CFU / mL) every day for 21 days.

[0065] (5) Index analysis:

[0066] During the experiment, the body weight of the rats was measured weekly, and the diet and water intake records were recorded. The dynamic body weight data of the rats in each group are as Figure 1 shown. As Figure 1 can be seen, after the gavage ended, the body weight of the rats in each group began to increase. After the experiment ended, the body weight of the rats in each group increased to about 300 g, indicating that hyperuricemia had no significant effect on the body weight of the rats, and the intervention of the probiotic agent also had no significant effect.

[0067] (5.2) Effect of probiotic intervention on serum uric acid level in hyperuricemic rats:

[0068] After the last gavage ended, the rats were fasted for 12 h, blood was collected from the tip of the tail, centrifuged at 3500 r / min at 4℃ for 20 min, and the serum was taken into an EP tube. A uric acid assay kit was used to detect the serum uric acid content; the serum uric acid content of the rats in each group is as Figure 2 shown.

[0069] Figure 2 The results showed that compared with the blank group, the uric acid level in the model group was much higher than that in the blank group; compared with the model group, after the intervention of each group of probiotics, the uric acid level in the serum of the rats decreased to varying degrees. The uric acid content in the BC99 group, Compound Group 1, and Compound Group 2 was significantly lower than that in the model group, and the reduction of uric acid level in Compound Group 1 and Compound Group 2 was the most significant, indicating that the two cooperate and promote each other in reducing the serum uric acid of hyperuricemic rats.

[0070] (5.3) Effect of probiotic intervention on the activity of serum xanthine oxidase (XOD) in hyperuricemic rats:

[0071] After the last gavage, the rats were fasted for 12 h, blood was collected from the tip of the tail, centrifuged at 3500 r / min at 4 °C for 20 min, and the serum was taken into an EP tube. An XOD activity assay kit was used to detect the XOD activity; the XOD content in the serum of rats in each group was as Figure 3 shown.

[0072] Uric acid in the human body is mainly decomposed from nucleic acids and other purine compounds metabolized by cells and purines in food through the action of enzymes. Uric acid is a component of nucleic acids, that is, the final product of the catabolism of adenine and guanine in the human body. Hypoxanthine and xanthine are the direct precursors of uric acid. Under the action of xanthine oxidase (XOD), hypoxanthine is oxidized to xanthine, and xanthine is oxidized to uric acid. Figure 3 The results showed that compared with the blank group, the XOD activity in the model group was higher than that in the blank group; compared with the model group, after intervention with each group of probiotics, the XOD activity in the serum of rats decreased to varying degrees, thereby reducing uric acid production and reducing further harm to the body. The XOD activity in the serum of rats in the BC99 group, the composite 1 group, and the composite 2 group was significantly lower than that in the model group, and the decrease in the XOD activity in the serum of rats in the composite 1 group and the composite 2 group was the most significant, indicating that the two cooperate and promote each other in reducing the XOD activity in the serum of hyperuricemic rats.

[0073] (5.4) Effects of probiotic intervention on the levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in the serum of hyperuricemic rats:

[0074] After the last gavage, the rats were fasted for 12 h, blood was collected from the tip of the tail, centrifuged at 3500 r / min at 4 °C for 20 min, and the serum was taken into an EP tube. The contents were detected respectively by referring to the methods of the AST and ALT detection kits; the AST and ALT contents in the serum of rats in each group were as Figure 4 、 Figure 5 shown.

[0075] Studies have shown that uric acid is produced in the liver, muscle and adipose tissues, and the liver damage in patients with hyperuricemia is also serious. Aspartate aminotransferase (AST) is mainly distributed in the myocardium, followed by tissues such as the liver, skeletal muscle and kidneys. Normally, the AST content in the serum is low, but when the corresponding cells are damaged, the cell membrane permeability increases, and the AST in the cytoplasm is released into the blood, so its serum concentration can increase. Alanine aminotransferase (ALT) is a biological enzyme, mainly present in the cytoplasm of hepatocytes, and the intracellular concentration is 1000 - 3000 times higher than that in the serum. As long as 1% of the hepatocytes are damaged, the serum enzyme can be doubled. Therefore, alanine aminotransferase (ALT) and aspartate aminotransferase (AST) are recommended by the World Health Organization as the most sensitive detection indicators for liver function damage.

[0076] From Figure 4 、 Figure 5It can be seen that compared with the blank group, the contents of AST and ALT in the model group were much higher than those in the blank group; compared with the model group, after the intervention of each group of probiotics, the contents of AST and ALT in the serum of rats decreased to varying degrees. The contents of AST and ALT in the serum of rats in the BC99 group, the composite 1 group, and the composite 2 group were lower than those in the model group, and the contents of AST and ALT in the serum of rats in the composite 1 group and the composite 2 group decreased most significantly, indicating that the two cooperate and promote each other in reducing the contents of AST and ALT in the serum of hyperuricemic rats.

[0077] (5.5)Effect of probiotic intervention on the levels of serum inflammatory factors in hyperuricemic rats:

[0078] After the last gavage, the rats were fasted for 12 h, anesthetized by intraperitoneal injection of 4% chloral hydrate solution, blood was collected from the eyeballs, centrifuged at 3500 r / min at 4℃ for 20 min, and the serum was taken into an EP tube. The contents were detected respectively according to the methods of the TNF-α, IL-6, and IL-1β detection kits; the contents of serum inflammatory factors in rats of each group were as Figures 6 - 8 shown.

[0079] It can be Figures 6 - 8 seen that compared with the blank group, the levels of TNF-α, IL-6, and IL-1β in the model group were higher than those in the blank group; compared with the model group, after the intervention of each group of probiotics, the levels of TNF-α, IL-6, and IL-1β in the serum of rats decreased to varying degrees. The levels of TNF-α, IL-6, and IL-1β in the serum of rats in the BC99 group, the composite 1 group, and the composite 2 group decreased more significantly, and the composite 1 group and the composite 2 group were better, indicating that the two have a synergistic effect in reducing the levels of serum inflammatory factors in hyperuricemic rats.

[0080] (5.6)Effect of probiotic intervention on the levels of serum creatinine and urea nitrogen in hyperuricemic rats:

[0081] After the last gavage, the rats were fasted for 12 h, anesthetized by intraperitoneal injection of 4% chloral hydrate solution, blood was collected from the eyeballs, centrifuged at 3500 r / min at 4℃ for 20 min, and the serum was taken into an EP tube. The contents were detected respectively according to the methods of the serum creatinine and urea nitrogen detection kits; the contents of serum creatinine and urea nitrogen in rats of each group were as Figures 9 - 10 shown.

[0082] Long-term hyperuricemia will further aggravate renal function damage, specifically manifested as a further increase in creatinine and urea nitrogen. It can be Figures 9 - 10It can be seen that compared with the blank group, the contents of serum creatinine and urea nitrogen in the model group were higher than those in the blank group; compared with the model group, after the intervention of each group of probiotics, the contents of serum creatinine and urea nitrogen in rats were improved to varying degrees. The reduction effects of serum creatinine and urea nitrogen contents in the BC99 group, the composite 1 group, and the composite 2 group were more significant, and the composite 1 group and the composite 2 group were superior, indicating that the two had synergistic effects in reducing the levels of serum creatinine and urea nitrogen.

[0083] The applicant declares that the technical solution of the present invention is illustrated by the above embodiments, but the present invention is not limited to the above embodiments, that is, it does not mean that the present invention must rely on the above embodiments to be implemented. Those skilled in the art should understand that any improvement of the present invention, the equivalent substitution of each raw material of the product of the present invention, the addition of auxiliary components, the selection of specific methods, etc. all fall within the protection scope and the disclosure scope of the present invention.

[0084] The preferred embodiments of the present invention have been described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the scope of the technical concept of the present invention, various simple modifications can be made to the technical solution of the present invention, and these simple modifications all belong to the protection scope of the present invention.

[0085] In addition, it should be noted that in the above specific embodiments, the various specific technical features described can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not separately describe various possible combination methods.

Claims

1. Use of a strain combination consisting of Bacillus coagulans BC99 and Bifidobacterium bifidum BBi32 in the preparation of a probiotic preparation for assisting in reducing uric acid levels; The Bacillus coagulans BC99 is the Bacillus coagulans with the preservation number of CGMCC No. 21801 Bacillus coagulans Strain BC99; The Bifidobacterium bifidum BBi32 is the Bifidobacterium bifidum with the preservation number of CGMCC No. 16923 Bifidobacterium bifidum strain BBi32; The viable count ratio of the BC99 strain to the BBi32 strain is 1:10 - 10:

1.

2. The application according to claim 1, wherein The dosage form of the probiotic preparation includes solution, powder, tablet, granule or capsule.

3. The application according to claim 1, characterized in that, The viable count of the probiotic preparation is not less than 1×10 9 CFU / mL or 1×10 9 CFU / g.

4. The application according to claim 1, characterized in that When the dosage form of the probiotic preparation is a solution, it is prepared by the following method: Inoculate the relevant strains into a culture medium for activation and fermentation culture to obtain a fermentation broth; after centrifuging the fermentation broth, resuspend it with a solvent to obtain a bacterial suspension or a mixed bacterial suspension; When the dosage form of the probiotic agent is a freeze-dried powder, it is prepared by the following method: Inoculate the relevant strains into a culture medium for activation and fermentation culture to obtain a fermentation broth; after centrifuging the fermentation broth, mix it with a cryoprotectant and then perform freeze-drying to obtain a bacterial powder or a mixed bacterial powder.

5. The application according to claim 1, wherein The probiotic preparation also contains excipients; the excipients include any one or a combination of at least two of fillers, binders, wetting agents, disintegrants, emulsifiers, solubilizing agents, solubilizers, osmotic pressure regulators, colorants, pH regulators, antioxidants, bacteriostatic agents or buffers.

6. Use of a strain combination consisting of Bacillus coagulans BC99 and Bifidobacterium bifidum BBi32 in the preparation of a probiotic preparation for improving hyperuricemia; The Bacillus coagulans BC99 is the Bacillus coagulans with the preservation number of CGMCC No. 21801 Bacillus coagulans Strain BC99; The Bifidobacterium bifidum BBi32 is the Bifidobacterium bifidum with the preservation number of CGMCC No. 16923 Bifidobacterium bifidum BBi32 strain; The viable count ratio of the BC99 strain to the BBi32 strain is 1:10 - 10:

1.

7. The application according to claim 6, wherein The viable count of the probiotic preparation is not less than 1×10 9 CFU / mL or 1×10 9 CFU / g.

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