CN117882774A

CN117882774A - Composition with uric acid reducing effect and application thereof

Info

Publication number: CN117882774A
Application number: CN202410281949.7A
Authority: CN
Inventors: 毛学英; 许雅茹; 姜雨辰
Original assignee: Yantai Yuanlide Marine Biological Co ltd; China Agricultural University
Current assignee: Yantai Yuanlide Marine Biological Co ltd; China Agricultural University
Priority date: 2024-03-13
Filing date: 2024-03-13
Publication date: 2024-04-16
Anticipated expiration: 2044-03-13

Abstract

The invention relates to a composition with uric acid reducing effect and application thereof. Among them, the composition III with the best uric acid reducing effect is: 10 parts of sea cucumber peptide, 50 parts of celery extract, 460 parts of skim milk powder, 70 parts of chrysanthemum extract, 10 parts of chitosan oligosaccharide, 2 parts of luteolin and 1 part of quercetin. The invention provides scientific basis for the combined intervention and alleviation of hyperuricemia diseases of sea cucumber peptide, celery extract, skim milk powder, chrysanthemum extract, chitosan oligosaccharide, luteolin and quercetin, and the realization of functional food, special medical food, drug development and industrial production.

Description

Composition with uric acid reducing effect and application thereof

Technical Field

The invention belongs to the technical field of medical health products, and particularly relates to a composition with uric acid reducing effect and application thereof.

Background

Hyperuricemia is a chronic disease, which refers to a disease in which purine metabolism disorder or uric acid excretion decreases in the body, resulting in elevated uric acid in serum. According to the research of large epidemiology, hyperuricemia is one of common diseases in the modern society, is a risk factor of metabolic diseases such as gout, uric acid kidney stones, cardiovascular and cerebrovascular diseases (such as hypertension), diabetes, and the like, has very high incidence rate in middle-aged and elderly people, and the number of patients is greatly increased along with the adjustment of the dietary structure of people. In humans, elevated blood uric acid levels are associated with abnormal nucleic acid metabolism and reduced renal excretion in the body, and Xanthine Oxidase (XOD), a key enzyme regulating uric acid production, is widely present in a variety of organisms and can act to reduce serum uric acid in the body by catalyzing the oxidation of hypoxanthine to xanthine and further to uric acid, inhibiting the activity of XOD. Therefore, maintaining uric acid levels in human serum is extremely important for human health, and XOD can be a key target for the treatment of hyperuricemia.

The uric acid lowering drugs on the market at present have great side effects such as allergy, kidney function injury and the like, and are mainly aimed at gout patients, but have no good effect and safety on asymptomatic hyperuricemia patients, and can effectively prevent and treat hyperuricemia. The celery extract is a natural plant component extracted from fresh celery, and has rich antioxidants and nutrients. The main components of the food comprise flavonoid compounds, vitamins, minerals, cellulose and the like. The celery extract has wide application prospect in the fields of health products, foods and the like, and has the effects of resisting aging, resisting inflammation, reducing blood pressure, improving digestion and the like. Celery extracts are attracting attention in the health field due to their natural ingredients and multifunctional properties. Sea cucumber can be used for continuing aging, eliminating fatigue, improving immunity, enhancing disease resistance, is rich in more than 50 natural precious active substances such as proteins, minerals, vitamins and the like, and 18 amino acids contained in sea cucumber can be used for enhancing the metabolic function of tissues, enhancing the activity of organism cells and improving the immunity of human bodies. Therefore, the celery extract and the sea cucumber peptide are combined, so that the composition has both functionality and nutrition, and has great market prospect as an effective and safe daily drink for treating hyperuricemia.

Disclosure of Invention

In order to overcome the defects in the prior art, the invention provides a composition with uric acid reducing effect and application thereof.

In one aspect, the invention provides a composition for relieving hyperuricemia, which consists of sea cucumber peptide, celery extract, skim milk powder, chrysanthemum extract, chitosan oligosaccharide, luteolin and quercetin;

the weight parts of the sea cucumber peptide are within the range of 10-30;

the weight part of the celery extract is within the range of 10-50;

the weight parts of the skim milk powder are in the range of 350-460;

the weight part of the chrysanthemum extract is in the range of 70-100;

the weight part of the chitosan oligosaccharide is within the range of 10-50;

the weight part of luteolin is in the range of 2-5;

the content of luteolin in luteolin is 90% -95%;

the content of quercetin in the luteolin is 95% -98%.

The weight part of the quercetin is in the range of 1-2;

the weight ratio of the preferred composition is 10:50:460:70:10:2:1.

The sea cucumber peptide is prepared by the following steps: (1) Homogenizing sea cucumber flesh to obtain sea cucumber homogenate, and shearing and homogenizing the sea cucumber homogenate to obtain sea cucumber homogenate; (2) Regulating the pH value of the sea cucumber homogenized liquid to 7.0, and freeze-drying the obtained sea cucumber homogenized liquid to obtain freeze-dried powder for later use; (3) Preparing a sea cucumber freeze-dried powder into a 5% solution, regulating the pH value to 8.0, adding alkaline protease, compound protease and papain at 55 ℃ for enzymolysis, wherein the total enzyme activity of the three proteases is 3000U, the adding ratio of the alkaline protease to the compound protease to the papain is 1:1:1, and the sea cucumber proteolytic liquid is obtained after enzymolysis for 1 h; (4) Regulating the pH value of the obtained sea cucumber protein enzymolysis liquid to 7.0, and freeze-drying the enzymolysis liquid to obtain sea cucumber peptide powder; and (5) separating, purifying and identifying the sequence to determine the sea cucumber peptide.

The sea cucumber peptide sequence comprises EKFPPPM (SEQ ID NO. 1), PPLVKPW (SEQ ID NO. 2) and KDLGGI (SEQ ID NO. 3).

The invention adopts uric acid, creatinine and urea nitrogen level analysis, and proves that the composition can relieve hyperuricemia and renal function injury. The test results show that the composition relieves the symptoms of patients with hyperuricemia by changing the relative abundance of the flora in the intestinal tract. The composition can change intestinal flora and intestinal flora metabolites of rats.

In another aspect, the present invention provides a composition for alleviating hyperuricemia, which is synergistic by the synergistic effect of sea cucumber peptide and celery extract.

The optimal proportion of the synergistic effect of the sea cucumber peptide and the celery extract is 1:5.

On the other hand, the invention provides the application of the two compositions in preparing products for relieving hyperuricemia and the application thereof in preparing functional formulas for lowering uric acid of middle-aged and elderly people, foods for special medical purposes and medicines.

The hyperuricemia is caused by a disorder of purine metabolism or uric acid excretion itself.

Compared with the prior art, the invention has the beneficial effects that:

according to the invention, celery extract, sea cucumber peptide, chrysanthemum extract, chitosan oligosaccharide, luteolin and quercetin are compounded, mixed for 15-30 min in a double-helix conical mixer, so that a solid beverage with uric acid reducing activity is obtained, and the uric acid reducing activity of the composition is determined by effectively inhibiting xanthine oxidase activity. The results show that the celery extract and the sea cucumber peptide can synergistically exert effects, and 10 parts by weight of the sea cucumber peptide, 50 parts by weight of the celery extract, 460 parts by weight of skim milk powder, 70 parts by weight of the chrysanthemum extract, 10 parts by weight of chitosan oligosaccharide, 2 parts by weight of luteolin and 1 part by weight of quercetin are the optimal proportion of the composition.

Drawings

FIG. 1 shows the results of uric acid reduction and renal function impairment in rats from the compositions. A is uric acid lowering result; b is serum creatinine, C is urea nitrogen.

FIG. 2 is a graph showing the effect of the composition on rat serum and liver XOD activity and ADA activity. A is serum XOD activity; b is serum ADA activity; c is liver XOD activity; d is liver ADA activity.

FIG. 3 is a photograph of kidney pathology tissue of rats with composition III.

FIG. 4 shows the effect of composition III on the alpha-diversity of the intestinal flora of rats (A) Shannon index (B) Chao index (C) Simpson index (D) Ace index.

FIG. 5 includes a Venn plot of composition III (A) versus HUA rat intestinal flora, and the number of OUT (B) from different treatment groups.

FIG. 6 is a graph showing the effect of composition III on the composition of intestinal flora at the portal level in rats, wherein (A) is the relative abundance of the flora at the portal level for the different treatment groups and (B) the relative abundance of the different flora in the treatment groups.

FIG. 7 is a graph showing the effect of composition III on the composition of the intestinal flora of rats at the family level, wherein (A) is the relative abundance of the flora at the family level for the different treatment groups and (B) is the relative abundance of the different flora in the treatment groups.

FIG. 8 is a graph showing the effect of composition III on the composition of intestinal flora on the genus level for rats, wherein (A) is the relative abundance of the flora on the genus level for the different treatment groups and (B) the relative abundance of the different flora in the treatment groups.

FIG. 9 is a PCoA analysis of HUA rat intestinal flora.

FIG. 10 shows NMDS analysis of HUA rat intestinal flora.

FIG. 11 is a graph of composition III versus short chain fatty acid content of rats, wherein (A) is acetic acid, (B) is propionic acid, (C) is butyric acid, (D) is isobutyric acid, and (E) is valeric acid.

Detailed Description

The objects and functions of the present invention and methods for achieving these objects and functions will be elucidated with reference to exemplary comparative examples and embodiments. However, the present invention is not limited to the exemplary embodiments disclosed below; this may be implemented in different forms. The essence of the description is merely to aid one skilled in the relevant art in comprehensively understanding the specific details of the invention. Hereinafter, the weight portion refers to the concentration of the same weight component, and 1 weight portion is 10 mg.

The formula of the composition with uric acid reducing effect is as follows:

the composition I comprises the following raw materials in parts by weight: 10 parts of sea cucumber peptide, 10 parts of celery extract, 460 parts of skim milk powder, 70 parts of chrysanthemum extract, 10 parts of chitosan oligosaccharide, 2 parts of luteolin and 1 part of quercetin.

The composition II comprises the following raw materials in parts by weight: 10 parts of sea cucumber peptide, 30 parts of celery extract, 460 parts of skim milk powder, 70 parts of chrysanthemum extract, 10 parts of chitosan oligosaccharide, 2 parts of luteolin and 1 part of quercetin.

The composition III comprises the following raw materials in parts by weight: 10 parts of sea cucumber peptide, 50 parts of celery extract, 460 parts of skim milk powder, 70 parts of chrysanthemum extract, 10 parts of chitosan oligosaccharide, 2 parts of luteolin and 1 part of quercetin.

The composition IV comprises the following raw materials in parts by weight: 20 parts of sea cucumber peptide, 50 parts of celery extract, 460 parts of skim milk powder, 70 parts of chrysanthemum extract, 10 parts of chitosan oligosaccharide, 2 parts of luteolin and 1 part of quercetin.

The composition V comprises the following raw materials in parts by weight: 30 parts of sea cucumber peptide, 50 parts of celery extract, 460 parts of skim milk powder, 70 parts of chrysanthemum extract, 10 parts of chitosan oligosaccharide, 2 parts of luteolin and 1 part of quercetin.

The raw materials of the composition VI comprise the following components in parts by weight: 10 parts of sea cucumber peptide, 50 parts of celery extract, 350 parts of skim milk powder, 70 parts of chrysanthemum extract, 10 parts of chitosan oligosaccharide, 2 parts of luteolin and 1 part of quercetin.

The raw materials of the composition VII comprise the following components in parts by weight: 10 parts of sea cucumber peptide, 50 parts of celery extract, 460 parts of skim milk powder, 100 parts of chrysanthemum extract, 10 parts of chitosan oligosaccharide, 2 parts of luteolin and 1 part of quercetin.

The composition VIII comprises the following raw materials in parts by weight: 10 parts of sea cucumber peptide, 50 parts of celery extract, 460 parts of skim milk powder, 70 parts of chrysanthemum extract, 50 parts of chitosan oligosaccharide, 2 parts of luteolin and 1 part of quercetin.

The composition IX comprises the following raw materials in parts by weight: 10 parts of sea cucumber peptide, 50 parts of celery extract, 460 parts of skim milk powder, 70 parts of chrysanthemum extract, 10 parts of chitosan oligosaccharide, 5 parts of luteolin and 1 part of quercetin.

The composition X comprises the following raw materials in parts by weight: 10 parts of sea cucumber peptide, 50 parts of celery extract, 460 parts of skim milk powder, 70 parts of chrysanthemum extract, 10 parts of chitosan oligosaccharide, 2 parts of luteolin and 2 parts of quercetin.

Mixing sea cucumber peptide, celery extract, skim milk powder, chrysanthemum extract, chitosan oligosaccharide, luteolin and quercetin in a double-helix conical mixer for 15-30 min to obtain solid beverage with uric acid reducing activity.

Specific comparisons are shown below:

comparative example 1: 10 parts of sea cucumber peptide, 460 parts of skim milk powder, 70 parts of chrysanthemum extract, 10 parts of chitosan oligosaccharide, 2 parts of luteolin and 1 part of quercetin;

comparative example 2: 50 parts of celery extract, 460 parts of skim milk powder, 70 parts of chrysanthemum extract, 10 parts of chitosan oligosaccharide, 2 parts of luteolin and 1 part of quercetin;

comparative example 3: 10 parts of sea cucumber peptide, 50 parts of celery extract, 70 parts of chrysanthemum extract, 10 parts of chitosan oligosaccharide, 2 parts of luteolin and 1 part of quercetin;

comparative example 4: 10 parts of sea cucumber peptide, 50 parts of celery extract, 460 parts of skim milk powder, 10 parts of chitosan oligosaccharide, 2 parts of luteolin and 1 part of quercetin;

comparative example 5: 10 parts of sea cucumber peptide, 50 parts of celery extract, 460 parts of skim milk powder, 70 parts of chrysanthemum extract, 2 parts of luteolin and 1 part of quercetin;

comparative example 6: 10 parts of sea cucumber peptide, 50 parts of celery extract, 460 parts of skim milk powder, 70 parts of chrysanthemum extract, 10 parts of chitosan oligosaccharide and 1 part of quercetin;

comparative example 7: 10 parts of sea cucumber peptide, 50 parts of celery extract, 460 parts of skim milk powder, 70 parts of chrysanthemum extract, 10 parts of chitosan oligosaccharide and 2 parts of luteolin;

comparative example 8: 460 parts of skim milk powder, 70 parts of chrysanthemum extract, 10 parts of chitosan oligosaccharide, 2 parts of luteolin and 1 part of quercetin;

comparative example 9: 50 parts of celery extract, 70 parts of chrysanthemum extract, 10 parts of chitosan oligosaccharide, 2 parts of luteolin and 1 part of quercetin;

comparative example 10:50 parts of celery extract, 460 parts of skim milk powder, 10 parts of chitosan oligosaccharide, 2 parts of luteolin and 1 part of quercetin;

comparative example 11: 50 parts of celery extract, 460 parts of skim milk powder, 70 parts of chrysanthemum extract, 2 parts of luteolin and 1 part of quercetin;

comparative example 12: 50 parts of celery extract, 460 parts of skim milk powder, 70 parts of chrysanthemum extract, 10 parts of chitosan oligosaccharide and 1 part of quercetin;

comparative example 13: 50 parts of celery extract, 460 parts of skim milk powder, 70 parts of chrysanthemum extract, 10 parts of chitosan oligosaccharide and 2 parts of luteolin;

comparative example 14: 10 parts of sea cucumber peptide, 70 parts of chrysanthemum extract, 10 parts of chitosan oligosaccharide, 2 parts of luteolin and 1 part of quercetin;

comparative example 15: 10 parts of sea cucumber peptide, 460 parts of skim milk powder, 10 parts of chitosan oligosaccharide, 2 parts of luteolin and 1 part of quercetin;

comparative example 16: 10 parts of sea cucumber peptide, 460 parts of skim milk powder, 70 parts of chrysanthemum extract, 2 parts of luteolin and 1 part of quercetin;

comparative example 17: 10 parts of sea cucumber peptide, 460 parts of skim milk powder, 70 parts of chrysanthemum extract, 10 parts of chitosan oligosaccharide and 1 part of quercetin;

comparative example 18: 10 parts of sea cucumber peptide, 460 parts of skim milk powder, 70 parts of chrysanthemum extract, 10 parts of chitosan oligosaccharide and 2 parts of luteolin;

comparative example 19: 10 parts of sea cucumber peptide, 50 parts of celery extract, 10 parts of chitosan oligosaccharide, 2 parts of luteolin and 1 part of quercetin;

comparative example 20: 10 parts of sea cucumber peptide, 50 parts of celery extract, 70 parts of chrysanthemum extract, 2 parts of luteolin and 1 part of quercetin;

comparative example 21: 10 parts of sea cucumber peptide, 50 parts of celery extract, 70 parts of chrysanthemum extract, 10 parts of chitosan oligosaccharide and 1 part of quercetin;

comparative example 22: 10 parts of sea cucumber peptide, 50 parts of celery extract, 70 parts of chrysanthemum extract, 10 parts of chitosan oligosaccharide and 2 parts of luteolin;

comparative example 23: 10 parts of sea cucumber peptide, 50 parts of celery extract, 460 parts of skim milk powder, 2 parts of luteolin and 1 part of quercetin;

comparative example 24: 10 parts of sea cucumber peptide, 50 parts of celery extract, 460 parts of skim milk powder, 10 parts of chitosan oligosaccharide and 1 part of quercetin;

comparative example 25: 10 parts of sea cucumber peptide, 50 parts of celery extract, 460 parts of skim milk powder, 10 parts of chitosan oligosaccharide and 2 parts of luteolin;

comparative example 26: 10 parts of sea cucumber peptide, 50 parts of celery extract, 460 parts of skim milk powder, 70 parts of chrysanthemum extract and 1 part of quercetin;

comparative example 27: 10 parts of sea cucumber peptide, 50 parts of celery extract, 460 parts of skim milk powder, 70 parts of chrysanthemum extract and 2 parts of luteolin;

comparative example 28: 10 parts of sea cucumber peptide, 50 parts of celery extract, 460 parts of skim milk powder, 70 parts of chrysanthemum extract and 10 parts of chitosan oligosaccharide;

comparative example 29: 70 parts of chrysanthemum extract, 10 parts of chitosan oligosaccharide, 2 parts of luteolin and 1 part of quercetin;

comparative example 30: 460 parts of skim milk powder, 10 parts of chitosan oligosaccharide, 2 parts of luteolin and 1 part of quercetin;

comparative example 31: 460 parts of skim milk powder, 70 parts of chrysanthemum extract, 2 parts of luteolin and 1 part of quercetin;

comparative example 32: 460 parts of skim milk powder, 70 parts of chrysanthemum extract, 10 parts of chitosan oligosaccharide and 1 part of quercetin;

comparative example 33: 460 parts of skim milk powder, 70 parts of chrysanthemum extract, 10 parts of chitosan oligosaccharide and 2 parts of luteolin;

comparative example 34: 10 parts of chitosan oligosaccharide, 2 parts of luteolin and 1 part of quercetin;

comparative example 35: 70 parts of chrysanthemum extract, 2 parts of luteolin and 1 part of quercetin;

comparative example 36: 70 parts of chrysanthemum extract, 10 parts of chitosan oligosaccharide and 1 part of quercetin;

comparative example 37: 70 parts of chrysanthemum extract, 10 parts of chitosan oligosaccharide and 2 parts of luteolin;

comparative example 38: 2 parts of luteolin and 1 part of quercetin;

comparative example 39: 10 parts of chitosan oligosaccharide and 1 part of quercetin;

comparative example 40: 10 parts of chitosan oligosaccharide and 2 parts of luteolin;

comparative example 41: 1 part by weight of quercetin;

comparative example 42: 2 parts by weight of luteolin;

comparative example 43: 10 parts of sea cucumber peptide, 50 parts of honeysuckle extract, 460 parts of skim milk powder, 70 parts of chrysanthemum extract, 10 parts of chitosan oligosaccharide, 2 parts of luteolin and 1 part of quercetin;

comparative example 44: 10 parts of sea cucumber peptide, 50 parts of sour cherry powder, 460 parts of skim milk powder, 70 parts of chrysanthemum extract, 10 parts of chitosan oligosaccharide, 2 parts of luteolin and 1 part of quercetin;

comparative example 45: 10 parts of sea cucumber peptide, 50 parts of celery seed extract, 460 parts of skim milk powder, 70 parts of chrysanthemum extract, 10 parts of chitosan oligosaccharide, 2 parts of luteolin and 1 part of quercetin;

comparative example 46: 60 parts of celery extract, 460 parts of skim milk powder, 70 parts of chrysanthemum extract, 10 parts of chitosan oligosaccharide, 2 parts of luteolin and 1 part of quercetin.

EXAMPLE 1 screening of composition formulation raw materials

Uric Acid (UA) is mainly controlled by purine metabolism-related enzymes during in vivo production, where XOD is a key enzyme that regulates uric acid production. XOD has a primary function of catalyzing oxidation of hypoxanthine to xanthine and further oxidation to uric acid, and its substrates include purines, pyrimidines, pterins, aromatic heterocycles, certain aldehydes, and the like. Therefore, the production of uric acid can be inhibited by inhibiting the activity of XOD, thereby having the effect of reducing uric acid activity. The experiment adopts the method for measuring the inhibition rate of the XOD to characterize the uric acid reducing activity of the sample, and comprises the following specific steps: taking 250 mu L of sample solution to be detected and 250 mu L of XOD, adding the sample solution to a 2 mL centrifuge tube, incubating for 10 min at 37 ℃, adding 750 mu L of xanthine solution to start the reaction, continuing incubating for 15min at 37 ℃, and adding 400 mu L of 1M hydrochloric acid to stop the reaction after the reaction is finished. Positive and negative controls were made with 5. Mu.g/mL allopurinol and 50 mM Tris-HCl buffer, respectively. The sample was passed through a 0.22 μm PES membrane. HPLC conditions were column: c18 column (250X 4.6 mm, 5 μm); mobile phase: 85% NH 10 mM ₄ H ₂ PO ₄ An aqueous solution and 15% methanol; flow rate: 0.8 mL/min; UV:290 nm. A50. Mu.M uric acid solution was used as a standard to determine the retention time of uric acid. Quantitative calculations were performed with corresponding peak areas based on retention time.

P(XOD)=(c _Control -c _sample )/c _Control ×100%

According to the experimental method, the XOD inhibitory activity of single components of resveratrol, luteolin, quercetin, chitosan oligosaccharide, skim milk powder, honeysuckle extract, sour cherry powder, celery seed extract, celery extract, sea cucumber peptide and chrysanthemum extract is measured, so that the raw materials of the composition formula with uric acid reducing activity are screened.

The sea cucumber peptide is prepared by the following steps: (1) Homogenizing sea cucumber flesh to obtain sea cucumber homogenate, and shearing and homogenizing the sea cucumber homogenate to obtain sea cucumber homogenate; (2) Regulating the pH value of the sea cucumber homogenized liquid to 7.0, and freeze-drying the obtained sea cucumber homogenized liquid to obtain freeze-dried powder for later use; (3) Preparing a sea cucumber freeze-dried powder into a 5% solution, regulating the pH value to 8.0, adding alkaline protease, compound protease and papain at 55 ℃ for enzymolysis, wherein the total enzyme activity of the three proteases is 3000U, the adding ratio of the alkaline protease to the compound protease to the papain is 1:1:1, and the sea cucumber proteolytic liquid is obtained after enzymolysis for 1 h; (4) Regulating the pH value of the obtained sea cucumber protein enzymolysis liquid to 7.0, and freeze-drying the enzymolysis liquid to obtain sea cucumber peptide powder; (5) The sea cucumber peptide sequence is determined by separation and purification, and the specific sequences comprise EKFPPPM (SEQ ID NO. 1), PPLVKPW (SEQ ID NO. 2) and KDLGGI (SEQ ID NO. 3). Namely, the sea cucumber peptide at least comprises one of the sea cucumber peptide sequences.

As shown in table 1, the screened raw materials 2 parts by weight of resveratrol, 2 parts by weight of luteolin, 1 part by weight of quercetin, 10 parts by weight of chitosan oligosaccharide, 10 parts by weight of skim milk powder, 50 parts by weight of honeysuckle extract, 50 parts by weight of sour cherry powder, 50 parts by weight of celery seed extract, 50 parts by weight of celery extract, 10 parts by weight of sea cucumber peptide, 70 parts by weight of chrysanthemum extract all have uric acid reducing activity, and meanwhile, factors such as biological activity function, cost, taste and safety are required to be comprehensively considered. The method specifically comprises the aspects of nutritional value, functional components, interaction, price stability, seasonal fluctuation, storage stability, texture mouthfeel, aroma and taste, appearance color, food safety, allergy risk, disease prevention capability, sustainability, ecological friendliness, social responsibility and the like of raw materials. By comprehensively considering the factors, a food formula raw material selection standard meeting the requirements, high quality and safety can be prepared, and recommended intake and processing are considered, wherein sea cucumber peptide, celery extract, skim milk powder, chrysanthemum extract, chitosan oligosaccharide, luteolin and quercetin are selected as raw materials in the formula for researching the specific addition of the formula.

Table 1 xanthine oxidase inhibition ratio of formulation raw materials

EXAMPLE 2 uric acid-lowering Activity test of compositions

The XOD inhibitory activity of the 46 compositions of comparative example and compositions i-x was measured by the inhibition rate test method for determining XOD mentioned in example 1 to characterize the uric acid lowering activity effect of the compositions, and the results are shown in table 2. Comparison of comparative examples 1-42 with the results of composition III shows that each of the components of composition III, component to component, exert an XOD inhibitory activity; comparative examples 43-45 the 50 parts by weight of celery extract in composition III was replaced with the same parts by weight of honeysuckle extract, sour cherry powder, celery seed extract, respectively, and the results showed that 50 parts by weight of celery extract plays an important role in composition III. In contrast, comparative example 46 shows that 60 parts by weight of celery extract replaces 10 parts by weight of sea cucumber peptide and 50 parts by weight of celery extract in the original composition III, and the XOD inhibition activity is reduced, which indicates that there is a synergistic effect between sea cucumber peptide and celery extract. The uric acid reducing activity of the composition formula added with the celery extract or the sea cucumber peptide is greatly improved, and the uric acid reducing activity is far greater than the XOD inhibitory activity when the celery extract and the sea cucumber peptide exist respectively, so that the celery extract and the sea cucumber peptide are further shown to synergistically exert the uric acid reducing effect. The compositions IV, V, VII, IX and X show higher XOD inhibition activity, but sea cucumber peptide, luteolin, chitosan oligosaccharide and quercetin are all expensive raw materials, the addition amount of the raw materials is not too much, and the increase is not obvious although the addition amount is increased according to the result; in addition, as the overall taste of the product is affected by too high celery extract and too much intake is not suitable for human body, the experiment group is evaluated according to the sensory evaluation standard of table 3, and the result table 2 shows that the score of the composition III is 26.9, and the sensory evaluation result is excellent, so that the uric acid reducing activity of the composition III is best, and the ratio of the sea cucumber peptide, the celery extract, the skim milk powder, the chrysanthemum extract, the chitosan oligosaccharide, the luteolin and the quercetin is 10:50:460:70:10:2:1.

TABLE 2 Effect of formulation raw materials on uric acid lowering Activity

TABLE 3 sensory evaluation criteria

Example 3 animal experiments demonstrate the uric acid lowering efficacy of the compositions

32 SD male rats (body weight 200 g + -20 g), 23+ -3deg.C were selected; 65.+ -. 5% humidity, light-dark cycle 12 h, keep ventilation good and rats were kept for 1 week to fit the laboratory environment before the experiment began. Rats were randomly divided into NC, HUA, allopurinol, and formula groups (8 rats per group). Then, NC rats are filled with gastric distilled water, other rats are filled with 1.5g/kg BW of potassium gastroxymate, the model is built 1 time a day for 7 continuous days, and when the concentration of uric acid in serum is more than 110 mu mol/L, the model building is successful.

HUA group, allopurinol group and formula group were administered 1h after the administration of gastric lavage, and rats were respectively given gastric distilled water, positive drug and uric acid lowering composition III. Wherein rats in NC group and HUA group only had distilled water, allopurinol group lavage positive drug allopurinol, dosage was 20mg/kg BW, formulation group lavage per day example 2 uric acid lowering product 500mg/kg BW.

All rats were examined daily for body weight, diet and health. Retroorbital plexus blood samples were collected by intraperitoneal injection of 3% sodium pentobarbital (30 mg/kg BW) under anesthesia. Blood samples were centrifuged at 3000 Xg for 15min at 4℃to obtain serum for further physiological analysis. Serum samples were collected 3 times on days 7, 14 and 21 of administration, rats were sacrificed on day 28, blood, liver and kidney were collected and stored at-80 ℃ for detection. Kidney and liver metrics are based on fresh body weight: kidney or liver index (mg/g) =kidney or liver body weight (mg)/body weight (g).

And measuring uric acid level in the blood sample on the 7 th day, wherein the serum uric acid concentration is more than 110 mu mol/L, and the modeling is successful. Starting to perfuse the allopurinol and the formulation composition. Uric acid levels in serum were measured in 14 and 21, and after the rats were sacrificed, blood was collected to measure uric acid, creatinine and urea nitrogen in serum, as measured using commercial test kits. XOD and Adenosine Deaminase (ADA) activity in serum and liver was determined.

As shown in FIG. 1A, it can be seen that uric acid levels in hyperuricemia rats were all greater than 110. Mu. Mol/L after one week of gastric lavage, i.e., hyperuricemia modeling was considered successful. After three weeks of intervention, normal rat serum uric acid in NC group was 83.83 + -3.32 μmol/L, but serum uric acid in HUA group hyperuricemia rat reached 203+ -5.9 μmol/L, and rat uric acid content in formula group and allopurinol group was significantly reduced, wherein allopurinol group serum uric acid was reduced to 88.33+ -2.94 μmol/L, and gradually recovered to normal level compared with NC group. In the formula set, the serum uric acid is reduced to 113+/-4.09 mu mol/L, which indicates that the composition III has the potential of developing into an anti-hyperuricemia candidate drug.

In practice, serum creatinine and urea nitrogen concentrations are commonly used to characterize renal function status. As shown in fig. 1B and C, HUA group mice had significantly elevated serum creatinine and urea nitrogen levels compared to NC group. The urea nitrogen and creatinine levels of the formulation were significantly lower than those of the HUA group, and the results indicate that supplementing the compositions provided by the present study can alleviate hyperuricemia and associated kidney function impairment.

Effects of the composition on rat serum XOD activity and ADA activity, the results are shown in figure 2A, B. The serum XOD activity and ADA activity of the hyperuricemia rats in the HUA group are remarkably increased compared with those of the normal rats in the NC group, and after the allopurinol group is infused with the stomach-positive medicament, no remarkable difference exists between the serum XOD activity and ADA activity and the NC group, and the normal level is recovered. After the gastric lavage composition, the serum XOD activity and ADA activity of the formulation group were significantly reduced compared to the HUA group, which had therapeutic effects on serum XOD and ADA of hyperuricemia rats, although unlike allopurinol. These results indicate that in hyperuricemia rats, serum XOD activity and ADA activity are enhanced, leading to increased uric acid production, thereby inducing the occurrence of hyperuricemia. The composition can reduce the activity of serum XOD and ADA, thereby reducing the generation of uric acid by inhibiting the activity of uric acid synthesis key enzyme, and further relieving the symptoms of hyperuricemia rats.

As shown in FIG. 2C, D, the effect of the composition on liver XOD activity and ADA activity of rats was 15.15+ -0.45U/g prot in NC group normal rats, 17.34+ -0.62U/g prot in normal rats, 29.79 + -0.21U/g prot in HUA group hyperuricemia rats and 24.50+ -0.18U/g prot in normal rats, and significant decrease in liver XOD activity and ADA activity in hyperuricemia rats after administration of allopurinol and composition, inhibiting the activity of uric acid generating key enzymes in liver.

The histopathological image of the kidney is shown in fig. 3, the HUA group has some pathological changes, the glomerulus at the black arrow is atrophic, the tubular at the yellow arrow is dilated, the cell brush border is shed, and the like, but under the action of allopurinol, the kidney of the rat is improved, and compared with the NC group, the kidney glomerulus is plump and the cell compact gap is small. After the stomach is irrigated, compared with the hyperuricemia rats in the HUA group, the kidneys of the rats are also obviously improved. These results indicate that the formulated composition is capable of ameliorating the damage to renal function in hyperuricemic rats.

Example 4 effect of formulation group on composition of intestinal flora species of HUA rats.

To explore the effect of the product on the diversity and richness of the intestinal flora of HUA rats, the a-diversity of the intestinal flora of rats was analyzed. The study selects the diversity of shannon indexes and Simpson reaction microbial communities, the larger the shannon indexes are, the smaller the Simpson indexes are, the higher the diversity of sample microbial communities is represented, the richness of the chao indexes and Ace reaction microbial communities is selected, and the larger the chao indexes and Ace indexes are, the higher the richness of the sample microbial communities is represented. As can be seen from fig. 4 (a) and (C), compared with NC group, the shannon index and Simpson index of the HUA rat intestinal flora are both significantly decreased (p < 0.05), indicating that hyperuricemia affects the diversity of the rat intestinal flora, and the intestinal microbiology of hyperuricemia patients is less diverse, and the intestinal microecology is unbalanced; compared with the NC group, the allopurinol group has no significant difference (p > 0.05) between the shannon index and the Simpson index of the intestinal flora of the rats, which indicates that the allopurinol as a positive drug can promote the diversity of the intestinal flora of the rats suffering from hyperuricemia, and the effect of the formula group is less obvious than that of the allopurinol, but can promote the shannon index and the Simpson index, and can improve the diversity of the intestinal flora of the rats to a certain extent. Compared with the NC group, the Chao index and the Ace index of the HUA rat intestinal flora are both obviously reduced (p < 0.05), which indicates that hyperuricemia can affect the richness of the rat intestinal flora; compared with NC group, allopurinol group has no significant difference (p > 0.05) between the chao index and Ace index of rat intestinal flora, which indicates that the positive drug allopurinol can promote the richness of rat intestinal flora with hyperuricemia, while formula group has less obvious effect than allopurinol, but can promote the chao index and Ace index, and improve the richness of rat intestinal flora to a certain extent. Taken together, it is shown that the product formulation group can improve intestinal flora of hyperuricemia rats.

Biological barriers play an important role in the intestinal health of the human body. Changes in the structure of the intestinal microbiota affect the integrity of the intestinal barrier. More and more studies indicate that hyperuricemia has a significant impact on the composition of the intestinal microbiota, and that there is a two-way relationship between hyperuricemia and intestinal microbiota. A significant increase in uric acid levels in the body can cause changes in the intestinal microbiota. For statistical analysis of the similarity of the species composition of the intestinal flora of each group of rats, venn plots were selected to show OTUs common and unique to each group of rats. As shown in fig. 5, there were 261 total OTUs for the four groups of rats, 153 total OTUs for NC group rats, 51 total OTUs for HUA group rats, 26 total OTUs for allopurinol group rats, 28 total OTUs for formula group rats, and 564, 424, 441 and 424 total OTUs for the four groups of rats, respectively.

The composition of the intestinal flora of each group of rats at the portal level is shown in fig. 6, the intestinal flora of four groups of rats mainly consists of fimicites (thick-walled fungus phylum), the relative abundance of bacterioidota (bacteroida) of the HUA group is significantly increased (p < 0.05), but the relative abundance of bacterioidota (bacteroida) is significantly decreased (p < 0.05) after the allopurinol group is intragastrically, and the formulation group and the allopurinol group have no significant difference (p > 0.05) compared to NC group of mice. While the relative abundance of actioniobacteria (actinomycota) in HUA group was significantly reduced (p < 0.05) compared to NC group mice, the relative abundance of actioniobacteria (actinomycota) was significantly increased (p < 0.05) after allopurinol group gavage, and the relative abundance of actioniobacteria (actinomycota) with HUA group after formula group gavage product was also significantly increased (p < 0.05). Bactoidota (Bacteroides) is involved in bile acid metabolism in vivo, converts intestinal-bound bile acid into unbound bile acid, and further affects the expression of XOD and uric acid production in the liver.

The composition of the intestinal flora of each group of rats at the family level is shown in fig. 7, hyperuricemia rats of HUA group have significantly increased relative abundance of Lactobacillaceae (lactylidae) and Lachnospiraceae (coryneform clostridiaceae) compared to NC group normal rats (p < 0.05), but the relative abundance of Lactobacillaceae (lactylidae) and Lachnospiraceae (coryneform) is significantly reduced after the allopurinol group is intragastized (p < 0.05), and the formulation group and allopurinol group have no significant difference (p > 0.05). Whereas relative abundance of Akkermansiaceae (Akkermansiaceae) and peptostacoccaceae (peptostrecoccaceae) was significantly reduced (p < 0.05) compared to NC group mice, relative abundance of Akkermansiaceae (Akkermansiaceae) and peptostacoccaceae (peptostaceae) was significantly increased (p < 0.05) after allopurinol group gavage, and relative abundance of Akkermansiaceae (Akkermansiaceae) and peptostacoccaceae (peptostaceae) was also significantly increased (p < 0.05) after formula group gavague product.

Effect of the product on composition at the genus level of the intestinal flora of HUA rats. The composition of the intestinal flora of each group of rats at the genus level is shown in fig. 8, the hyperuricemia rats of the HUA group significantly decreased (p < 0.05) relative abundance of Lactobacillus (Lactobacillus), blautia (blauja), bacilli and paralacteoides (parabacillus) compared to NC group normal rats, the relative abundance of Lactobacillus (Lactobacillus), blauja (bacillus) and paralacteoides (parabacillus) in the formula group also significantly increased and higher than that of the other group (p < 0.05), but there was no significant difference in the relative abundance of Lactobacillus (Lactobacillus), blauja (b acteoides), bacteoides (bacillus) and paraacteoides (parabacillus) in the formula group for the group of Bactoides (bacillus) and paraacteoides (parabacillus) and their respective abundance of the other group (p < 0.05). Whereas relative abundance of Akkermansia (Akkermansia) and romidepa (Luo Long butcher) was significantly reduced (p < 0.05) in HUA compared to NC group mice, relative abundance of Akkermansia (Akkermansia) and romidepa (Luo Long butcher) was significantly increased (p < 0.05) after allopurinol group gavage, and relative abundance of Akkermansia (Akkermansia) and romidepa (Luo Long butcher) was also significantly increased (p < 0.05) compared to HUA after formulation group gavage. The purine metabolism, amino acid metabolism and glycolytic pathways in the intestinal tract of patients suffering from hyperuricemia are all significantly changed, so that bacteria such as Lactobacillus, bactoides and Parabacterides are involved in the processes, and the relative abundance of the bacteria is reduced after the formulation is infused into the stomach, which indicates that the formulation can alleviate the corresponding symptoms of patients suffering from hyperuricemia by changing the relative abundance of related flora in the intestinal tract.

The composition can reduce purine absorption in intestinal tract, promote uric acid metabolism in intestinal tract, and relieve intestinal inflammatory reaction by increasing probiotic ratio. Finally, the product formulation reduces the proportion of pathogenic bacteria in the intestinal tract.

Example 5 Effect of composition formulation group on beta-diversity of HUA rat intestinal flora

HUA rat intestinal flora principal coordinate analysis. Principal co-ordinates analysis (PCoA) can be used to study the similarity or difference of the composition of the sample colony, and find out the potential Principal components affecting the composition difference of the sample colony by dimension reduction; PCoA is plotted based on a selected distance matrix. The PCoA analysis was performed on the OTU composition of the intestinal flora of each group of rats, and the results are shown in fig. 9. As can be seen from the graph, there is a distinct cluster of different groups of rat intestinal flora compositions, indicating that there is a difference in the composition of the individual groups of rat intestinal flora. NC and HUA group mice, allopurinol group and formula group rat intestinal flora can be clearly distinguished along the PC1 axis; NC and HUA groups of mice, allopurinol groups and formula groups of rats intestinal flora can be distinguished to some extent along the PC2 axis; indicating that allopurinol and the formula can greatly change intestinal flora of rats.

Non-metric multidimensional scaling analysis (Nonmetric multidimensional scale, NMDS) can reduce the dimensions of data by pipelining, thereby locating, analyzing and categorizing, reflecting the differences between different samples in the form of points, reflecting the degree of difference between samples by the point-to-point distance. The results of NMDS analysis of the OTU composition of each group of rat intestinal flora are shown in fig. 10, and similar to the results of PCoA analysis above, there is a distinct cluster of the composition of the intestinal flora of different groups of rats, further indicating that there is a difference in the composition of the intestinal flora of each group of rats.

Short chain fatty acids (Short Chain Fatty Acids, SCFAs) are metabolites of the intestinal flora and have proven to be critical in maintaining intestinal morphology and function. In FIG. 11, the individual concentrations of five SCFAs were determined by gas chromatography-mass spectrometry. Acetic acid, propionic acid, butyric acid, isobutyric acid and valeric acid were significantly reduced on average in the model group. And after the positive medicines allopurinol and the experimental formula are administrated, the levels of acetic acid, propionic acid, butyric acid, isobutyric acid and valeric acid are obviously improved compared with the levels of the model group. Furthermore, it can be seen from the results that although the experimental group had not increased levels of propionic acid, butyric acid, isobutyric acid and valeric acid as compared to the positive control group, acetic acid, propionic acid, butyric acid, isobutyric acid and valeric acid were improved on average, which was beneficial to maintain the intestinal morphology and function to some extent.

The above description is only of the preferred embodiments of the present invention and is not intended to limit the present invention, but various modifications and variations can be made to the present invention by those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A composition for relieving hyperuricemia is characterized by comprising sea cucumber peptide, celery extract, skim milk powder, chrysanthemum extract, chitosan oligosaccharide, luteolin and quercetin;

the weight parts of the sea cucumber peptide are within the range of 10-30;

the weight part of the celery extract is within the range of 10-50;

the weight parts of the skim milk powder are in the range of 350-460;

the weight part of the chrysanthemum extract is in the range of 70-100;

the weight part of the chitosan oligosaccharide is within the range of 10-50;

the weight part of luteolin is in the range of 2-5;

the weight part of the quercetin is in the range of 1-2;

preferably, the weight ratio of the sea cucumber peptide, the celery extract, the skim milk powder, the chrysanthemum extract, the chitosan oligosaccharide, the luteolin and the quercetin in the composition is 10:50:460:70:10:2:1.

2. The composition of claim 1, wherein the sea cucumber peptide is derived from a proteolytic enzyme of sea cucumber.

3. The composition of claim 1, wherein the sequence of the sea cucumber peptide comprises EKFPPPM, PPLVKPW, KDLGVLI.

4. The composition of claim 1, wherein the composition inhibits xanthine oxidase activity.

5. A composition for alleviating hyperuricemia according to claim 1, wherein the composition is synergistic in effect by sea cucumber peptide and celery extract.

6. The composition of claim 5, wherein the optimum ratio of the synergistic effect of the sea cucumber peptide and the celery extract is 1:5.

7. Use of a composition according to any one of claims 1-4 or 5-6 for the preparation of a product for alleviating hyperuricemia caused by a disorder of purine metabolism or uric acid excretion itself.

8. Use of a composition according to any one of claims 1-4 or 5-6 for the preparation of a hyperuricemia-relieving product comprising sea cucumber peptide.

9. Use of a composition according to any one of claims 1-4 or 5-6 for the preparation of functional formulas for lowering uric acid in the middle-aged and elderly, foods for special medical use, pharmaceutical products.

10. Use of the composition according to any one of claims 1-4 or 5-6 for the preparation of functional formulas, special medical use foods, medicines for middle-aged and elderly people containing sea cucumber peptides for lowering uric acid.