A composition for reducing uric acid and application thereof

Abstract

The present application relates to the technical field of reducing uric acid, and discloses a composition capable of reducing uric acid, which comprises 20-30 parts of chicory extract, 15-20 parts of coix seed powder, 10-20 parts of sunflower disc extract, 9-15 parts of galangal extract, 10-15 parts of polygonatum extract and 7-15 parts of xanthine oxidase inhibiting peptide; the xanthine oxidase inhibiting peptide is extracted by enzymolysis of bonito processing byproducts. The composition has a high inhibiting effect on XOD activity, can reduce the levels of creatinine and urea nitrogen, realizes uric acid reduction, and has the effects of inhibiting renal tubular expansion and reducing renal interstitial inflammatory cell infiltration, and has the effect of protecting kidney. Meanwhile, the bonito processing byproducts are utilized by preparing XOD inhibiting active polypeptide, a new utilization way of bonito processing byproducts is developed, and the utilization value of bonito processing byproducts is improved.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of reducing uric acid, and particularly relates to a composition capable of reducing uric acid and application. BACKGROUND

[0002] Uric acid is the final product of purine metabolism in the human body. When the amount of uric acid generated by the body is much greater than the amount of excretion, the content of uric acid in the blood continues to rise to form hyperuricemia, which causes recurrent inflammation, kidney disease, cardiovascular and cerebrovascular diseases, gout and other diseases. The reasons for abnormal uric acid production are various, including excessive intake of exogenous purines, excessive production of endogenous purines, and reduced kidney function. Excessive intake of exogenous purines can be regulated by diet. Excessive production of endogenous purines and reduced kidney function often mean that the body is diseased, making it more difficult to handle. Excessive production of endogenous purines is mainly caused by abnormal enzyme systems, such as increased xanthine oxidase activity and excessive phosphoribosyl pyrophosphate synthetase activity.

[0003] The common method for treating hyperuricemia at present is to take chemical drugs such as allopurinol and Febuxostat to reduce the amount of uric acid generated, but such drugs can cause corresponding adverse reactions, such as mild rash (MPE), severe skin adverse reactions (SCAR), abnormal liver function, gastrointestinal reactions and rashes. Studies have shown that certain animal polypeptides, such as tuna polypeptides, can reduce uric acid levels by inhibiting XOD activity, have the characteristics of low preparation cost, high safety and easy absorption, and have become a research hotspot in the field of reducing uric acid. However, the internal causes of high uric acid are diverse, and inhibiting XOD activity does not always lead to a synchronous reduction in uric acid levels, so it is not enough to only focus on inhibiting XOD activity, and it needs to be considered from multiple aspects. Chinese patent CN111704650B discloses extracting two polypeptides with anti-uric acid effect from bonito meat, and compounding them with pueraria powder, chrysanthemum powder, coix seed powder and gaoliangjiang powder to form a composite preparation, which has good XOD activity inhibition and uric acid reduction effect. Ren Jiaoyan of South China Institute of Technology extracted bonito peptides from bonito meat and prepared a uric acid-reducing solid beverage product by compounding pueraria extract, which can reduce the uric acid value of patients with uric acid and has entered industrialization. This idea of compounding animal polypeptides and plant extracts is expected to become a safer and more effective way to reduce uric acid.

[0004] Skipjack belongs to low value tuna, and a large amount of processing by-products will be produced in the process of processing production, such as internal organs, dark meat, fish skin and fish bones, etc., accounting for about 50-70% of the weight. The annual processing capacity of skipjack in Zhoushan and Ningbo area is in the forefront in China, and the processing by-products of skipjack are up to 50 tons per day. These by-products are usually processed into cheap feed, and the utilization value is low, which causes a large amount of waste of valuable nutrients and functional components. Because the types and contents of amino acids or functional polypeptides contained in different parts of skipjack are different, the effects are also different, and currently there is no report on extracting active substances that can inhibit XOD activity and reduce uric acid level from skipjack processing by-products, and there is no report on realizing the high-efficiency uric acid-reducing effect by combining the extracted products with traditional Chinese medicine ingredients. SUMMARY

[0005] The purpose of the present application is to provide a composition for reducing uric acid, which utilizes the active peptides extracted from skipjack processing by-products and the plant extracts to cooperate from multiple aspects to achieve the purpose of reducing uric acid level, and at the same time, opens up a new way for the utilization of skipjack processing by-products and improves the utilization value of skipjack processing by-products.

[0006] The present application provides the following technical solutions:

[0007] A composition for reducing uric acid, comprising the following components by weight:

[0008] Chicory extract 20-30 parts, yiyi ren powder 15-20 parts, sunflower disc extract 10-20 parts, huangjiang extract 9-15 parts, yuzhu extract 10-15 parts, xanthine oxidase inhibiting peptide 3-8 parts;

[0009] The xanthine oxidase inhibiting peptide is extracted by enzymolysis of skipjack processing by-products.

[0010] The polypeptides extracted from skipjack processing by-products are reasonably compounded with plant extracts in the present application, and the uric acid-reducing effect is achieved through synergistic effect, specifically:

[0011] The XOD inhibiting peptide with XOD inhibiting activity is obtained by enzymatic hydrolysis of the processing by-product of the bonito, and the enzyme used can be selected from the group consisting of flavor protease, trypsin, papain, neutral protease, bromelain, alkaline protease and aquatic protease. On this basis, the medicinal properties of the chicory extract are bitter and salty, and it is generally used for diuresis, swelling, heat-clearing and detoxification. Modern pharmacological studies have found that it has obvious effects on hyperuricemia. It can inhibit the expression of intestinal transport protein CNT2, inhibit the intake of intestinal purine nucleosides, reduce uric acid levels, and increase the clearance rate of uric acid in the kidney. It can also reduce the protein expression of the high-risk factor of hyperuricemia, i.e., the kidney Glut9, and inhibit the reabsorption of uric acid in the kidney, so as to promote the excretion of uric acid in the kidney and achieve the purpose of reducing uric acid. The chlorogenic acid, chicoric acid and aesculin in the chicory are closely related to the effect of reducing uric acid. The coix seed can drain water and remove dampness and wind. Clinical pharmacology has summarized that the coix seed has good effects of removing dampness and purging turbidity, and it is a good auxiliary drug that can promote the excretion of the kidney and reduce the uric acid content in the body. At the molecular level, it also has the effects of affecting DNA binding transcription factors and cysteine-type endopeptidase activity. In addition, the ferulic acid, coumaric acid and chlorogenic acid in the coix seed can bind with XOD and inhibit the activity of XOD. The chlorogenic acid rich in the sunflower disc extract further enhances the effect of the coix seed. The alkaloids contained in the sunflower disc can specifically bind with uric acid to form a sunflower alkaloid uric acid complex to eliminate uric acid. The flavones in the sunflower disc can effectively scavenge oxygen free radicals in the body, prevent nucleic acid oxidation, reduce purine content, and also reduce blood sugar and monoglyceride, reduce capillary fragility, increase permeability to reduce the pressure on kidney cells and allow more uric acid to pass through.

[0012] In this way, the above components synergistically cooperate from multiple aspects to reduce creatinine, uric acid nitrogen, inhibit XOD activity, excrete uric acid and block the excessive production of uric acid, so as to reduce the uric acid level. Further experiments have shown that it can also inhibit the dilation of the renal tubule and relieve the inflammatory cell infiltration of the renal interstitium, so as to achieve the effect of protecting the kidney. At the same time, it also opens up a new way for the utilization of the processing by-product of the bonito and improves the utilization value of the processing by-product of the bonito.

[0013] Each plant extract used in the above composition is a water extract or an alcohol extract. It can be purchased or self-made after being extracted with water or ethanol, concentrated and spray dried. Taking the water extraction method as an example, the raw material is washed, crushed and sieved through a 100-mesh sieve. Then it is mixed with water at a mass ratio of 1:20, and hot water is used for extraction at 85℃ for 5 times. The extract is cooled, concentrated and spray dried in vacuum to obtain the product.

[0014] As a preferred embodiment of the present application, the preparation process of the xanthine oxidase inhibiting peptide is as follows:

[0015] The pH value and temperature of the slurry of the processing byproduct of the bonito are adjusted, and trypsin or aquatic protease is added for enzymatic hydrolysis. After the enzymatic hydrolysis, the supernatant is separated, and the supernatant is separated and dried to obtain the xanthine oxidase inhibiting peptide.

[0016] Currently, flavor protease and other enzymes are often used to hydrolyze the bonito fish meat to prepare the XOD inhibiting peptide, which has a better enzymatic hydrolysis effect, and the XOD inhibiting rate of the obtained enzymatic hydrolysate is high. However, the processing byproduct of the bonito is derived from the internal organs, dark meat, fish skin and fish bones, etc., and has more complex components, large differences in the types and contents of amino acids and active peptides compared with the bonito fish meat. In our screening experiment, the XOD inhibiting rate of the polypeptide product obtained by treating the processing byproduct of the bonito with flavor protease and other enzymes is not high. Although the degree of hydrolysis is very high when the processing byproduct of the bonito is treated with animal protease, the content of small molecules in the enzymatic hydrolysate is high, but the obtained polypeptide does not have XOD inhibiting activity, which indicates that the types and abilities of the biological activity of the polypeptide product are related to the used enzymes. After analysis, the XOD inhibiting rate of the polypeptide obtained by treating the processing byproduct of the bonito with trypsin and aquatic protease is high, and trypsin is better. Therefore, the polypeptide prepared by trypsin or aquatic protease is selected for compounding with plant extracts.

[0017] As a preferred embodiment of the present application, the pH range of the enzymatic hydrolysis is 6.3-8.2, and the enzymatic hydrolysis temperature range is 40-55°C.

[0018] As a preferred embodiment of the present application, the processing byproduct of the bonito is also pre-enzymatically hydrolyzed with animal protease, and then trypsin or aquatic protease is added for enzymatic hydrolysis.

[0019] In further research, it is found that the polypeptide product obtained by using animal protease in combination with trypsin can further improve the uric acid lowering activity of the composition. Aquatic protease has a similar degree of hydrolysis as animal protease, and the XOD inhibiting activity of the polypeptide product corresponding to the aquatic protease is higher, but the combination of aquatic protease and trypsin does not significantly improve the reduction of uric acid level, which may be because, on the one hand, there is a certain synergistic effect between different proteases, and the spatial structure of proteins and polypeptides is complex. The high degree of hydrolysis of animal protease makes the proteins in the processing byproduct of the bonito decompose into more small molecular weight peptide segments, thereby exposing more trypsin treatment sites, so that the effect is better when trypsin is used for treatment. On the other hand, there is a synergistic effect between the prepared polypeptide components and plant extracts.

[0020] As a preferred embodiment of the present application, the amount of enzyme added is 2-5wt% of the mass of the slurry.

[0021] As a preferred embodiment of the present application, in the slurry, the mass fraction of the processing byproduct of the bonito is 15-40wt%.

[0022] As a preferred embodiment of the present application, the tuna processing by-products are one or more of the following: tuna processing generated viscera, dark meat, fish skin and fish bones.

[0023] The above composition is used for preparing a medicine for treating reducing uric acid.

[0024] The composition of the present application has the effects of inhibiting renal tubular expansion and reducing renal interstitial inflammatory cell infiltration.

[0025] The beneficial effects of the present application are as follows:

[0026] In the composition of the present application, the extracted polypeptides of tuna processing by-products are reasonably compounded with plant extracts such as chicory extract, coix seed powder, sunflower disc extract, galangal extract, polygonatum extract, etc., which have high XOD activity inhibition effect, can reduce the levels of creatinine and urea nitrogen, achieve the reduction of blood uric acid, and can inhibit renal tubular expansion and reduce renal interstitial inflammatory cell infiltration, and have the effect of protecting the kidney. At the same time, the tuna processing by-products are utilized by preparing XOD inhibitory active polypeptides, which opens up a new way of utilizing tuna processing by-products and improves the utilization value of tuna processing by-products. BRIEF DESCRIPTION OF DRAWINGS

[0027] Figure 1 is a nutritional component ratio chart of the tuna processing by-products used in the present application.

[0028] Figure 2 is the degree of hydrolysis (A) and solid content (B) of the enzymatic products of different proteases in Example 1.

[0029] Figure 3 is a chart of the in vitro XOD inhibition rate of the enzymatic products of different proteases in Example 1.

[0030] Figure 4 is a comparison chart of the serum uric acid content (A), plasma creatinine content (B), blood urea nitrogen content (C), and XOD activity (D) of mice in different groups in Example 3.

[0031] Figure 5 is a kidney tissue section chart of the normal group (A), model group (B), positive group (C), high-dose group (D), and medium-dose group (E) of mice. DETAILED DESCRIPTION

[0032] The specific embodiments of the present application are further described below.

[0033] Unless otherwise specified, the raw materials used in the present application can be purchased from the market or commonly used in the art. Unless otherwise specified, the methods in the following examples are conventional methods in the art. Unless otherwise specified, the raw materials used in the present application can be purchased from the market or commonly used in the art. Unless otherwise specified, the methods in the following examples are conventional methods in the art.

[0034] The skipjack tuna processing byproducts used in this invention consist of skipjack tuna viscera, dark meat, skin, and bones produced during processing, and are provided by Zhejiang Xingye Group.

[0035] The inventors analyzed the nutritional components and amino acid composition of skipjack tuna processing byproducts, as detailed below.

[0036] Nutritional composition: Moisture was determined by atmospheric pressure heating drying; protein was determined by semi-micro Kjeldahl nitrogen determination; fat was determined by Soxhlet extraction; ash was determined by ashing; and total sugar was determined by Fehling's volumetric method. Results are as follows: Figure 1 As shown.

[0037] from Figure 1 It can be seen that skipjack tuna processing byproducts are rich in protein and also relatively high in fat. It should be noted that because different methods are used to determine different nutrients, the total percentage of each nutrient exceeding 100% is within the normal range of experimental error.

[0038] Amino acid composition: Take 100g of bonito processing by-product sample, add 6mol / L HCl, purge with nitrogen, heat seal, and hydrolyze in an oven at 110℃ for 24h. After deacidification, dilute with water to an appropriate concentration and determine with an automatic amino acid analyzer. The results are shown in Table 1.

[0039] Table 1. Amino acid composition of skipjack tuna processing byproducts (g / 100g)

[0040]

[0041] Note: Tryptophan is destroyed under strong acid, so the results do not include tryptophan. * indicates an essential amino acid.

[0042] As shown in the table, a total of 16 amino acids were detected in the skipjack tuna processing byproducts (see Table 1), including 7 essential amino acids. The amino acids with the highest content were glutamic acid (2.37%) and aspartic acid (1.89%), which are related to umami flavor. Lysine was also relatively abundant (1.75%), while methionine was the least abundant amino acid (0.56%). The total amino acid content of the skipjack tuna processing byproducts was 19.5% of the sample mass. The ratios of essential amino acids to total amino acids and essential amino acids to non-essential amino acids were 0.43 and 0.76, respectively, which are higher than the FAO / WHO recommended values ​​of 0.40 and 0.60, indicating that the protein in the skipjack tuna processing byproducts is of high quality.

[0043] The following examples and comparative examples all use skipjack tuna processing byproducts from the above-mentioned sources.

[0044] Example 1: Screening of proteases

[0045] 1) Papain and flavor protease (Zhejiang Yinuo Biotechnology Co., Ltd.); trypsin and neutral protease (Henan Yangshao Biochemical Engineering Co., Ltd.); bromelain, alkaline protease, and pepsin (Huamao Shuanghui Industrial (Group) Co., Ltd. Biochemical Pharmaceutical Plant); animal protease and aquatic protease (Nanning Dongheng Huadao Biotechnology Co., Ltd.) were selected respectively to enzymatically hydrolyze skipjack tuna processing by-products under their respective recommended optimal pH and temperature conditions. Specific parameters are shown in Table 2.

[0046] Table 2 Enzymatic hydrolysis conditions for different proteases

[0047]

[0048] The enzymatic hydrolysis process of the above protease is as follows:

[0049] Skipjack tuna processing byproducts were mixed with water to obtain a slurry. After adjusting the pH and temperature of each protease to a suitable level, the mixture was kept for 30 minutes. The corresponding protease was then added for enzymatic hydrolysis. After enzymatic hydrolysis, the final enzymatic hydrolysate was centrifuged at 10,000 r / min for 15 minutes. The upper oil film and bottom residue were discarded, and the clear liquid in the middle was retained and spray-dried.

[0050] 2) Determination of Xanthine Oxidase (XOD) Activity Inhibition Rate

[0051] (1) Solution preparation:

[0052] Preparation of 0.2 M, pH 7.5 phosphate buffer (PBS): Weigh 4.56 g of dipotassium hydrogen phosphate and 2.71 g of potassium dihydrogen phosphate and dilute to 100 mL of distilled water respectively, and mix well at a ratio of 80:20.

[0053] Preparation of 1.5 mM xanthine solution: Weigh 0.0228 g xanthine powder, dissolve it in a trace amount of 4 mol / L NaOH, and then make up to 100 mL with buffer solution.

[0054] Preparation of 0.0125 U / mL XOD solution: Take 5 U of stock solution, take 400 μL of stock solution and dilute to 50 mL with buffer solution, set aside;

[0055] (2) Determination of XOD inhibition rate:

[0056] Add 40 µL of 0.0125 U / mL XOD solution and 50 µL of polypeptide supernatant to the reaction tube, shake well and incubate at 25 °C for 10 min. Add 50 µL of 1.5 mM xanthine solution, shake well and incubate at 25 °C for 30 min. Measure the absorbance value (OD) at a wavelength of 290 nm.

[0057] The formula for calculating the XOD inhibition rate is as follows:

[0058] ,

[0059] The test group is added with XOD solution and polypeptide supernatant; the test control group is replaced with 40 μL PBS instead of 40 μL XOD solution; the blank group is replaced with 50 μL PBS instead of 50 μL polypeptide supernatant; and the blank control is replaced with 50 μL PBS instead of 50 μL polypeptide supernatant and 40 μL PBS instead of 40 μL XOD solution.

[0060] The test takes allopurinol (2 μg / mL) as a positive control, and each sample is tested in triplicate, and the results are shown in the attached Figure 2 .

[0061] As can be seen from Figure 2 , animal proteases, trypsin and aquatic proteases have high degrees of hydrolysis, achieving full hydrolysis of the processing by-products of bonito to obtain low-molecular-weight polypeptide products. According to the existing research results, XOD inhibits polysaccharides with low molecular weight, and it is predicted that the enzymatic products of the three enzymes should have higher XOD inhibitory activity.

[0062] But as can be seen from Figure 3 , compared with the XOD inhibition rate (27.66%) of the positive control allopurinol (2 μg / mL), the XOD inhibition rate (51.64%) of the trypsin hydrolysate is the strongest, followed by the aquatic protease, and the hydrolysates of animal protease and pepsin have no XOD inhibitory effect. The reason may be that trypsin can break the peptide bond formed by the carboxyl group of Arg and Lys, and the unique cation and hydrogen bond characteristics of arginine-rich peptides are conducive to binding with the flavin adenine dinucleotide (FAD) in the six binding sites of the XOD crystal structure, forming a non-competitive and irreversible inhibitory effect, thereby inhibiting the activity of XOD, and the bonito processing by-products of the present application are rich in arginine.

[0063] Example 2: Uric acid-lowering and XOD activity inhibitory effects of the compositions

[0064] 1) Different methods are used to hydrolyze the polypeptides (I-VIII) obtained from the processing by-products of bonito, and different compositions (I-VIII) are obtained by compounding the polypeptides with plant extracts. The weight amounts of the components in the compositions are as follows:

[0065] 25 parts of chicory extract, 12 parts of huangjiang extract, 12 parts of polygonatum extract, 15 parts of sunflower disc extract, 18 parts of coix seed powder, and 5 parts of polypeptide, wherein:

[0066] Polypeptide I: polypeptide product prepared by trypsin in Example 1;

[0067] Polypeptide II: polypeptide product prepared by aquatic protease in Example 1;

[0068] Polypeptide III: The polypeptide product prepared from the animal protease in Example 1;

[0069] Polypeptide IV: Unlike the polypeptide I obtained in Example 1, animal protease was first added to the slurry for enzymatic hydrolysis, and after heating to inactivate it, trypsin was added to continue enzymatic hydrolysis to obtain the polypeptide product.

[0070] Polypeptide V: Unlike the polypeptide I obtained in Example 1, trypsin was first added to the slurry for enzymatic hydrolysis, and after heating to inactivate it, animal protein was added for further enzymatic hydrolysis to obtain the polypeptide product.

[0071] Polypeptide VI: Unlike Polypeptide I in Example 1, animal protease and trypsin were added to the slurry for enzymatic hydrolysis.

[0072] Polypeptide VII: Unlike Polypeptide II in Example 1, the polypeptide product obtained by first adding animal protease to the slurry for enzymatic hydrolysis, heating to inactivate it, and then adding aquatic protease for further enzymatic hydrolysis.

[0073] Polypeptide VIII: Unlike the polypeptide I obtained in Example 1I, the aquatic protease was first added to the slurry for enzymatic hydrolysis, and after heating to inactivate it, trypsin was added to continue enzymatic hydrolysis to obtain the polypeptide product.

[0074] All extracts were extracted with water;

[0075] 2) Tests on uric acid reduction and XOD inhibition performance

[0076] Eight-week-old mice were randomly divided into 14 groups: a normal control group (Control), a model group (MG), and a positive control group (PC).

[0077] Experimental groups I–VIII and control groups I–III. Except for the normal group, which was administered distilled water by gavage, the other groups were administered adenine and ethambutol hydrochloride by gavage to establish the uric acid model, respectively, at a dose of 1 g / kg (once daily). The treatment continued until the uric acid content reached 140 mol / L. The positive control group was administered the positive control drug allopurinol tablets (Guangdong Bidi Pharmaceutical Co., Ltd.) by gavage at a dose of 10 mg / kg. Experimental groups I–VIII were administered their corresponding compositions I–VIII by gavage, and the control groups were administered their corresponding control substances I–III by gavage, at a dose of 500 mg / kg (medium dose). One hour after gavage, adenine and ethambutol hydrochloride were administered by gavage again. After three consecutive days, blood was collected from the mice's eyeballs, and the serum uric acid level and XOD activity were tested using a kit (Nanjing Jiancheng Bioengineering Institute). The results are shown in Table 2.

[0078] Among them, the composition used in control group I was a pure plant extract, the composition used in control group II was composed of 12 parts of kudzu root powder added to composition I, and the composition used in control group III was composed of 12 parts of kudzu root powder replacing the galangal extract in composition I.

[0079] Table 2 XOD activity and blood uric acid level of each group

[0080]

[0081] From the above table, compared with the normal group, the liver xanthine oxidase activity of the model group mice was significantly increased, which accelerated the formation of uric acid in the body of mice, and the modeling was successful. By analyzing the XOD activity level and blood uric acid level of each experimental group, control group and positive group, it can be found that in most cases, the lower the XOD activity level, the lower the blood uric acid content, but there are also cases where the XOD level is low and the blood uric acid content is high, such as the positive group and the experimental group I, and the experimental group IV and the experimental group VIII, etc. The XOD activity reduction level and the blood uric acid reduction level are not always synchronized, and the reason may be that the content of blood uric acid is affected by many factors, and XOD activity is only one of the factors.

[0082] By comparing experimental groups I to VIII and control group I, it can be known that the composition obtained by compounding the polypeptide hydrolyzed by trypsin and aquatic protease with plant extract can further reduce the XOD activity and blood uric acid level, but the compounding effect of the polypeptide hydrolyzed by aquatic protease is relatively weak. In addition, the effect of using two different enzymes to prepare polypeptide, first animal protease hydrolysis, and then trypsin hydrolysis, is the best, which is better than using trypsin alone. This may be due to the higher combination of animal protein and trypsin in the enzymolysis of bonito processing by-products, and on the other hand, the reduction of blood uric acid is affected by many factors, such as in experimental group VIII, first aquatic protease hydrolysis and then trypsin hydrolysis, the XOD activity inhibition effect is the best, but the blood uric acid level is still higher than that of experimental group IV, and the combination of polypeptide hydrolysis product and plant extract needs to be considered. By comparing experimental group I and control groups II to III, it can be known that the selection of components of plant extract in the composition also affects the overall effect, such as in control group II, the introduction of pueraria powder does not significantly change the overall effect, while in control group III, the use of pueraria powder instead of high-quality ginger extract, although pueraria powder also contains flavonoids, the overall effect has declined, which reflects the combination within the composition.

[0083] Overall, under the determined plant extract, the compounding of polypeptide hydrolyzed by trypsin or aquatic protease can further improve the level of reducing uric acid, and has economic nature, and the compounding of polypeptide prepared by first animal protease and then trypsin hydrolysis can further obviously improve the effect.

[0084] Example 3 Dose effect of composition I

[0085] Based on the mouse experiment in Example 2, further adopt high dose of composition I to gavage model mice, the gavage amount is 1000 mg / kg, that is high dose group, compared with the medium dose group corresponding to experimental group I in Example 2.

[0086] 1) Serum uric acid, XOD activity and creatinine, urea nitrogen levels:

[0087] Blood was taken from the eyeball, and the kit (Nanjing Jiancheng Biological Engineering Institute) was used to test the blood uric acid, XOD activity, creatinine and urea nitrogen levels, and the results are shown in Figure 4

[0088] As shown in Figure 4 (A), the serum uric acid content of the modeling group was significantly higher than that of the normal group, indicating that the modeling was successful. Compared with the modeling group, the serum uric acid levels of the positive control and experimental groups were significantly reduced and lower than that of the normal group, indicating that allopurinol, high-dose gout soup and medium-dose gout soup can significantly reduce the serum uric acid of rats. Among them, high-dose and medium-dose gout soup showed a dose-negative correlation trend on the serum uric acid level of mice, that is, the higher the dose level of gout soup, the lower the content of serum uric acid, and the better the treatment effect. In addition, the serum uric acid level of mice treated with high-dose gout soup was lower than that of the positive group, indicating that the regulation effect of high-dose group on serum uric acid of mice was better than that of allopurinol, while the regulation effect of medium-dose gout soup was equivalent to that of allopurinol.

[0089] As shown in Figure 4 (B), compared with the normal group, gavage of adenine and ethambutol hydrochloride significantly increased the plasma creatinine level of mice. The positive control and experimental groups can significantly reduce the plasma urea nitrogen level of mice. Among them, the ability of mice to reduce plasma creatinine in the experimental group was positively correlated with the dose of the group formula, and the higher the dose of the group formula, the stronger the ability to reduce plasma creatinine. At the same time, the urea nitrogen levels of mice treated with two groups of formulas were lower than that of the allopurinol treatment group, indicating that the regulation ability of the group formula on the plasma creatinine of mice was better than that of allopurinol in this model.

[0090] As shown in Figure 4 (C), compared with the normal group, the blood urea nitrogen level of the mice gavaged with adenine and ethambutol hydrochloride increased, indicating that the glomerular filtration ability of mice was blocked. The blood urea nitrogen level of the mice treated with allopurinol group was slightly higher than that of the modeling group, indicating that allopurinol could not regulate the blood urea nitrogen level of mice. Both high-dose and medium-dose groups of formulas can reduce the blood urea nitrogen level of mice, but the relationship with the dose is not obvious. In addition, compared with the normal group, the blood urea nitrogen level of the mice treated in the experimental group is still high, which may be due to the irreversible damage to the glomerular filtration ability of mice. The formula can treat to a certain extent, but it cannot completely restore its function.

[0091] As shown in Figure 4 ​(D) showed that compared with the normal group, the liver xanthine oxidase activity of the model group mice was significantly increased, which accelerated the formation of uric acid in the mice. Compared with the model group, the liver xanthine oxidase activity of the positive group and the experimental group mice was reduced, and the XOD activity level of the two groups of gout soup with high and medium doses was higher than that of allopurinol group, indicating that in this model, the inhibition ability of the prescription on the liver XOD activity of mice is better than that of allopurinol, and the inhibition effect is not obviously related to the dose.

[0092] 2) Kidney tissue pathological changes of mice

[0093] The mice in each group in Example 3 were sacrificed, and the kidneys were taken to make sections, and after staining, the results were observed as shown in Figure 5

[0094] As shown in Figure 5 (A), the glomerulus and renal tubule structure of the normal group mice was normal; the normal group kidney showed that the cytoplasm of renal tubular epithelial cells was rich, the cell nucleus was large and loose, the tubular lumen was small, the interstitial area was small, the tubules and tubules were "back to back", and no interstitial widening was observed, but there was obvious inflammatory cell infiltration in the interstitium.

[0095] As shown in Figure 5 (B), a large number of renal tubular epithelial cells (TEC) apoptosis, necrosis, exfoliation or vacuolar degeneration were observed in the model group, highly dilated renal tubules coexisted with atrophic renal tubules, there were a large number of golden yellow purine metabolite crystals deposited in the renal interstitium, and there were protein casts in the renal tubules. The renal interstitium was severely fibrotic and proliferative, and a large number of inflammatory cells infiltrated the renal interstitium, and the inflammatory cells wrapped the purine metabolite crystals. The renal capsule cavity was dilated, and the renal tubular mesangial cells were slightly proliferated. The renal cortex was thin, the renal tubular epithelial cells were atrophic, the tubular lumen was dilated, the distal convoluted tubule and collecting duct were cystic dilatation, the renal interstitium was significantly widened in most fields, and significant fibroblast proliferation and fibrosis were observed in the interstitium.

[0096] As shown in Figure 5 (C), the renal tubule of the positive group was dilated, there were a small amount of golden yellow purine metabolite crystals, the TEC was slightly vacuolar degeneration, the renal interstitium was slightly fibrotic, and a small amount of inflammatory cells were observed, the renal capsule cavity was dilated, and the renal glomerular mesangial cells were slightly proliferated.

[0097] As shown in Figure 5 (D), the renal tubule of the high dose group was slightly dilated, a few were moderately dilated, the renal interstitium was slightly fibrotic, there were a small amount of golden yellow purine metabolite crystals in the renal interstitium, the renal capsule cavity was slightly dilated, and the renal glomerular mesangial cell proliferation was not obvious.

[0098] As shown in Figure 5 ​(E) showed that the medium dose group had moderate expansion of renal tubules, and renal tubular atrophy, TEC necrosis, exfoliation, a small amount of golden yellow purine metabolite crystal deposition in the renal interstitium, moderate fibrosis and a small amount of inflammatory cell infiltration in the interstitium, glomerular cystic cavity expansion, and mild proliferation of mesangial cells.

[0099] From the above results, it can be seen that the composition I can inhibit the expansion of renal tubules and reduce the inflammatory cell infiltration in the renal interstitium, and has a certain kidney protection effect.

Claims

1. A composition capable of reducing uric acid, characterized by comprising, The composition comprises the following components by weight: Chicory extract 20-30 parts, Galangal extract 9-15 parts, Polygonatum extract 10-15 parts, Sunflower disc extract 10-20 parts, Coix seed powder 15-20 parts, and xanthine oxidase inhibitory peptide 3-8 parts; The xanthine oxidase inhibitory peptide is extracted by adjusting the pH of the slurry of the processing by-product of the bonito and adding protease to enzymatically hydrolyze the processing by-product of the bonito; The protease is trypsin, animal protease, or aquatic protease; The plant extracts in the composition are obtained by water extraction.

2. The composition of claim 1, wherein, The preparation process of the xanthine oxidase inhibitory peptide is as follows: The pH and temperature of the slurry of the processing by-product of the bonito are adjusted, trypsin or aquatic protease is added for enzymatic hydrolysis, the supernatant is separated after the enzymatic hydrolysis, and the xanthine oxidase inhibitory peptide is obtained by drying the supernatant.

3. The composition of claim 2, wherein, The pH range of the enzymatic hydrolysis is 6.3-8.2, and the enzymatic hydrolysis temperature range is 40-55℃.

4. The composition of claim 2, wherein, The slurry of the processing by-product of the bonito is also pre-enzymatically hydrolyzed by animal protease, and then trypsin or aquatic protease is added for enzymatic hydrolysis.

5. The composition according to claim 2 or 3 or 4, characterized in that, The amount of the enzyme added is 2-5wt% of the mass of the slurry.

6. The composition of claim 2, wherein In the slurry, the mass proportion of the processing by-product of the bonito is 15-40wt%.

7. The composition according to claim 2 or 6, characterized in that, The processing by-product of the bonito is one or more of the following: internal organs, dark meat, fish skin, and fish bones produced during the processing of the bonito.

8. Use of the composition of any one of claims 1-7 in the preparation of a drug for reducing uric acid.

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