Preparation method and application of uric acid reducing composition

Abstract

The invention discloses a preparation method and application of a uric acid reducing composition, and belongs to the technical field of traditional Chinese medicines. The preparation method of the composition for reducing uric acid comprises the following steps: mixing rhizoma smilacis glabrae powder, adenophora stricta powder, common yam rhizome powder, green tea powder, folium sennae powder, cassia occidentalis powder and water to obtain a raw material solution, carrying out alcohol extraction on the raw material solution, dissolving the extract in water, inoculating microorganisms, carrying out anaerobic fermentation, and carrying out concentration, freeze-drying and other steps on the fermentation liquor to obtain the composition for reducing uric acid. Various traditional Chinese medicines are matched for use and verified to have the uric acid reducing effect, and the finally prepared composition has the better uric acid reducing effect through the steps of firstly carrying out alcohol extraction on the traditional Chinese medicines and then carrying out fermentation treatment on the obtained alcohol extract. The preparation method is simple and easy to implement, safe and environmentally friendly, has important significance on improvement or treatment of hyperuricemia, and has wide application prospects.

Description

Technical Field

[0001] The present invention belongs to the technical field of traditional Chinese medicine, and particularly relates to a preparation method and use of a uric acid-lowering composition. Background Art

[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 from the body through the kidneys. It plays a variety of beneficial roles in the human 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, and also participating in maintaining the acid-base balance of body fluids, affecting platelet function, regulating blood pressure, and protecting the nervous system, etc. However, when uric acid secretion is excessive, it will lead to a series of health problems, such as triggering gout, causing joint swelling, redness, heat and pain. Long-term hyperuricemia will also form uric acid kidney stones, affecting kidney function, and at the same time increasing the risk of cardiovascular diseases, being closely related to hypertension, coronary heart disease, etc., and also being closely related to important components of metabolic syndrome such as hypercoagulable state of blood, hyperglycemia, hyperlipidemia, etc. In view of these adverse consequences, reducing uric acid levels is particularly important. It can not only prevent gout attacks, relieve the pain of patients, but also protect kidney function, reduce the risk of cardiovascular diseases, improve the quality of life and life expectancy of patients, and has a positive impact on overall metabolic health.

[0003] The treatment of hyperuricemia or gout usually requires long-term use of uric acid-lowering drugs, and the existing drugs have side effects that limit their extensive and long-term use. Therefore, the development of drugs with low toxicity and side effects that can be used for a long time and at the same time have a significant improvement or treatment effect on hyperuricemia has become a research and development hotspot. Traditional Chinese medicine is a natural treasure house for discovering drugs for the prevention and treatment of hyperuricemia and gout that can be used for a long time, and probiotic fermentation is also one of the effective strategies to improve drug efficacy. Summary of the Invention

[0004] In view of the problems existing in the prior art, the present invention provides a preparation method and use of a uric acid-lowering composition with relatively significant effects, low side effects and simple and easy operation.

[0005] In order to achieve the above object, the present invention adopts the following technical solutions:

[0006] A preparation method of a uric acid-lowering composition, comprising the following steps:

[0007] S1: Mix rhizoma smilacis glabrae powder, adenophora stricta powder, rhizoma dioscoreae powder, green tea powder, folium sennae powder, semen cassiae powder with an ethanol-aqueous solution of 40 - 80 v / v%, perform alcohol extraction for 8 - 12 h, filter to obtain an alcohol extract, and lyophilize the alcohol extract to obtain an extract, wherein the alcohol extraction temperature is 25 - 35°C;

[0008] S2: Prepare the extract into a 6 - 10 wt% mixture with sterile deionized water, sterilize the mixture, inoculate the mixture with mixed bacteria and then perform anaerobic fermentation for 24 - 48 h. Among them, the fermentation temperature is 35 - 38 °C, and the fermentation pH is 5 - 7;

[0009] S3: After the fermentation in S2 ends, obtain the fermentation broth, perform ultrasonic lysis on the fermentation broth, filter it, and then obtain the fermentation filtrate through irradiation sterilization. Concentrate and lyophilize the fermentation filtrate to obtain the uric acid - lowering composition;

[0010] Preferably, in step S1, the mass ratio of the glabrous greenbrier rhizome powder, adenophora root powder, common yam rhizome powder, green tea powder, senna leaf powder, and cassia seed powder is 1:1 - 3:2 - 4:3 - 5:4 - 8:2 - 4.

[0011] Preferably, in step S1, the mass ratio of the total mass of the glabrous greenbrier rhizome powder, adenophora root powder, common yam rhizome powder, green tea powder, senna leaf powder, and cassia seed powder to the ethanol - aqueous solution is 1:8 - 10.

[0012] Preferably, in step S1, the mass ratio of the glabrous greenbrier rhizome powder, adenophora root powder, common yam rhizome powder, green tea powder, senna leaf powder, and cassia seed powder is 1:2:3:4:6:3.

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

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

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

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

[0017] The present invention includes six plant raw materials, namely, smilax glabra powder, adenophora stricta powder, rhizoma dioscoreae powder, green tea powder, folium sennae powder, and semen cassiae powder. Among them, smilax glabra powder, folium sennae powder, and semen cassiae powder have good diuretic effects, can increase urine volume, promote the excretion of uric acid, and reduce the accumulation of uric acid in the body. Components such as tea polyphenols in green tea powder have antioxidant and metabolic regulatory effects, which can further improve the uric acid metabolism environment. Adenophora stricta powder and rhizoma dioscoreae powder can regulate the body's metabolic function, have a positive impact on the metabolic process of uric acid, and help slow down the generation rate of uric acid. Rhizoma dioscoreae powder and folium sennae powder can regulate the intestinal flora, maintain the balance of the intestinal microecosystem, help improve the intestinal metabolism ability of uric acid, and further promote the excretion of uric acid. The six raw materials work together through different pathways, starting from multiple aspects such as promoting uric acid excretion, regulating uric acid metabolism, and improving the intestinal environment, to form a synergistic effect and jointly reduce the uric acid level in the body. The effects of each raw material complement each other. The combination of the diuretic effects of smilax glabra powder and folium sennae powder and the antioxidant effect of green tea powder can enhance the effect of reducing uric acid. The combination of the metabolic regulatory effects of adenophora stricta powder and rhizoma dioscoreae powder and the diuretic effect of semen cassiae powder further improves the uric acid metabolism efficacy.

[0018] The Lactobacillus plantarum (preservation number: CGMCC NO.26508) and Lactobacillus fermentum (CGMCC NO.16754) used in the present invention are both anaerobic fermentation strains that are already available in the prior art and can be used in food. The inventor of the present invention accidentally found through a large number of fermentation bacteria screenings in the early stage that when the above two strains are used for mixed fermentation, more anthraquinones and flavonoids can be obtained than single strains or other strain combinations under the same fermentation environment and / or the same amount of bacterial strains used.

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

[0020] (1) Through the continuous steps of first alcohol extraction and then fermentation with a specific probiotic combination, the present invention makes the fermented product have as many different types of active and effective components as possible, 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 the present invention are all non-toxic or low-toxic substances, and there is no residue after volatilization, ensuring the safety of the human body.

[0022] (3) The combined use of various traditional Chinese medicines in the present invention has been verified to have a certain uric acid-lowering effect. The inventor further makes the finally prepared composition have a better uric acid-lowering effect through the steps of first extracting the active substances of the traditional Chinese medicine and then fermenting the obtained extract, which is of great significance for improving or treating hyperuricemia. Description of the Drawings

[0023] Figure 1: Effects of the mixed brewing solution of Smilax glabra Roxb. powder, Adenophora stricta Miq. powder, Dioscorea opposita Thunb. powder, green tea powder, Senna leaf powder, and Cassia obtusifolia L. powder on physiological and biochemical indexes of hyperuricemia mice. (A) Establishment of hyperuricemia animal model. (B) Body weight. (C) Serum uric acid level. (D) Serum urea nitrogen level. (E) Liver XOD activity. (F) Liver index. (G) Kidney index. All data are expressed as mean ± SEM (n = 6). a-c indicate significant differences in mice under different indexes, P < 0.05.

[0024] Figure 2 : RT-qPCR detection Figure 1 Expression of renal and small intestinal uric acid transporter mRNAs in mice of each group. Kidney (A) URAT1, (B) GLUT9, (C) MRP4, and small intestine (D) ABCG2. All data are expressed as mean ± SEM (n = 6). a-b indicate significant differences in mice under different indexes, P < 0.05. Specific implementation manners

[0025] To better understand the present invention, the present invention will be further described below in conjunction with specific embodiments. The terms used in the embodiments are for describing specific specific implementation manners and do not constitute a limitation on the protection scope of the present invention.

[0026] For the experimental methods without specific conditions noted in the following embodiments, they are generally carried out according to conventional conditions or according to the conditions recommended by the manufacturer. The materials, reagents, etc. used, unless otherwise specified, can be obtained from commercial channels.

[0027] The water described in the present invention, unless otherwise specified, is deionized water.

[0028] According to the previous research of the inventors, taking Smilax glabra Roxb. powder, Adenophora stricta Miq. powder, Dioscorea opposita Thunb. powder, green tea powder, Senna leaf powder, and Cassia obtusifolia L. powder ( the main components of Changrun Tea) as raw materials, the concentrated solution obtained by hot water brewing and concentration has a significant effect on reducing uric acid. For details, see Figure 1-2 , where Figure 1 The Control group was intragastrically administered with drinking water, the HUA (hyperuricemia model) group was intragastrically administered with drinking water, the HUA-L group was intragastrically administered with the concentrated solution at 0.325 mg / g.BW / day, the HUA-M group was intragastrically administered with the concentrated solution at 0.650 mg / g.BW / day, and the HUA-H group was intragastrically administered with the concentrated solution at 1.300 mg / g BW / day. The extract was intragastrically administered at a standard volume of 0.1 mL / 10 g BW.

[0029] The experimental procedure is as shown in Figure 1 A. Figure 1 B records the changes in the body weight of mice during the feeding process, and there are no significant statistical differences among the groups. Figure 1C- Figure 1 The results showed that the uric acid level in the model group of mice was significantly higher than that in the normal group (increased by 61.9%), indicating that the hyperuricemia model was successfully established. Low-dose, medium-dose, and high-dose tea water (concentrations were 0.0325 g / mL, 0.065 g / mL, and 0.130 g / mL respectively) all significantly reduced the serum uric acid level in mice. Compared with the normal group, the blood urea nitrogen (BUN) level in the model group of mice was significantly increased, indicating that the drug effectively induced hyperuricemia and renal insufficiency in mice. In addition, the low-dose, medium-dose, and high-dose groups of tea extract all restored the BUN level in mice to the normal level. From the experimental results of uric acid and blood urea nitrogen in mice, there was no significant statistical difference in the uric acid-lowering effect of low, medium, and high-dose tea water on hyperuricemic mice. Compared with the normal group, the activity of xanthine oxidase (XOD) in the liver of the model group of mice was significantly enhanced, increasing by 49.5%, indicating that tea water treatment could significantly inhibit the activity of XOD enzyme and restore the liver XOD enzyme activity of hyperuricemic mice to the level of normal mice, suggesting that Changrun tea water could reduce the production of uric acid by inhibiting the key enzyme for uric acid synthesis in the liver. In addition, the liver index and kidney index of the model group of mice were increased to a certain extent compared with the normal group, but the difference in the kidney index was not significant, indicating that there may be lesions in the organs of drug-induced hyperuricemic mice.

[0030] Further exploration of the regulatory effect of the aforementioned raw material composition on uric acid transporters in the kidneys and intestines of mice showed that ( Figure 2 ) the mRNA expression levels of URAT1 and GLUT9 genes in the kidneys of the model group were significantly up-regulated compared with the normal group. Compared with the model group, the mRNA expression levels of URAT1 and GLUT9 genes in the low-dose tea treatment group of mice were significantly down-regulated, that is, Changrun tea water could inhibit the reabsorption of uric acid by the kidneys by inhibiting the expression of URAT1 and GLUT9 genes. The expression level of MRP4 in the kidneys of the model group was significantly down-regulated compared with the normal group, and the expression level of MRP4 in the low-dose tea treatment group was significantly higher than that in the model group and had no statistical difference from the normal group, indicating that tea water could promote the excretion of uric acid by up-regulating the expression of MRP4 gene in HUA mice. Figure 2 D showed that compared with the normal group, the mRNA expression level of ABCG2 in the small intestine of the model group was significantly decreased, and the mRNA expression level of ABCG2 in the low-dose tea treatment group was significantly higher than that in the model group and had no statistical difference from the normal group, indicating that the sample could promote the excretion of uric acid in the intestines of mice by up-regulating the mRNA expression of ABCG2 in HUA mice.

[0031] Based on the above preliminary research results, in the present invention, further exploration was carried out using rhizoma smilacis glabrae powder, adenophora stricta powder, rhizoma dioscoreae powder, green tea powder, folium sennae powder, and semen cassiae powder as raw materials. First, the active substances were extracted, and then the obtained extract was fermented, in order to further enhance its uric acid-lowering effect.

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

[0033] A uric acid-lowering composition, and its preparation method comprises the following steps:

[0034] Composition 1:

[0035] Step 1: Weighing: Weigh and mix the raw materials of smilax glabra powder, adenophora stricta powder, rhizoma dioscoreae powder, green tea powder, senna leaf powder, and cassia seed powder in a mass ratio of 1:2:3:4:6:3 for standby.

[0036] Step 2: Ethanol extraction: Mix the prepared mixed powder with 65 v / v% ethanol-aqueous solution in a mass ratio of 1:9, perform ethanol extraction at 30 °C for 10 h, then filter, and lyophilize the ethanol extract to obtain an extract.

[0037] Step 3: Anaerobic fermentation: Use sterile deionized water to prepare the extract into a mixed solution containing 8 wt% extract. After sterilizing the mixed solution, inoculate a mixed bacterium composed of Lactobacillus plantarum CGMCC NO.26508 and Lactobacillus fermentum CGMCC NO.16754 accounting for 6 wt% of the mixed solution in an anaerobic environment, and ferment at 37 °C for 36 h. After fermentation, a fermentation broth is obtained. Among them, in the mixed bacterium, the mass ratio of Lactobacillus plantarum CGMCC NO.26508 to Lactobacillus fermentum CGMCC NO.16754 is 1:1.6, and the viable cell counts of Lactobacillus plantarum CGMCC NO.26508 and Lactobacillus fermentum CGMCC NO.16754 used are both 1×10 6 CFU / g; the initial pH of fermentation is 6.

[0038] Step 4: Ultrasonic lysis: Perform ultrasonic lysis on the fermentation broth at an ultrasonic frequency of 30 kHz for 45 min, then filter and irradiate for sterilization to obtain a fermentation filtrate. Concentrate the fermentation filtrate to 30% of the original volume and lyophilize 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 different technical parameters are shown in Table 1 below.

[0040] Table 1 Technical parameters

[0041]

[0042]

[0043] Composition ①: Different from Composition 1, Lactobacillus plantarum CGMCC No.31000 is used to replace Lactobacillus plantarum CGMCC NO.26508, and the other conditions and parameters are the same as those of Composition 1.

[0044] Composition ②: Different from Composition 1, Lactobacillus fermentum CGMCC No. 25306 is used to replace Lactobacillus fermentum CGMCC NO. 16754, and the remaining conditions and parameters are the same as those of Composition 1.

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

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

[0047] Composition ⑤: Different from Composition 1, the rhizome of glabrous greenbrier powder, the root of straight ladybell powder, and the rhizome of Chinese yam powder are missing, and the missing parts by mass are supplemented with an equal amount of sterile water, and the remaining conditions and parameters are the same as those of Composition 1.

[0048] Composition ⑥: Different from Composition 1, the fermentation raw materials used are the rhizome of glabrous greenbrier powder, the root of straight ladybell powder, the rhizome of Chinese yam powder, green tea powder, folium sennae powder, and semen cassiae powder, and their mass ratio is 1:3:2:3:4:6, and the remaining conditions and parameters are the same as those of Composition 1.

[0049] Composition ⑦: Different from Composition 1, water of the same mass is used to replace 65 v / v% ethanol-aqueous solution, and the remaining conditions and parameters are the same as those of Composition 1.

[0050] Example 1: Determine the contents of total anthraquinones and total flavonoids in Compositions 1-3, Compositions ①-⑦, and the control

[0051] Take Brand Changrun Tea as the control; measure the contents of total anthraquinones and total flavonoids in Compositions 1-3, Compositions ①-⑦, and the control respectively, and the results are shown in Table 2.

[0052] Preparation method of the control: Use boiling water to brew. Take a bag of commercially available Changrun Tea, brew it with 400 mL of 100 °C drinking water for 30 min to obtain the tea soup, freeze-dry the tea soup to obtain the freeze-dried powder of the control, and use the freeze-dried powder of the control as the sample for determination.

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

[0054] The determination method of the content of total flavonoids is carried out according to the method recorded in "DB 34 / T 2743-2016 Determination of Total Flavonoids in Sophora japonica and Its Products - Spectrophotometry".

[0055] Table 2 Determination results of the content

[0056]

[0057] According to Table 2:

[0058] 1) By comparing the experimental results of Compositions 1-3 and Compositions ①-②, it can be seen that Lactobacillus plantarum and Lactobacillus fermentum of specific strains can produce synergistic promotion during fermentation, and the contents of total anthraquinones and total flavonoids in the prepared compositions are higher, producing unexpected technical effects.

[0059] 2) By comparing the experimental results of Compositions 1-3 and 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 contents of total anthraquinones and total flavonoids in the composition, and only when the mass ratio of Lactobacillus plantarum to Lactobacillus fermentum is in the range of 1:1.5 - 1.7, the contents of total anthraquinones and total flavonoids in the prepared composition are relatively high.

[0060] 3) By comparing the experimental results of Compositions 1-3 and Compositions ⑤-⑦, it can be seen that the traditional Chinese medicine components, their ratios, and the extraction methods all have obvious effects on the contents of total anthraquinones and total flavonoids in the composition. The above experimental results show that: after alcohol extraction and fermentation with the composite bacteria composed of the specific strains of the present invention, the contents of total flavonoids and total anthraquinones in the finally prepared composition have both increased.

[0061] Example 2 Animal Toxicological Safety Test

[0062] 1) Materials and Methods

[0063] 1. Test substances: Samples of Compositions 1 to 3 in Table 1; Prepared into a 1 g / mL mixed solution with drinking water.

[0064] 2. Acute toxicity experiment: Select 60 Kunming mice with a body weight of 20 ± 2 g, half male and half female; Environment: Temperature 23 ± 1°C, humidity 54 ± 2%.

[0065] Take the concentrated liquid of the samples of Compositions 1-3 and orally administer it to the mice by gavage once. The gavage volume is 0.2 mL / 10 g b.w., and the equivalent dose is 20.0 g / kg b.w. Fast the mice for 16 hours before gavage, and continuously observe for two weeks after gavage, record the poisoning manifestations and death conditions of the mice, as shown in Table 3.

[0066] Table 3 Acute Toxicity Test of Mice

[0067] Group Gender Route Dose (g / kg b.w.) Number of deaths (animals) MTD (g / kg b.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, it can be seen that: during the test, no obvious abnormalities were found in the mice, and no deaths occurred, indicating that Compositions 1-3 in the present 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 at 5 days post-fertilization (5 dpf) were randomly selected and placed in 6-well plates, with 30 zebrafish treated in each well (experimental group). The composition sample was administered in water (concentrations are shown in Tables 4 and 5). A normal control group and a model control group were simultaneously set up, and the volume of each well was 3 mL. Except for the normal control group, potassium oxonate and sodium xanthine were administered in water to the other experimental groups so that the concentration of potassium oxonate in the water was 1.95 mg / mL and the concentration of sodium xanthine was 0.0875 mg / mL to establish a zebrafish hyperuricemia model. After treatment at 28 °C for 1 day, the MTC of the sample on the model zebrafish was measured.

[0081] Evaluation of uric acid-lowering efficacy:

[0082] Wild-type AB strain zebrafish at 5 dpf were randomly selected and placed in 6-well plates, with 30 zebrafish treated in each well (experimental group). The composition sample was administered in water (concentrations are shown in Tables 4 and 5). The positive control was allopurinol at 136 μg / mL. A normal control group and a model control group were simultaneously set up, and the volume of each well was 3 mL. Except for the normal control group, potassium oxonate and sodium xanthine were administered in water to the other experimental groups so that the concentration of potassium oxonate in the water was 1.95 mg / mL and the concentration of sodium xanthine was 0.0875 mg / mL to establish a zebrafish hyperuricemia model. After treatment at 28 °C for 1 day, data were collected using the AmplexTM Red Uric Uric Acid Kit with a multifunctional microplate reader software, and the fluorescence value of zebrafish uric acid was analyzed. The uric acid-lowering efficacy of the sample was evaluated based on the statistical analysis results of this index (see Table 5). The statistical processing results were expressed as mean ± SE. Statistical analysis was performed using SPSS 26.0 software, and p < 0.05 indicated that the difference was statistically significant.

[0083] Results:

[0084] MTC (minimum toxic concentration): Under the experimental conditions of this study, the MTC of the uric acid-lowering efficacy of Composition 1, Composition 2, and Composition 3 > 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 of this study, Composition 1, Composition 2, and Composition 3 have 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] According to the results in Table 5, compared with the model control group, Compositions 1-3 all have a significant uric acid-lowering effect. Among them, in the dose range of 500-2000 μg / mL, the uric acid-lowering effect is positively correlated with the dose. Compared with the uric acid decline rate of 1000-2000 μg / mL, the uric acid decline rate of 500-1000 μg / mL is higher. Therefore, a dose of 1000 μg / mL is selected as the optimal concentration.

[0093] By comparing the results of Compositions ①-⑦ and Composition 1, it can be seen that at a dose of 1000 μg / mL, Composition 1 has the best effect. By comparing the results of Composition 1 and Compositions 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 the present invention; and by comparing the results of Composition 1 and Compositions ③-④, it can be seen that the mass ratio of Lactobacillus plantarum to Lactobacillus fermentum selected in the present invention also significantly affects the uric acid-lowering effect of the composition.

[0094] By comparing the results of Composition 1 and Compositions ⑤-⑥, it can be seen that the fermentation raw materials in the present invention also have a certain synergistic effect, and the composition obtained within the range of the plant raw materials and their mass parts ratio selected in the present invention has the best uric acid-lowering effect; and by comparing the results of Composition 1 and Composition ⑦, it can be seen that the alcohol extraction method selected in the present invention also helps to enhance the uric acid-lowering effect of the composition.

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

[0096] The above detailed description is a specific description of one feasible embodiment of the present invention. This embodiment is not intended to limit the patent scope of the present invention. Any equivalent implementation or modification without departing from the present invention shall be included within the scope of the technical solution of the present invention.

Claims

1. A method for preparing a uric acid-lowering composition, characterized in that: The method comprises the following steps: S1: Mix smilax glabra powder, adenophora powder, Chinese yam powder, green tea powder, senna leaf powder, and cassia tiliaceus powder with 40-80 v / v% ethanol-water solution, perform alcohol extraction for 8-12 hours, filter to obtain an alcohol extract, freeze-dry the alcohol extract to obtain an extract, wherein the alcohol extraction temperature is 25-35°C; S2: using sterile deionized water to prepare the extract into a 6-10 wt% mixed solution, sterilizing the mixed solution, inoculating the mixed solution with mixed bacteria and then anaerobically fermenting it for 24-48 hours, wherein the fermentation temperature is 35-38° C. and the fermentation pH is 5-7; S3: After the fermentation in S3 is completed, a fermentation liquid is obtained, the fermentation liquid is subjected to ultrasonic lysis, filtered, the filtrate is collected and sterilized to obtain a fermentation filtrate, and the fermentation filtrate is concentrated and freeze-dried to obtain a uric acid-lowering composition; The mixed bacteria inoculated in step S2 are composed of Lactobacillus plantarum with a preservation number of CGMCC NO.26508 and Lactobacillus fermentum with a preservation number of CGMCC NO.16754; the mass ratio of Lactobacillus plantarum to Lactobacillus fermentum is 1:1.5-1.7; the total addition amount of Lactobacillus plantarum and Lactobacillus fermentum is 5-8wt% of the mixed solution; The mass ratio of the smilax glabra powder, adenophora root powder, Chinese yam powder, green tea powder, senna leaf powder and cassia seed powder in step S1 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 Chinese yam powder, adenophora root powder, Chinese yam powder, green tea powder, senna leaf powder and cassia seed powder is 1:2:3:4:6:

3.

3. The method for preparing the uric acid-lowering composition according to claim 1, characterized in that: The mass ratio of the plant lactobacillus to the fermented lactobacillus is 1:1.5-1.

6.

4. The method for preparing the uric acid-lowering composition according to claim 1, characterized in that: The mass ratio of Lactobacillus plantarum to Lactobacillus fermentum is 1:1.

6.

5. The method for preparing the uric acid-lowering composition according to claim 1, characterized in that: In the step S1, the mass ratio of the total mass of the smilax glabra powder, adenophora radix powder, yam powder, green tea powder, senna leaf powder, and cassia toad powder to the ethanol-water solution is 1:8-10.

6. The method for preparing the uric acid-lowering composition according to claim 1, characterized in that: The sterilization described in S2 and S3 is irradiation sterilization.

7. A uric acid-lowering composition prepared according to the method for preparing a uric acid-lowering composition according to any one of claims 1 to 6.

8. Use of the uric acid lowering composition as claimed in claim 7 in the preparation of uric acid lowering products.

9. The use according to claim 8, characterized in that: The dosage form of the uric acid lowering product is any one of tea, paste, tablet, capsule, pill, granule or oral solution.

10. The use according to claim 8, characterized in that: The dosage form of the uric acid lowering product is tea.

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