Pediococcus acidilactici SWU-HX39 capable of relieving hyperuricemia

Abstract

The pediococcus acidilactici SWU-HX39 with the uricemia reducing function is classified and named as pediococcus acidilactici, and is preserved in the China General Microbiological Culture Collection Center (CGMCC), the preservation number is CGMCC No.33055, the preservation date is December 13, 2024, and the preservation address is No.3, Yard 1, Beichen West Road, Chaoyang District, Beijing. The pediococcus acidilactici SWU-HX39 disclosed by the invention can be used for relieving kidney injury caused by hyperuricemia; synthesis of uric acid can be reduced, the concentration of uric acid in blood can be reduced, the excretion capacity of the body to the uric acid is improved, reabsorption of the body to the uric acid is reduced, and the effect of reducing the uric acid is achieved through two ways; in addition, the SWU-HX39 can be used for reducing the expression of LPS (Lipopolysaccharide), and reducing the expression of inflammatory factors IL-1beta, IL-6 and TNF (Tumor Necrosis Factor) alpha at the same time. The traditional Chinese medicine composition not only has the effects of resisting inflammation and relieving liver inflammation, but also can improve the SCFAs content, especially propionic acid, in the body of a hyperuricemia patient, so that the uric acid level is reduced, and the abnormal uric acid content is improved.

Description

Technical Field

[0001] The present invention relates to the field of microbial technology, and particularly relates to a Pediococcus acidilactici SWU-HX39 with the effect of alleviating hyperuricemia. Background Art

[0002] Hyperuricemia is a disease caused by purine metabolism disorders, mainly manifested as increased uric acid synthesis in the body or decreased renal uric acid excretion, or both. This disease is not only closely related to gout, but also may cause systemic diseases such as cardiovascular diseases, kidney diseases and metabolic syndrome. Its development is often accompanied by metabolic disorders, especially diseases involving the liver, kidneys and intestines, reflecting complex physiological and pathological processes. Research shows that the role of the liver and kidneys in purine metabolism directly affects the accumulation of uric acid. When the blood uric acid level is too high, uric acid crystals and precipitates may cause complications such as gout, especially in joints and cartilage. Therefore, gout patients urgently need effective measures to reduce the body's uric acid level.

[0003] The methods for treating hyperuricemia mainly include two approaches: drug treatment and non-drug treatment. Commonly used drugs for drug treatment include uricosuric drugs such as benzbromarone, or drugs that inhibit uric acid synthesis such as allopurinol. However, drug treatment may be accompanied by clinical side effects and economic burdens. In contrast, non-drug treatment, by reducing the intake of high-purine foods and supplementing probiotics or fermented foods that help degrade uric acid, shows novelty and potential, and has become a promising alternative or adjuvant treatment strategy.

[0004] Probiotics are a type of bacteria beneficial to the health of the host. Research shows that lactic acid bacteria can alleviate hyperuricemia by inhibiting the activity of xanthine oxidase; the intake of Lactobacillus can promote the transformation of compounds in the intestines of hyperuricemic mice and reduce purine levels. In addition, it also reduces the purine concentration in the liver and inhibits the activity of xanthine oxidase. There is a large amount of clinical and epidemiological evidence indicating that certain probiotics can reduce serum uric acid levels, but the uric acid-lowering effects and mechanisms of different probiotics vary. The application of probiotics in the treatment of hyperuricemia is still in its infancy. Due to the large variety of probiotics, with different uric acid-lowering mechanisms of action, intestinal colonization ability and potential pathogenicity, the uric acid-lowering effects exerted in the body also vary significantly. Therefore, there is an urgent need to seek lactic acid bacteria that are safe and can very effectively reduce serum uric acid concentration. Summary of the Invention

[0005] The object of the present invention is to provide a Pediococcus acidilactici with the effect of alleviating hyperuricemia. Through animal experiments, its effects of reducing serum uric acid concentration, anti-inflammation, regulating the intestinal microbiota, promoting uric acid excretion and improving kidney metabolism are confirmed.

[0006] Another object of the present invention is to provide the application of the above-mentioned Pediococcus acidilactici.

[0007] The object of the present invention is achieved by the following technical solutions: A Pediococcus acidilactici SWU-HX39 with the function of reducing hyperuricemia, characterized in that: the classification name of the Pediococcus acidilactici is Pediococcus acidilactici ( Pediococcus acidilactici ) SWU-HX39, which is deposited in the General Microbiology Center of the China Committee for Culture Collection of Microorganisms, with the deposit number of CGMCC No. 33055, the deposit date of December 13, 2024, and the deposit address of No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0008] Pediococcus acidilactici SWU-HX39 effectively inhibits the activity of XOD in the liver, and at the same time reduces the levels of creatinine and urea nitrogen in the serum of mice, thereby reducing the uric acid content in the blood of patients.

[0009] In addition, Pediococcus acidilactici SWU-HX39 can also promote the excretion of uric acid by patients, and at the same time inhibit and reduce the reabsorption of uric acid by the body.

[0010] Pediococcus acidilactici SWU-HX39 can relieve hyperuricemia by regulating LPS and liver inflammation, and has an obvious anti-inflammatory effect.

[0011] Pediococcus acidilactici SWU-HX39 can also regulate the level of the body's intestinal microbiota and increase the content of SCFAs in the body.

[0012] The above-mentioned Pediococcus acidilactici SWU-HX39 is used in the preparation of a drug for regulating and reducing uric acid.

[0013] The above-mentioned Pediococcus acidilactici SWU-HX39 is used in the preparation of a food for assisting in uric acid regulation.

[0014] The above-mentioned Pediococcus acidilactici SWU-HX39 is used in the preparation of an anti-inflammatory drug.

[0015] The above-mentioned Pediococcus acidilactici SWU-HX39 is used in the preparation of a drug or food for regulating intestinal flora.

[0016] The present invention has the following technical effects: The Pediococcus acidilactici SWU-HX39 of the present invention can relieve kidney damage caused by hyperuricemia; it can also reduce the synthesis of uric acid, lower the uric acid concentration in the blood, at the same time improve the body's uric acid excretion ability, and reduce the body's reabsorption of uric acid, achieving the effect of reducing uric acid through both methods; in addition, SWU-HX39 can reduce the expression of LPS, and at the same time reduce the expression of inflammatory factors IL-1β, IL-6, TNF-α. That is, it has the effects of anti-inflammation and reducing liver inflammation, and can also increase the content of SCFAs in patients with hyperuricemia, especially propionic acid, thereby reducing the uric acid level and improving the abnormal uric acid content. Brief Description of the Drawings

[0017] Figure 1 : Pictures of the colony morphology and staining characteristics of SWU-HX39

[0018] Figure 2 : Results of the simulated gastrointestinal fluid tolerance test of SWU-HX39

[0019] Figure 3 : Graph of the body weight changes of hyperuricemic mice intervened by SWU-HX39

[0020] Figure 4 : Graph of the changes in kidney index of hyperuricemic mice intervened by SWU-HX39

[0021] Figure 5 : HE staining of kidney tissue sections of hyperuricemic mice intervened by SWU-HX39

[0022] Figure 6 : Changes in the contents of UA (uric acid), CRE (creatinine), BUN (blood urea nitrogen) in the serum of hyperuricemic mice intervened by SWU-HX39 and the activity levels of XOD (xanthine oxidase) in the liver and serum

[0023] Figure 7 : Graph of the changes in TNF-α, IL-1β, IL-6 and LPS levels in hyperuricemic mice intervened by SWU-HX39

[0024] Figure 8 : Column graph of the relative species abundances at the genus level of hyperuricemic mice intervened by SWU-HX39

[0025] Figure 9 : Content of SCFAs in cecal contents after intervention with SWU-HX39

[0026] Figure 10 : Expression of genes related to renal uric acid transport in hyperuricemic mice intervened by SWU-HX39 Detailed Embodiments

[0027] The present invention will be specifically described below through examples. It is necessary to point out here that the following examples are only used to further illustrate the present invention and should not be construed as limiting the protection scope of the present invention. Those skilled in the art can make some non-essential improvements and adjustments to the present invention according to the above content of the present invention.

[0028] Example 1 Isolation, screening and identification of strains: Multiple lactic acid bacteria samples with excellent acid resistance were isolated from cheese and pickles.

[0029] The strain was inoculated into fresh MRS liquid medium at 2% (v / v) and incubated at 37 °C for 24 h. 2 mL of bacterial suspension was taken and centrifuged at 4000 r / min for 10 min. The suspension was washed three times with 1 mL of 0.85% NaCl solution. The concentration of the bacterial suspension was adjusted to 10 using a bacterial turbidity meter. 9 cfu / mL. Take 1 mL of bacterial suspension, centrifuge at 4000 r / min for 10 min, discard the supernatant, resuspend the cells in 750 μL of 1mmol / L inosine-guanosine solution, and incubate at 37 °C for 120 min. After incubation, centrifuge at 4000 r / min for 10 min, take the supernatant, and mix the supernatant with 0.1 M HClO 4 Add HClO in a volume ratio of 9:1 4 After mixing, filter the solution with a 0.22 μm filter membrane for testing.

[0030] The residual inosine and guanosine content in the solution was determined using a high performance liquid chromatography (HPLC) system. Chromatographic conditions: reverse phase column ThermoFisher-C18 (4.6 mm×250 mm, 5 μm), mobile phase 10 mmol / L potassium dihydrogen phosphate solution (pH 5.0) and methanol (volume ratio 90:10), flow rate 1 mL / min, column temperature 25°C, wavelength 254 nm, elution time 30 min. The degradation rate was calculated according to the formula.

[0031] Degradation rate = ×100% Where C 0 : Initial concentration of guanosine (inosine) standard solution, mmol / L; C: Residual concentration of guanosine (inosine) standard solution after reaction, mmol / L.

[0032] A strain with excellent degradation ability for inosine and guanosine was obtained through computational screening. The degradation rates of inosine and guanosine were 82.75%±4.74 and 78.80%±4.28, respectively.

[0033] The purified strain was taken out of the glycerol tube and inoculated into fresh MRS liquid medium at 2% (v / v) and cultured at 37°C for 24 h. After two generations of activation, a single colony was streaked on the MRS plate with a sterile inoculation loop, and the purified isolate was examined under a microscope and Gram staining was performed. The results were as follows: Figure 1 shown.

[0034] The strain was further identified by 16S rDNA sequencing through multiple plate isolations. The 16S rDNA of the strain was amplified by PCR and sequenced, and the alignment analysis was carried out using the BLAST (Basic Local Alignment Search Tool) program in the NCBI database. After sequence comparison, it was identified as Pediococcus acidilactici.

[0035] This strain was named SWU-HX39 and deposited in the General Microbiology Center of the China Microbial Culture Collection Center. The deposit number is CGMCC No. 33055, the deposit date is December 13, 2024, and the deposit address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0036] For the simulated gastrointestinal fluid tolerance of strain SWU-HX39: Simulated gastric fluid: Pepsin was added to sterile PBS (0.1 mol / L, pH 2.5) to a final concentration of 3 g / L; simulated intestinal fluid: Trypsin was added to sterile PBS (0.1 mol / L, pH 8.0) to a final concentration of 1 g / L. The lactic acid bacteria activated for two generations were cultured overnight, washed twice with sterile saline (0.85% NaCl, w / v) and resuspended, and then inoculated into the simulated gastric fluid to make the final concentration of the bacterial solution 108 CFU / mL. Incubate at 37 °C for 3 h. At 0 and 3 h, the bacterial solution was serially diluted 10-fold with sterile saline, and 100 μL of the appropriate dilution was spread on MRS plates and cultured at 37 °C for 48 h, and then the viable bacteria count was calculated. Then 1 mL of the culture was inoculated into 9 mL of simulated intestinal fluid and incubated at 37 °C for 8 h. At 0, 2, 4, and 8 h, the bacterial solution was serially diluted 10-fold with sterile saline, and 100 μL of the appropriate dilution was spread on MRS plates and cultured at 37 °C for 48 h, and then the viable bacteria count was calculated. The survival rate of lactic acid bacteria in the simulated gastrointestinal fluid was calculated according to the following formula:

[0037] Nt, the viable bacteria count of lactic acid bacteria at different times after treatment with simulated gastrointestinal fluid; N0, the initial viable bacteria count of lactic acid bacteria.

[0038] The results are as Figure 2 shown. Pediococcus acidilactici SWU-HX39 has excellent tolerance in simulated gastric fluid and simulated intestinal fluid.

[0039] Hemolytic activity test: Pediococcus acidilactici SWU-HX39 was inoculated on Columbia agar plates containing 5% sterile defibrinated sheep blood and cultured at 37 °C for 48 h. Hemolytic activity is divided into three types: α-hemolysis (green area), β-hemolysis (clear and transparent area), and γ-hemolysis (no hemolysis area, i.e., negative). The experimental results showed that Pediococcus acidilactici SWU-HX39 had no hemolytic activity, demonstrating its safety.

[0040] Example 2 Animal experiments were conducted to test the effects and uric acid-lowering mechanism of Pediococcus acidilactici SWU-HX39 (abbreviated as HX39 in the accompanying drawings): SPF-grade male Kunming mice, 6 weeks old, weighing 27 ± 2 g, were purchased from Hunan Slack Jingda Experimental Animal Co., Ltd. The animal laboratory was of SPF clean grade with a standard pathogen-free environment. The feeding conditions were an environmental temperature of 25 ± 1 °C, humidity of 60 ± 10%, and a light / dark cycle of 12 / 12. The feed was ordinary growth feed, and the bedding was wood chips. All mice could drink water freely, and all operations strictly adhered to the experimental ethical principles and requirements. The mice were normally fed for 7 days to allow environmental adaptation and then divided into a blank group, a model group, an HX39 group, and a positive control group (allopurinol group), with 8 mice in each group. An adenine combined with potassium oxonate combined model of chronic renal failure was selected and improved based on the results of previous preliminary experiments. Finally, a suspension of 75 mg / kg adenine and 220 mg / kg potassium oxonate was used for gavage, and the solvent was 0.5% sodium carboxymethylcellulose solution. The gavage volume was 200 μL, and gavage was performed at 9:00 am every day. The HX39 group was given 100 μL of a suspension of the SWU-HX39 strain at a concentration of 10 9 CFU to the mice 1 h after gavage. The allopurinol group was fed 8 mg / kg of allopurinol, while the control group and the model group only received normal saline solution. After the gavage ended on the same day, the remaining drugs were directly discarded without recovery to avoid affecting the experimental results due to repeated freezing and thawing. During the 24-day animal experiment, the mice were weighed and observed at 11:00 am every day.

[0041] Before dosing on the 24th day, the mice were fasted for 16 h without water. (1) Their fresh urine and feces were collected and quickly placed on ice, and then immediately transferred to a -80 °C refrigerator for storage. (2) The mice were subjected to enucleation of the eyeballs, and blood was collected from the orbital region. The collected blood was centrifuged at 3500 r / min for 15 min at 4 °C, and the serum was stored in a -80 °C refrigerator for later use. (3) The mice were sacrificed by cervical dislocation, and dissected to obtain the liver, kidneys, small intestine, colon contents, and cecal contents.

[0042] (1) Determination of organ index: Mouse organ tissues need to be rinsed in physiological saline pre-cooled at 4°C to remove blood and adherent connective tissues. After drying with filter paper, weigh them and calculate the organ index according to the following formula:

[0043] During the experiment, the fur of the mice in the normal group was shiny, their excretory function was normal, and their body weight increased steadily, showing a good physiological state. On the contrary, the mice in the model group showed dull fur luster, weakened activity, and abnormally increased urine output. As the experiment progressed, the body weight of the mice in this group showed a significant decline in the later stage, accompanied by a significant increase in water intake and urine output, aggravated difficulty in defecation, and obvious signs of hair erection, which reflected the poor health of the mice. As Figure 3 shown, after supplementing with Pediococcus acidilactici SWU-HX39, the trend of weight loss was effectively curbed. At the end of the experiment, there was no significant difference in the body weight of the mice in the HX39 group and the positive drug control group.

[0044] The changes in organ index are as Figure 4 shown. Compared with the control group, the kidney index of the mice in the model group was significantly increased (P<0.0001), indicating that the kidneys might be congested or edematous, confirming that the modeling agent had obvious toxic and side effects on the kidneys of mice. However, after intervention with Pediococcus acidilactici, the kidney index of the mice showed a significant decrease (P<0.05), indicating that Pediococcus acidilactici SWU-HX39 could effectively relieve the kidney injury induced by the modeling agent.

[0045] (2)Histological analysis Mouse kidneys were immersed in tissue fixative for 24 h, dehydrated step by step with ethanol of different concentrations, then cleared with xylene, and embedded in paraffin to make sections. The sections were dewaxed with xylene I and xylene II for 5 min respectively, and then treated with an ethanol gradient (absolute ethanol for 5 min, 95% ethanol for 2 min, 80% ethanol for 2 min, 70% ethanol for 2 min), and distilled water for 2 min. The dewaxed tissue sections were stained with hematoxylin staining solution for 20 min, and then rinsed with tap water. Next, use the differentiating solution for 30 s, and soak the tissue samples in water for 15 min. Then stain with eosin staining solution for 30 s and rinse with tap water. After the samples were soaked in water for 5 min, dehydrated with an ethanol gradient, cleared with xylene, and sealed with neutral gum. Finally, observe and take pictures with an optical microscope.

[0046] From Figure 5It can also be seen that in the histological analysis of the kidneys, the surface color of the kidneys of the control group mice was dark red, the cortical layer thickness was moderate, and all the internal tissue structures of the kidneys were in a normal state. In contrast, in the kidneys of the model group, a large amount of uric acid crystals were deposited in the renal interstitium and renal tubules, with focal mild atrophy of the glomeruli, and obvious disorder of the glomerular and renal tubular structures (black arrows); the lumens of the renal tubules were significantly dilated, the arrangement of epithelial cells was significantly disordered, and the basement membrane was intact (green arrows); the blood vessels in the interstitium were moderately dilated and congested (red arrows); and a large number of lymphocytes infiltrated (blue arrows). However, after administration of Pediococcus acidilactici SWU-HX39, the symptoms of kidney disease were significantly alleviated, the symptoms of nephropathy in mice were all reduced, and their morphology was more inclined to that of the blank group. Compared with the normal control group, the color of the kidneys in the positive control group was significantly lighter, and there was also a significant tendency for the kidney volume to increase, that is, there were also certain degrees of lesions in the kidneys. This result not only verified the protective effect of Pediococcus acidilactici SWU-HX39 on the kidneys of hyperuricemia mice, but also provided morphological evidence for its potential kidney health maintenance function.

[0047] (3)Determination of biochemical indicators (UA, CRE, and BUN): Commercially available detection kits (Nanjing Jiancheng Institute, China) were used to detect the contents of UA (uric acid), CRE (creatinine), BUN (blood urea nitrogen) in the serum, and the XOD (xanthine oxidase) levels in the serum and liver. The determination was carried out according to the instructions of the kits. Eight samples were selected from each group for independent determination.

[0048] A typical sign of hyperuricemia is an increase in serum uric acid levels, usually accompanied by corresponding increases in creatinine and blood urea nitrogen levels. According to Figure 6 the data shown, compared with the control group, the serum uric acid, creatinine, and blood urea nitrogen concentrations of the model group mice were significantly increased (P < 0.0001), and the model group was 3.6 times that of the control group, which marked the successful construction of the hyperuricemia model. However, under the intervention of Pediococcus acidilactici HX39, the serum uric acid concentration of the mice decreased significantly (P < 0.0001), and the serum uric acid concentration in the HX39 group decreased by (49.8%) compared with the model group. At the same time, the levels of creatinine (P < 0.001) and blood urea nitrogen (P < 0.01) also decreased significantly. The changes in these biochemical indicators were consistent with the previous kidney section observations and kidney index measurement results. This effect may be attributed to the ability of the strain SWU-HX39 to reduce the levels of precursor substances related to uric acid synthesis, thereby reducing the production of uric acid and alleviating the symptoms of hyperuricemia in mice.

[0049] Xanthine oxidase XOD, as a key enzyme in uric acid synthesis, its activity in the liver will increase correspondingly when the uric acid level rises. As Figure 6As shown, compared with the control group, the XOD activities in the livers and sera of the model group mice were significantly increased (P < 0.01). After supplementing with SWU-HX39, the XOD activities in the liver and serum decreased respectively (P < 0.05 and P < 0.01). These results indicate that the effect of Pediococcus acidilactici SWU-HX39 in alleviating hyperuricemia in mice is partly attributed to its ability to inhibit XOD activity, thereby reducing the synthesis of uric acid in the body.

[0050] (4)Determination of anti-inflammatory effect: Lipopolysaccharide (LPS) in the sera of mice was determined by ELISA to evaluate typical inflammatory cytokines in the sera, so as to evaluate the damage caused by HUA-induced renal fibrosis in mice (IL-1β, TNF-α and IL-6).

[0051] The results are as Figure 7 shown. Among the pro-inflammatory cytokines in the sera of the model group mice compared with the control group, the contents of tumor necrosis factor TNF-α, interleukin IL-1β, IL-6 and LPS were all significantly increased, showing obvious abnormal cytokine levels. The intervention of Pediococcus acidilactici SWU-HX39 effectively reduced the contents of TNF-α, IL-1β, IL-6 and LPS in the sera of hyperuricemia mice, and the reduction rates were 18.78%, 27.06%, 44.48% and 30.47% respectively. Compared with the positive drug group, Pediococcus acidilactici SWU-HX39 showed comparable effects in regulating the levels of these cytokines, and there was no statistically significant difference between the two groups (P > 0.05). This indicates that Pediococcus acidilactici SWU-HX39 has the ability to improve cytokine imbalance in hyperuricemia mice, down-regulate the levels of pro-inflammatory factors in the body, so as to achieve an anti-inflammatory effect.

[0052] (5)Analysis of intestinal flora Mouse feces were taken out of the -80 °C refrigerator and thawed. Total genomic DNA of the microbial community was extracted according to the instructions of the E.Z.N.A.® soil DNA kit (Omega Bio-tek, Norcross, GA, U.S.). The quality of the extracted genomic DNA was detected by 1% agarose gel electrophoresis, and the DNA concentration and purity were measured using NanoDrop2000. Using the extracted DNA as a template, the V3-V4 variable region of the 16S rRNA gene was amplified by PCR using the upstream primer 338F (5’-ACTCCTACGGGAGGCAGCAG-3’) and the downstream primer 806R (5’-GGACTACHVGGGTWTCTAAT-3’) carrying Barcode sequences. The PCR products were recovered using 2% agarose gel, and the recovered products were detected and quantified using a DNA gel recovery and purification kit. Then use NEXTFLEX The Rapid DNA-Seq Kit was used to construct a library for the purified PCR products, and sequencing was performed using the Illumina Nextseq 2000 platform. Fastp and FLASH were used to perform quality control and splicing on the original sequencing sequences. Based on the default parameters, the DADA2 plugin in the Qiime2 pipeline was used to denoise the optimized sequences after quality control and splicing. Based on the Sliva16S rRNA gene database, the Naive bayes classifier in Qiime2 was used to perform a series of statistical or visualization analyses on the ASVs.

[0053] The Bacteroidetes is a key member of the human gut microbiota, which is crucial for maintaining the health of the body and is deeply involved in multiple life processes such as immune enhancement, nutritional metabolism regulation, and antibacterial and antiviral defenses. Studies have shown that tryptophanase has been found in Bacteroidetes, and the relative abundance of the Bacteroidetes is closely related to the level of the tryptophanase gene. Tryptophan metabolites, as ligands, can activate the aryl hydrocarbon receptor signal in various diseases such as inflammation, chronic kidney disease, and cancer. Moreover, tryptophan metabolism derived from gut microbiota has been shown to mediate kidney injury through the activation of the AHR receptor signal. Compared with the blank group, the abundance of the Bacteroidetes in the gut microbiota of the model group mice was significantly increased (p < 0.05), while the abundances of the Firmicutes and Bifidobacterium ( Patescibacteria ) were significantly decreased (p < 0.05 and P < 0.001), respectively. However, the intervention of Pediococcus acidilactici SWU-HX39 effectively reversed these adverse changes.

[0054] At the genus level, in-depth analysis was performed on the genera with significant differences among the groups. The results are as Figure 8 shown. Compared with the blank group, the model group significantly increased the relative abundances of the genus Bacteroides (P < 0.01) and the genus Parabacteroides (P < 0.05), and decreased the relative abundances of the genus Eubacterium / Eubacterium and the genus Ruminococcus. The intervention of Pediococcus acidilactici SWU-HX39 significantly decreased the relative abundances of the genus Bacteroides (P < 0.05) and the genus Parabacteroides, while increasing the relative abundances of the genus Eubacterium / Eubacterium and the genus Ruminococcus. The genus Ruminococcus is the main beneficial species inhabiting the cecum and colon, which can degrade various fibers and polysaccharides and produce short-chain fatty acids (SCFAs). Studies have shown that the abundance of SCFA-producing bacteria (such as Ruminococcus) in hyperuricemic mice is significantly decreased, accompanied by kidney injury. These results indicate the effectiveness and potential of the intervention of Pediococcus acidilactici SWU-HX39 in effectively regulating the gut microbiota level, correcting the imbalance of the gut microbiota, and improving hyperuricemia in mice.

[0055] (6) Determination of short-chain fatty acids (SCFAs): Take 50 mg of cecal contents in a 2 mL centrifuge tube, dissolve it by shaking with saturated NaCl until there are no obvious lumps, and add H 2 SO 4 to acidify. After shaking and mixing evenly, extract short-chain fatty acids with anhydrous ether. After extracting at 4°C for 45 min, centrifuge at 12,000 r / min for 15 min and take the supernatant. Use anhydrous sodium sulfate to remove the residual water in the supernatant, centrifuge again, take the upper ether phase, filter it through a nylon membrane, add it to a brown injection vial, and analyze the content of its short-chain fatty acids (acetic acid, propionic acid, isobutyric acid, butyric acid, isovaleric acid, valeric acid) by GC.

[0056] The beneficial effects of SCFAs on human metabolism have been widely recognized, including improving glucose homeostasis, enhancing glucose tolerance, and increasing insulin sensitivity. This study focused on the effect of Pediococcus acidilactici SWU-HX39 on the concentration of SCFAs in the cecal contents of hyperuricemic mice. After testing, compared with the blank group, the concentrations of acetic acid, propionic acid, butyric acid, and total acid in the model group of mice were significantly decreased (P < 0.001). The contents of acetic acid, propionic acid, isobutyric acid, butyric acid, isovaleric acid, and valeric acid in the HX39 group of mice increased to varying degrees compared with the model group. Among them, the concentrations of acetic acid, propionic acid, and butyric acid increased extremely significantly. The specific experimental results are as Figure 9 shown. Pediococcus acidilactici SWU-HX39 can significantly increase the content of SCFAs in hyperuricemic mice, especially in the increase of propionic acid concentration (P < 0.0001). This indicates that under the intervention of SWU-HX39, the SCFAs level in hyperuricemic mice increased to varying degrees. SWU-HX39 promoted the increase of SCFA level in hyperuricemic mice by increasing the abundance of bacteria producing SCFAs, thereby reducing the uric acid level and improving the abnormal uric acid content. This result is mutually confirmed with the results of intestinal flora structure analysis, further confirming the positive role of Pediococcus acidilactici SWU-HX39 in regulating the intestinal microecology and metabolic health of hyperuricemic mice. Although the SCFAs content in the positive control group increased compared with the model group, it did not reach the level of the HX39 group.

[0057] (7) Analysis of the gene expression of uric acid transporters in mouse kidneys The maintenance of the steady state of in vivo uric acid concentration depends on the dynamic balance between its generation and excretion. In the human physiological mechanism, the kidneys are responsible for about 2 / 3 of the uric acid excretion task. In this common metabolic disorder of hyperuricemia, 90% of the cases are attributed to the defect of the urate excretion mechanism. It is reported that ABCG2, SLC22A6, and SLC17A1 genes play important roles in the excretion process of uric acid, while the GLUT9 gene is closely related to the reabsorption process of uric acid.

[0058] As Figure 10As shown, compared with the control group, the expression levels of the three genes related to uric acid excretion, ABCG2, SLC22A6, and SLC17A1, in the kidneys of mice in the model group were significantly down-regulated (P < 0.0001), and the expression level of the GLUT9 gene was significantly up-regulated (P < 0.05). This indicates that their uric acid excretion ability was severely inhibited and the reabsorption of uric acid increased, further exacerbating the increase in blood uric acid levels. After intervention with Pediococcus acidilactici SWU-HX39, the expression levels of ABCG2, SLC22A6, and SLC17A1 genes were significantly up-regulated, and the expression level of the GLUT9 gene was effectively decreased (P < 0.01). The overall change trend was similar to that of the positive control group, but allopurinol showed a more significant effect in reducing the expression of GLUT9. In summary, Pediococcus acidilactici SWU-HX39 had a positive regulatory effect on the uric acid homeostasis of hyperuricemia model mice at the kidney level by regulating the expression of genes related to uric acid excretion and reabsorption.

Claims

1. A strain of Pediococcus acidilactici SWU-HX39 having the function of reducing uricemia, characterized in that: The classification name of the lactic acid Pediococcus is Pediococcus acidilactici ( Pediococcus acidilactici )SWU-HX39, deposited in the General Microbiology Center of China Microorganism Culture Collection Administration, with the deposit number CGMCC No.33055, the deposit date is December 13, 2024, and the deposit address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

2. Use of Pediococcus acidilactici SWU-HX39 as claimed in claim 1 in the preparation of a medicament for lowering uric acid.

3. Use of Pediococcus acidilactici SWU-HX39 as claimed in claim 1 in preparing food for assisting uric acid regulation.

4. Use of Pediococcus acidilactici SWU-HX39 as claimed in claim 1 in the preparation of anti-inflammatory drugs.

5. Use of Pediococcus acidilactici SWU-HX39 as claimed in claim 1 in the preparation of medicines or foods for regulating intestinal flora.

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