CN115715796A - Composition capable of reducing uric acid and application - Google Patents

Abstract

The invention relates to the technical field of uric acid reduction, and discloses a composition capable of reducing uric acid, which comprises, by weight, 20-30 parts of a chicory extract, 15-20 parts of coix seed powder, 10-20 parts of a sunflower disc extract, 9-15 parts of a galangal extract, 10-15 parts of a polygonatum odoratum extract, and 7-15 parts of xanthine oxidase inhibitory peptide; the xanthine oxidase inhibitory peptide is extracted from by-product of bonito processing by enzymolysis. The composition has higher XOD activity inhibition effect, can reduce creatinine and urea nitrogen levels, realize uric acid reduction, inhibit renal tubular dilation and reduce renal interstitial inflammatory cell infiltration, and has the effect of protecting the kidney. Meanwhile, the skipjack processing by-product is utilized by preparing XOD inhibitory activity polypeptide, so that a new way for utilizing the skipjack processing by-product is developed, and the utilization value of the skipjack processing by-product is improved.

Description

Composition capable of reducing uric acid and application

Technical Field

The invention relates to the technical field of uric acid reduction, and particularly relates to a composition capable of reducing uric acid and application thereof.

Background

Uric acid is the end product of the metabolism of human purines. When the uric acid production of the body is far greater than the excretion, the uric acid content in the blood is continuously increased to form hyperuricemia, which causes recurrent inflammation, nephropathy, cardiovascular and cerebrovascular diseases, gout and other diseases. The causes of abnormal uric acid production are various, including excessive exogenous purine intake, excessive endogenous purine production, renal function decline, and the like. Excessive exogenous purine intake can be regulated by attention to diet. Excessive endogenous purine production, reduced renal function, etc. often mean pathological changes in the body and increased difficulty in handling. The excessive endogenous purine production is mainly caused by enzyme system abnormality, such as xanthine oxidase activity increase, phosphoribosyl pyrophosphate synthetase activity increase, and the like.

At present, the common method for treating hyperuricemia is to take Allopurinol, febuxostat and other chemical drugs to reduce the production of uric acid, but the drugs can cause corresponding adverse reactions, such as mild rash (MPE), severe skin adverse reaction (SCAR), abnormal liver function, gastrointestinal reaction, rash and the like. Research shows that some animal polypeptides, such as tuna polypeptides, can reduce uric acid levels by inhibiting XOD activity, have the characteristics of low preparation cost, high safety, easy absorption and the like, and become a research hotspot in the field of uric acid reduction. However, the intrinsic causes of hyperuricemia are various, and inhibition of XOD activity does not always bring about a simultaneous decrease in uric acid level, so that attention is not paid to inhibition of XOD activity, and it is necessary to consider from many aspects. Chinese patent CN111704650B discloses two polypeptides with anti-uric acid effect extracted from bonito meat, and compounded with radix Puerariae powder, flos Chrysanthemi powder, coicis semen powder, and rhizoma Alpiniae Officinarum powder to form a compound preparation with good XOD activity inhibiting and uric acid reducing effects. The preparation method comprises the steps of extracting bonito peptide from bonito fish meat by delightful science of university of south China, compounding the bonito peptide with a radix puerariae extract to prepare a solid beverage product for reducing uric acid, and reducing uric acid value of uric acid patients, wherein the solid beverage product is already industrialized. The idea of compounding animal polypeptide and plant extract is expected to become a safer and more effective way for reducing uric acid.

Bonito belongs to low-value tuna, and a large amount of processing byproducts such as viscera, dark meat, fish skin, fish bones and the like are generated in the processing production process, and account for about 50-70% of the weight. The annual processing amount of bonito in Zhoushan and Ningbo areas is in the front of China, the processing amount of the bonito is as high as 50 tons/day, the byproducts are generally processed into cheap feed, the utilization value is low, and a large amount of precious nutrients and functional components are wasted. Because the types and the contents of amino acids or functional polypeptides contained in different parts of bonito are different, the effects are also different, at present, active substances which can inhibit XOD activity and reduce uric acid level are extracted from bonito processing byproducts, and no research report that the effect of efficiently reducing uric acid is realized by matching the extracted products with traditional Chinese medicine components is found.

Disclosure of Invention

The invention aims to provide a composition capable of reducing uric acid, which is prepared by compounding active peptide extracted from skipjack processing byproducts with plant extracts and combining the active peptide and the plant extracts from various aspects to achieve the aim of reducing uric acid level, and simultaneously, develops a new path for utilizing the skipjack processing byproducts and improves the utilization value of the skipjack processing byproducts.

The invention provides the following technical scheme:

the composition capable of reducing uric acid comprises the following components in parts by weight:

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 3-8 parts of xanthine oxidase inhibitory peptide;

the xanthine oxidase inhibitory peptide is extracted from a by-product of bonito processing by enzymolysis.

The invention reasonably compounds the polypeptide extracted from skipjack processing byproducts and plant extracts, and achieves the effect of reducing uric acid through synergistic interaction, and specifically comprises the following steps:

obtaining xanthine oxidase inhibitory peptide with XOD inhibitory activity by enzymolysis of bonito processing byproduct with specific enzyme, and reducing XOD activity, wherein the enzyme can be selected from flavourzyme, trypsin, papain, neutral protease, bromelain, alkaline protease, and aquatic protease. On the basis, the chicory extract is bitter in taste and salty in traditional Chinese medicine property, is generally used for inducing diuresis to reduce edema, clearing heat and removing toxicity, and modern pharmacological research finds that the chicory extract has an obvious curative effect on hyperuricemia, inhibits the intake of purine nucleosides in the intestinal tract by inhibiting the expression of an intestinal transport protein CNT2 to reduce the uric acid level, can increase the clearance rate of renal uric acid, reduces the protein expression of a hyperuricemia high-risk factor renal Glut9, and inhibits the renal uric acid reabsorption so as to promote the renal uric acid excretion to achieve the purpose of reducing uric acid, and chlorogenic acid, chicoric acid, aesculin and chicory in the chicory have close relation with the uric acid reducing effect; coix seed has the effects of inducing diuresis, removing dampness, and dispelling wind. Clinical pharmacology concludes that the coix seed has good effects of clearing damp and purging turbidity, is a good adjuvant drug, can assist in promoting renal excretion and reducing the content of uric acid in a body, has the effects of influencing the activities of DNA (deoxyribonucleic acid) combined transcription factors and cysteine type endopeptidases at a molecular level, and ferulic acid, coumaric acid, chlorogenic acid and the like in the coix seed can be combined with XOD (X-ray diffraction) to inhibit the activity of the XOD. The chlorogenic acid in the sunflower disc extract further enhances the effect of the coix seeds. The alkaloid contained in the sunflower disc can be specifically combined with uric acid to form a sunflower alkali-uric acid compound, so that uric acid is eliminated; the flavone in the sunflower disc can effectively remove oxygen free radicals in vivo, prevent nucleic acid from being oxidized, reduce purine content, simultaneously reduce blood sugar and monoglyceride, reduce capillary fragility, and increase permeability to relieve renal cell pressure and permeate more uric acid; the total phenol and flavone components in the galangal extract and the polygonatum extract can inhibit the activity of XOD, simultaneously reduce the expression of URAT1 and GLUT9 proteins, reduce the urea nitrogen and creatinine level and have the obvious effect of reducing uric acid.

Therefore, through the synergistic cooperation of the components from multiple surfaces, the effects of reducing creatinine and uric acid nitrogen, inhibiting XOD activity, discharging uric acid, blocking excessive production of uric acid and the like are achieved, the uric acid level is reduced, and further experiments prove that the composition can also inhibit renal tubular dilatation, relieve the infiltration of renal interstitial inflammatory cells, achieve the kidney protection effect, simultaneously develop a new way for utilizing skipjack processing byproducts, and improve the utilization value of the skipjack processing byproducts.

The plant extracts used in the composition are water extracts or alcohol extracts of the plants, and are obtained by water or ethanol extraction, concentration and spray drying, and can be purchased or made by self. The self-made method is, for example, a water extraction method, and the preparation method comprises the steps of cleaning and crushing raw materials, sieving the raw materials by a 100-mesh sieve, mixing the raw materials with water according to the mass ratio of 1.

As a preferred aspect of the present invention, xanthine oxidase inhibitory peptides are prepared as follows:

adjusting pH and temperature of slurry of bonito by-product, adding trypsin or aquatic protease for enzymolysis, separating clear liquid, and separating and drying the clear liquid to obtain xanthine oxidase inhibitory peptide.

At present, flavourzyme and the like are mostly adopted to carry out enzymolysis on bonito and fish meat to prepare XOD inhibitory peptide, so that the XOD inhibitory peptide has a good enzymolysis effect, and the XOD inhibitory rate of an obtained enzymolysis product is high. However, the by-products of bonito processing are derived from viscera, dark flesh, skin, bones, etc., and the components are more complicated than those of bonito meat, and the types and contents of amino acids and active peptides contained therein are greatly different. In screening experiments, the XOD inhibition rate of the obtained polypeptide product is not high when the skipjack processing byproducts are treated by flavourzyme and the like. And the animal protease is adopted to treat the bonito processing by-product, although the hydrolysis degree is very high, the small molecule content in the enzymolysis product is high, the obtained polypeptide does not have XOD inhibitory activity, which shows that the biological activity type and capability of the polypeptide product have relevance to the enzyme used. After analysis, the polypeptide XOD inhibition rate is high when the bonito processing by-products are treated by trypsin and aquatic protease, wherein the trypsin is preferred. Therefore, the polypeptide prepared by trypsin or aquatic protease is selected to be compounded with the plant extract.

Preferably, the pH range of the enzymolysis is 6.3-8.2, and the enzymolysis temperature range is 40-55 ℃.

Preferably, the method further comprises subjecting the slurry obtained as a by-product of bonito processing to preliminary enzymatic hydrolysis with an animal protease, and then adding trypsin or an aquatic protease to the slurry for enzymatic hydrolysis.

In further research, animal protease and trypsin are used together, and the obtained polypeptide product can further improve the uric acid reducing activity of the composition. The aquatic protease and the animal protein have similar hydrolysis degrees, the XOD inhibitory activity of a corresponding polypeptide product of the aquatic protease is higher, but the uric acid level is not obviously reduced and improved after the aquatic protein is matched with the trypsin, which is probably because certain synergistic matching exists among different proteases, the spatial structures of the protein and the polypeptide are complex, and the high hydrolysis degree of the animal protease is utilized to decompose a bonito processing byproduct protein to obtain more peptide fragments with small molecular weight, so that more trypsin treatment sites are exposed, and the effect is better after the trypsin treatment is used; on the other hand, there is a synergistic effect between the prepared polypeptide component and the plant extract.

Preferably, the enzyme is added in an amount of 2 to 5wt% based on the mass of the slurry.

In the present invention, the slurry preferably contains a by-product of bonito processing in an amount of 15 to 40wt%.

As a preferable aspect of the present invention, the by-product of bonito processing is one or more of viscera, dark flesh, skin, and bone produced by bonito processing.

The composition is applied to the preparation of medicines, health products or foods for reducing uric acid or inhibiting xanthine oxidase activity.

The composition can be used for preparing medicines, health products or foods for inhibiting renal tubule dilatation.

The composition is applied to preparing medicines, health products or foods for reducing infiltration of renal interstitial inflammatory cells.

The composition of the invention has the functions of inhibiting tubular dilatation and reducing infiltration of renal interstitial inflammatory cells.

The invention has the following beneficial effects:

in the composition, the polypeptide extracted from the skipjack processing by-product is reasonably compounded with plant extracts such as a chicory extract, a coix seed powder, a sunflower disc extract, a galangal extract, a polygonatum odoratum extract and the like, so that the composition has a higher XOD activity inhibition effect, can reduce creatinine and urea nitrogen levels, realizes reduction of hematuria water, can inhibit renal tubular dilation and reduce renal interstitial inflammatory cell infiltration, and has a kidney protection effect. Meanwhile, the skipjack processing by-product is utilized by preparing XOD inhibitory activity polypeptide, so that a new way for utilizing the skipjack processing by-product is developed, and the utilization value of the skipjack processing by-product is improved.

Drawings

FIG. 1 is a graph showing the ratio of the nutrient components of a by-product of bonito processing used in the present application.

FIG. 2 shows the degree of hydrolysis (A) and the solids content (B) of the enzymatic products of the different proteases of example 1.

FIG. 3 is a graph of the in vitro XOD inhibition of the enzymatic products of the different proteases of example 1.

FIG. 4 is a graph comparing the serum uric acid content (A), the plasma creatinine content (B), the blood urea nitrogen content (C), and the XOD activity (D) of different groups of mice in example 3.

FIG. 5 is a kidney tissue section of the mouse normal group (A), model group (B), positive group (C), high dose group (D), and medium dose group (E).

Detailed Description

The following further describes the embodiments of the present invention.

The starting materials used in the present invention are commercially available or commonly used in the art unless otherwise specified, and the methods in the following examples are conventional in the art unless otherwise specified.

The by-product of bonito processing used in the present invention is composed of viscera, dark flesh, skin, and bones produced during bonito processing, and is provided by the Shijiang industry group.

The present inventors analyzed the nutrient content and amino acid composition of bonito processing by-products as follows.

The nutrient components are as follows: the water was dried by heating at normal pressure, the protein was subjected to a semi-trace kjeldahl method, the fat was subjected to a soxhlet extraction method, the ash was subjected to an ashing method, and the total sugar was subjected to a fielin's volumetric method, the results being shown in fig. 1.

As can be seen from fig. 1, the skipjack processing by-product was rich in protein and relatively high in fat. The total content of the nutrient components exceeds 100% due to different measurement methods, and falls within the range of normal experimental error.

Amino acid composition: taking 100g of a sample of bonito processing byproducts, adding 6mol/L HCl, charging nitrogen, performing hot melt sealing, hydrolyzing in an oven at 110 ℃ for 24h, deacidifying, fixing the volume of water to an appropriate amount, and measuring with an amino acid automatic analyzer, wherein the results are shown in Table 1.

TABLE 1 skipjack processing by-product amino acid composition (g/100 g)

Note: tryptophan is destroyed under strong acid, so the results do not include tryptophan. * Represents essential amino acids.

As can be seen from the table, 16 amino acids were detected from the skipjack processing by-products (see table 1), and 7 essential amino acids were detected. The most abundant amino acids were glutamic acid (2.37%) and aspartic acid (1.89%) associated with umami taste, and in addition, lysine (1.75%) was also high, and the least abundant amino acid was methionine (0.56%). The total amount of amino acids as a by-product of bonito processing was 19.5% by mass of the sample, wherein the ratios of essential amino acids/total amino acids and essential amino acids/non-essential amino acids were 0.43 and 0.76, respectively, which were higher than FAO/WHO recommended values of 0.40 and 0.60, indicating that the protein as a by-product of bonito processing was a high-quality protein.

The following examples and comparative examples each used bonito processing by-products from the above sources.

Example 1 screening of proteases

1) Selecting papain and flavourzyme (Zhejiang Yinuo Biotechnology Co., ltd.) respectively; trypsin, neutral protease (hengshao biochemical engineering limited, hennan); bromelain, alcalase, pepsin (Hua, shinkawa industries, ltd, biochemicals); animal protease and aquatic protease (Nanning Donghuangtong Huadao biological Co.) are used for enzymolysis of bonito processing by-products under respective recommended optimum pH and temperature conditions, and specific parameters are shown in Table 2.

TABLE 2 enzymatic conditions for different proteases

The enzymolysis process of the protease comprises the following steps:

mixing the skipjack processing by-product with water, homogenizing to obtain slurry, adjusting pH and temperature of protease, maintaining for 30min, adding corresponding protease for enzymolysis, centrifuging the final stage enzymolysis solution at 10000r/min for 15min, discarding the upper oil film and the bottom residue, retaining the middle clear solution, and spray drying.

2) Measurement of inhibition ratio of Xanthine Oxidase (XOD) Activity

(1) Preparing a solution: preparation of 0.2M, pH 7.5.5 Phosphate Buffered Saline (PBS): weighing 4.56g of dipotassium hydrogen phosphate and 2.71g of monopotassium phosphate, respectively fixing the volume to 100mL of distilled water, and uniformly mixing with a water solution of 80;

1.5mM xanthine solution preparation: weighing 0.0228g of xanthine powder, dissolving the xanthine powder with trace 4mol/L of NaOH, and then diluting to 100mL with buffer solution for later use;

0.0125U/mL XOD solution preparation: 5U of stock solution, taking 400 mu L of stock solution, and diluting to 50mL with buffer solution for later use;

(2) Determination of XOD inhibition:

adding 40 μ L of 0.0125U/mL XOD solution and 50 μ L of polypeptide clear solution into a reaction tube, shaking uniformly, incubating at 25 deg.C for 10min, adding 50 μ L of 1.5mM xanthine solution, shaking uniformly, incubating at 25 deg.C for 30min, and measuring the absorbance OD at 290 nm.

The XOD inhibition rate is calculated according to the following formula:

wherein, the test group is a group added with XOD solution and polypeptide clear liquid; the test control group uses 40 mu LPBS to replace 40 mu LXOD solution; blank group 50 u L PBS instead of 50 u L polypeptide clear solution; blank controls 50. Mu.L of polypeptide supernatant was replaced with 50. Mu.L of LPBS and 40. Mu.L of LPBS was replaced with 40. Mu.L of LXOD solution.

Allopurinol (2 mug/mL) is used as a positive control in the test, and each sample is simultaneously subjected to 3 parallel tests, and the results are shown in the attached figure 2 in the specification.

As can be seen from fig. 2, animal protease, trypsin, aquatic protease, and the like have a high degree of hydrolysis, and complete hydrolysis of bonito processing by-products is achieved, resulting in a low molecular weight polypeptide product. According to the indication of the existing research results, the XOD inhibits the polysaccharide with low molecular weight, and the enzymolysis products of the three enzymes are predicted to have higher XOD inhibitory activity.

However, as can be seen from fig. 3, the inhibition ratio of the obtained bonito by-product after hydrolysis with different proteases was the strongest (51.64%) compared to the inhibition ratio of positive control allopurinol (2 μ g/mL) by XOD (27.66%), and the second is the production of aquatic product protease, which is the hydrolysate of animal protease and pepsin, which has no inhibition effect on XOD. The reason may be that trypsin can break peptide bonds formed by Arg and Lys carboxyl groups, and the specific cation and hydrogen bond characteristics of the arginine-rich peptide are favorable for combining with Flavin Adenine Dinucleotide (FAD) in 6 combining sites of the XOD crystal structure to form a non-competitive and irreversible inhibition effect, so that the XOD activity is inhibited, while the skipjack processing by-product of the application has abundant arginine.

Example 2 inhibitory Effect of the composition on uric acid-lowering and XOD Activity

1) Polypeptide (I-VIII) obtained by enzymolysis of skipjack processing byproducts by different methods is respectively compounded with plant extracts to obtain different compositions (I-VIII), wherein the compositions comprise the following components in parts by weight:

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

polypeptide I: the polypeptide product prepared by trypsin in example 1;

polypeptide II: the polypeptide product produced by the protease in water in example 1;

polypeptide III: the polypeptide product produced by the animal protease of example 1;

polypeptide IV: different from the polypeptide I obtained in the embodiment 1, the animal protease is firstly added into the slurry for enzymolysis, and the polypeptide product is obtained by adding the trypsin for continuous enzymolysis after heating and inactivation;

a polypeptide V: different from the polypeptide I obtained in the embodiment 1, firstly adding trypsin into the slurry for enzymolysis, heating and inactivating, and then adding animal protein for continuous enzymolysis to obtain a polypeptide product;

polypeptide VI: different from the polypeptide I obtained in the example 1, animal protease and trypsin are added into the slurry for enzymolysis; polypeptide VII: different from the polypeptide II obtained in the embodiment 1, the animal protease is firstly added into the slurry for enzymolysis, and the aquatic protease is added after heating and inactivation for continuous enzymolysis to obtain a polypeptide product;

a polypeptide VIII: different from the polypeptide I obtained in the embodiment 1I, the polypeptide I is obtained by firstly adding water into the serous fluid for enzymolysis by protease, heating and inactivating the serous fluid, then adding trypsin for continuous enzymolysis to obtain a polypeptide product;

all extracts are extracted with water;

2) Uric acid lowering and XOD inhibition performance test

Mice (8 weeks old) were randomly divided into 14 groups, normal (Control), model (MG), positive (PC), experimental I-VIII, and Control I-III. Except for the normal group of gavage distilled water, other groups are respectively gavage adenine and ethambutol hydrochloride for molding, the gavage amount is 1g/kg (1 time per day), when the treatment is continuously carried out until the uric acid content reaches 140mol/L, the positive group is gavage positive medicament allopurinol tablets (Guangdong Pedy pharmaceutical industry Co., ltd.), the gavage amount is 10mg/kg, the experimental groups I to VIII are respectively gavage corresponding compositions I to VIII, the control groups are respectively gavage corresponding controls I to III, the gavage amount is 500mg/kg (middle dose), the gavage 1h is followed by continuous gavage of adenine and ethambutol hydrochloride, after 3 days of continuous execution, the eyeballs of the mice are blood-sampled, and the hematuria level and XOD activity are tested by using a kit (Nanjing institute of Biotechnology), and the results are shown in Table 2.

Wherein the composition used in the control group I is pure plant extract, and the composition used in the control group II is prepared by adding 12 parts of radix Puerariae powder into the composition I; the composition used in control group III was prepared by replacing galangal rhizome extract in composition I with 12 parts of kudzu root powder.

TABLE 2 XOD Activity and blood uric acid levels of the groups

As can be seen from the above table, compared with the normal group, the liver xanthine oxidase activity of the mice in the molding group is obviously increased, the formation of uric acid in the bodies of the mice is accelerated, and the molding is successful. By analyzing the XOD activity level and the blood uric acid level of each experimental group, the control group and the positive group, it can be found that the lower the XOD activity level is, the lower the blood uric acid level is in most cases, but there are cases where the XOD level is low and the blood uric acid level is conversely 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 decrease level and the blood uric acid decrease level are not always synchronized because the blood uric acid level is influenced by various factors, and the XOD activity is only one of the factors.

As can be seen by comparing the experimental groups I to VIII with the control group I, the composition obtained by compounding the plant extract with the polypeptide hydrolyzed by the trypsin and the aquatic protease can further reduce the XOD activity and the level of the hematuric acid, but the improvement of the compounding effect of the polypeptide enzymolysis product after the aquatic protease is hydrolyzed is relatively weak. In addition, two different enzymes are adopted to prepare the polypeptide, so that the hydrolysis effect of animal protease is optimal firstly, and the hydrolysis effect of trypsin is optimal later, which is superior to that of the single use of the trypsin. This is probably due to the fact that the animal protein and trypsin have a higher complexing effect on the enzymolysis of bonito processing by-products on the one hand, and on the other hand, the reduction of serum uric acid is influenced by various factors, for example, in experimental group VIII, the inhibition effect of XOD activity is the best after the enzymolysis of aquatic product proteolysis and then the enzymolysis of trypsin, but the serum uric acid level is still higher than that in experimental group IV, and the complexing effect of polypeptide enzymolysis products and plant extracts needs to be considered. However, as can be seen from the comparison between the experimental group I and the control groups II to III, the selection of the components of the plant extract in the composition also affects the overall effect, for example, after the pueraria powder is additionally introduced into the control group II, the overall effect is not significantly changed, and for the control group III, after the pueraria powder is used to replace the galangal extract, although the pueraria powder also contains flavone, the overall effect is reduced to some extent, which reflects the coordination effect inside the composition.

In a whole, under the condition of a determined plant extract, polypeptide compounding prepared by enzymolysis of trypsin or aquatic protease is selected, so that the uric acid level can be further reduced, the economy is realized, and the action effect can be further obviously improved by adopting the polypeptide compounding prepared by enzymolysis of animal protease and trypsin.

EXAMPLE 3 dose Effect of composition I

On the basis of the experiment of the mice in the example 2, the high-dose composition I is further used for intragastric administration of the model mice, the intragastric administration amount is 1000mg/kg, and the high-dose group is compared with the medium-dose group corresponding to the experimental group I in the example 2.

1) Blood uric acid, XOD activity and creatinine, uric acid nitrogen levels:

blood is taken from eyeballs, and the kit (Nanjing institute of bioengineering) is used for testing blood uric acid, XOD activity, creatinine and urea nitrogen level, and the result is shown in figure 4.

As shown in FIG. 4 (A), the serum uric acid content of the molding group is significantly higher than that of the normal group, indicating that the molding is successful. Compared with the model group, the serum uric acid levels of the positive control and the experimental group are obviously reduced and are lower than those of the normal group, which shows that allopurinol and high-dose gout Shang Hezhong dose gout Shang Junke obviously reduce the serum uric acid of the rat. The gout decoction in the high-dose and medium-dose groups has a dose negative correlation trend on the blood uric acid level of mice, namely the higher the dose level of the gout decoction is, the lower the blood uric acid content is, and the better the treatment effect is. In addition, the serum uric acid level of the mice in the high-dose gout decoction is lower than that in the positive group, which shows that the regulation effect of the high-dose formula on the serum uric acid of the mice in the model is better than that of allopurinol, and the medium-dose gout decoction is equivalent to that of allopurinol.

As shown in FIG. 4 (B), gavage of adenine and ethambutol hydrochloride significantly increased plasma creatinine levels in mice compared to the normal group. The positive group and the experimental group can obviously reduce the plasma urea nitrogen level of mice. In the experimental group, the plasma creatinine reduction capability of the mice is positively correlated with the dosage of the formula, and the plasma creatinine reduction capability is stronger when the dosage of the formula is high. Meanwhile, the urea nitrogen level of mice treated by the two groups of formulas is lower than that of the other purine treatment group, which shows that the regulation capability of the formulas on mouse plasma creatinine is better than that of the other purine in the model.

As shown in FIG. 4 (C), blood urea nitrogen levels of mice that were gastroplasted with adenine and ethambutol hydrochloride were elevated as compared with the normal group, indicating that the glomerular filtration ability of the mice was impaired. The blood urea nitrogen level of the mice treated with the allopurin group was slightly elevated compared to the level of the adult group, indicating that allopurin could not regulate the blood urea nitrogen level of the mice. Both the high and middle dose groups of the formula 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 by the experimental group is still higher, the irreversible damage to the glomerular filtration capability of the mice is probably caused, and the formula can be treated to a certain extent but cannot completely recover the functions.

As shown in FIG. 4 (D), the liver xanthine oxidase activity of the mice in the model-making group was significantly increased, and the formation of uric acid in the mice was accelerated, as compared with the normal group. Compared with the model-making group, the activities of the xanthine oxidase in the livers of the mice in the positive group and the experimental group are reduced, and the level of the activity of the gout decoction in the two groups with high and medium doses for reducing XOD is higher than that of the purine group, which shows that the inhibition capability of the formula in the model on the XOD activity in the livers of the mice is better than that of the purine group, and the inhibition effect and the dose relation are not obvious.

2) Pathological change of kidney tissue of mouse

The mice of example 3 were sacrificed, the kidneys were removed and sectioned, and the results of observation after staining are shown in FIG. 5.

As shown in fig. 5 (a), the glomerular and tubular structures of the normal group of mice were normal; the kidney in the normal group is characterized in that the cytoplasm of renal tubular epithelial cells is rich, the cell nucleus is large and loose, the tubular cavity is small, the interstitial area is small, the canals and the tubules are back to back, interstitial broadening is not seen, but obvious inflammatory cell infiltration exists in the interstitium.

As shown in fig. 5 (B), the model group showed a large amount of apoptosis, necrosis, shedding or vacuolar degeneration of Tubular Epithelial Cells (TEC), coexistence of highly dilated tubules and atrophic tubules, large amount of golden yellow purine metabolite crystal deposition in the tubules and renal interstitium, and protein casts in the tubules. The renal interstitium is heavily fibrotic and hyperplastic, the renal interstitium can be infiltrated by a large amount of inflammatory cells, and the inflammatory cells wrap purine metabolite crystals. The renal capsule cavity expands and the renal tubular mesangial cells slightly proliferate. The renal cortex becomes thin, the renal tubular epithelial cells are atrophied, the tubular cavity is expanded, the distal tubule and the collecting duct are saccular expansion, the renal interstitium is obviously widened in most visual fields, and obvious fibroblast proliferation and fibrosis are seen in the interstitium.

As shown in FIG. 5 (C), the positive group had tubular dilatation, small amount of crystallization of golden yellow purine metabolites, slight vacuolar denaturation of TEC, mild fibrosis of renal interstitium, small amount of inflammatory cell infiltration, renal vesicle cavity dilatation, and mild proliferation of mesangial cells.

As shown in fig. 5 (D), the high dose group had mild tubular dilation, few moderate dilations, mild fibrosis of the renal interstitium, a small amount of golden yellow purine metabolite crystals in the renal interstitium, mild dilatation of the renal cystic space, and insignificant proliferation of mesangial cells.

As shown in fig. 5 (E), the medium dose group has moderate dilation of renal tubules, and renal tubular atrophy, TEC necrosis, shedding, small amount of golden yellow purine metabolite crystal deposition in renal interstitium, interstitial moderate fibrosis with small amount of inflammatory cell infiltration, dilatation of renal glomerular bursa cavity, and mild proliferation of renal glomerular mesangial cells.

From the above results, it can be seen that the composition I can inhibit the dilation of renal tubules and reduce the infiltration of renal interstitial inflammatory cells, and has a certain kidney-protecting effect.

Claims (10)

1. The composition capable of reducing uric acid is characterized by comprising the following components in parts by weight:

20-30 parts of chicory extract, 9-15 parts of galangal extract, 10-15 parts of polygonatum extract, 10-20 parts of sunflower disc extract, 15-20 parts of coix seed powder and 3-8 parts of xanthine oxidase inhibitory peptide;

the xanthine oxidase inhibitory peptide is extracted from a by-product of bonito processing by enzymolysis.

2. The composition of claim 1, wherein the xanthine oxidase inhibitor peptide is prepared by the following steps:

adjusting pH and temperature of slurry of bonito by-product, adding trypsin or aquatic protease for enzymolysis, separating clear liquid, and separating and drying the clear liquid to obtain xanthine oxidase inhibitory peptide.

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

4. The composition of claim 2, further comprising subjecting a slurry obtained by producing bonito to preliminary digestion with an animal protease, and then adding trypsin or an aquatic protease to the slurry.

5. A composition according to claim 2 or 3 or 4, characterised in that the enzyme is added in an amount of 2 to 5% by weight of the mass of the slurry.

6. The composition as claimed in claim 2, wherein the slurry contains the by-product of bonito processing in an amount of 15 to 40wt%.

7. The composition as described in claim 2 or 6, wherein the by-product of bonito processing is one or more of viscera, dark flesh, skin and bone of bonito processing.

8. Use of the composition according to any one of claims 1 to 7 for the preparation of a medicament, health product or food for lowering uric acid or inhibiting xanthine oxidase activity.

9. Use of the composition according to any one of claims 1 to 7 for the preparation of a medicament, health product or food for inhibiting tubular dilatation.

10. Use of the composition according to any one of claims 1 to 7 for the preparation of a medicament, health product or food for reducing infiltration of renal interstitial inflammatory cells.

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