A method for preparing a composition for reducing uric acid and use thereof

Abstract

The application discloses a preparation method of a uric acid reducing composition and application thereof, and belongs to the technical field of traditional Chinese medicines. The preparation method of the uric acid reducing composition comprises the following steps: alcohol extraction is performed on raw material liquid obtained by mixing smilax glabra powder, adenosma powder, dioscorea opposita powder, green tea powder, senna leaf powder and grass cassia powder and water, and then the extract is dissolved in water and subjected to anaerobic fermentation by using microorganisms, so that the fermentation liquid is obtained after concentration, freeze-drying and other steps. The multiple traditional Chinese medicines are used in cooperation and have been verified to have the uric acid reducing effect. Further, the steps of performing alcohol extraction on the traditional Chinese medicines first and then performing fermentation treatment on the obtained alcohol extract make the finally prepared composition have better uric acid reducing effect. Moreover, the preparation method is simple and easy to implement, safe and environment-friendly, has important significance for improving or treating hyperuricemia, and has a wide application prospect.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine technology, specifically relating to a method for preparing a uric acid-lowering composition and its uses. Background Technology

[0002] Uric acid is the end product of purine metabolism in the human body. Under normal circumstances, it dissolves in the blood and is excreted through the kidneys. It plays many beneficial roles in the body, such as acting as an antioxidant to scavenge free radicals, protecting cells from oxidative stress damage (accounting for 60% of the total antioxidant capacity of plasma), maintaining fluid acid-base balance, influencing platelet function, regulating blood pressure, and protecting the nervous system. However, excessive uric acid secretion can lead to a series of health problems, such as triggering gout, causing redness, swelling, heat, and pain in the joints. Long-term high uric acid can also lead to uric acid kidney stones, affecting kidney function, and increasing the risk of cardiovascular disease. It is closely related to hypertension, coronary heart disease, and other conditions, and is also closely associated with important components of metabolic syndrome such as hypercoagulability, hyperglycemia, and hyperlipidemia. Given these adverse consequences, lowering uric acid levels is particularly important. It can not only prevent gout attacks and alleviate patient suffering, but also protect kidney function, reduce the risk of cardiovascular disease, improve patients' quality of life and life expectancy, and have a positive impact on overall metabolic health.

[0003] Treatment of hyperuricemia or gout typically requires long-term use of uric acid-lowering drugs, but the side effects of existing drugs limit their widespread and long-term use. Therefore, developing drugs with fewer side effects that can be used long-term while significantly improving or treating hyperuricemia has become a research hotspot. Traditional Chinese medicine is a natural treasure trove for discovering long-term medications for the prevention and treatment of hyperuricemia and gout, and probiotic fermentation is also an effective strategy to improve efficacy. Summary of the Invention

[0004] This invention addresses the problems existing in the prior art by providing a method for preparing a uric acid-lowering composition with significant effects, low side effects, and simple operation, as well as its applications.

[0005] To achieve the above objectives, the present invention adopts the following technical solution:

[0006] A method for preparing a uric acid-lowering composition includes the following steps:

[0007] S1: Mix Smilax glabra powder, Adenophora stricta powder, Dioscorea opposita powder, green tea powder, Senna leaf powder, and Cassia tora powder with 40-80 v / v% ethanol-water solution and extract with alcohol for 8-12 hours. Filter to obtain alcohol extract, freeze-dry alcohol extract to obtain extract. The alcohol extraction temperature is 25-35℃.

[0008] S2: Prepare a 6-10 wt% mixture of extract using sterile deionized water, sterilize the mixture, inoculate the mixture with mixed bacteria, and then anaerobic ferment for 24-48 h. The fermentation temperature is 35-38℃ and the fermentation pH is 5-7.

[0009] S3: After the fermentation of S2 is completed, the fermentation broth is obtained. The fermentation broth is subjected to ultrasonic lysis, filtered, and then sterilized by irradiation to obtain the fermentation filtrate. The fermentation filtrate is concentrated and freeze-dried to obtain the uric acid-lowering composition.

[0010] Preferably, the mass ratio of Smilax glabra powder, Adenophora stricta powder, Dioscorea opposita powder, green tea powder, Senna leaf powder, and Cassia tora powder in step S1 is 1:1-3:2-4:3-5:4-8:2-4.

[0011] Preferably, in step S1, the total mass ratio of Smilax glabra powder, Adenophora stricta powder, Dioscorea opposita powder, green tea powder, Senna leaf powder, and Cassia tora powder to the ethanol-water solution is 1:8-10.

[0012] Preferably, the mass ratio of Smilax glabra powder, Adenophora stricta powder, Dioscorea opposita powder, green tea powder, Senna leaf powder, and Cassia tora powder in step S1 is 1:2:3:4:6:3.

[0013] Preferably, the mixed bacteria inoculated in step S2 consists of *Lactobacillus plantarum* with accession number CGMCC NO.26508 and *Lactobacillus fermentum* with accession number CGMCC NO.16754; the mass ratio of *Lactobacillus plantarum* to *Lactobacillus fermentum* is 1:1.5-1.7; and the total amount of *Lactobacillus plantarum* and *Lactobacillus fermentum* added is 5-8 wt% of the mixture.

[0014] Preferably, the mass ratio of *Lactobacillus plantarum* and *Lactobacillus fermentum* in step S2 is 1:1.5-1.6.

[0015] Preferably, the mass ratio of *Lactobacillus plantarum* and *Lactobacillus fermentum* in step S2 is 1:1.6.

[0016] A uric acid-lowering tea preparation comprising the uric acid-lowering composition prepared by the aforementioned method.

[0017] This invention comprises six plant-based ingredients: Smilax glabra powder, Adenophora stricta powder, Dioscorea opposita powder, green tea powder, Senna leaf powder, and Cassia tora powder. Among these, Smilax glabra powder, Senna leaf powder, and Cassia tora powder have excellent diuretic effects, increasing urine output, promoting uric acid excretion, and reducing uric acid accumulation in the body. The polyphenols and other components in green tea powder have antioxidant and metabolic-regulating effects, further improving the uric acid metabolic environment. Adenophora stricta powder and Dioscorea opposita powder can regulate human metabolic function, positively impacting the uric acid metabolism process and helping to slow down uric acid production. Dioscorea opposita powder and Senna leaf powder can regulate intestinal flora. Maintaining the balance of the intestinal microecology helps improve the intestine's ability to metabolize uric acid, further promoting uric acid excretion. The six ingredients work together through different pathways, promoting uric acid excretion, regulating uric acid metabolism, and improving the intestinal environment, forming a synergistic effect to jointly reduce the level of uric acid in the body. The effects of each ingredient complement each other. The diuretic effect of Smilax glabra powder and Senna leaf powder combined with the antioxidant effect of green tea powder can enhance the uric acid-lowering effect. The metabolic regulation effect of Adenophora stricta powder and Dioscorea opposita powder combined with the diuretic effect of Cassia tora powder further improve the efficacy of uric acid metabolism.

[0018] The *Lactobacillus plantarum* (CGMCC NO. 26508) and *Lactobacillus fermentum* (CGMCC NO. 16754) used in this invention are both existing anaerobic fermentation strains that can be used in food. During extensive screening of fermentation strains, the inventors unexpectedly discovered that using the aforementioned two strains for mixed fermentation yields more anthraquinones and flavonoids than using a single strain or other combinations of strains under the same fermentation environment and / or strain dosage conditions.

[0019] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0020] (1) The present invention uses a series of steps, namely alcohol extraction followed by fermentation with a specific combination of probiotics, to make the fermented product have more different types of active ingredients, thereby enhancing the uric acid-lowering effect of the fermented product.

[0021] (2) The solvents used in the extraction process and the necessary substances used in the fermentation process of this invention are all non-toxic or low-toxic substances, and there are no residues after volatilization, which ensures safety to the human body.

[0022] (3) The combination of various Chinese medicines in this invention has been verified to have a certain uric acid lowering effect. Furthermore, the inventors have further improved the composition by first extracting the active substances of the Chinese medicines and then fermenting the obtained extracts. The composition has been experimentally verified to have a better uric acid lowering effect, which is of great significance for improving or treating hyperuricemia. Attached Figure Description

[0023] Figure 1Effects of a mixture of Smilax glabra powder, Adenophora stricta powder, Dioscorea opposita powder, green tea powder, Senna leaf powder, and Cassia tora powder on physiological and biochemical indicators in mice with hyperuricemia. (A) Establishment of an animal model of hyperuricemia. (B) Body weight. (C) Serum uric acid level. (D) Serum urea nitrogen level. (E) Hepatic XOD activity. (F) Liver index. (G) Kidney index. All data are expressed as mean ± SEM (n = 6). ac indicates significant differences in mice across different indicators, P < 0.05.

[0024] Figure 2 RT-qPCR detection Figure 1 Expression of uric acid transporter mRNA in the kidney and small intestine of mice in each group. Kidney (A) URAT1, (B) GLUT9, (C) MRP4 and small intestine (D) ABCG2. All data are expressed as mean ± SEM (n=6). ab indicates significant differences in mice under different indicators, P<0.05. Detailed Implementation

[0025] To better understand the present invention, the present invention will be further described below with reference to specific embodiments. The terminology used in the embodiments is for describing specific implementation schemes and does not constitute a limitation on the scope of protection of the present invention.

[0026] Unless otherwise specified, experimental methods in the following examples are generally performed under standard conditions or as recommended by the manufacturer. All materials and reagents used are commercially available unless otherwise specified.

[0027] Unless otherwise specified, all water mentioned in this invention is deionized water.

[0028] According to the inventor's preliminary research, the formula contains Smilax glabra powder, Adenophora stricta powder, Dioscorea opposita powder, green tea powder, Senna leaf powder, and Cassia tora powder. The main ingredient of Changrun Tea is used as a raw material. The concentrated solution obtained by brewing with hot water and then concentrating it has a significant effect on lowering uric acid. See details below. Figure 1-2 ,in Figure 1 The control group was administered drinking water by gavage; the HUA (hyperuricemia model) group was administered drinking water by gavage; the HUA-L group was administered a concentrated extract at a dose of 0.325 mg / g BW / day; the HUA-M group was administered a concentrated extract at a dose of 0.650 mg / g BW / day; and the HUA-H group was administered a concentrated extract at a dose of 1.300 mg / g BW / day. The extract was administered by gavage at a standard volume of 0.1 mL / 10 g BW.

[0029] Experimental procedures as follows Figure 1 As shown in Figure A. Figure 1 B recorded the changes in mouse body weight during feeding, and there were no statistically significant differences between the groups. Figure 1C- Figure 1 The results showed that the uric acid level in the model group mice was significantly higher than that in the normal group (increased by 61.9%), indicating that the hyperuricemia model was successfully established. Low, medium, and high doses of tea (concentrations of 0.0325 g / mL, 0.065 g / mL, and 0.130 g / mL, respectively) all significantly reduced serum uric acid levels in mice. Compared with the normal group, the blood urea nitrogen (BUN) level in the model group mice was significantly increased, indicating that the drug effectively induced hyperuricemia and renal insufficiency in mice. In addition, low, medium, and high doses of tea extract all restored BUN levels in mice to normal levels. The experimental results on uric acid and BUN in mice showed no statistically significant difference in the uric acid-lowering effect of low, medium, and high doses of tea in mice with hyperuricemia. Compared with the normal group, the activity of xanthine oxidase (XOD) in the liver of mice in the model group was significantly enhanced, increasing by 49.5%, indicating that tea treatment can significantly inhibit XOD enzyme activity and restore the liver XOD enzyme activity of mice with hyperuricemia to the level of normal mice. This suggests that Changrun Tea can reduce uric acid production by inhibiting key enzymes in uric acid synthesis in the liver. In addition, the liver and kidney indices of mice in the model group were increased to some extent compared with those in the normal group, but the difference in kidney index was not significant, suggesting that there may be lesions in the organs of mice with drug-induced hyperuricemia.

[0030] Further investigation into the regulatory effects of the aforementioned raw material composition on uric acid transport proteins in the mouse kidney and intestine revealed that ( Figure 2 The mRNA expression levels of URAT1 and GLUT9 genes in the kidneys of the model group were significantly upregulated compared to the normal group. Compared to the model group, the mRNA expression levels of URAT1 and GLUT9 genes in the low-dose tea treatment group were significantly downregulated, indicating that Changrun tea can inhibit renal reabsorption of uric acid by suppressing URAT1 and GLUT9 gene expression. The expression level of MRP4 in the kidneys of the model group was significantly downregulated compared to the normal group, while the MRP4 expression level in the low-dose tea treatment group was significantly higher than that in the model group, with no statistically significant difference compared to the normal group. This suggests that tea can promote uric acid excretion by upregulating MRP4 gene expression in HUA mice. Figure 2 D indicates that, compared with the normal group, the expression level of ABCG2 mRNA in the small intestine of the model group was significantly reduced, while the expression level of ABCG2 mRNA in the low-dose tea treatment group was significantly higher than that in the model group, and there was no statistical difference with the normal group. This suggests that the sample can promote the excretion of uric acid in the intestine of HUA mice by upregulating the expression of ABCG2 mRNA.

[0031] Based on the previous research results, this invention further explores the use of Smilax glabra powder, Adenophora stricta powder, Dioscorea opposita powder, green tea powder, Senna leaf powder, and Cassia tora powder as raw materials. The active substances are first extracted from these materials, and then the obtained extracts are fermented to further enhance their uric acid-lowering effects.

[0032] Example 1: Preparation of a uric acid-lowering composition

[0033] A uric acid-lowering composition, the preparation method of which includes the following steps:

[0034] Composition 1:

[0035] Step 1: Weighing: Weigh out the powders of Smilax glabra, Adenophora stricta, Dioscorea opposita, green tea, Senna leaf, and Cassia tora in a mass ratio of 1:2:3:4:6:3 and mix them together for later use.

[0036] Step 2: Ethanol extraction: The prepared mixed powder was mixed with 65 v / v% ethanol-water solution at a mass ratio of 1:9, and extracted with ethanol at 30°C for 10 h. Then the mixture was filtered and the ethanol extract was freeze-dried to obtain the extract.

[0037] Step 3: Anaerobic Fermentation: The extract was prepared into a mixture containing 8 wt% extract using sterile deionized water. After sterilization, the mixture was inoculated with a 6 wt% mixture of *Lactobacillus plantarum* CGMCC NO. 26508 and *Lactobacillus fermentum* CGMCC NO. 16754 under anaerobic conditions. Fermentation was carried out at 37°C for 36 hours. The fermentation broth was obtained after fermentation. The mass ratio of *Lactobacillus plantarum* CGMCC NO. 26508 to *Lactobacillus fermentum* CGMCC NO. 16754 in the mixture was 1:1.6, and the viable count of both *Lactobacillus plantarum* CGMCC NO. 26508 and *Lactobacillus fermentum* CGMCC NO. 16754 was 1 × 10⁻⁶. 6 CFU / g; fermentation start pH was 6.

[0038] Step 4: Ultrasonic lysis: The fermentation broth is ultrasonically lysed at a frequency of 30 kHz for 45 min, then filtered and sterilized by irradiation to obtain the fermentation filtrate. The fermentation filtrate is concentrated to 30% of its original volume and freeze-dried to obtain the uric acid-lowering composition.

[0039] The preparation steps of compositions 2-3 are the same as those of composition 1, and the specific technical parameters are shown in Table 1 below.

[0040] Table 1 Technical Parameters

[0041]

[0042]

[0043] Composition ①: Unlike composition 1, Lactobacillus plantarum CGMCC No. 31000 is used instead of Lactobacillus plantarum CGMCC NO. 26508. All other conditions and parameters are the same as those of composition 1.

[0044] Composition ②: Unlike composition 1, Lactobacillus fermentum CGMCC No. 25306 is used instead of Lactobacillus fermentum CGMCC No. 16754. All other conditions and parameters are the same as those of composition 1.

[0045] Composition ③: Unlike composition 1, the mass ratio of Lactobacillus plantarum to Lactobacillus fermentum is 1:2, and the other conditions and parameters are the same as those of composition 1.

[0046] Composition ④: Unlike composition 1, the mass ratio of Lactobacillus plantarum to Lactobacillus fermentum is 1:1, and the other conditions and parameters are the same as those of composition 1.

[0047] Composition ⑤: Unlike composition 1, it lacks Smilax glabra powder, Adenophora stricta powder, and Dioscorea opposita powder. The missing mass fractions are made up with an equal amount of sterile water. All other conditions and parameters are the same as those of composition 1.

[0048] Composition ⑥: Unlike Composition 1, the fermentation raw materials used are Smilax glabra powder, Adenophora stricta powder, Dioscorea opposita powder, green tea powder, Senna leaf powder, and Cassia tora powder, with a mass ratio of 1:3:2:3:4:6. The other conditions and parameters are the same as those of Composition 1.

[0049] Composition ⑦: Unlike Composition 1, the 65 v / v ethanol-water solution is replaced with the same mass of water, and the other conditions and parameters are the same as those of Composition 1.

[0050] Example 1: Determination of total anthraquinone and total flavonoid content in compositions 1-3, compositions ①-⑦, and comparative examples.

[0051] Pick The brand Changrun Tea was used as a comparative example; the total anthraquinone content and total flavonoid content in compositions 1-3, compositions ①-⑦, and the comparative example were measured respectively, and the results are shown in Table 2.

[0052] Comparative preparation method: Take one bag of commercially available Changrun Tea and brew it with 400mL of 100℃ drinking water for 30min to obtain tea soup. Freeze-dry the tea soup to obtain comparative freeze-dried powder. Use the comparative freeze-dried powder as a sample for determination.

[0053] The test method for total anthraquinone content refers to "Determination of Total Anthraquinone in Health Foods" (Ye Bisha et al., Chinese Journal of Health Laboratory Technology, DOI: 10.3969 / j.issn.1004-8685.2007.05.027).

[0054] The total flavonoid content was determined according to the method described in DB 34 / T 2743-2016 Determination of Total Flavonoid Content in Sophora japonica buds and their products by spectrophotometry.

[0055] Table 2 Results of content determination

[0056]

[0057] According to Table 2:

[0058] 1) Comparing the experimental results of compositions 1-3 with those of compositions ①-②, it can be seen that specific strains of Lactobacillus plantarum and Lactobacillus fermentum can synergistically promote fermentation, resulting in higher contents of total anthraquinones and total flavonoids in the prepared compositions, producing unexpected technical effects.

[0059] 2) Comparing the experimental results of compositions 1-3 with those of compositions ③-④, it can be seen that the mass ratio of Lactobacillus plantarum to Lactobacillus fermentum in the mixed bacteria also significantly affects the content of total anthraquinones and total flavonoids in the composition. Only compositions prepared with a mass ratio of Lactobacillus plantarum to Lactobacillus fermentum of 1:1.5-1.7 have relatively high contents of total anthraquinones and total flavonoids.

[0060] 3) Comparing the experimental results of compositions 1-3 with those of compositions ⑤-⑦, it can be seen that the components and ratios of the traditional Chinese medicine, as well as the extraction method, all have a significant impact on the content of total anthraquinones and total flavonoids in the compositions. The above experimental results indicate that after alcohol extraction and fermentation with a compound microbial strain composed of the specific strains of this invention, the content of total flavonoids and total anthraquinones in the final prepared compositions is increased.

[0061] Example 2: Animal Toxicological Safety Test

[0062] 1) Materials and Methods

[0063] 1. Test substance: Samples of compositions 1 to 3 in Table 1; prepared as a 1 g / mL mixture using drinking water.

[0064] 2. Acute toxicity test: Sixty Kunming mice weighing 20±2g were selected, half male and half female; environment: temperature 23±1℃, humidity 54±2%.

[0065] The concentrated sample solutions of compositions 1-3 were administered orally to mice once by gavage at a volume of 0.2 mL / 10 g b.w., equivalent to a dose of 20.0 g / kg bw. Mice were fasted for 16 hours before gavage and observed for two weeks afterward. The toxic symptoms and mortality of the mice were recorded, as shown in Table 3.

[0066] Table 3 Acute toxicity test in mice

[0067] Group gender way Dosage (g / kgb.w.) Number of deaths (animals) MTD (g / kgb.w.) Composition 1 male Oral 20.0 0 ＞20.0 Composition 1 female Oral 20.0 0 ＞20.0 Composition 2 male Oral 20.0 0 ＞20.0 Composition 2 female Oral 20.0 0 ＞20.0 Composition 3 male Oral 20.0 0 ＞20.0 Composition 3 female Oral 20.0 0 ＞20.0

[0068] According to the results in Table 3, no obvious abnormalities were observed in the mice during the experiment, and none of them died, indicating that compositions 1-3 in this invention have no acute toxicity.

[0069] Example 3 Animal Efficacy Test

[0070] Preparation of Test Sample Solution

[0071] Test groups: Composition 1-3 and Composition ①-⑦, with deionized water as the solvent;

[0072] Normal control group: Deionized water.

[0073] Positive control group: Allopurinol, batch number: MKCV7617, Sigma, with DMSO as the solvent.

[0074] Experimental animals:

[0075] Zebrafish were all raised in fish culture water at 28°C (water quality: 200 mg of instant sea salt was added to every 1 L of reverse osmosis water, with a conductivity of 500 μS / cm; pH of 7.0; hardness of 80 mg / L CaCO3). The zebrafish used in this experiment were from Hangzhou Huante Biotechnology Co., Ltd. The experimental animal production license: SCXK(Zhe)2022-0003. The experimental environment used in this experiment was provided by Hangzhou Huante Biotechnology Co., Ltd. The experimental animal use license: SCXK(Zhe)2022-0004.

[0076] Instruments, consumables and reagents:

[0077] Dissecting microscope (SZX7, OLYMPUS, Japan); Precision electronic balance (CP214, OHAUS, USA); CCD camera (VertA1, Shanghai Tusen Vision Technology Co., Ltd., China); 96-well microplate (Costar, China); Multifunctional microplate reader (SPARK, TECAN, Austria); 6-well plate (Zhejiang Beilanbo Biotechnology Co., Ltd., China).

[0078] Potassium oxonate (batch number H2405012, Shanghai Aladdin Biochemical Technology Co., Ltd., China); Sodium xanthine (batch number X3627-1G, Sigma, USA); Amplex RedUric (uric acid kit) (batch number 2810925, Thermo Fisher Scientific (China) Co., Ltd., USA); Dimethyl sulfoxide (DMSO, batch number 20220614, Sinopharm Chemical Reagent Co., Ltd., China).

[0079] Detection method:

[0080] Wild-type AB strain zebrafish, 5 days post-fertilization (5 dpf), were randomly selected and placed in 6-well plates, with 30 zebrafish treated in each well (experimental group). Water-soluble administration of the combined sample (concentrations shown in Tables 4 and 5) was performed. A normal control group and a model control group were also included, with a well volume of 3 mL. Except for the normal control group, all other experimental groups were treated with water-soluble potassium oxonate and sodium xanthine to establish a zebrafish hyperuricemia model with a potassium oxonate concentration of 1.95 mg / mL and a sodium xanthine concentration of 0.0875 mg / mL. After treatment at 28℃ for 1 day, the MTC of the samples in the model zebrafish was measured.

[0081] Evaluation of uric acid-lowering efficacy:

[0082] Wild-type AB strain zebrafish (5 dpf) were randomly selected and placed in 6-well plates, with 30 zebrafish treated in each well (experimental group). The aqueous solution of the treatment composition (concentrations shown in Tables 4 and 5) was administered. Allopurinol 136 μg / mL was used as a positive control. A normal control group and a model control group were also included, with a volume of 3 mL per well. Except for the normal control group, all other experimental groups were treated with potassium oxonate and sodium xanthine in aqueous solution to establish a zebrafish hyperuricemia model with a potassium oxonate concentration of 1.95 mg / mL and a sodium xanthine concentration of 0.0875 mg / mL. After treatment at 28℃ for 1 day, data were collected using the Amplex™ Red Uric uric acid kit and a multi-functional microplate reader. The uric acid fluorescence value of the zebrafish was analyzed, and the statistical analysis results were used to evaluate the uric acid-lowering efficacy of the samples (see Table 5). Statistical results are expressed as mean ± SE. Statistical analysis was performed using SPSS 26.0 software; p < 0.05 indicated statistical significance.

[0083] result:

[0084] MTC (Minimum Toxic Concentration): Under the experimental conditions, the uric acid-lowering efficacy (MTC) of compositions 1, 2, and 3 was >2000 μg / mL. See Table 4 for details.

[0085] Table 4 MTC Detection

[0086]

[0087]

[0088] Evaluation of uric acid-lowering efficacy: Under the experimental conditions, compositions 1, 2, and 3 all exhibited uric acid-lowering efficacy. See Table 5 for details.

[0089] Table 5 Evaluation of uric acid-lowering efficacy

[0090]

[0091] Note: Compared with the model control group, *p<0.05, **p<0.01, ***p<0.001

[0092] As shown in Table 5, compositions 1-3 all exhibited significant uric acid-lowering effects compared to the model control group. Within the dosage range of 500-2000 μg / mL, the uric acid-lowering effect was positively correlated with the dosage. Compared to the uric acid-lowering rate of 1000-2000 μg / mL, the uric acid-lowering rate of 500-1000 μg / mL was higher. Therefore, a dosage of 1000 μg / mL was selected as the optimal concentration.

[0093] Comparing the results of compositions ①-⑦ with those of composition 1, it can be seen that composition 1 has the best effect at a dose of 1000 μg / mL. Compared with the results of compositions 1 and ①-②, it can be seen that there is a significant synergistic effect between *Lactobacillus plantarum* CGMCC NO.26508 and *Lactobacillus fermentum* CGMCC NO.16754 selected in this invention. Furthermore, comparing the results of composition 1 with those of ③-④, it can be seen that the mass ratio of *Lactobacillus plantarum* to *Lactobacillus fermentum* selected in this invention also significantly affects the uric acid-lowering effect of the composition.

[0094] Comparing the results of composition 1 with those of compositions ⑤-⑥, it can be seen that the fermentation raw materials in this invention also have a certain synergistic effect. The composition obtained from the plant raw materials and their mass fractions selected in this invention has the best uric acid lowering effect. Furthermore, comparing the results of composition 1 with those of composition ⑦, it can be seen that the alcohol extraction method selected in this invention also helps to enhance the uric acid lowering effect of the composition.

[0095] The experimental results also show that the uric acid-lowering effect of each composition is positively correlated with the total flavonoid and total anthraquinone content, indicating that the above components play a crucial role in the composition's uric acid-lowering effect.

[0096] The above detailed description is a specific description of one of the feasible embodiments of the present invention. This embodiment is not intended to limit the patent scope of the present invention. All equivalent implementations or modifications that do not depart from the present invention should be included within the scope of the technical solution of the present invention.

Claims

1. A method of preparing a hypouricemic composition, characterized by, The method comprises the following steps: S1: mixing the smilax glabra, adiantium capillus-veneris, dioscorea opposita, green tea powder, senna leaf powder, cassia tora powder with 40-80v / v% ethanol-water solution, alcohol extraction for 8-12h, filtering to obtain alcohol extract, freeze-drying the alcohol extract to obtain the extract, wherein the alcohol extraction temperature is 25-35℃; S2: using sterile deionized water to prepare the extract into a 6-10wt% mixture, sterilizing the mixture, inoculating the mixture with mixed bacteria, and anaerobic fermentation for 24-48h, wherein the fermentation temperature is 35-38℃ and the fermentation pH is 5-7; S3: obtaining the fermentation broth after the fermentation of S3 is completed, ultrasonic lysis of the fermentation broth, filtering, sterilizing the collected filtrate to obtain the fermentation filtrate, and obtaining the uric acid-lowering composition by concentrating and freeze-drying the fermentation filtrate; In step S2, the mixed bacteria inoculated are composed of Lactobacillus plantarum with preservation number CGMCC NO.26508 and Lactobacillus fermentum with preservation number CGMCC NO.16754; the mass ratio of the Lactobacillus plantarum and the Lactobacillus fermentum is 1:1.5-1.7; and the total amount of the Lactobacillus plantarum and the Lactobacillus fermentum added is 5-8wt% of the mixture. In step S1, the mass ratio of the smilax glabra, adiantium capillus-veneris, dioscorea opposita, green tea powder, senna leaf powder, and cassia tora powder is 1:1-3:2-4:3-5:4-8:2-4.

2. The method for preparing the uric acid-lowering composition according to claim 1, characterized in that, The mass ratio of the smilax glabra, adiantium capillus-veneris, dioscorea opposita, green tea powder, senna leaf powder, and cassia tora powder is 1:2:3:4:6:

3.

3. The method of claim 1, wherein the composition is prepared by mixing the uric acid-lowering agent and the pharmaceutically acceptable carrier. The mass ratio of the Lactobacillus plantarum and the Lactobacillus fermentum is 1:1.5-1.

6.

4. The method of claim 1, wherein the composition is prepared by mixing the uric acid-lowering agent and the pharmaceutically acceptable carrier. The mass ratio of the Lactobacillus plantarum and the Lactobacillus fermentum is 1:1.

6.

5. The method for preparing the uric acid-lowering composition according to claim 1, characterized in that, In step S1, the mass ratio of the total amount of the smilax glabra, adiantium capillus-veneris, dioscorea opposita, green tea powder, senna leaf powder, and cassia tora powder to the ethanol-water solution is 1:8-10.

6. The method of claim 1, wherein the composition is prepared by the steps of: The sterilization in S2 and S3 is radiation sterilization.

7. The uric acid-lowering composition prepared by the method of any one of claims 1-6.

8. The use of the uric acid-lowering composition of claim 7 in the preparation of a uric acid-lowering drug.

9. Use according to claim 8, characterized in that, The dosage form of the uric acid-lowering drug is any one of a tea, a paste, a tablet, a capsule, a pill, a granule, or an oral liquid.

10. Use according to claim 9, characterized in that, The dosage form of the uric acid-lowering drug is a tea.

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