Disclosure of Invention
The invention aims to provide a composition which has no toxic or side effect and can be taken for a long time and is beneficial to reducing uric acid level of hyperuricemia patients, so as to solve the problem that the current western medicine treatment can cause damage to liver and kidney, and solve the problems that the traditional Chinese medicine treatment extraction process is complex, the taste is poor, the decocting traditional Chinese medicine procedure is complex, the patients are difficult to take continuously, and the like.
To achieve the technical purpose of the invention, the invention provides a composition for relieving hyperuricemia and a preparation method thereof. The composition can effectively reduce uric acid, urea nitrogen and creatinine levels in serum of hyperuricemia mice, improve renal function of the hyperuricemia mice, and simultaneously reduce four levels of total cholesterol, triglyceride, high-density fat and lipid of low-density fat in serum of the hyperuricemia mice, and has lipid-lowering effect. The composition provided by the invention is rich in protein and dietary fiber, low in fat, good in brewing property, and good in application prospect in preparation of medicines or medicine and food homologs for relieving hyperuricemia.
The invention provides a composition for relieving hyperuricemia, which consists of 40% of polished round-grained rice, 5-10% of poria cocos, 5-10% of Chinese yam, 5-10% of lily, 5-10% of jujube, 5-10% of chicken's gizzard-membrane, 5-10% of fried coix seed, 1-2% of rose and the balance of skim milk powder in percentage by mass. Preferably, the composition comprises 40% of polished round-grained rice, 7% of poria cocos, 8% of Chinese yam, 10% of lily, 6% of jujube, 8% of chicken's gizzard-skin, 7% of stir-fried coix seed, 1.5% of rose and the balance of skim milk powder in percentage by mass.
Further, in the composition for relieving hyperuricemia provided by the invention, the polished round-grained rice is selenium-enriched polished round-grained rice, and the selenium content of the selenium-enriched polished round-grained rice is preferably 18.4 mug/100 g.
The present invention also provides a method of preparing a composition for alleviating hyperuricemia, comprising: cleaning raw materials (except skim milk powder) of the composition for relieving hyperuricemia, drying at 70 ℃ until the water content is 9%, crushing, sieving with a 50-mesh sieve, mixing and ball milling at a ball milling ratio of 1:10, ball milling at 230r/min for 12h, parching for 5min, and adding the skim milk powder to obtain the composition for relieving hyperuricemia.
Also, the invention claims the use of said composition for the preparation of a medicament for alleviating hyperuricemia. The medicine can effectively reduce uric acid, urea nitrogen and creatinine levels in serum, improve renal function and simultaneously has the lipid-lowering effect. The medicine disclosed by the invention is used for relieving hyperuricemia and is suitable for people suffering from hyperuricemia.
Furthermore, the composition, medicine or medicine-food homologous (composition) for relieving hyperuricemia provided by the invention can effectively reduce uric acid, urea nitrogen and creatinine levels in serum of hyperuricemia mice, improve renal functions of the hyperuricemia mice, and simultaneously reduce four levels of total cholesterol, triglyceride, high-density fat and low-density fat in serum of the hyperuricemia mice, thereby having lipid-lowering efficacy.
Furthermore, the composition for relieving hyperuricemia provided by the invention is rich in protein, dietary fiber, low in fat and good in brewing.
Further, the composition, medicament or pharmaceutical/food source (composition) of the present invention may be prepared into a dosage form for convenient administration, including any of tablets, capsules, powders, granules or pills, according to methods well known to those skilled in the art. Particularly preferably, the composition, medicament or pharmaceutical-food homologous (composition) according to the present invention is prepared as a powder.
Compared with the prior art, the invention has the following beneficial effects or advantages:
The composition for relieving hyperuricemia provided by the invention can effectively reduce uric acid, urea nitrogen and creatinine levels in serum of hyperuricemia mice through multi-component compounding, improve renal functions of the hyperuricemia mice, and simultaneously reduce four levels of total cholesterol, triglyceride, high-density fat and low-density fat in serum of the hyperuricemia mice, has lipid-lowering efficacy, and has better effects in protecting kidneys and lipid-lowering and liver-protecting aspects.
The composition for relieving hyperuricemia prepared by the invention has dose effect in the intervention effect of reducing the level of uric acid, urea nitrogen and creatinine caused by hyperuricemia and reducing the level of total cholesterol, triglyceride, high-density fat and lipid of low-density fat in serum of hyperuricemia mice, namely the intervention effect of the composition on the level of uric acid, urea nitrogen and creatinine caused by hyperuricemia and the level of total cholesterol, triglyceride, high-density fat and lipid of low-density fat in serum is more obvious along with the increase of the dose.
The composition for relieving hyperuricemia, which is prepared by the invention, is rich in protein and dietary fiber, low in fat, easy to disperse in liquid, good in brewing property, convenient to take, and has a good application prospect in preparing medicines or medicine and food homologs for relieving hyperuricemia.
Detailed Description
The following describes the technical aspects of the present invention with reference to examples, but the present invention is not limited to the following examples. The experimental methods and the detection methods in each embodiment are conventional methods unless otherwise specified; the food materials, unless otherwise specified, are commercially available.
Example 1
The present example provides a method for preparing a composition for alleviating hyperuricemia, comprising the following steps:
Cleaning raw materials of the composition for relieving hyperuricemia, drying at 70 ℃, measuring the water content of the raw materials to be 9%, crushing the dried raw materials, sieving with a 50-mesh sieve, mixing and ball milling, ball milling the mixture for 12 hours at a ball milling ratio of 1:10 and 230r/min to obtain a mixture, and adding skim milk powder to obtain the composition for relieving hyperuricemia.
Example 2
This example provides specific components and amounts of a composition for alleviating hyperuricemia, prepared in the same manner as in example 1.
Test group 1 formulation: the composition for relieving hyperuricemia comprises 40g of selenium-enriched polished round-grained rice, 7g of poria cocos, 8g of Chinese yam, 10g of lily, 6g of jujube, 8g of chicken's gizzard-skin, 7g of fried coix seed, 1.5g of rose and 12.5g of skimmed milk powder per 100g of composition.
Test group 2 formulation: the composition for relieving hyperuricemia comprises 40g of selenium-enriched polished round-grained rice, 5g of poria cocos, 10g of Chinese yam, 5g of lily, 10g of jujube, 10g of chicken's gizzard-skin, 5g of fried coix seed, 1g of rose and 14g of defatted milk powder per 100g of composition.
Test group 3 formulation: the composition for relieving hyperuricemia comprises 40g of selenium-enriched polished round-grained rice, 10g of poria cocos, 5g of Chinese yam, 8g of lily, 5g of jujube, 5g of chicken's gizzard-skin, 10g of fried coix seed, 2g of rose and 15g of defatted milk powder per 100g of composition.
Control group 1 formulation (without Poria cocos in test group 1): the composition for relieving hyperuricemia comprises 40g of selenium-enriched polished round-grained rice, 8g of Chinese yam, 10g of lily, 6g of jujube, 8g of chicken's gizzard-membrane, 7g of fried coix seed, 1.5g of rose and 19.5g of skimmed milk powder per 100g of the composition.
Control group 2 formulation (lilies in test group 1 replaced with lilyturf root): the composition for relieving hyperuricemia comprises 40g of selenium-enriched polished round-grained rice, 7g of poria cocos, 8g of Chinese yam, 10g of dwarf lilyturf tuber (the drug property of dwarf lilyturf tuber is similar to that of lily), 6g of Chinese date, 8g of chicken's gizzard-membrane, 7g of fried coix seed, 1.5g of rose and 12.5g of skimmed milk powder per 100g of composition.
Control group 3 formulation: commercially available compositions (available from Kirschner trade Co., st.) for alleviating hyperuricemia.
The selenium-enriched polished round-grained rice used in this example was purchased from Qingya agricultural technologies Co., ltd. In Jiujinai county, and the selenium content was 18.4. Mu.g/100 g.
Example 3
This example provides a nutritional profile, amino acid profile and a brewing profile of the composition for alleviating hyperuricemia prepared in test group 1 and the composition for alleviating hyperuricemia commercially available in control group 3.
1. Analysis of nutritional ingredients
The composition for alleviating hyperuricemia prepared in test group 1 was subjected to nutritional analysis and compared with a commercially available composition for alleviating hyperuricemia. Ash content determination: the burning method is referred to GB 5009.4-2016 "determination of ash in food"; total protein determination: kjeldahl method, refer to GB 5009.5-2016 "determination of protein in food"; crude fat measurement: soxhlet extraction, reference GB 5009.6-2016 "determination of fat in food"; sodium content determination: reference GB 5009.91-2017 "determination of sodium in food"; measuring the content of dietary fiber: the enzyme weight method is referred to GB/T5009.88-2008 'determination of dietary fiber in food'; carbohydrate content: and adding the contents of water, ash, protein, fat and sodium. The measurement results are shown in Table 1.
Table 1: analysis of nutritional ingredients
As can be seen from the results in Table 1, the composition for hyperuricemia group prepared by the test group 1 can provide energy, protein, fat, dietary fiber and carbohydrate required by human body, meets the requirement of balanced nutrition, is rich in protein and dietary fiber and has low fat and low carbon water compared with the composition for alleviating hyperuricemia, and can be seen from nutrition, has the effects of helping healthy diet and slimming of hyperuricemia group, and has a certain alleviating effect on diabetes and hyperlipidemia caused by hyperuricemia.
2. Amino acid species analysis
The composition for alleviating hyperuricemia prepared in test group 1 was subjected to amino acid type analysis and compared with the composition for alleviating hyperuricemia commercially available in control group 3. Amino acid composition analysis: accurately weighing a certain amount of sample in a hydrolysis tube, adding HCl (10 mL,6.0 mol/L), vacuumizing, filling nitrogen, sealing the hydrolysis tube after repeating for three times, hydrolyzing (110 ℃ for 22 h), cooling, filtering, taking 2mL of filtrate in a colorimetric tube, performing water bath (60 ℃ for 10 min), drying under reduced pressure, adding a proper amount of HCl (0.02 mol/L), and standing for 30min to obtain a liquid to be tested. Sample measurement: the absorbance at 570nm and 440nm was measured using a mixed amino acid standard working solution injected into an amino acid automatic analyzer, referring to JJG1064-2011 amino acid analyzer assay protocol and instrument instructions, and the measurement results are shown in table 2.
Table 2: analysis of amino acid composition
As can be seen from the results of Table 2, the total amino acid amount (TAA) of the composition for alleviating hyperuricemia prepared by the present invention is 9.08%, which is higher than that of the commercially available composition by 8.58%, which is consistent with the results of the protein content measurement of Table 1. From the amino acid composition, 16 amino acids were detected in both compositions, 7 of the essential amino acids, no tryptophan, 9 of the unnecessary amino acids, and the respective amino acids were close in terms of ratio. Among the 16 amino acids detected, the glutamic acid content was the highest, the average was 1.70g/100g, aspartic acid (0.84 g/100 g) and leucine (0.79 g/100 g) times, and the methionine content was the lowest, the average was 0.035g/100g.
The ratio of total essential amino acid content to total amino acid content (EAA/TAA) of the high-quality protein should be 0.4, and the ratio of essential amino acid to non-essential amino acid (EAA/NEAA) should be 0.6. The composition for alleviating hyperuricemia prepared by the invention is 0.37, the commercial composition is 0.34, and neither nutritional composition is achieved, which is mainly related to the undetermined tryptophan content of the essential amino acid. Research shows that tryptophan content in polished round-grained rice can reach 129mg/g, so that EAA/TAA of the composition for hyperuricemia group prepared by the invention can be higher than 0.4.
It was further found that the EAA/NEAA of the composition for hyperuricemia group prepared according to the present invention was 0.58, close to 0.6, and thus, the composition for hyperuricemia group prepared according to the present invention belongs to a high quality protein resource as a whole.
3. Brewing analysis
The compositions for alleviating hyperuricemia prepared in the test group 1 and the control groups 1 to 2 were subjected to the brewing analysis and compared with the composition for alleviating hyperuricemia commercially available in the control group 3.
(1) Determination of Water solubility index and Water absorption index
3G samples (W 0) were weighed accurately and placed into centrifuge tubes (W 1) and 30 mL distilled water was added and stirred until thoroughly mixed. The sample was placed in a 30 ℃ constant temperature water bath for 30min a with continuous slow stirring clockwise. After stirring, the sample was placed in a centrifuge and centrifuged at 4000 r/min for 20: 20 min. The centrifuged supernatant was poured into an evaporation dish of known weight (W 3), dried and weighed (W 2). Meanwhile, the centrifuge tube and the precipitate were weighed (W 4). Water Solubility Index (WSI), water Absorbency Index (WAI) were calculated.
WSI=(W4-W1)×100%/W0
WAI=(W2-W3)×100%/W0
(2) Determination of the centrifugal precipitation Rate
Sample 1g was taken and mixed with 5g of 80℃distilled water, 21 g (M 1) was taken from the well mixed sample and placed in a centrifuge tube, centrifuged at 4000 r/min for 30 min, the supernatant was decanted off and the mass of the remaining precipitate (M 2) was weighed. The centrifuge Sediment Ratio (SR) was calculated.
SR=M2×100%/M1
(3) Determination of dispersibility
A 5g sample was weighed and recorded to the nearest 0.001 g mass, placed in a 50mL beaker and distilled water at 60 ℃ was added at 30 mL. After stirring 0.5 min with a glass rod in the same direction, standing for 5: 5min, and adding 30: 30 mL distilled water at 60 ℃ again for dilution. The diluted sample was sieved through a 50 mesh sieve, carefully rinsed with distilled water, the unsieved agglomerate was collected, dried in an oven at 105 ℃ for 6 h, and the dried sample was cooled and weighed. The dispersity index is more than or equal to 95 percent; 90% -95% is good; 80% -90% is better; less than or equal to 80 percent. 3 parallel experiments were performed to reduce experimental error. A Dispersion Index (DI) is calculated.
DI=[1-m/M(1-w)]×100%
Wherein: m is the mass/g of the dried agglomerate; m is the total mass/g of the sample; w is the moisture content in the sample/%.
(4) Determination of caking Rate
Accurately weighing 5g samples in a beaker, adding water 25 mL at 60 ℃, stirring 5 min, and standing for 30 s. Taking a 50-mesh screen, filtering, washing the agglomerate with deionized water, drying in a drying oven 1 h to constant weight, cooling to room temperature by a cooler, and weighing. Subtracting the mass of the screen gives the mass of the agglomerate (Tian Wenjing et al 2021). The caking ratio (AR) was calculated.
AR=(m2-m1)/m×100%
Wherein: m is the mass/g of the sample; m 1 is screen mass/g; m 2 is the mass/g of screen and agglomerate after constant weight.
(5) Determination of wettability
200ML of deionized water (50 ℃) was added to a 250mL beaker, and a 0.5 g sample was accurately weighed and evenly spread on the water surface, and the time required for powder addition to complete sedimentation was accurately recorded.
The measurement results are shown in Table 3.
Table 3: brewing analysis
The reconstitution of the composition determines its product stability. As can be seen from the results of Table 3, the composition for alleviating hyperuricemia prepared according to the present invention has a water solubility index of 19.34%, which is 17.84% higher than that of the commercially available composition, and also 18.35% higher than that of the composition of comparative example 1 and 17.33% higher than that of the composition of comparative example 2. The water absorption index 372.5% and the centrifugal precipitation rate 54.43% of the composition prepared by the invention are lower than those of the composition sold in the market and the compositions prepared by comparative examples 1-2, so that the composition for relieving hyperuricemia prepared by the invention has better product stability than the composition sold in the market, the starch is fully gelatinized, and the macromolecular nutrient substances can be efficiently degraded.
Example 4
This example provides scanning electron microscopy and fourier infrared spectroscopy of a composition for alleviating hyperuricemia prepared in test group 1 and a commercially available composition for alleviating hyperuricemia of control group 3.
1. Scanning electron microscope analysis
After crushing and screening the raw materials, placing the materials on a loading platform according to the instrument requirements. After the metal spraying, the samples were photographed at an accelerating potential of 5.0kV, and the shapes of the samples were analyzed at multiples of 150, 500, 2000 and 5000, and scanning electron microscopy was performed using Zeiss genmini a, and the results are shown in fig. 1.
As can be seen from FIG. 1, the composition for alleviating hyperuricemia prepared by the present invention has a loose structure and appears as a flat sheet. The reason for this phenomenon may be that starch is gelatinized after the ultra-micro treatment, and part of the starch is degraded to generate oligosaccharides such as maltose, dextrin and the like, so that the content of micromolecular starch is increased, and the water solubility of the product is enhanced; the protein is denatured, so that protease inhibitor factors such as trypsin inhibitor, tannic acid and the like and anti-nutritional factors lose activity, a fluffy structure with more enzyme fulcrums is formed, the protein is partially degraded, the content of free amino acid is increased, the digestion and absorption rate of the protein can be improved, and the water absorption property, the water solubility and the like of the nutritional composition suitable for hyperuricemia groups prepared by the invention are facilitated.
2. Fourier infrared spectroscopy
Uniformly mixing a sample and potassium bromide according to the mass ratio of 1:100, pressing into a sheet, detecting the sample by adopting a Fourier transform infrared spectrometer Siemens femto 35, and measuring the wavelength range of 4000-400 cm -1, wherein the scanning times are 32 times. The resolution is 4 cm -1. And taking air as a background to perform spectrum acquisition. The analysis results are shown in FIG. 2.
As can be seen from fig. 2, the composition for alleviating hyperuricemia prepared by the present invention has a c=c stretching vibration peak at 1642cm -1, and a stronger c—o stretching vibration peak at 1076cm -1, which is generated due to the breaking of α -1,4 glycosidic bond in starch, probably due to the extrusion by ball milling, the crystal structure in starch material is destroyed, the melting phenomenon of the crystalline part occurs, and the hydrophilic groups such as hydroxyl group, ether group, carboxyl group, etc. in starch molecule are exposed, and form a high viscosity single-phase uniform dispersion system together with water molecules, so that the powder has good adjustability.
Example 5
This example provides four level determinations of acute toxicity test, kidney and liver protection test on hyperuricemia mice, and lipid in mouse serum for compositions for alleviating hyperuricemia prepared in test group 1 and for compositions for alleviating hyperuricemia commercially available in control group 3.
1. Acute toxicity test
The experimental mice (C57 male mice, weight 18+ -2 g) were randomly divided into three groups of male and female halves, each group being 4 male mice and 4 female mice, and the compositions for alleviating hyperuricemia prepared in the experimental groups 1 to 3 were fed respectively, and the volume was 2g/10g of the weight, closely observed for 8 hours and recorded, and further observed for 7 days. In 7 days, the mice have no death, active behaviors, normal ingestion, bright hair color, no loose dirt, no syncope and saliva, no abnormal secretion in eyes, mouth, nose, ears and perianal, no tail and toe break. 5 animals were selected from the 3 groups, and the major organs such as heart, liver, lung, stomach and thymus were seen with naked eyes without abnormal changes, indicating that the compositions of the test groups 1-3 were all free of acute toxicity.
2. Kidney and liver protection test in mice
C57 male mice were randomly divided into 7 groups, namely a normal control group (NC), a model control group (MC), an allopurinol group (10 mg/kg) (AP), a composition high-dose group (RH) for alleviating hyperuricemia, a composition medium-dose group (RM) for alleviating hyperuricemia, a composition low-dose group (RL) for alleviating hyperuricemia, and a control 1 group (HG). The composition prepared in RH group gastric lavage test group 1 has a gastric lavage dosage of 3600mg/kg; the composition prepared in RM gastric lavage test group 1 was at a gastric lavage dose of 2100mg/kg; the composition prepared in the RL group gastric lavage test group 1 has a gastric lavage dosage of 900mg/kg; the commercial composition for relieving hyperuricemia of HG group lavage control group 3, the dosage of lavage is 900mg/kg; the blank control group, the model control group and the allopurinol group are given with physiological saline; each treatment group was continuously intragastrically for 14 days. Suspending Potassium Oxazinate (PO) in 0.5% sodium carboxymethylcellulose (CMC-Na) water solution to obtain potassium oxazinate solution, continuously injecting and administering the potassium oxazinate solution into MC, AP, RH, RM, RL and HG groups of mice per day in an intraperitoneal mode on 15-21 days of intragastric administration, injecting the potassium oxazinate solution into the mice per day for 1h on an empty stomach of 9 points per day, and then injecting the potassium oxazinate solution into the mice for 1h, and then irrigating the same volume of CMC-Na of 0.5% of the same volume of the NC group and the MC group respectively. Mice were sacrificed 3h after the last dose, liver and kidneys were immediately separated in an ice bath, eyeball blood was taken, and blood samples were centrifuged at 5000 r/min for 10 minutes at 4 ℃ to obtain serum samples. Serum, liver, one kidney were stored at-80 ℃ until assayed. The other kidney was stored in paraformaldehyde at 4 ℃. The uric acid, urea nitrogen (BUN) and Creatinine (CRE) levels in the serum of mice were measured using Chemray800,800 full-automatic biochemical analyzer (available from Shenzhen Lei Du technologies Co., ltd.) and the measurement results are shown in FIGS. 3-5.
As can be seen from FIG. 3, the combination of uricase inhibitor PO caused hyperuricemia in mice, which is manifested as a significant increase in serum uric acid levels. Uric acid levels in serum were significantly elevated in model mice compared to control mice (P < 0.001). This indicates that hyperuricemia was successfully modeled. Serum uric acid levels were significantly reduced by 94.71% (P < 0.001) in the positive drug group compared to the model group. Compositions for alleviating hyperuricemia serum uric acid levels were significantly reduced by 55.75%, 57.38% and 64.42% (P < 0.001) in the high, medium and low dose groups of mice. The HG group was reduced by about 44.87% (P < 0.001) compared to the model group. Composition for alleviating hyperuricemia the serum uric acid levels were significantly reduced (P < 0.05) in the high dose group compared to the low dose group. The test groups of the compositions prepared in comparative examples 1 and 2 were less effective in lowering uric acid levels in the serum of mice than the high, medium and low dose groups, and were less effective in lowering abnormally elevated uric acid levels in the serum of hyperuricemia mice. Experimental results show that the composition for relieving hyperuricemia can effectively reduce the abnormally elevated uric acid level in serum of hyperuricemia mice.
As can be seen from fig. 4 and 5, both BUN and CRE levels in the serum of mice in the model group were significantly increased (P <0.001 or P < 0.01) compared to the control group; compared to the model group, the BUN levels in the serum of mice in the different dose groups were reduced by 25.02%, 20.51% and 25.49% (P <0.05 or P < 0.01), respectively. CRE levels were reduced by 17.15%, 23.44% and 22.63% (P <0.05 or P < 0.01), respectively. BUN and CRE levels in HG group decreased by 22.48% and 21.42% (P <0.01 or P < 0.001), respectively, while positive drugs had no significant improvement on both. The test groups of the compositions prepared in comparative examples 1 and 2 were inferior in the effect of reducing BUN and CRE contents in serum of mice to those of high, medium and low dose groups, and were inferior in the effect of improving renal function levels in hyperuricemia mice. Experimental results show that the composition for relieving hyperuricemia can effectively improve the renal function level of hyperuricemia mice.
3. Determination of lipid four-item level in mouse serum
The total serum cholesterol (TC), triglyceride (TG), high density fat (HDL-C), low density fat (LDL-C) levels in the serum of mice were measured using Chemray800,800 full-automatic biochemical analyzer, and the measurement results are shown in FIGS. 6-9.
From FIGS. 6-9, the serum lipid levels were reduced to a different extent in the model group compared to the blank group. Compositions for alleviating hyperuricemia the serum LDL-C levels in mice in the different dose groups were elevated by 21.05%, 30.07% and 36.80% (P <0.05 or P <0.01 or P < 0.001), respectively, compared to the model group. The TC, TG content of the medium and high dose groups were significantly reduced by 9.27%, 15.89% (P <0.05 or P < 0.001) and 35.52% and 37.89% (P < 0.001), respectively, compared to the model group. HDL-C was 13.35% reduced in the high dose group compared to the model group (P < 0.001). Compositions for alleviating hyperuricemia high dose groups showed significantly reduced TC and HDL-C (P < 0.05) compared to the low dose group. LDL-C in HG group was significantly elevated 33.07% (P < 0.01). The test groups of the compositions prepared in comparative examples 1 and 2 were inferior to the high, medium and low dose groups in terms of the reduction effect of TC content, TG content and HDL-C content in the serum of mice, and the inferior to the high, medium and low dose groups in terms of the improvement effect of LDL-C level in the serum of mice, and had poorer lipid-lowering functions. Experimental results show that the composition for relieving hyperuricemia can effectively reduce four lipid levels in serum of hyperuricemia mice, has lipid-lowering efficacy, and has a certain treatment effect on hyperuricemia caused by hyperuricemia.
4. Determination of Xanthine Oxidase (XOD) and Adenosine Deaminase (ADA) activity in mouse liver
The results of the assay are shown in fig. 10 and 11, and the model control group ADA and XOD activities were significantly higher than the normal control group (P < 0.01). ADA activity was significantly inhibited in the AP, RL, RM and RH groups compared to the model control group (P <0.01 or P < 0.001). XOD activity was also significantly inhibited in the AP, RL, RM and RH groups compared to the model control group (P < 0.05). The test groups of compositions prepared in comparative examples 1 and2 were perfused, and the mice livers had higher ADA and XOD activity than the high, medium and low dose groups, with poorer uric acid lowering effect. XOD activity of HG group and ADA play an important role in purine metabolism. Experimental results show that the composition for relieving hyperuricemia can achieve the effect of reducing uric acid by inhibiting XOD and ADA.
5. Renal histopathological assay
After kidney tissue was fixed in 4% paraformaldehyde solution (4% pfa solution) for 48 h hours, gradient ethanol was used for dehydration, transparency, paraffin impregnation, embedding, paraffin section (2 μm), hematoxylin-eosin staining (HE staining) and periodic acid-schiff staining (PAS staining), and after mounting, the images were collected by an image acquisition system (fig. 12).
As can be seen from fig. 12, the glomeruli of the normal control group were normal in size and shape, and their boundaries were clear. The control group of hyperuricemia model can be seen with tubular lumen expansion, loose epithelial cell edema and interstitial cell edema. The allopurinol medicine group can be seen as epithelial cell edema, renal interstitial vasodilation congestion and blood cell infiltration near the renal hila. Compared with the model group, the glomerulus of each treatment group is nearly normal in size and shape. The kidney pathology structure of the high-dose test group of the composition for relieving hyperuricemia is normal, the glomerulus is uniform in size, normal in shape and clear in boundary. The results show that the food composition for relieving hyperuricemia can improve kidney injury of mice.
Example 6
This example provides sensory evaluation and formula optimization of compositions for alleviating hyperuricemia
Sensory evaluation method: 20 specially trained evaluators (10 men and 10 women with ages of 18-25 years) are selected to form a sensory evaluation group, the prepared composite rice thin is brewed with hot water at 100 ℃ according to a ratio of 1:3, and the evaluators perform sensory evaluation according to sensory evaluation criteria (table 4).
Table 4: sensory scoring criteria
A single factor test and an orthogonal test are used to determine the optimal formulation of the composition for alleviating hyperuricemia using the sensory evaluation score as an indicator. Based on a single factor test, an L18 (37) orthogonal test (table 5) is designed, and a sensory evaluation score is taken as an investigation index to obtain an orthogonal test result (table 6), so that an optimal formula is screened.
Table 5: orthogonal test design table
Table 6: results of the orthogonal test
As shown in Table 6, the optimal formula comprises 40% of polished round-grained rice, 7% of poria cocos, 8% of Chinese yam, 10% of lily, 6% of jujube, 8% of chicken's gizzard-skin, 7% of stir-fried coix seed, 1.5% of rose and 12.5% of skim milk powder. And (3) performing verification tests on the optimal formula, wherein the sensory score of the optimal combination is 95 and higher than that of each test result in the orthogonal test, so that the sensory quality of the formula determined by the orthogonal test is stable and feasible.
The present invention may be better implemented as described above, and the above examples are merely illustrative of preferred embodiments of the present invention and not intended to limit the scope of the present invention, and various changes and modifications made by those skilled in the art to the technical solution of the present invention should fall within the scope of protection defined by the present invention without departing from the spirit of the design of the present invention.