CN117783405A - Screening and application of uric acid and blood glucose reducing active ingredients in Opuntia Dillenii fruit purified extract - Google Patents
Screening and application of uric acid and blood glucose reducing active ingredients in Opuntia Dillenii fruit purified extract Download PDFInfo
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- CN117783405A CN117783405A CN202311700274.7A CN202311700274A CN117783405A CN 117783405 A CN117783405 A CN 117783405A CN 202311700274 A CN202311700274 A CN 202311700274A CN 117783405 A CN117783405 A CN 117783405A
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- glu
- xod
- purified extract
- alpha
- uric acid
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- 238000012216 screening Methods 0.000 title claims abstract description 23
- LEHOTFFKMJEONL-UHFFFAOYSA-N Uric Acid Chemical compound N1C(=O)NC(=O)C2=C1NC(=O)N2 LEHOTFFKMJEONL-UHFFFAOYSA-N 0.000 title claims abstract description 21
- 239000008280 blood Substances 0.000 title claims abstract description 19
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- TVWHNULVHGKJHS-UHFFFAOYSA-N Uric acid Natural products N1C(=O)NC(=O)C2NC(=O)NC21 TVWHNULVHGKJHS-UHFFFAOYSA-N 0.000 title claims abstract description 18
- 229940116269 uric acid Drugs 0.000 title claims abstract description 18
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Abstract
The invention provides screening and application of uric acid and blood glucose reducing active ingredients in a cactus fruit purified extract, wherein the screening method comprises the following steps: after incubating the purified extract of opuntia ficus-indica with XOD/a-Glu, bioaffinity ultrafiltration separates the ligand-XOD/a-Glu inhibitor complex from unbound complex, followed by analytical identification and quantification of the ligand compound released from the ligand-XOD/a-Glu inhibitor complex by the UPLC-QTRAP-MS/MS method. According to the invention, the XOD and alpha-Glu inhibitors are screened and identified from the Opuntia Dillenii fruit purified extract for the first time by combining bioaffinity ultrafiltration with UPLC-QTRAP-MS/MS technology, and binding sites and related action mechanisms of the inhibitors and enzymes are explained according to molecular docking, so that theoretical and technical support is provided for preparing natural medicines for reducing uric acid and reducing blood glucose, and the method has a great application prospect.
Description
Technical Field
The invention belongs to the technical field of extraction of plant active ingredients, and relates to screening and application of uric acid and blood glucose reducing active ingredients in a cactus fruit purified extract.
Background
Hyperuricemia (HUA) is a metabolic abnormality syndrome caused by purine metabolic disturbance, and is a major risk factor for gout and also a major risk factor for obesity, hypertension, atherosclerosis, dyslipidemia, cardiovascular diseases, chronic kidney disease and diabetes. Xanthine Oxidase (XOD) is a key molybdenum-containing enzyme involved in catabolism of purine nucleic acids and catalyzes oxidation of Xanthine to uric acid, while producing Reactive Oxygen Species (ROS), and XOD inhibitors reduce serum uric acid concentration by inhibiting uric acid synthesis, and are useful in preventing hyperuricemia. Allopurinol, an XOD inhibitor, is the most commonly prescribed drug for the treatment of gout, but it is associated with hypersensitivity reactions.
Diabetes (Diabetes mellitus, DM) is a chronic metabolic disease caused by abnormal carbohydrate metabolism, postprandial hyperglycemia can cause severe damage, dysfunction and failure of multiple organs and tissues with progressive metabolic complications. Alpha-glucosidase (alpha-Glu) catalyzes the hydrolysis of dietary carbohydrates and converts them to monosaccharides, which are then absorbed in the jejunum, and is prone to cause diabetes and other related diseases. Whereas alpha-Glu inhibitors delay glucose absorption, reduce postprandial blood glucose and insulin levels, typical alpha-Glu inhibitors, such as acarbose and miglitol, also cause gastrointestinal side effects.
Natural products with specific active frameworks, active molecules and good biological activity play an important role in the long history of human resistance to metabolic diseases, and many enzyme inhibitors have been found and isolated from extracts of fruits, vegetables and herbs. Previous research reports indicate that polyphenols such as quercetin, kaempferol, isorhamnetin, rutin, hyperoside and quercetin are isolated and identified from Flos Sophorae Immaturus as potential inhibitors of XOD. In addition, rutin, isoquercitrin, chlorogenic acid, quercetin and cinnamic acid are screened and identified from Pyrus pyrifolia Fruit to have a certain inhibition effect on alpha-Glu. Thus, natural products extracted from plants can act as potent inhibitors of XOD and alpha-Glu.
Cactus fruit (OFI), also known as Rosa roxburghii, is a plant native to Mexico and belongs to the Cactaceae family. OFI not only has a great deal of functions, nutrition and biological activity, but also has social, agricultural and ecological benefits. The reported researches on uric acid and blood glucose reducing related components in the cactus fruit are less, wherein the effective components and related action mechanisms of the XOD and alpha-Glu inhibitor are not reported yet, and the cactus fruit is not fully developed and utilized in uric acid and blood glucose reducing activity.
Disclosure of Invention
The invention aims to provide screening and application of uric acid and blood glucose reducing active ingredients in the cactus fruit purified extract, and the screening of uric acid and blood glucose reducing potential inhibitors from the cactus fruit purified extract is carried out for the first time, so that theoretical and technical support is provided for preparing natural medicines for reducing uric acid and blood glucose, and the application prospect is great.
In order to achieve the above purpose, the invention adopts the following technical scheme:
the invention provides a screening method of uric acid and blood glucose reducing active ingredients in a cactus fruit purified extract, which comprises the following steps: after incubating the purified extract of opuntia ficus-indica with XOD/a-Glu, bioaffinity ultrafiltration separates the ligand-XOD/a-Glu inhibitor complex from unbound complex, followed by analytical identification and quantification of the ligand compound released from the ligand-XOD/a-Glu inhibitor complex by the UPLC-QTRAP-MS/MS method.
Preferably, 10mg/mL of the purified extract of Opuntia Dillenii fruit is mixed with 5U/mL of an XOD solution having a pH of 7.4 and 10U/mL of an alpha-Glu solution having a pH of 6.8 in a volume ratio of 1:2 incubated in a 30kDa ultrafiltration tube at 37℃for 30 min.
Preferably, the ultrafiltration is performed by centrifugation at 12000 Xg for 10 minutes using an ultrafiltration tube, and the enzyme-bound material in the ultrafiltration tube is eluted by washing the compound not specifically bound to XOD/α -Glu three times with 0.2M/0.1M phosphate buffer and then dissociating with 90% methanol to obtain the eluent.
Preferably, the bioaffinity of the inhibitor and XOD/α -Glu is calculated according to the following formula:
wherein C1, C2 and C0 represent the concentration of the selected compound in the purified extracts of Opuntia Dillenii having activity XOD/alpha-Glu, inactivated XOD/alpha-Glu and no XOD/alpha-Glu, respectively.
Preferably, the target compound is chromatographed on a Waters UPLC liquid chromatography column, with phase a being a 0.1% formic acid in water and phase B being acetonitrile at a flow rate of 0.8mL/min, with a gradient elution procedure of: 0-5 min, 0-15% A; 5-20 min, 15-25% A; 20-40 min, 25-50% A; 40-55 min, 50-80% A; 55-60 min,15% A, column temperature set at 40 ℃, autosampler temperature set at 4 ℃ and sample injection amount 2 μl.
Preferably, mass spectrometry is performed in a multi-reaction monitoring mode with ion source parameters as follows: ionSpray Voltage: +5500/-4500V,Curtain Gas:35psi,Temperature:400 ℃, ion Source Gas 1:60psi,Ion Source Gas 2:60psi,DP: +100V.
Preferably, the purified extract of Opuntia Dillenii is screened for seven XOD inhibitor components: dihydromyricetin, kaempferide, methyl gallate, isorhamnetin, chlorogenic acid, naringenin, dihydroquercetin and eight alpha-Glu inhibitor components: kaempferide, isorhamnetin, astragalin, rutin, isoquercitrin, naringin, chlorogenic acid, and dihydroquercetin.
Preferably, the purified extract of opuntia ficus-indica is prepared by the following method: removing seeds of fresh picked cactus fruit, vacuum freeze drying, grinding into powder, sieving with 60 mesh sieve, fully mixing fruit powder with 60% ethanol, ultrasonic-assisted extraction, centrifuging to collect supernatant, re-extracting residue once according to the same method, combining the supernatants extracted twice, concentrating the supernatant with a rotary evaporator, filling a chromatographic column with AB-8 macroporous adsorption resin for purification, washing effluent with distilled water until colorless, eluting with 95% ethanol, concentrating the eluent eluted with ethanol, and freeze drying to obtain purified polyphenol extract.
The invention also provides application of the uric acid and blood glucose reducing active ingredient obtained by the screening method in preparation of uric acid and blood glucose reducing products.
Preferably, the product comprises a medicament.
According to the invention, the XOD and the alpha-Glu inhibitor are screened and identified from the Opuntia Dillenii fruit purified extract for the first time by combining bioaffinity ultrafiltration with UPLC-QTRAP-MS/MS technology, in an ultrafiltration system, the mixture of the target object and the extract passes through an ultrafiltration membrane, natural product components specifically combined with high molecular weight target object molecules are trapped on the membrane, and unbound mixture components are removed, so that the method is simple, reliable and quick, the binding site between the inhibitor and enzyme and related action mechanism are further clarified according to molecular butt joint, theoretical and technical support is provided for preparing natural medicines or health care products for reducing uric acid and reducing blood glucose, further development and utilization of the Opuntia Dillenii fruit are facilitated, and the method has great application prospect.
Drawings
FIG. 1 shows the antioxidant activity and enzyme inhibition IC of the purified extract of Opuntia Dillenii fruit of the present invention 50 Values.
FIG. 2 is a chromatogram of a bioaffinity ultrafiltration screening enzyme inhibitor of the present invention.
FIG. 3 shows the degree of biological binding of the ultrafiltration-screened enzyme inhibitors of the invention and the standard enzyme inhibition IC 50 Values.
FIGS. 4 and 5 are surface and 2D graphs of the interface between the inhibitor of the invention and xanthine oxidase molecules.
FIGS. 6 and 7 are surface and 2D graphs of the interface of an inhibitor of the invention with an alpha-glucosidase molecule.
Detailed Description
In order to more clearly illustrate the present invention, the present invention will be described in further detail below with reference to examples and with reference to the accompanying drawings. It is to be understood by persons skilled in the art that the following detailed description is illustrative and not restrictive, and that this invention is not limited to the details given herein.
Examples
1. Preparation of purified extract of Opuntia Dillenii fruit
Removing seeds from fresh picked cactus fruits, vacuum freeze-drying, grinding into powder, sieving with a 60-mesh sieve, fully mixing the powder with 60% ethanol (W/v=1:30), carrying out ultrasonic auxiliary extraction for 20min at 40 ℃ and 320W, centrifuging for 10min at 4 ℃ and 10000g, collecting supernatant, re-extracting residues once according to the same method, and combining the supernatants extracted from the two steps. After concentrating the supernatant by rotary evaporator, purifying by packing the column with AB-8 macroporous adsorbent resin, washing the effluent with distilled water until colorless, and eluting with about 2BV 95% ethanol. Finally, concentrating the eluent eluted by ethanol by using a rotary evaporator, and obtaining the purified polyphenol extract after freeze drying.
2. Determination of total phenol content
Total polyphenol content was determined by Folin-Ciocalteu colorimetry and expressed as milligrams of Gallic Acid Equivalent (GAE) per gram of sample in Dry Mass (DM) in mg GAE/g DW.
The total polyphenol content of the extract purified by macroporous resin is 355.03mg GAE/g DW, which is improved by about 86 times compared with the crude extract, thus indicating that the cactus fruit extract has rich bioactive substances.
3. Determination of antioxidant Activity
ABTS according to DPPH + Four different antioxidant methods of CUPRAC, FRAR were used to evaluate the antioxidant activity of the purified extracts of opuntia ficus-indica. Trolox was used as a positive control and a standard curve was drawn, DPPH/ABTS + The scavenging activity values are expressed in. Mu. Mol Trolox (TE)/g DM samples.
Wherein: ac is DPPH/ABTS + Absorbance of the solution and sample solvent mixture, ai is absorbance of the DPPH/ABTS solution and sample solution mixture, aj is absorbance of the solvent and sample solution mixture.
Iron ion reduction capacity (FRAP) and copper ion reduction capacity (CUPRAC) were expressed in. Mu. Mol Trolox (TE)/g DW with Trolox as positive control and standard curves were drawn.
The health benefits of opuntia ficus-indica polyphenols may depend on their antioxidant and free radical scavenging activity. The results of the four antioxidant activity measurements of the extracted ABTS+ and FRAP, DPPH, CUPRAC are shown in figure 1 (left graph), which shows that the extract has a certain antioxidant activity, wherein the influence of FRAP (1091.40 mmol TE/100g DW) is most remarkable.
4. XOD and alpha-Glu inhibition activity assay
mu.L of the sample was taken in a 96-well plate, 50. Mu.L of XOD solution (0.1U) was added, and after shaking for 10s, incubated at 37℃for 5min. Then 150. Mu.L of xanthine solution (0.2 mM) was added, the absorbance was measured at 290nm at intervals of 20s, the change in absorbance was recorded over 10min, and the sample solution was replaced with 50. Mu.L of phosphate buffer (0.2M, pH 7.5) in the blank. With allopurinol as a positive control, XOD inhibition activity was calculated according to the following formula:
wherein, (dA/dt) blank: blank reaction rate; (dA/dt) sample: reaction rate of the sample.
Dissolving 100 μL 1U/ml α -Glu in 0.1M phosphate buffer (PBS, pH 6.8), incubating with 100 μL polyphenol extract at 37deg.C for 10min, adding 100 μL 5mM PNPG solution, reacting at 37deg.C for 20min, adding 500 μL 1M Na 2 CO 3 The solution terminated the reaction. Absorbance was measured at 405 nm. The alpha-Glu inhibitory activity was calculated using acarbose and PBS as positive and blank controls according to the following formula:
wherein A1, A0, B1 and B0 represent absorbance of the blank test group (containing PBS buffer and enzyme), the blank control group (containing PBS buffer only), the sample test group (containing sample extract, PBS buffer solution and enzyme) and the sample control group (containing sample extract and PBS buffer), respectively.
The inhibitory activity of the extract on XOD and α -Glu was evaluated by in vitro enzyme inhibition experiments, as shown in fig. 1 (right panel), the extract was purified on XOD (IC 50 = 199.00 μg/mL) and α -Glu (IC 50 159.67 μg/mL), wherein allopurinol and acarbose are positive controls for XOD and α -Glu, respectively, the extract has better inhibitory activity on α -Glu than acarbose (IC) 50 ＝345.67μg/mL)。
5. Screening and identification of potential inhibitors
Screening the extract for XOD and alpha-Glu inhibitors, comprising the following steps: mu.L of 10mg/mL of the extract and 200. Mu.L of XOD (5U/mL, pH 7.4)/alpha-Glu (10U/mL, pH 6.8) were incubated in a 30kDa ultrafiltration tube at 37℃for 30 minutes, then the reaction was centrifuged, and finally the ligand-XOD/alpha-Glu inhibitor complex was captured at 37℃and centrifuged at 12000 Xg for 10 minutes. The ligand was then released from the inhibitor complex by three washes with 200 μl of 0.2M/0.1M phosphate buffer, followed by final addition of 200 μl of 90% methanol for dissociation, and repeated twice. The released ligand was quantified by UPLC-QTRAP-MS/MS. The same ultrafiltration step was performed with inactivated XOD/alpha-Glu as negative control. The bioaffinity of the inhibitor and XOD/alpha-Glu was calculated according to the formula.
Wherein C1, C2 and C0 represent the concentration of the selected compound in the active XOD/alpha-Glu, inactive XOD/alpha-Glu and OFI extract without XOD/alpha-Glu, respectively.
6. UPLC-QTRAP-MS/MS assay
The present application uses an EXION LC System (SCIEX) ultra high performance liquid chromatograph to chromatographically separate the target compound by a Waters UPLC liquid chromatography column (Waters Acquity UPLC HSS T, 1.8 μm, 2.1X100 mm). The phase A of liquid chromatography is aqueous solution containing 0.1% formic acid, the phase B is acetonitrile with the flow rate of 0.8mL/min, and the gradient elution procedure is as follows: 0-5 min, 0-15% A; 5-20 min, 15-25% A; 20-40 min, 25-50% A; 40-55 min, 50-80% A; 55-60 min,15% A. The column temperature was set at 40℃and the autosampler temperature was set at 4℃with a sample loading of 2. Mu.L.
Sciex QTrap 6500 Mass Spectrometry Instrument parameter set up:
the present application uses a SCIEX 6500qtrap+ triple quadrupole mass spectrometer equipped with a IonDrive Turbo V ESI ion source for mass spectrometry in multi-reaction monitoring (MRM) mode. The ion source parameters are as follows: ionSpray Voltage: +5500/-4500V,Curtain Gas:35psi,Temperature:400 ℃, ion Source Gas 1:60psi,Ion Source Gas 2:60psi,DP: +100V.
To better determine uric acid-and blood glucose-lowering components of purified OFI extracts, the present application employs a non-targeted metabonomics approach based on UPLC-MS. The qualitative identification of the substance can be performed by comparing the substance database with standards such as retention time, ionic mass to charge ratio, etc. As shown in fig. 2, 16 chemical components were identified in total in the purified OFI extract. Comprising 5 phenolic acids and derivatives thereof (peaks 1,2,3,8, 12) and 11 flavonoids (peaks 4-7, 9-11, 13-16). Wherein 5 phenolic acids and derivatives thereof are identified as chlorogenic acid, methyl gallate, gentisic acid, ferulic acid and methyl caffeate respectively, and peaks 4-7, 9-11 and 13-16 are identified as dihydromyricetin, isoquercitrin, rutin, kaempferide, kaempferol-3-O-rutinoside, dihydroquercetin, astragalin, quercetin, naringenin, kaempferol and isorhamnetin respectively.
Seven XOD inhibitor components and eight alpha-Glu inhibitor components were separately screened from the purified extract by bioaffinity ultrafiltration screening (fig. 2). Analysis of the bioaffinity of the identified isolated inhibitors to the enzyme revealed that the affinities to XOD were, in order, dihydromyricetin (53.13%), kaempferide (49.21%), methyl gallate (28.19%), isorhamnetin (13.30%), chlorogenic acid (7.56%), naringenin (6.10%), dihydroquercetin (3.25%). The affinity with alpha-Glu is sequentially kaempferide (54.75%), isorhamnetin (24.07%), astragalin (19.00%), rutin (17.31%), isoquercitrin (16.06%), naringenin (9.55%), chlorogenic acid (7.85%), dihydroquercetin (3.23%). The results show that dihydromyricetin and kaempferide have a stronger affinity for XOD, while kaempferide and isorhamnetin have a greater affinity for alpha-Glu than other inhibitors. And there are significant differences in affinity between different compounds and enzymes, which may be due to the different competitive binding relationships between the bioactive component and the different enzymes.
To further analyze the affinity and correlation between the two enzyme inhibitors, the inhibitory activity of a single inhibitor standard on both enzymes was determined separately (fig. 3). The results also show that the inhibitors of both enzymes are screened to have certain inhibition effect on the inhibitors, which indicates that the ultrafiltration screening technology can be used for rapidly screening natural inhibitors.
7. Molecular docking
The ligands and proteins required for molecular docking are prepared using AutoDock Vina software (http:// Vina. Scripps. Edu /), and the crystal structure of the target protein is pre-treated, including removing hydrogenation, modifying amino acids, optimizing energy and adjusting force field parameters, before meeting the low energy conformation of the ligand structure. And finally, carrying out molecular docking on the target structure and the active ingredient structure, carrying out docking by using vina in pyrx software, carrying out visual analysis on the target structure by using Pymol, and carrying out visual analysis on the target structure and the active ingredient structure by using a Discovery Studio 2020 Client in a 2D map.
To elucidate the mechanism of inhibition of potential inhibitors and enzymes, a molecular docking technique was used for validation. It is generally believed that when the molecular docking binding energy is less than 0, two molecules have spontaneous binding ability, and when the molecular docking binding energy is less than-5.0 kcal/mol, two molecules have better binding activity. The docking binding energy of the seven inhibitors and the XOD is within the range of-10.6 to-7.4 kcal/mol, which is less than-5.0 kcal/mol, indicating that the inhibitors all show good binding capacity. In the butt joint of eight inhibitors and alpha-Glu, the butt joint binding energy is in the range of-8.4 to-6.9 kcal/mol, and the screened inhibitors have inhibition effect on the alpha-Glu. Further analysis found that the screening inhibitors were mainly docked to the active center of the enzyme by hydrogen bonding, hydrophobic interactions, etc. to inhibit the activity of the enzyme (fig. 4-7). From the analysis, it is clear that the active compounds screened by ultrafiltration can be well inserted into the active pocket of the key enzyme and exhibit significant interactions with the key amino acid residues of the enzyme.
Screening results show that the main types of the two enzyme inhibitors are phenolic compounds and flavonoid compounds, and the compounds are used as active ingredients of natural sources, have good biological activity and antioxidant activity, and can be used for preparing medicines and health care products for resisting hyperuricemia or diabetes.
The foregoing description of the preferred embodiments of the invention is not intended to limit the invention to the precise form disclosed, and any such modifications, equivalents, and alternatives falling within the spirit and scope of the invention are intended to be included within the scope of the invention.
Claims (10)
1. The screening method of uric acid and blood sugar reducing active ingredients in the purified extract of the cactus fruit comprises the following steps: after incubating the purified extract of opuntia ficus-indica with XOD/a-Glu, bioaffinity ultrafiltration separates the ligand-XOD/a-Glu inhibitor complex from unbound complex, followed by analytical identification and quantification of the ligand compound released from the ligand-XOD/a-Glu inhibitor complex by the UPLC-QTRAP-MS/MS method.
2. The screening method according to claim 1, wherein 10mg/mL of the purified extract of opuntia ficus-indica is mixed with 5U/mL of XOD solution at pH 7.4 and 10U/mL of α -Glu solution at pH 6.8, respectively, in a volume ratio of 1:2 incubated in a 30kDa ultrafiltration tube at 37℃for 30 min.
3. The method according to claim 2, wherein the ultrafiltration is performed by centrifugation at 12000 Xg for 10 minutes using an ultrafiltration tube, the compound not specifically bound to XOD/α -Glu is washed three times with 0.2M/0.1M phosphate buffer, and the enzyme-bound substance in the ultrafiltration tube is eluted by dissociation of 90% methanol to obtain an eluate.
4. The screening method of claim 1, wherein the bioaffinity of the inhibitor and XOD/α -Glu is calculated according to the following formula:
wherein C1, C2 and C0 represent the concentration of the selected compound in the purified extracts of Opuntia Dillenii having activity XOD/alpha-Glu, inactivated XOD/alpha-Glu and no XOD/alpha-Glu, respectively.
5. The method of claim 1, wherein the target compound is chromatographed on a Waters UPLC liquid chromatography column, wherein the phase a is an aqueous solution containing 0.1% formic acid, the phase B is acetonitrile at a flow rate of 0.8mL/min, and the gradient elution procedure is: 0-5 min, 0-15% A; 5-20 min, 15-25% A; 20-40 min, 25-50% A; 40-55 min, 50-80% A; 55-60 min,15% A, column temperature set at 40 ℃, autosampler temperature set at 4 ℃ and sample injection amount 2 μl.
6. The method of claim 5, wherein the mass spectrometry is performed in a multi-reaction monitoring mode with ion source parameters as follows: ionSpray Voltage: +5500/-4500V,Curtain Gas:35psi,Temperature:400 ℃, ion Source Gas 1:60psi,Ion Source Gas 2:60psi,DP: +100V.
7. The screening method of claim 1, wherein seven XOD inhibitor components are screened from the purified extract of opuntia ficus-indica: dihydromyricetin, kaempferide, methyl gallate, isorhamnetin, chlorogenic acid, naringenin, dihydroquercetin and eight alpha-Glu inhibitor components: kaempferide, isorhamnetin, astragalin, rutin, isoquercitrin, naringin, chlorogenic acid, and dihydroquercetin.
8. The screening method according to claim 1, wherein the purified extract of opuntia ficus-indica is prepared by the following method: removing seeds of fresh picked cactus fruit, vacuum freeze drying, grinding into powder, sieving with 60 mesh sieve, fully mixing fruit powder with 60% ethanol, ultrasonic-assisted extraction, centrifuging to collect supernatant, re-extracting residue once according to the same method, combining the supernatants extracted twice, concentrating the supernatant with a rotary evaporator, filling a chromatographic column with AB-8 macroporous adsorption resin for purification, washing effluent with distilled water until colorless, eluting with 95% ethanol, concentrating the eluent eluted with ethanol, and freeze drying to obtain purified polyphenol extract.
9. Use of the uric acid-lowering and blood glucose-lowering active ingredient obtained by the screening method of any one of claims 1 to 8 in the preparation of uric acid-lowering and blood glucose-lowering products.
10. The use according to claim 9, wherein the product comprises a medicament.
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