A uric acid-lowering probiotic complex and its application

Abstract

The present invention relates to a uric acid-lowering probiotic complex and its application. The probiotic complex comprises Lactobacillus reuteri FPH2951 and Bifidobacterium breve FPHC4024; wherein Lactobacillus reuteri FPH2951 is classified as Lactobacillus reuteri and has a deposit number of GDMCC No. 62946; and Bifidobacterium breve FPHC4024 is classified as Bifidobacterium breve and has a deposit number of GDMCC No. 62945. The probiotic complex of the present invention can significantly reduce serum uric acid (UA), creatinine (Cr), and blood urea nitrogen (BUN) levels and increase urine excretion of creatinine (Cr) and blood urea nitrogen (BUN) through the synergistic effect of Lactobacillus reuteri FPH2951 and Bifidobacterium breve FPHC4024, and has broad application prospects.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedicine, and particularly relates to a uric acid-lowering probiotic complex and applications thereof. Background Art

[0002] Hyperuricemia (HUA) is an idiopathic inflammatory disease characterized by long-term disturbances in purine metabolism and persistently elevated serum uric acid (UA) concentrations. In recent years, with the continuous improvement of living standards and changes in dietary patterns, particularly the increasing intake of high-purine foods such as seafood, meat, and beer, the incidence of HUA has been increasing annually, with a trend toward younger patients. Although HUA is not fatal, when uric acid concentrations exceed saturation, UA crystals may deposit, triggering a sterile inflammatory response in local tissues, increasing the burden of complications and leading to tissue damage such as gout, atherosclerosis, type 2 diabetes, vascular damage, and chronic kidney disease. Ultimately, gout can develop. Furthermore, hyperuricemia is associated with a variety of common metabolic diseases, such as diabetes, hypertension, and dyslipidemia, making it the fourth most common "hyper" after hypertension, hyperglycemia, and hyperlipidemia.

[0003] Existing treatments for HUA include both pharmacological and non-drug therapies, but all are associated with adverse reactions. Clinically used drugs primarily include uricosuric drugs, uricosuric inhibitors, and alkaline agents, which alleviate the symptoms of hyperuricemia by increasing uric acid excretion, inhibiting uric acid production, or alkalizing urine. However, these drugs all suffer from low tolerability, high dependency, and numerous complications and side effects. Regarding dietary control, simply restricting purine intake is extremely difficult to alleviate hyperuricemia, as most foods contain purines due to their presence in plant and animal cells. Furthermore, restricting purine intake can easily lead to nutritional imbalances. Given the shortcomings and deficiencies of existing treatments, there is an urgent need for an effective, stable, safe, and convenient HUA treatment strategy.

[0004] As a normal flora in the human intestine, lactic acid bacteria can regulate metabolic processes such as blood lipids and blood sugar in the human body. Lactic acid bacteria are non-toxic, harmless, and have no side effects. They are increasingly valued in the clinical treatment and prevention of metabolic intestinal diseases, and have shown broad clinical application prospects and market potential in effectively regulating gastrointestinal metabolic functions. For example, in the Chinese authorized invention patent CN118222464B, a plant lactobacillus and its application in uric acid-lowering products are disclosed. This strain can effectively inhibit the increase in serum uric acid, blood urea nitrogen, and serum creatinine levels, reduce the occurrence of hyperuricemia; maintain normal serum lipid levels; improve oxidative stress damage; inhibit the secretion of inflammatory cytokines, promote the expression of anti-inflammatory factors, and have a protective effect on the liver and colon; regulate and improve intestinal flora imbalance. Provide technical support for the preparation of functional foods, probiotic preparations, fermented products, and drugs that lower uric acid. For example, China's authorized invention patent CN118147019B discloses fermented Lactobacillus mucinus GS31 and its uric acid-lowering application. The fermented Lactobacillus mucinus can significantly lower uric acid levels, promote the excretion of excess uric acid in the body, and reduce the production of uric acid in the body, further alleviating damage to kidney function, and reducing inflammatory factors in the body. It has a certain improvement and preventive effect on hyperuricemia and related diseases caused by hyperuricemia.

[0005] The applicant's previously authorized invention patent CN116970530B discloses a Bifidobacterium breve FPHC4024 for preventing and treating diarrhea in children and its application, which belongs to the field of probiotic technology. The Bifidobacterium breve FPHC4024 is classified and named Bifidobacterium breve, with a deposit number of GDMCC NO.62945 and a deposit date of November 3, 2022. The depository is the Guangdong Provincial Microbial Culture Collection Center, address: 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, Institute of Microbiology, Guangdong Academy of Sciences, Postal Code: 510070. The invention provides a Bifidobacterium breve FPHC4024 that can cause the absorption of excess water in the colon, and has a good effect in preventing, alleviating and treating diarrhea in children, with the advantages of good efficacy, quick effect, safety, no side effects, and a short treatment cycle. However, the invention patent does not disclose that it can lower uric acid.

[0006] The applicant's prior invention patent application CN117018043A discloses a probiotic strain and its application, a composition for preventing and treating diarrhea in children, which belongs to the field of probiotic technology. The probiotic strain is Lactobacillus reuteri FPH2951, classified and named Lactobacillus reuteri, with a deposit number of GDMCC NO.62946 and a deposit date of November 3, 2022. The depository is the Guangdong Provincial Microbial Culture Collection Center, address: 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, Institute of Microbiology, Guangdong Academy of Sciences, Postal Code: 510070. The probiotics of this invention can play a good role in preventing, alleviating and treating diarrhea in children, and have the advantages of good efficacy, quick effect, short treatment cycle, etc., and have broad application prospects. However, this invention patent does not disclose that it can reduce uric acid.

[0007] To this end, the inventors of the present application conducted further research and found that the above-mentioned strains can reduce uric acid, thereby completing the present invention. Summary of the Invention

[0008] To address the deficiencies of the prior art, the present invention proposes a uric acid-lowering probiotic complex and its application. Specifically, to achieve the objectives of the present invention, the present invention intends to adopt the following technical solutions:

[0009] One aspect of the present invention relates to a uric acid-lowering probiotic complex, characterized in that the probiotic complex comprises Lactobacillus reuteri FPH2951 and Bifidobacterium breve FPHC4024; wherein Lactobacillus reuteri FPH2951 is classified as Lactobacillus reuteri, with a preservation number of GDMCC NO.62946; Bifidobacterium breve FPHC4024 is classified as Bifidobacterium breve, with a preservation number of GDMCC NO.62945.

[0010] In a preferred embodiment of the present invention, the uric acid-lowering probiotic complex further comprises prebiotics.

[0011] In a preferred embodiment of the present invention, the ratio of the number of Lactobacillus reuteri FPH2951 to that of Bifidobacterium breve FPHC4024 is 1-2:1-2; preferably 1-1.25:1-1.25.

[0012] Another aspect of the present invention relates to the use of the above-mentioned uric acid-lowering probiotic complex in the preparation of a medicament for lowering uric acid.

[0013] Preferably, the drug has at least one of the following effects: reducing the uric acid level in hyperuricemic mice, specifically reducing the levels of uric acid (UA), creatinine (Cr) and urea nitrogen (BUN) in serum, and increasing the excretion of creatinine (Cr) and urea nitrogen (BUN) in urine. Beneficial effects

[0014] The probiotic complex of the present invention, through the synergistic action of Lactobacillus reuteri FPH2951 and Bifidobacterium breve FPHC4024, can significantly reduce the levels of uric acid (UA), creatinine (Cr), and blood urea nitrogen (BUN) in serum, and increase the excretion of creatinine (Cr) and blood urea nitrogen (BUN) in urine, and has broad application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1 : Microscope photos of Lactobacillus reuteri FPH2951.

[0016] Figure 2 : Microscopic photograph of Bifidobacterium breve FPHC4024. DETAILED DESCRIPTION

[0017] To further understand the present invention, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the embodiments described are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0018] Unless otherwise specified, all reagents involved in the embodiments of the present invention are commercially available products and can be purchased through commercial channels.

[0019] Example 1

[0020] Cultivating probiotics:

[0021] Lactobacillus reuteri FPH2951 is a Gram-positive rod-shaped bacterium that does not produce spores. When cultured anaerobically at 36°C for 2 days on MRS (lactic acid bacteria culture medium shown in Table 1), the colonies are milky white, round, convex, with smooth surfaces and neat edges. Microscopic photos are shown below. Figure 1 shown.

[0022] Table 1: Culture medium of Lactobacillus reuteri FPH2951

[0023]

[0024] Bifidobacterium breve FPHC4024 is a Gram-positive rod-shaped bacterium that does not form spores. After anaerobically incubating on MRS (lactic acid bacteria culture medium shown in Table 2) plates at 37°C for 2 days, the colonies were milky white, round, convex, with smooth surfaces and neat edges. Microscopic photos are shown below. Figure 2 shown.

[0025] Table 2: Culture medium for Bifidobacterium breve FPHC4024

[0026]

[0027] Experimental methods:

[0028] Seventy male SPF Balb / c mice weighing 18-20 g were housed at 20-25°C, with a relative humidity of (50±5)% and a 12-h / 12-h photoperiod. They were randomly divided into seven groups of 10 mice each: a normal control group (blank control), a model group, a positive control group, a control experimental group (single strain), and an experimental group (mixed strain group). ① Blank control group: fed with normal feed, and the solvent was gavaged alone; ② Model group: given potassium oxonate (250 mg / kg, the solvent was 5% CMC-Na aqueous solution); ③ Positive control group: given potassium oxonate (250 mg / kg, the solvent was 5% CMC-Na aqueous solution) + allopurinol (5 mg / kg); ④ Control experimental group 1 (i.e., Bifidobacterium breve FPHC4024 group): given potassium oxonate (250 mg / kg, the solvent was 5% CMC-Na aqueous solution) + Bifidobacterium breve FPHC4024 group (1.8×10 10 CFU / kg); ⑤ Control experimental group 2 (i.e., Lactobacillus reuteri FPH2951 group): administered with potassium oxonate (250 mg / kg, solvent: 5% CMC-Na aqueous solution) + Lactobacillus reuteri FPH2951 group (1.8×10 10 CFU / kg); ⑥ Experimental group 1 (i.e., mixed strain group): administered with potassium oxonate (250 mg / kg, solvent: 5% CMC-Na aqueous solution) + Lactobacillus reuteri FPH2951 (1.0×10 10 CFU / kg) + Bifidobacterium breve FPHC4024 group (0.8×10 10 CFU / kg); ⑦ Experimental group 2 (i.e., mixed strain group): administered with potassium oxonate (250 mg / kg, solvent: 5% CMC-Na aqueous solution) + Lactobacillus reuteri FPH2951 (0.8×10 10 CFU / kg) + Bifidobacterium breve FPHC4024 group (1.0×10 10 CFU / kg)

[0029] All groups, except the blank control group, received the above-mentioned doses via gavage. The blank group received an equal volume of vehicle (5% CMC-Na aqueous solution). All groups received a gavage volume of 0.3 mL / 100 g once daily for 14 days. After 14 days of fasting for 12 hours, blood was collected from the eyeballs and centrifuged at 3000 rpm for 20 minutes to separate serum. Urine was collected from cages for 24 hours. Serum and urine uric acid (UA), creatinine (Cr), and serum urea nitrogen (BUN) levels were measured using commercially available kits. (The UA, BUN, and creatinine (Cr) kits were all purchased from Solarbio.)

[0030] Table 3: Effects of each experimental group on UA, Cr, and BUN in hyperuricemia mice (n=10)

[0031]

[0032] As shown in Table 1, the levels of UA, BUN, and Cr in the serum of each group were compared. After oral administration of potassium oxonate, the UA level of the model group mice was significantly higher than that of the normal group (P<0.05), indicating that the hyperuricemia mouse model was successfully established.

[0033] After 14 days of treatment with the mixed strain, serum uric acid levels in mixed strain experimental groups 1 and 2 were significantly reduced (P < 0.05) compared with the model group and control experimental groups 1 and 2. Serum creatinine and urea nitrogen levels in mixed strain experimental groups 1 and 2 were also significantly reduced (P < 0.05) compared with the model group and control experimental groups 1 and 2. The changes in urea nitrogen and creatinine indicate that the mixed strain has a strong protective effect on the kidneys through the synergistic effect of the strains. At the same time, the level of creatinine excreted in urine is close to that of the normal group.

[0034] The above describes the preferred embodiments of the present invention, which is not intended to limit the present invention. Those skilled in the art may make improvements and changes to the embodiments disclosed herein without departing from the scope and spirit of the present invention.

Claims

1. Use of a uric acid-lowering probiotic complex in the preparation of a medicament for lowering uric acid, characterized in that: The probiotics in the probiotic complex are Lactobacillus reuteri FPH2951 and Bifidobacterium breve FPHC4024; wherein the classification name of Lactobacillus reuteri FPH2951 is Lactobacillus reuteri. Limosilactobacillus reuteri , the deposit number is GDMCCNO.62946; Bifidobacterium breve FPHC4024 is classified and named Bifidobacterium breve Bifidobacterium breve , the deposit number is GDMCC NO.62945; the quantity ratio of Lactobacillus reuteri FPH2951 and Bifidobacterium breve FPHC4024 is 1-2:1-2.

2. The use according to claim 1, wherein the uric acid-lowering probiotic complex further comprises prebiotics.

3. The use according to claim 1, wherein the ratio of the number of Lactobacillus reuteri FPH2951 to that of Bifidobacterium breve FPHC4024 is 1-1.25:1-1.25.

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