A composition for reducing uric acid and preventing and treating gout and its application in the preparation of drugs for reducing uric acid and preventing and treating gout

Abstract

The present invention discloses a composition, which comprises mixed extract A and mixed extract B, and the mass ratio of mixed extract A to mixed extract B is 1:4 - 4:1; wherein mixed extract A is prepared from 20 - 40 parts by weight of Phyllanthus emblica, 10 - 30 parts by weight of honeysuckle, 5 - 15 parts by weight of fresh Imperata cylindrica root, and 5 - 15 parts by weight of dandelion; mixed extract B is prepared from 5 - 15 parts by weight of Cassia obtusifolia, 20 - 40 parts by weight of Cynara scolymus, 5 - 15 parts by weight of Lilium brownii, and 10 - 30 parts by weight of Cornus officinalis; when the composition is applied to the preparation of drugs for reducing uric acid and preventing and treating gout, the experimental results show that the composition can significantly inhibit uric acid production, promote uric acid excretion, thereby reducing blood uric acid, and at the same time has the effects of significantly reducing gouty joint swelling, inhibiting joint inflammation and oxidative stress state. In addition, the composition of the present invention is derived from medicinal and edible homologous plant raw materials, and is safe, convenient to eat and has no toxic side effects.

Description

Technical Field

[0001] The present invention relates to the technical field of uric acid lowering and gout prevention and treatment, and in particular to a uric acid lowering and gout prevention and treatment composition and an application of the composition in the preparation of a uric acid lowering and gout prevention and treatment medicine. Background Art

[0002] Uric acid is a product of purine metabolism in the human body. When uric acid is overproduced or cannot be excreted normally, it accumulates in the body, leading to hyperuricemia. Excessive uric acid accumulation in the body can not only cause stones, kidney damage, and cardiovascular damage, but also, when it accumulates in large quantities in joints, it can cause gout, making it one of the main causes of gout. Gout is an inflammatory disease caused by the accumulation of urate in the joints and surrounding tissues, characterized by severe pain and inflammation. Hyperuricemia and gout have become common diseases worldwide. Statistics show that the global prevalence of gout is increasing, particularly in developed countries. In the United States, the prevalence of gout is approximately 3%, and the prevalence is also gradually increasing in Asian countries. In China, hyperuricemia and gout have become a serious public health problem. According to statistics, the prevalence of hyperuricemia in China has exceeded 10%, and the number of gout patients has exceeded 10 million. Although there are currently several drugs available on the market for the treatment of hyperuricemia and gout, such as benzbromarone, which promotes uric acid excretion; allopurinol and febuxostat, which inhibit uric acid synthesis; and colchicine and nonsteroidal anti-inflammatory drugs, which are used during acute gout attacks, these uric acid-lowering gout treatments, while effective, can also cause serious side effects, such as elevated transaminases, kidney stones, jaundice, and cardiovascular problems. Therefore, there is an urgent need to develop safe and effective methods for lowering uric acid and preventing gout. In China, while many traditional Chinese herbal medicines may be effective in treating hyperuricemia and gout, they often suffer from shortcomings such as slow onset and weak efficacy. Furthermore, some Chinese herbal medicines often have certain toxic side effects and require specialized medical guidance for use, significantly limiting their application. Therefore, how to screen Chinese herbal medicines from the vast sea of Chinese herbal medicines for those with minimal or no toxic side effects and the ability to lower uric acid and prevent gout, how to maximize the availability of these active ingredients, and how to scientifically combine them to enhance their efficacy are pressing challenges. Summary of the Invention

[0003] Therefore, to solve the above problems, the present invention provides a composition for reducing uric acid and preventing and treating gout, its preparation method and application. The raw materials selected in the composition of the present invention are all plants listed in the medicine and food homology catalog of our country, which are safe for consumption. Through scientific proportioning and using special preparation methods and processes, a composition is prepared. It has been confirmed by animal experiments that the composition prepared by the present invention can significantly inhibit uric acid production, promote uric acid excretion, thereby reducing blood uric acid. At the same time, it has the effects of significantly reducing the swelling of gout joints, inhibiting joint inflammation and oxidative stress state, and the composition prepared by the present invention has better effects compared with the traditional preparation method.

[0004] To achieve the above object, the present invention provides the following technical solutions:

[0005] The composition of the present invention includes mixed extract A and mixed extract B, and the mass ratio of mixed extract A to mixed extract B is 1:4 - 4:1; wherein mixed extract A is prepared from 20 - 40 parts by weight of Phyllanthus emblica, 10 - 30 parts by weight of Lonicera japonica, 5 - 15 parts by weight of fresh Imperata cylindrica roots, and 5 - 15 parts by weight of Taraxacum officinale; mixed extract B is prepared from 5 - 15 parts by weight of Cassia obtusifolia, 20 - 40 parts by weight of Cynara scolymus, 5 - 15 parts by weight of Lilium brownii, and 10 - 30 parts by weight of Cornus officinalis.

[0006] The preparation of the above composition is as follows

[0007] (1) Respectively crush Phyllanthus emblica, Lonicera japonica, fresh Imperata cylindrica roots, and Taraxacum officinale, and the particle size after crushing is 30 - 50 mesh. Mix them, mix mixture A with water, and the material - liquid ratio of mixture A to water is g:mL = 1:5 - 1:10. Under stirring, add food - grade cellulase and hemicellulase, incubate at a constant temperature for 2 - 6 h at the optimal temperature of the enzyme, then boil for 30 - 60 min, cool and filter to obtain extract 1 (the filter residue after filtration is extracted 1 - 2 times repeatedly), extract the filter residue with edible alcohol 2 - 3 times to obtain extract 2, combine extract 1 and extract 2, and spray - dry to obtain mixed extract A;

[0008] The addition amount of the food - grade cellulase is 0.5 - 5% of the mass of mixture A, the addition amount of the food - grade hemicellulase is 0.1% - 1.5% of the mass of mixture A, the volume concentration of the edible alcohol is 50 - 90%; the material - liquid ratio of the filter residue to the edible alcohol is g:mL = 1:3 - 1:6; the stirring speed is 100 - 200 rpm / min;

[0009] (2) Crush Cassia obtusifolia, Cynara scolymus, Lilium brownii, and Cornus officinalis respectively. The particle size after crushing is 20 - 40 mesh. Mix them. Mix mixture B with water. The material - liquid ratio of mixture B to water is 1 g:5 - 10 mL. Under stirring, first add food - grade cellulase, incubate at the optimal temperature of the enzyme for 3 - 7 h, then add a mixed enzyme of α - amylase and β - amylase, continue to incubate at the optimal temperature of the enzyme for 4 - 8 h, then boil for 60 - 90 min, cool and filter to obtain extract 3 (the filter residue after filtration is extracted 1 - 2 times repeatedly). Extract the filter residue with edible alcohol 2 - 3 times to obtain extract 4. Combine extract 3 and extract 4, and spray - dry to obtain mixed extract B;

[0010] The addition amount of the food - grade cellulase is 0.5 - 5% of the mass of mixture B, the addition amount of the mixed enzyme is 0.1% - 0.5% of the mass of mixture B, the mass ratio of α - amylase to β - amylase is 4:1 - 1:4, the volume concentration of edible alcohol is 70 - 100%, and the material - liquid ratio of the filter residue to edible alcohol is 1:5 - 1:8; The stirring speed is 100 - 200 rpm / min;

[0011] (3) Mix the mixed extract A obtained in step (1) with the mixed extract B obtained in step (2) to obtain a composition for reducing uric acid and preventing and treating gout;

[0012] The present invention applies the above - mentioned composition to the preparation of a drug for reducing uric acid and preventing and treating gout. The ingredient (or active ingredient) of the drug is the above - mentioned composition, and one or more pharmaceutically acceptable excipients can also be added to improve the absorption effect of the drug or facilitate its use, such as making it into capsules, pills, powders, tablets, granules, oral liquids, injections, etc., that is, making it into a suitable dosage form for use.

[0013] The composition for reducing uric acid and preventing and treating gout provided by the present invention can significantly inhibit uric acid production, promote uric acid excretion, and at the same time can significantly reduce gouty joint swelling, inhibit joint inflammation and reduce the oxidative stress state of joint tissues, thereby exerting the efficacy of reducing uric acid and preventing and treating gout.

[0014] Compared with the prior art, the present invention has the following advantages:

[0015] 1. Compared with general traditional preparation methods, the composition prepared by the present invention has a more significant and better effect;

[0016] 2. The composition of the present invention is all derived from raw materials of medicinal and edible homologous plants, is safe and convenient to eat, and has no toxic and side effects. Description of the Drawings

[0017] Figure 1 Shows the results of the effects of different compositions on the activity of xanthine oxidase in the liver of hyperuricemic mice. All values in the figure are expressed as mean ± S.E. (n = 10); Different letters in the bar graph indicate significant differences ( p < 0.05);

[0018] Figure 2 Shows the results of the effects of different compositions on the mRNA transcription of the ABCG2 gene in the kidneys of hyperuricemic mice fed a high-purine diet. All values in the figure are expressed as mean ± S.E. ( n = 10), and different letters in the bar graph indicate significant differences ( p < 0.05);

[0019] Figure 3 Shows the results of the effects of different compositions on the serum uric acid levels of hyperuricemic mice fed a high-purine diet. All values in the figure are expressed as mean ± S.E. ( n = 10), and different letters in the bar graph indicate significant differences ( p < 0.05);

[0020] Figure 4 Shows the effects of different compositions on the ankle joint swelling rate at different time periods after sodium urate injection into the joints of rats. All values in the figure are expressed as mean ± S.E. ( n = 8), and different letters in the same bar graph indicate significant differences ( p < 0.05);

[0021] Figure 5 Shows the results of the effects of different compositions on the ankle joint morphology and HE staining of synovial tissues in gouty rats induced by sodium urate. A: Intuitive view of the rat ankle joint; B: Results of HE staining sections of synovial tissues at 200 times magnification;

[0022] Figure 6 Shows the effects of different compositions on the levels of SOD, GSH, MDA, IL-1β, IL-6, and TNF-α in the ankle joint tissues of gouty rats induced by sodium urate. All values are expressed as mean ± S.E. ( n = 8), and different letters in the bar graph indicate significant differences ( p < 0.05). Detailed implementation methods

[0023] The following are the implementation examples of the present invention. The described examples are for further describing the present invention rather than limiting it. The specific experimental procedures and their results described in the examples are only used to illustrate the present invention. The methods in the examples are conventional methods unless otherwise specified, and the reagents used are conventional commercially available reagents or reagents prepared according to conventional methods;

[0024] Example 1: Preparation of Composition I by the method of the present invention

[0025] 1. Respectively, pulverize 30 parts by weight of Phyllanthus emblica, 20 parts by weight of honeysuckle, 10 parts by weight of fresh Imperata cylindrica roots, and 10 parts by weight of dandelion to a particle size of 40 mesh. After mixing, mix mixture A with drinking water at a material-liquid ratio of g:mL of 1:6, and add food-grade cellulase and hemicellulase at 150 rpm / min (the addition amount of food-grade cellulase is 1.5% of the mass of mixture A, and the addition amount of food-grade hemicellulase is 0.5% of the mass of mixture A). Incubate at a constant temperature of 50 °C for 5 h, then boil for 45 min and cool to room temperature, filter to obtain the filtrate, re-extract the filter residue according to the above steps 2 times, collect and combine the filtrates to obtain extract 1; after repeated extraction, extract the filter residue with edible alcohol (volume concentration of 80%, material-liquid ratio of g:mL of 1:5) 3 times, collect and combine the extracts to obtain extract ②, combine extract 1 and extract 2, and spray-dry to obtain mixed extract A;

[0026] 2. Respectively, pulverize 10 parts by weight of Cassia obtusifolia, 30 parts by weight of Cynara scolymus, 10 parts by weight of Lilium brownii, and 20 parts by weight of Cornus officinalis to 40 mesh. After mixing, mix mixture B with drinking water at a material-liquid ratio of g:mL of 1:8, and first add food-grade cellulase under stirring at 180 rpm / min (the addition amount of food-grade cellulase is 2% of the mass of mixture B). Incubate at a constant temperature of 50 °C for 4 h, then add a mixed enzyme of α-amylase and β-amylase (the addition amount of the mixed enzyme is 0.3% of the mass of mixture B, and the mass ratio of α-amylase to β-amylase is 3:2), and continue to incubate at a constant temperature of 5 °C for 6 h, then boil for 75 min, cool and filter to obtain the filtrate, repeat the above enzyme extraction steps 2 times, collect and combine the filtrates to obtain extract 3; after repeated extraction, extract the filter residue with 90% edible alcohol 3 times (material-liquid ratio of g:mL of 1:7), collect and combine the extracts to obtain extract 4, and finally combine extract 3 and extract 4 and spray-dry to obtain mixed extract B;

[0027] 3. Mix mixed extract A and mixed extract B evenly according to a mass ratio of 3:2 to obtain the composition Ⅰ of the present invention.

[0028] Example 2: Prepare composition Ⅱ by traditional method

[0029] 1. Respectively, pulverize 30 parts by weight of Phyllanthus emblica, 20 parts by weight of honeysuckle, 10 parts by weight of fresh Imperata cylindrica roots, and 10 parts by weight of dandelion into 40 mesh, mix, and then stir and mix the mixture with drinking water at a material-liquid ratio of g:mL of 1:6 (150 rpm / min), then boil for 45 minutes, cool to room temperature and filter to obtain the filtrate, and repeat the above steps 2 times; then extract the filter residue with 80% edible alcohol at a material-liquid ratio of g:mL of 1:5 3 times, and after combining all the filtrates, concentrate and spray-dry to obtain mixed extract A;

[0030] 2. Respectively pulverize 10 parts by weight of cassia seed, 30 parts by weight of artichoke, 10 parts by weight of lily, and 20 parts by weight of cornel to 40 mesh. After mixing, stir and mix the mixture with drinking water at a material-liquid ratio of g:mL of 1:8 (180 rpm / min), then boil for 75 minutes, cool to room temperature and filter to obtain a filtrate, and repeat the above steps for re-extraction 2 times; then extract the filter residue with 90% edible alcohol at a material-liquid ratio of g:mL of 1:7 for 3 times. After combining all the filtrates, concentrate and spray-dry to obtain a mixed extract B;

[0031] 3. Mix the mixed extract A and the mixed extract B evenly according to a mass ratio of 3:2 to obtain Composition II.

[0032] Example 3: Experiment on the efficacy of different compositions in preventing and treating hyperuricemia in mice induced by a high-purine diet

[0033] 1. Construction and grouping of animal models

[0034] A mouse hyperuricemia model was established by using a high-purine yeast diet combined with intraperitoneal injection of potassium oxonate. Through animal model experiments, the effects of different compositions on blood uric acid, xanthine oxidase activity in the liver, and mRNA transcription of the uric acid transporter ABCG2 gene in the kidneys of hyperuricemic mice were observed. The specific methods are as follows:

[0035] Purchase 40 SPF-grade male Kunming mice (20g ± 10%) from Hunan Slack Jingda Experimental Animal Co., Ltd. Before the start of the experiment, randomly divide the 40 mice into 4 groups, with 10 mice in each group, namely: normal control group (Group C), hyperuricemia model group (Group M), Composition II experimental group (Group Y), and Composition I experimental group (Group X); after the start of the experiment, Group C and Group M were given intragastric administration of normal saline every day, Group Y was given intragastric administration of Composition II every day, with a gavage dose of 600 mg / kg b.w, and Group X was given intragastric administration of Composition I prepared by the method of the present invention every day, with a gavage dose of 600 mg / kg b.w; except for Group C which ate normal feed, all the experimental mice in the other three groups ate high-purine yeast feed and were given intragastric administration of potassium oxonate (200 mg / kg b.w). After continuous feeding for 3 months, all the mice were fasted but not water-deprived for 12 hours, then anesthetized, and blood was collected and centrifuged to separate and collect plasma for subsequent index measurement; then the liver and kidney tissues of the mice were taken out, rinsed clean with normal saline, frozen in liquid nitrogen and stored at -80°C for standby.

[0036] 2. Determination of xanthine oxidase activity in the liver

[0037] Xanthine oxidase is a key enzyme for uric acid production in the body, mainly present in the liver. The increase in its activity is one of the main causes of hyperuricemia. Therefore, inhibiting the activity of xanthine oxidase in liver tissue to reduce uric acid can effectively prevent hyperuricemia. Detection was carried out using a xanthine oxidase kit (Nanjing Jiancheng), and the test method was carried out with reference to the kit instructions;

[0038] The results are shown in Figure 1 , it can be seen from the figure that compared with the normal control group (Group C), the xanthine oxidase activity in the liver of mice in the high-purine diet plus oxonic acid potassium group, that is, the model group (Group M), increased significantly ( p < 0.05), and compared with Group M, both Composition Ⅰ and Composition Ⅱ obtained by the two preparation methods could effectively inhibit the activity of xanthine oxidase. However, compared with Composition Ⅱ (Group Y) prepared by the general traditional preparation method, Composition Ⅰ (Group X) prepared by the method of the present invention had a better inhibitory effect on the activity of xanthine oxidase, and there was no significant difference between it and the normal control group ( p < 0.05), while the xanthine oxidase activity in Group Y was still significantly higher than that in the normal control group ( p < 0.05).

[0039] 3. Determination of relative mRNA expression level of uric acid transporter ABCG2 gene in the kidney

[0040] Uric acid excretion disorder in the kidney is another main cause of the onset of hyperuricemia. In the kidney, ABCG2 protein is a key protein for uric acid transport and excretion. Real-time fluorescence quantitative PCR was used to detect the mRNA transcription level of the ABCG2 protein gene in the kidney. The specific method steps are as follows:

[0041] The primers for ABCG2 protein gene and reference protein GAPDH gene were designed by Shanghai Sangon Biological Engineering Co., Ltd. according to the literature, specifically: ABCG2-F is 5’-CACTGACCCTTCCATCCTCTTC-3’, ABCG2-R is 5’-GCCCTGTTTAGACATCCTTTTCA-3’; GAPDH-F is 5’-AGGTCGGTGTGAACGGATTTG -3’, GAPDH-R is 5’-TGTAGACCATGTAGTTGAGGTCA-3’. Total RNA of the kidney was extracted using the RNAprep Pure Tissues kit provided by Tiangen Biochemical Technology (Beijing) Co., Ltd., reverse transcribed into cDNA using Tiangen Biochemical's Fast King RT kit, and then real-time fluorescence quantitative PCR was performed using Tiangen Biochemical's SuperReal PreMix Plus (SYBR Green) kit on an Applied Biosystems StepOnePlus™ RT-PCR device. It was pre-denatured at 95°C for 30 s, then denatured at 95°C for 10 s, annealed at 50°C for 20 s, and extended at 60°C for 30 s, with a total of 40 cycles; the data was analyzed using ABI StepOnePlus™ software 2.3, and the gene transcription level was obtained by normalizing the data with the reference GAPDH gene.

[0042] The results are shown in Figure 2 , and it can be seen from the figure that compared with the normal control group (Group C), the mRNA transcription level of ABCG2 gene in the kidneys of mice in the high-purine diet plus oxonic acid potassium group, namely the model group (Group M), decreased significantly ( p < 0.05). Compared with Group M, both Composition Ⅰ and Composition Ⅱ prepared by the two preparation methods could increase the mRNA transcription level of this gene. However, compared with Composition Ⅱ (Group Y) prepared by the general traditional preparation method, Composition Ⅰ (Group X) prepared by the method of the present invention had a better effect on enhancing the mRNA transcription level of ABCG2 gene, and there was no significant difference between it and the normal control group ( p < 0.05). Although Composition Ⅱ (Group Y) could also increase the mRNA transcription level of this gene, the improvement effect was limited and was still significantly lower than that of the normal control group ( p < 0.05).

[0043] 4. Determination of blood uric acid content in plasma

[0044] The main symptom of hyperuricemia is that the uric acid content in the blood exceeds the normal standard. It was detected using a uric acid detection kit (Nanjing Jiancheng), and the test method was carried out according to the kit instructions;

[0045] The results are shown inFigure 3 , it can be seen from the figure that compared with the normal control group (Group C), the uric acid content in the blood of mice in the high-purine diet plus potassium oxonate group, namely the model group (Group M), increased significantly, doubling ( p <0.05), and compared with Group M, both Composition Ⅰ and Composition Ⅱ prepared by the two preparation methods could reduce the uric acid content in the blood of mice to a certain extent. However, Composition Ⅱ (Group Y) prepared by the general traditional preparation method had a weaker effect on reducing blood uric acid. Compared with Composition Ⅰ (Group Y), Composition Ⅰ (Group X) prepared by the method of the present invention had a better and more obvious effect on reducing the blood uric acid content in mice. After 3 months of consumption, it could effectively prevent the increase of blood uric acid, and there was no significant difference from the normal control group ( p <0.05).

[0046] Example 4: Efficacy experiment of different compositions in preventing and treating sodium urate-induced gout in rats

[0047] 1. Construction and grouping of animal models

[0048] A rat gout model was established by injecting sodium urate solution into the ankle joint cavity to observe the effects of different compositions in preventing and treating sodium urate-induced gout in rats. The specific method is as follows:

[0049] Thirty-two SPF-grade male SD rats (200 g ± 5%) were purchased from Hunan Slack Jingda Experimental Animal Co., Ltd. Before the start of the experiment, the 32 rats were randomly divided into 4 groups (8 rats in each group). Each group was the normal control group (Group C), the gout model group (Group M), the Composition Ⅱ experimental group (Group Y), and the Composition Ⅰ experimental group (Group X). After the start of the experiment, Group C and Group M were given intragastric administration of normal saline every day. Group Y was given intragastric administration of Composition Ⅱ prepared by the traditional preparation method every day, and the intragastric administration dose was 400 mg / kg b.w. Group X was given intragastric administration of Composition Ⅰ prepared by the method of the present invention every day, and the intragastric administration dose was 400 mg / kg b.w. After 28 days of intragastric administration, sodium urate was ultrasonically dissolved in sterile normal saline at a concentration of 30 mg / mL. After the rats were anesthetized with isoflurane, 100 μL of sodium urate solution was injected into the ankle joint cavity of each rat in Group M, Group Y, and Group X, and the same dose of sterile normal saline was injected into Group C. The ankle circumference was recorded at 4 h, 8 h, 12 h, 24 h, and 48 h after injection. All rats were fasted for 12 h after 48 hours of injection, and after being anesthetized and sacrificed with isoflurane, joint tissues 0.5 cm above and below the ankle joint were collected. A part of the joint tissues was fixed with 4% paraformaldehyde for use as ankle synovial tissue sections, and the remaining joint tissues were stored at -80 °C for subsequent determination of biochemical indexes.

[0050] 2. Measurement results of the swelling rate of rat ankles

[0051] During the acute attack of gout, the main manifestations are joint swelling and pain. The diameter of the rat ankle joint before and after injection of sodium urate was measured using a vernier caliper, and the swelling rate was calculated. Figure 4 The swelling rate of the rat ankle joint at different time periods after injection of sodium urate; as can be seen from the figure, 4 hours after injection, compared with the normal control group C, the ankle joints of the other three groups of rats showed very significant swelling ( p <0.05), among which the swelling of group M and group Y was the most severe, and there was no significant difference ( p >0.05), while the swelling of group X was significantly lighter ( p <0.05). 8 hours after injection of sodium urate, the swelling rate of the ankle joints of each group of rats further increased, and the pattern was the same as that at 4 hours. From 12 hours, the swelling rate of each group began to decline, but there was still no obvious difference in the swelling rate between the composition II (group Y) prepared by the traditional preparation method and the model group M ( p >0.05), while the swelling rate of group X was significantly lower than that of the other two groups ( p <0.05). After 24 hours, the swelling rate of the ankle joints of each group further decreased, and the situation among the groups was the same as that at 12 hours. When 48 hours after injection of sodium urate, the swelling rate of the ankle joint of the rats in the model group M was the most severe, followed by group Y, and although the swelling rate of group X was still higher than that of the normal control group C, it was the lightest among the three groups ( p <0.05). These results indicate that the composition II prepared by the traditional preparation method has no obvious anti-swelling effect in the early and middle stages of gout attack, but only has a certain anti-swelling effect in the later stage, while the composition I of the present invention can significantly inhibit the joint swelling rate throughout the acute attack period of gout, and can effectively reduce the severity of gout attack.

[0052] 3. Pathological results of rat ankle joint synovial tissue

[0053] The attack of gout is caused by joint inflammation after urate crystals accumulate in the joints to a certain extent. Figure 5 It is the direct view of the rat ankle joint induced by sodium urate and the results of pathological analysis of synovial tissue. Through the direct view of the joint ( Figure 5 A), it can be clearly shown that there is no obvious swelling in the ankle joint of group C, while the ankle joint tissue of group M is significantly swollen, and the joint swelling of group Y and group X has been improved to a certain extent, especially group X, and the joint swelling has been improved particularly significantly, and there is basically no obvious difference from the normal control group C. From the HE histopathological section ( Figure 5 B), it can be seen that the morphological structure of the synovial tissue in group C is normal, while a large number of inflammatory cell infiltrations appear in the synovial tissue of group M, and the inflammatory cells in the synovial tissue of group Y and group X have been significantly improved, especially group X, and there is no obvious inflammatory cell infiltration phenomenon in the synovial tissue, which is basically the same as that of the normal control group C.

[0054] 4. Determination Results of Tissue Biochemical Indexes and Inflammatory Factors

[0055] The amounts of superoxide dismutase (SOD), glutathione (GSH), and malondialdehyde (MDA) were detected using a biochemical index kit, and the test method was carried out according to the kit instructions; the detection of inflammatory factors interleukin (IL)-1β, IL-6, and tumor necrosis factor-α (TNF-α) was performed using an ELISA kit (Shanghai Enzyme-linked Biotechnology Co., Ltd.), and the method was carried out according to the kit instructions;

[0056] Figure 6 For the determination results of the biochemical indexes and inflammatory factors in the ankle joint tissues of gouty rats induced by sodium urate, it can be seen from Figure 6 that compared with the normal control group C, the levels of SOD and GSH in the ankle joint tissues of the model group M rats were significantly decreased, while the level of MDA was significantly increased ( p <0.05), indicating that the oxidative stress level in the ankle joints of M-group rats was significantly increased, and the expressions of inflammation-related factors IL-6, IL-1β, and TNF-α were up-regulated; both Composition Ⅰ and Composition Ⅱ prepared by the two preparation methods could improve the oxidative stress level of the rats' ankle joints to a certain extent and down-regulate the expression of inflammatory factors. However, generally speaking, the effect of the Composition Ⅰ of the present invention in reducing uric acid and preventing and treating gout was significantly better than that of Composition Ⅱ prepared by the traditional method.

[0057] In summary, it can be determined that the composition for reducing uric acid and preventing and treating gout prepared by the present invention can significantly inhibit uric acid production, promote uric acid excretion, thereby reducing blood uric acid, and at the same time has the effects of significantly reducing gouty joint swelling, inhibiting joint inflammation and oxidative stress state, and the effect of the composition of the present invention is better than that of the general traditional preparation method. Therefore, the composition for reducing uric acid and preventing and treating gout prepared by the present invention can be applied to the development of drugs for preventing and treating hyperuricemia and gout diseases.

Claims

1. A composition for reducing uric acid and preventing and treating gout, characterized in that, It is composed of mixed extract A and mixed extract B, and the mass ratio of mixed extract A to mixed extract B is 1:4 - 4:1; wherein mixed extract A is prepared from 20 - 40 parts by weight of Phyllanthus emblica, 10 - 30 parts by weight of honeysuckle, 5 - 15 parts by weight of fresh Imperata cylindrica roots, and 5 - 15 parts by weight of dandelion; mixed extract B is prepared from 5 - 15 parts by weight of Cassia obtusifolia, 20 - 40 parts by weight of Cynara scolymus, 5 - 15 parts by weight of Lilium brownii, and 10 - 30 parts by weight of Cornus officinalis.

2. The composition according to claim 1, wherein: For mixed extract A, Phyllanthus emblica, honeysuckle, fresh Imperata cylindrica roots, and dandelion are respectively pulverized and mixed. The mixture A is mixed with water, and under stirring, food - grade cellulase and hemicellulase are added. After incubation reaction at a constant temperature, it is boiled and then cooled and filtered to obtain extract 1; the filter residue is extracted with edible alcohol to obtain extract 2, and extract 1 and extract 2 are combined and spray - dried to obtain the product.

3. The composition according to claim 2, characterized in that: The particle size of the raw materials after pulverization is 30 - 50 mesh. The addition amount of food - grade cellulase is 0.5% - 5% of the mass of mixture A, and the addition amount of food - grade hemicellulase is 0.1% - 1.5% of the mass of mixture A. The constant - temperature incubation is carried out at the optimal temperature of the enzyme for 2 - 6 h, the boiling time is 30 - 60 min, and the enzyme extraction is carried out 2 - 3 times to obtain extract 1; the volume concentration of edible alcohol is 50% - 90%, and the edible - alcohol extraction is carried out 2 - 3 times to obtain extract 2.

4. The composition according to claim 1, characterized in that: For mixed extract B, Cassia obtusifolia, Cynara scolymus, Lilium brownii, and Cornus officinalis are respectively pulverized and mixed. The mixture B is mixed with water, and under stirring, food - grade cellulase is first added. After incubation reaction at a constant temperature, a mixed enzyme of α - amylase and β - amylase is added, and the incubation reaction is continued at a constant temperature. Then it is boiled, cooled and filtered to obtain extract 3, the filter residue is extracted with edible alcohol to obtain extract 4, and extract 3 and extract 4 are combined and spray - dried to obtain the product.

5. The composition according to claim 4, characterized in that: The particle size of the raw materials after pulverization is 20 - 40 mesh. The addition amount of food - grade cellulase is 0.5% - 5% of the mass of mixture B, the addition amount of the mixed enzyme is 0.1% - 0.5% of the mass of mixture B, and the mass ratio of α - amylase to β - amylase is 4:1 - 1:

4. The constant - temperature incubation of mixture B with food - grade cellulase is carried out at the optimal temperature of the enzyme for 3 - 7 h; the constant - temperature incubation after adding the mixed enzyme is carried out at the optimal temperature of the enzyme for 4 - 8 h, the boiling time is 60 - 90 min, and the enzyme extraction is carried out 2 - 3 times to obtain extract 3; the volume concentration of edible alcohol is 70% - 100%, and the edible - alcohol extraction is carried out 2 - 3 times to obtain extract 4.

6. Use of the composition according to any one of claims 1 - 5 in the preparation of a drug for reducing uric acid and preventing and treating gout.

7. The application according to claim 6, wherein: The drug includes pharmaceutically acceptable excipients, and the drug dosage form is a pharmaceutically suitable dosage form.

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