Application of sugarcane polyphenol extract in preparation of medicine for preventing and / or treating hyperuricemia

Abstract

The invention relates to an application of a sugarcane polyphenol extract in preparation of a medicine for preventing and / or treating hyperuricemia. The invention also discloses application of the sugarcane polyphenol extract in medicines or health care products for gout caused by hyperuricemia, chronic gout, liver injury and kidney injury. The technical problem to be solved is how to provide the sugarcane polyphenol extract, the sugarcane polyphenol extract contains at least 32 polyphenol monomers, the uric acid reducing effect is improved through the synergistic effect among the polyphenol monomers, and the sugarcane polyphenol extract can be used for preparing drugs or health care products for preventing and / or treating hyperuricemia.

Description

Technical Field

[0001] The present invention belongs to the technical field of pharmaceutical applications of sugarcane polyphenol extracts, and particularly relates to an application of the sugarcane polyphenol extracts in preparing a drug for preventing and / or treating hyperuricemia. Background Art

[0002] With the rapid development of the social economy and the significant improvement in people's quality of life, people's lifestyles and dietary patterns are quietly changing. Today, the intake of nutrients such as sugar, fat, and protein in the daily diet has increased significantly, which has directly led to the continued increase in the incidence of hyperuricemia.

[0003] The typical characteristic of hyperuricemia is an abnormally elevated blood uric acid level. In view of this, maintaining blood uric acid within the target range in the long term has become a core principle in the prevention and treatment of hyperuricemia. However, long-term reliance on uric acid-lowering drugs for treatment may not only induce potential risks such as liver and kidney damage, but may also increase the incidence of hypersensitivity reactions. In addition, patients often find it difficult to adhere to standardized treatment for a long time. These factors together constitute a major challenge in the treatment of hyperuricemia. Therefore, exploring ingredients with uric acid-lowering activity from natural resources has become the focus of current research.

[0004] At present, although the commonly used phenolic extracts on the market are expected to reduce uric acid, prevent and / or treat hyperuricemia, they are mostly limited to a single target and it is difficult to comprehensively regulate key links such as production, excretion and inflammation in the uric acid metabolic network. It is impossible to achieve efficient and precise intervention effects, and it is difficult to meet the comprehensive treatment needs under the complex pathological conditions of hyperuricemia. Summary of the Invention

[0005] The main purpose of the present invention is to provide a use of a sugarcane polyphenol extract in the preparation of a drug for preventing and / or treating hyperuricemia. The technical problem to be solved is how to provide a sugarcane polyphenol extract containing at least 32 polyphenol monomers, which improves the uric acid-lowering effect through the synergistic effect between the polyphenol monomers and can be used to prepare a drug or health product for preventing and / or treating hyperuricemia.

[0006] The purpose of the present invention and the technical problems solved are achieved by adopting the following technical solutions. According to the use of a sugarcane polyphenol extract proposed by the present invention in the preparation of a drug for preventing and / or treating hyperuricemia, the use of the sugarcane polyphenol extract in a drug or health product for gout, chronic gout, liver damage and kidney damage caused by hyperuricemia; chlorogenic acid, p-coumaric acid, vanillin, syringic acid, caffeic acid, isoerythrin, vitexin, tangerin, diosmin, naringenin, quercetin, rutin, isorhamnetin, catechin gallate, wheat flavonoids, biochanin A, umbelliferone, 5,4' -Dihydroxy-3,3'-dimethoxy-6,7-methylenedioxyflavone 4'-O-glucuronide, apigenin 7-O-glucuronide, rough ragweed, isoquercetin, psoralen, (-)-epigallocatechin, eriodictyol, proanthocyanidin trimer C1, 7,3',4'-trihydroxyflavone, 6"-O-malonyl glycitin, theaflavin 3'-O-gallate, myricetin, pinorexin, peony root, delphinidin 3-O-(6"-acetyl-galactoside).

[0007] Preferably, in the aforementioned application, the sugarcane polyphenol extract comprises, by mass percentage, chlorogenic acid 51.14%, p-coumaric acid 9.46%, vanillin 5.38%, syringic acid 3.69%, caffeic acid 3.23%, isorhizin 1.10%, vitexin 0.88%, dapoxetine 0.74%, diosmin 0.65%, naringenin 0.58%, quercetin 0.57%, rutin 0.54%, isorhamnetin 0.52%, catechin gallate 0.46%, wheat flavonoids 0.43%, biochanin A 0.34%, umbelliferone 0.27%, 5,4'-dihydroxy-3,3'-dimethoxy-6,7-methylenedioxyflavone 4'-O-glucuronide 0.19%, apigenin 7-O-glucuronide 0.16%, ragweed 0.15%, isoquercetin 0.12%, psoralen 0.11%, (-)-epigallocatechin 0.10%, eriodictyol 0.06%, proanthocyanidin trimer C1 0.05%, 7,3',4'-trihydroxyflavone 0.04%, 6"-O-malonyl glycitin 0.04%, theaflavin 3'-O-gallate 0.02%, myricetin 0.02%, pinoresin 0.01%, peonyin 0.01%, delphinidin 3-O-(6"-acetyl-galactoside) 0.003%, and the rest are active side effects.

[0008] Preferably, in the aforementioned application, the polyphenol content of the sugarcane polyphenol extract is 17.92-19.18 mgGAE / g.

[0009] Preferably, in the aforementioned application, the method for preparing the sugarcane polyphenol extract specifically comprises:

[0010] 1) At room temperature, weigh 100 mL of Mackey's terminal molasses, add water, mix, and then stir for 10 to 15 minutes to obtain a mixture A; the mixture A is stored at 26 to 28° C.; the volume of the mixture A is 200 mL; the Brix value of the mixture A is 48; and the pH value of the mixture A is 5.4 to 5.6;

[0011] 2) stirring the mixture A at 26-28° C. and slowly adding ethanol while continuously stirring during the addition of ethanol to obtain a mixture B; the ethanol concentration of the mixture B is 83% v / v;

[0012] 3) Continue stirring the mixture B until it becomes turbid and a black gel-like precipitate appears, then centrifuge at 4000 rpm and collect the supernatant to obtain a crude product; the volume of the crude product is 880 mL;

[0013] 4) heating the crude product in a water bath at 45° C. until no ethanol is contained in the crude product, thereby obtaining an extract; the extract has a Brix value of 64 to 65; the extract is dark or yellow in color and does not contain any particulate matter.

[0014] Preferably, in the aforementioned application, in step 1) and step 4), the Brix test method is: take 1 mL of sample, then add 1 mL of water to dilute, mix thoroughly, and then place a drop on an Ella refractometer to test the Brix.

[0015] The purpose of the present invention and the solution of its technical problems are also achieved by the following technical solutions. According to the present invention, a composition for preventing and / or treating hyperuricemia, gout, chronic gout, liver damage and kidney damage caused by hyperuricemia, the composition comprises a sugarcane polyphenol extract; the sugarcane polyphenol extract specifically comprises: chlorogenic acid, p-coumaric acid, vanillin, syringic acid, caffeic acid, isorhamnetin, vitexin, arachidonitrile, diosmin, naringenin, quercetin, rutin, isorhamnetin, catechin gallate, wheat flavonoids, biochanin A, umbelliferone, 5,4'-diol Hydroxy-3,3'-dimethoxy-6,7-methylenedioxyflavone 4'-O-glucuronide, apigenin 7-O-glucuronide, ragweed succourin, isoquercetin, psoralen, (-)-epigallocatechin, eriodictyol, proanthocyanidin trimer C1, 7,3',4'-trihydroxyflavone, 6"-O-malonyl glycitin, theaflavin 3'-O-gallate, myricetin, pinorexin, peonidin, delphinidin 3-O-(6"-acetyl-galactoside).

[0016] Preferably, the aforementioned composition, wherein the sugarcane polyphenol extract comprises, by mass percentage: chlorogenic acid 51.14%, p-coumaric acid 9.46%, vanillin 5.38%, syringic acid 3.69%, caffeic acid 3.23%, isorhizin 1.10%, vitexin 0.88%, dapoxetine 0.74%, diosmin 0.65%, naringenin 0.58%, quercetin 0.57%, rutin 0.54%, isorhamnetin 0.52%, catechin gallate 0.46%, wheat flavonoids 0.43%, biochanin A 0.34%, umbelliferone 0.27%, 5,4'-dihydroxy-3,3'-dimethoxy-6,7-methylenedioxyflavone 4'-O-glucuronide 0.19%, apigenin 7-O-glucuronide 0.16%, ragweed 0.15%, isoquercetin 0.12%, psoralen 0.11%, (-)-epigallocatechin 0.10%, eriodictyol 0.06%, proanthocyanidin trimer C1 0.05%, 7,3',4'-trihydroxyflavone 0.04%, 6"-O-malonyl glycitin 0.04%, theaflavin 3'-O-gallate 0.02%, myricetin 0.02%, pinoresin 0.01%, peonyin 0.01%, delphinidin 3-O-(6"-acetyl-galactoside) 0.003%, and the rest are active side effects.

[0017] Preferably, in the aforementioned composition, the polyphenol content of the sugarcane polyphenol extract is 17.92-19.18 mgGAE / g.

[0018] Preferably, the preparation method of the sugarcane polyphenol extract in the aforementioned composition specifically comprises:

[0019] 1) At room temperature, weigh 100 mL of Mackey's terminal molasses, add water, mix, and then stir for 10 to 15 minutes to obtain a mixture A; the mixture A is stored at 26 to 28° C.; the volume of the mixture A is 200 mL; the Brix value of the mixture A is 48; and the pH value of the mixture A is 5.4 to 5.6;

[0020] 2) stirring the mixture A at 26-28° C. and slowly adding ethanol while continuously stirring during the addition of ethanol to obtain a mixture B; the ethanol concentration of the mixture B is 83% v / v;

[0021] 3) Continue stirring the mixture B until it becomes turbid and a black gel-like precipitate appears, then centrifuge at 4000 rpm and collect the supernatant to obtain a crude product; the volume of the crude product is 880 mL;

[0022] 4) heating the crude product in a water bath at 45° C. until no ethanol is contained in the crude product, thereby obtaining an extract; the extract has a Brix value of 64 to 65; the extract is dark or yellow in color and does not contain any particulate matter.

[0023] Preferably, in the aforementioned composition, in step 1) and step 4), the Brix test method is: take 1 mL of sample, then add 1 mL of water to dilute, mix thoroughly, and then place a drop on an Ella refractometer to test the Brix.

[0024] By means of the above technical solution, the use of a sugarcane polyphenol extract proposed by the present invention in the preparation of a drug for preventing and / or treating hyperuricemia has at least the following advantages:

[0025] The present invention discloses an innovative application of sugarcane polyphenol extract. The disclosed sugarcane polyphenol extract is rich in 32 polyphenol monomer components with different structures. These polyphenol monomers have a significant synergistic effect and show a better uric acid-lowering effect under their joint action. Based on this characteristic, the sugarcane polyphenol extract shows broad application prospects and significant therapeutic effects in the development of compositions for preventing and / or treating hyperuricemia and its complications such as gout (including chronic gout), liver damage and kidney damage caused by it.

[0026] The sugarcane polyphenol extract of the present invention can also effectively inhibit the expression of inflammatory factors by regulating the PI3K / AKT / NF-κB signaling pathway, thereby alleviating the body damage caused by kidney inflammation in HUA hyperuricemia.

[0027] The above description is only an overview of the technical solution of the present invention. In order to more clearly understand the technical means of the present invention and implement it according to the contents of the specification, the following is a detailed description of the preferred embodiments of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 The test results of the total polyphenol content at different boiling times when the pH value is 4 according to the present invention;

[0029] Figure 2 The test results of the total polyphenol content at different boiling times when the pH value is 7 are shown in the present invention;

[0030] Figure 3 The test results of the half-maximal inhibitory concentration (IC50) of different types of polyphenols of the present invention on xanthine oxidase;

[0031] Figure 4 The effects of different types of polyphenols of the present invention on serum uric acid levels in hyperuricemic rats;

[0032] Figure 5The results are the test results of the effects of different doses of sugarcane polyphenols on the serum uric acid levels in hyperuricemia rats;

[0033] Figure 6 The test results of the effect of sugarcane polyphenols of the present invention on the expression of xanthine oxidase activity in the liver of hyperuricemia rats;

[0034] Figure 7 The test results of the effect of sugarcane polyphenols on the expression of URAT1 in the kidneys of hyperuricemia rats are shown in FIG.

[0035] Figure 8 The test results of the effect of sugarcane polyphenols on the expression of ABCG2 in the kidneys of hyperuricemia rats are shown in FIG.

[0036] Figure 9 The test results of the effect of sugarcane polyphenols on the expression of GLUT9 in the kidneys of hyperuricemia rats are shown in FIG.

[0037] Figure 10 The effect of sugarcane polyphenols of the present invention on the urea nitrogen content in serum;

[0038] Figure 11 The effect of sugarcane polyphenols of the present invention on the creatinine content in serum;

[0039] Figure 12 The effect of sugarcane polyphenols of the present invention on glomerular filtration rate;

[0040] Figure 13 The effect of sugarcane polyphenols of the present invention on alanine aminotransferase;

[0041] Figure 14 The effect of sugarcane polyphenols of the present invention on aspartate aminotransferase;

[0042] Figure 15 This is an H&E staining image of kidney sections stained with sugarcane polyphenols according to the present invention;

[0043] Figure 16 The effect of sugarcane polyphenols of the present invention on the IL-1β content in rat kidneys;

[0044] Figure 17 The effect of sugarcane polyphenols of the present invention on the IL-6 content in rat kidneys;

[0045] Figure 18 The effect of sugarcane polyphenols of the present invention on the TNF-α content in rat kidneys;

[0046] Figure 19 The effect of sugarcane polyphenols of the present invention on the expression of p-PI3K / PI3K pathway proteins in rat kidney;

[0047] Figure 20The effect of sugarcane polyphenols of the present invention on the expression of p-AKT / AKT pathway proteins in rat kidney;

[0048] Figure 21 This is the effect of sugarcane polyphenols of the present invention on the expression of p-NF-κB / NF-κB pathway proteins in rat kidney. DETAILED DESCRIPTION

[0049] To further illustrate the technical means and efficacy employed by the present invention to achieve its intended purpose, the following, in conjunction with the accompanying drawings and preferred embodiments, details the use of a sugarcane polyphenol extract in preparing a drug for preventing and / or treating hyperuricemia, including its specific implementation, structure, features, and efficacy. In the following description, different references to "one embodiment" or "embodiment" do not necessarily refer to the same embodiment. Furthermore, specific features, structures, or characteristics of one or more embodiments may be combined in any suitable manner.

[0050] The present invention proposes the use of a sugarcane polyphenol extract in the preparation of a drug for preventing and / or treating hyperuricemia, and the use of the sugarcane polyphenol extract in a drug or health product for gout, chronic gout, liver damage and kidney damage caused by hyperuricemia. The sugarcane polyphenol extract includes 32 polyphenol monomers.

[0051] Preferably, in the aforementioned application, the sugarcane polyphenol extract specifically comprises: chlorogenic acid, p-coumaric acid, vanillin, syringic acid, caffeic acid, isorhamnetin, vitexin, tannic acid flavonoids, diosmin, naringenin, quercetin, rutin, isorhamnetin, catechin gallate, wheat flavonoids, biochanin A, umbelliferone, 5,4'-dihydroxy-3,3'-dimethoxy-6,7-methylenedioxyflavone 4'-O -glucuronide, apigenin 7-O-glucuronide, rough ragweed, isoquercetin, psoralen, (-)-epigallocatechin, eriodictyol, proanthocyanidin trimer C1, 7,3',4'-trihydroxyflavone, 6"-O-malonyl glycitin, theaflavin 3'-O-gallate, myricetin, pinorexin, peonidin, delphinidin 3-O-(6"-acetyl-galactoside).

[0052] Preferably, in the aforementioned application, the sugarcane polyphenol extract comprises, by mass percentage, chlorogenic acid 51.14%, p-coumaric acid 9.46%, vanillin 5.38%, syringic acid 3.69%, caffeic acid 3.23%, isorhizin 1.10%, vitexin 0.88%, dapoxetine 0.74%, diosmin 0.65%, naringenin 0.58%, quercetin 0.57%, rutin 0.54%, isorhamnetin 0.52%, catechin gallate 0.46%, wheat flavonoids 0.43%, biochanin A 0.34%, umbelliferone 0.27%, 5,4'-dihydroxy-3,3'-dimethoxy-6,7-methylenedioxyflavone 4'-O-glucuronide 0.19%, apigenin 7-O-glucuronide 0.16%, ragweed 0.15%, isoquercetin 0.12%, psoralen 0.11%, (-)-epigallocatechin 0.10%, eriodictyol 0.06%, proanthocyanidin trimer C1 0.05%, 7,3',4'-trihydroxyflavone 0.04%, 6"-O-malonyl glycitin 0.04%, theaflavin 3'-O-gallate 0.02%, myricetin 0.02%, pinoresin 0.01%, peonyin 0.01%, delphinidin 3-O-(6"-acetyl-galactoside) 0.003%, and the rest are active side effects.

[0053] The present invention discloses the specific contents of the raw materials in the sugarcane polyphenol extract in terms of mass percentage, which are: chlorogenic acid (51.14%), p-coumaric acid (9.46%), vanillin (5.38%), syringic acid (3.69%), caffeic acid (3.23%), isorhamnetin (1.10%), vitexin (0.88%), tangerin (0.74%), diosmin (0.65%), naringenin (0.58%), quercetin (0.57%), rutin (0.54%), isorhamnetin (0.52%), catechin gallate (0.46%), wheat flavonoids (0.43%), biochanin A (0.34%), umbelliferone (0.27%), 5,4'-dihydroxy-3,3'-dimethoxy-6,7-methylenedioxy Flavonoid 4'-O-glucuronide (0.19%), apigenin 7-O-glucuronide (0.16%), ragweed glycoside (0.15%), isoquercetin (0.12%), psoralen (0.11%), (-)-epigallocatechin (0.10%), eriodictyol (0.06%), proanthocyanidin trimer C1 (0.05%), 7,3',4'-trihydroxyflavone (0.04%), 6"-O-malonyl glycitin (0.04%), theaflavin 3'-O-gallate (0.02%), myricetin (0.02%), pinoretin (0.01%), peonyin (0.01%), delphinidin 3-O-(6"-acetyl-galactoside) (0.003%), and the rest are active side effects.

[0054] It is particularly noteworthy that the active side effects disclosed in this invention are not impurities as traditionally defined in the prior art. In fact, these components are a class of polyphenols that are extremely rare and difficult to accurately measure due to the limited precision of detection methods. There may be hundreds of such polyphenols, and their difficulty in accurate detection is primarily due to the limitations of current detection methods, a situation widely recognized within the industry as a systematic error.

[0055] Preferably, in the aforementioned application, the polyphenol content of the sugarcane polyphenol extract is 17.92-19.18 mgGAE / g.

[0056] Preferably, in the aforementioned application, the method for preparing the sugarcane polyphenol extract specifically comprises:

[0057] 1) At room temperature, weigh 100 mL of Mackey's terminal molasses, add water, mix, and then stir for 10 to 15 minutes to obtain a mixture A; the mixture A is stored at 26 to 28° C.; the volume of the mixture A is 200 mL; the Brix value of the mixture A is 48; and the pH value of the mixture A is 5.4 to 5.6;

[0058] 2) stirring the mixture A at 26-28° C. and slowly adding ethanol while continuously stirring during the addition of ethanol to obtain a mixture B; the ethanol concentration of the mixture B is 83% v / v;

[0059] 3) Continue stirring the mixture B until it becomes turbid and a black gel-like precipitate appears, then centrifuge at 4000 rpm and collect the supernatant to obtain a crude product; the volume of the crude product is 880 mL;

[0060] 4) heating the crude product in a water bath at 45° C. until no ethanol is contained in the crude product, thereby obtaining an extract; the extract has a Brix value of 64 to 65; the extract is dark or yellow in color and does not contain any particulate matter.

[0061] Preferably, in the aforementioned application, in step 1) and step 4), the Brix test method is: take 1 mL of sample, then add 1 mL of water to dilute, mix thoroughly, and then place a drop on an Ella refractometer to test the Brix.

[0062] It is worth noting that the preparation method of the sugarcane polyphenol extract described in the present invention is a publicly available preparation method.

[0063] The present invention also provides a composition for preventing and / or treating hyperuricemia, gout, chronic gout, liver damage and kidney damage caused by hyperuricemia, wherein the composition comprises a sugarcane polyphenol extract; the sugarcane polyphenol extract comprises 32 polyphenol monomers.

[0064] Preferably, the aforementioned composition, wherein the sugarcane polyphenol extract specifically includes: chlorogenic acid, p-coumaric acid, vanillin, syringic acid, caffeic acid, isorhamnetin, vitexin, tangerin, diosmin, naringenin, quercetin, rutin, isorhamnetin, catechin gallate, wheat flavonoids, biochanin A, umbelliferone, 5,4'-dihydroxy-3,3'-dimethoxy-6,7-methylenedioxyflavone 4'- O-glucuronide, apigenin 7-O-glucuronide, ragweed radish, isoquercetin, psoralen, (-)-epigallocatechin, eriodictyol, proanthocyanidin trimer C1, 7,3',4'-trihydroxyflavone, 6"-O-malonyl glycitin, theaflavin 3'-O-gallate, myricetin, pinorexin, peonidin, delphinidin 3-O-(6"-acetyl-galactoside).

[0065] Preferably, in the aforementioned composition, the polyphenol content of the sugarcane polyphenol extract is 17.92-19.18 mgGAE / g.

[0066] Preferably, the aforementioned composition, wherein the sugarcane polyphenol extract comprises, by mass percentage: chlorogenic acid 51.14%, p-coumaric acid 9.46%, vanillin 5.38%, syringic acid 3.69%, caffeic acid 3.23%, isorhamnetin 1.10%, vitexin 0.88%, tannic acid 0.74%, diosmin 0.65%, naringenin 0.58%, quercetin 0.57%, rutin 0.54%, isorhamnetin 0.52%, catechin gallate 0.46%, wheat flavonoids 0.43%, biochanin A 0.34%, umbelliferone 0.27%, 5,4'-dihydroxy-3,3'-dimethoxy-6,7-methylenedioxyflavone 4'-O-glucuronide 0.19%, apigenin 7-O-glucuronide 0.16%, ragweed 0.15%, isoquercetin 0.12%, psoralen 0.11%, (-)-epigallocatechin 0.10%, eriodictyol 0.06%, proanthocyanidin trimer C1 0.05%, 7,3',4'-trihydroxyflavone 0.04%, 6"-O-malonyl glycitin 0.04%, theaflavin 3'-O-gallate 0.02%, myricetin 0.02%, pinoresin 0.01%, peonyin 0.01%, delphinidin 3-O-(6"-acetyl-galactoside) 0.003%, and the rest are active side effects.

[0067] Preferably, the preparation method of the sugarcane polyphenol extract in the aforementioned composition specifically comprises:

[0068] 1) At room temperature, weigh 100 mL of Mackey's terminal molasses, add water, mix, and then stir for 10 to 15 minutes to obtain a mixture A; the mixture A is stored at 26 to 28° C.; the volume of the mixture A is 200 mL; the Brix value of the mixture A is 48; and the pH value of the mixture A is 5.4 to 5.6;

[0069] 2) stirring the mixture A at 26-28° C. and slowly adding ethanol while continuously stirring during the addition of ethanol to obtain a mixture B; the ethanol concentration of the mixture B is 83% v / v;

[0070] 3) Continue stirring the mixture B until it becomes turbid and a black gel-like precipitate appears, then centrifuge at 4000 rpm and collect the supernatant to obtain a crude product; the volume of the crude product is 880 mL;

[0071] 4) heating the crude product in a water bath at 45° C. until no ethanol is contained in the crude product, thereby obtaining an extract; the extract has a Brix value of 64 to 65; the extract is dark or yellow in color and does not contain any particulate matter.

[0072] Preferably, in the aforementioned composition, in step 1) and step 4), the Brix test method is: take 1 mL of sample, then add 1 mL of water to dilute, mix thoroughly, and then place a drop on an Ella refractometer to test the Brix.

[0073] The present invention will be further described below with reference to specific embodiments, but this should not be construed as limiting the scope of protection of the present invention. Non-essential improvements and adjustments made to the present invention by those skilled in the art based on the above-mentioned contents of the present invention still fall within the scope of protection of the present invention.

[0074] Unless otherwise specified, the materials and reagents mentioned below are commercially available products familiar to those skilled in the art. Unless otherwise specified, the methods described are all well-known methods in the art. Unless otherwise defined, technical or scientific terms used shall have the same meanings as those commonly understood by those skilled in the art.

[0075] Example 1

[0076] A method for preparing a sugarcane polyphenol extract is as follows:

[0077] 1) Raw material preparation:

[0078] At room temperature (RT), 100 ml of Mackay terminal molass (140 g) was measured into a glass beaker. 100 ml of distilled water was then added and stirred manually with a glass stirring rod until most of the viscous molass was mixed with the water. The beaker was then placed on a magnetic stirrer and mixed for 10-15 minutes, maintaining the temperature at 26-28°C. The pH of the solution was 5.4-5.6. A 1 ml sample was removed to a final volume of 200 ml and diluted with 1 ml of water, mixed thoroughly, and then a drop was placed on an Ella refractometer, resulting in a Brix reading of 48.

[0079] 2) Extraction using AR ethanol (100% v / v):

[0080] The raw material (200 ml) was placed in a glass beaker on a magnetic stirrer and adjusted so that a clear vortex was formed. Ethanol was slowly added to the vortex to ensure rapid mixing of the raw material and ethanol, maintaining the temperature at 26-28°C. After approximately 30 minutes, 950 ml of ethanol was added, resulting in a final ethanol level of 83% v / v in the mixture. The mixture was stirred for an additional 30 minutes. During the addition of ethanol, multiple subsamples were collected to observe the change in solution color and the change in color of the precipitate as the percentage of ethanol increased.

[0081] 3) Recovery of 83% ethanol supernatant / extract:

[0082] The final mixture is turbid, with a solidified black, gelatinous precipitate at the bottom of the beaker. Collect the supernatant and centrifuge at 4000 rpm (2500 × g) for 5 minutes. Retain the clear, yellow supernatant and discard the black precipitate in the centrifuge tube. Centrifuge approximately 950 ml of the mixture, and the final volume of the recovered supernatant is 880 ml.

[0083] 4) Remove ethanol from the supernatant:

[0084] The ethanol was removed under vacuum using a Buchi rotary evaporator with a bath temperature of 45° C. The final concentrate (which did not contain any ethanol smell) had a volume of 62 ml and a Brix level of 64 to 65. Ethanol, its azeotrope and water were removed in the process.

[0085] 5) Bioactive extracts:

[0086] The final bioactive extract is dark / yellow in color, does not contain any particulate matter and has a sweet flavor similar to that of golden syrup or treacle.

[0087] Comparative Example 1

[0088] A method for preparing an apple polyphenol extract, comprising:

[0089] 1) Weigh 50 g of freeze-dried apple peel powder, add 500 mL of 80% v / v ethanol, and stir at 50°C for 4 h. Filter and centrifuge at 4000 rpm for 5 min, and collect the supernatant.

[0090] 2) The supernatant is subjected to rotary evaporation to remove the solvent, a small amount of distilled water is added to dissolve it, and then freeze-dried to obtain the apple polyphenol extract.

[0091] Comparative Example 2

[0092] A method for preparing a citrus polyphenol extract, comprising:

[0093] 1) Weigh 50 g of freeze-dried citrus peel powder, add 500 mL of 80% v / v ethanol, and stir at 50°C for 4 h. Filter and centrifuge at 4000 rpm for 5 min, and collect the supernatant.

[0094] 2) The supernatant is subjected to rotary evaporation to remove the solvent, a small amount of distilled water is added to dissolve it, and then freeze-dried to obtain the citrus polyphenol extract.

[0095] The stability tests of sugarcane polyphenol extract, apple polyphenol extract and citrus polyphenol extract were conducted as follows:

[0096] 1) Add 200 mL of distilled water to a conical flask, adjust the pH to 4, neutral (pH = 7), and wrap with tin foil to protect from light. Prepare several groups;

[0097] 2) Add 1.6g of sugarcane polyphenols, apple polyphenols, and citrus polyphenols to the pH-adjusted water to a concentration of 0.008g / mL.

[0098] 3) The resulting solution was placed in a boiling water bath with magnetic stirring. When the solution concentration reached 95°C, a sample was taken at time 0, and timing was started. 2 mL samples were taken at 0, 1, 5, 10, 30, and 60 minutes, respectively. The samples were labeled and tested.

[0099] 4) Three 200 μL aliquots of the sample were placed in 10 mL centrifuge tubes at each time point. 1.8 mL of distilled water was added, followed by 200 μL of Folin-phenol reagent, and the mixture was thoroughly mixed and allowed to rest for 5 minutes. Subsequently, 2 mL of 7% sodium carbonate and 800 μL of distilled water were added to each centrifuge tube, mixed thoroughly, and allowed to rest for 90 minutes. After the reaction, the absorbance was measured at 750 nm using a UV spectrophotometer. The total polyphenol content (as gallic acid) of the sugarcane polyphenol solutions treated at different times was calculated using a standard curve.

[0100] The embodiment and the comparative example were subjected to comparative experiments, and the experimental methods and results are as follows:

[0101] Boiling stability test:

[0102] 200mL of distilled water was added to a conical flask, and the pH was adjusted to acidic (pH=4) or neutral (pH=7), and the tin foil was wrapped to avoid light. 1.6g of sugarcane polyphenols, apple polyphenols, and citrus polyphenols were added to the water to adjust the pH to 0.008g / mL. The solution was placed in a boiling water bath and magnetically stirred at the same time. When the solution concentration reached 95°C, a sample was taken at this moment as time 0, and the timing was started. 2mL of samples were taken at 0, 1, 5, 10, 30, and 60min, and the samples were marked and tested. 200μL of the sample at each time point was taken three times and placed in a 10mL centrifuge tube. 1.8mL of distilled water was added, followed by 200μL of Folin phenol reagent, fully mixed, and allowed to stand for 5min. Then, 2mL of 7% sodium carbonate and 800μL of distilled water were added to each centrifuge tube, and the mixture was allowed to stand for 90min after fully mixing. After the reaction, the absorbance was measured at 750 nm using an ultraviolet spectrophotometer, and the total polyphenol content (in terms of gallic acid) of the sugarcane polyphenol solutions treated at different times was calculated according to the standard curve.

[0103] The test results are as follows:

[0104] Table 1: Verification of polyphenol boiling stability at different pH

[0105]

[0106] pass Figure 1 、 Figure 2 As can be seen from Table 1:

[0107] Experimental observations at a pH of 4 revealed that the polyphenol content of uncooked sugarcane polyphenols was 219.09 ± 4.17. After 60 minutes of cooking, the polyphenol content only slightly changed to 225.37 ± 0.70, maintaining a high degree of stability overall, with no significant decrease in total phenol content.

[0108] In contrast, apple polyphenols and citrus polyphenols showed a significant decrease in content after 5 minutes of boiling, dropping from an initial 222.96±3.26 and 225.31±3.01 to 197.84±2.59 and 198.14±2.27, respectively. As the boiling time was extended to 60 minutes, the content of these two polyphenols further decreased significantly, reaching as low as 142.69±3.66 and 123.44±2.83, respectively, significantly lower than the final content of sugarcane polyphenols, highlighting the excellent stability of sugarcane polyphenols under different pH conditions.

[0109] It is worth noting that when the boiling experiment was carried out at a pH value of 7, the results were consistent with those at a pH value of 4, further verifying the stability advantage of sugarcane polyphenols.

[0110] The sugarcane polyphenol extract was tested for its uric acid-lowering activity as follows:

[0111] In vitro experiments were conducted to determine the inhibitory effect of polyphenols on xanthine oxidase (XO), a key enzyme in uric acid production. Xanthine, a substrate for XO, was added to the reaction system, and the production of the reaction product (uric acid) was monitored to indirectly quantify XO's catalytic activity and analyze the inhibitory effect of polyphenols on XO.

[0112] In the in vivo experiment, the method of combined administration of hypoxanthine and potassium oxalate was used to establish a hyperuricemia model in rats. At the same time, the model rats were gavaged with sugarcane polyphenols and the characteristic component chlorogenic acid. The uric acid metabolism level, liver and kidney function level, uric acid metabolic target activity, kidney pathological changes, kidney inflammatory factor expression level and PI3K / AKT / NF-κB pathway protein expression were detected. Through multi-dimensional and multi-level data analysis, the uric acid-lowering activity of sugarcane polyphenols in vivo was determined.

[0113] The specific method is as follows:

[0114] Experimental Animals: Male Sprague Dawley rats (SPF grade, 6-8 weeks old, weighing 200±20g) were purchased from Beijing Weitonglihua Laboratory Animal Technology Co., Ltd. (Experimental Animal Production License No.: SCXK(Beijing)2021-0006). All rats were housed in a standardized environment, maintaining a constant temperature and humidity (room temperature 22-24°C, relative humidity 50-60%), and a standardized light cycle (12h / 12h light-dark cycle). Standard feed and water were available ad libitum. Rats were acclimated for one week before the experiment.

[0115] 1) After adaptive feeding, the experimental animals were randomly divided into 10 groups, with 8 rats in each group; specifically, blank group (Normal), sample control group (Sample control), model group (Mondel), low-dose sugarcane polyphenol group (SPL), medium-dose sugarcane polyphenol group (SPM), high-dose sugarcane polyphenol group (SPH), positive control chlorogenic acid group (CGA), vanillin group, apple polyphenol group, and citrus polyphenol group;

[0116] 2) First, a two-week sugarcane polyphenol nutritional intervention was conducted, with the recommended sugarcane polyphenol intake (10 g / day) converted to the rat feeding amount. All rats were gavaged with sugarcane polyphenols at the following doses: SPL group: 4.77 μg SP / g BW / d, SPM group: 9.54 μg SP / g BW / d, SPH group: 19.07 μg SP / g BW / d. The sample control group and SPH group received the same dose (19.07 μg SP / g BW / d). The CGA group, vanillin group, apple polyphenol group, and citrus polyphenol group all received 19.07 μg / g BW / d. The blank and model groups were gavaged with an equal amount of ultrapure water once daily for two weeks.

[0117] Table 2: Oral administration volume of different groups

[0118] Group gavage volume Group gavage volume SPL 4.77 μg SP / g BW / d Vanillin 19.07 μg / g BW / d SPM 9.54 μg SP / g BW / d Apple polyphenols 19.07 μg / g BW / d SPH 19.07 μg SP / g BW / d Citrus polyphenols 19.07 μg / g BW / d Sample control 19.07 μg SP / g BW / d Blank group 19.07 μg / g BW / d CGA 19.07 μg / g BW / d Model Group 19.07 μg / g BW / d

[0119] 3) Prepare modeling drugs:

[0120] Potassium oxonate (PO): 500 mg / kg;

[0121] Hypoxanthine (HX): 300 mg / kg;

[0122] Solvent: 0.5% sodium carboxymethylcellulose (CMC-Na);

[0123] 4) After 2 weeks of nutritional intervention, mice were gavaged with 500 mg / kg potassium oxonate (PO) combined with 300 mg / kg hypoxanthine (HX) to establish the model. The modeling drug was solubilized with 0.5% sodium carboxymethylcellulose (CMC-Na). The modeling drug was gavaged 2 hours after the nutritional intervention every day. The blank group and the sample control group were gavaged with the same amount of 0.5% CMC-Na.

[0124] 5) After the final oral administration of the modeling drug and each polyphenol group, the rats were fasted for 10 hours. Blood was collected from the abdominal aorta after euthanasia, and liver and kidney tissues were harvested intact. Unilateral kidneys were fixed in paraformaldehyde for pathological section preparation. The remaining organs were quickly frozen in liquid nitrogen and stored at -80°C for subsequent biochemical analysis.

[0125] It is worth noting that this experimental protocol complies with animal welfare and ethical requirements and was approved by the Animal Ethics Review Committee of PONY Testing Group Co., Ltd. (PONY-2023-FL-22).

[0126] Determination of xanthine oxidase inhibitory activity:

[0127] The catalytic activity of XO can be indirectly measured by the production of uric acid, a product, by adding xanthine, a substrate for XO, to the reaction system. Different concentrations of SP solutions, five polyphenol solutions (chlorogenic acid, vanillin, sugarcane polyphenols, apple polyphenols, and citrus polyphenols), and XO (0.01 U / mL) were prepared in a 0.1 M Tris-HCl buffer system (pH 6.8). 50 μL of SP or polyphenol monomers at different concentrations was added to a 96-well plate, followed by 50 μL of XO solution, mixed thoroughly, and incubated at 30°C for 5 minutes. The reaction was then initiated by the addition of 150 μL of xanthine solution (0.3 mM), and the absorbance at 292 nm was recorded for 600 seconds. The changes in absorbance in the presence and absence of the inhibitor were recorded as A and B, respectively. The inhibition rate of XO was calculated using the following formula:

[0128] XO enzyme relative activity (%) = A / B × 100%

[0129] Where A is the absorbance in the presence of an inhibitor, and B is the absorbance in the absence of an inhibitor.

[0130] The IC50 value of the inhibitor against XO can be obtained by fitting the inhibitor concentration and the inhibition rate.

[0131] The test results are as follows:

[0132] Table 3: Half-maximal inhibitory concentration (IC50) of different polyphenols on xanthine oxidase 50 )

[0133] Group <![CDATA[IC 50 (μg / mL)]]> Chlorogenic acid 314.84±18.01 Vanillin 458.52±17.24 Sugarcane polyphenols 335.36±18.04 Apple polyphenols 523.65±18.81 Citrus polyphenols 430.86±19.35

[0134] pass Figure 3As shown in Table 3, the inhibitory ability of sugarcane polyphenols, apple polyphenols, citrus polyphenols, chlorogenic acid and vanillin on xanthine oxidase (XO), a key enzyme in uric acid production, was determined by in vitro experiments. The results showed that the IC 50 The value was 335.36±18.04μg / mL, which was lower than the IC values of apple polyphenols (523.65±18.81μg / mL) and citrus polyphenols (430.86±19.35μg / mL). 50 The values indicate that at the same concentration, sugarcane polyphenols have the most significant inhibitory effect on xanthine oxidase compared with apple polyphenols and citrus polyphenols, indicating that they have the best inhibitory ability on the key enzyme of uric acid production.

[0135] Determination of uric acid content in rat serum

[0136] After rat plasma was allowed to stand at room temperature for 1 hour, it was centrifuged at 3500 rpm for 10 minutes to obtain serum. Rat serum UA levels were measured using a kit (enzyme colorimetric method). The procedure was as follows: distilled water, a standard, and 5 μL of each serum sample were added to a 96-well plate. Subsequently, 250 μL of a mixture containing peroxidase and uricase was added to each well. After mixing, the mixture was incubated at 37°C for 10 minutes. The absorbance at 510 nm was read using a microplate reader.

[0137] Table 4: Effects of different polyphenols on serum uric acid levels in hyperuricemic rats

[0138]

[0139] pass Figure 4 From Table 4 we can see that:

[0140] In vivo experiments, sugarcane polyphenols also showed the best uric acid-lowering activity at the same phenol content (e.g. Figure 4 The results of the in vitro experiments showed that sugarcane polyphenols showed a good uric acid-lowering effect both in vivo and in vitro.

[0141] At present, the general research consensus reached by those skilled in the art is that the amount of polyphenol added is positively correlated with the efficacy of reducing the serum uric acid level in hyperuricemic rats, that is, as the amount of polyphenol added increases, the uric acid-lowering effect gradually increases until it reaches the marginal effect. Figure 5Analysis of the data in Table 4 shows that serum uric acid levels gradually decreased in the low-dose sugarcane polyphenols group (SPL), the medium-dose sugarcane polyphenols group (SPM), and the high-dose sugarcane polyphenols group (SPH), reaching 77.80±19.37μmol / L, 46.84±15.97μmol / L, and 33.78±9.83μmol / L, consistent with this trend. However, further comparison of the experimental data between the high-dose sugarcane polyphenols group (SPH) and the chlorogenic acid group (CGA) revealed that serum uric acid levels in the high-dose sugarcane polyphenols group (SPH) were 33.78±9.83μmol / L and 52.37±12.07μmol / L, respectively. According to the prevailing logic, serum uric acid levels in the two groups should converge when the phenol content is equal. However, the experimental results showed that the SPH group had a significantly lower value than the CGA group. This phenomenon shows that when the phenol content is equal, the high-dose sugarcane polyphenol group (SPH) shows a better uric acid-lowering effect. This is attributed to the fact that sugarcane polyphenols are rich in polyphenol monomers, including 32 different types of polyphenol monomers. The synergistic effect between these polyphenol monomers makes its overall uric acid-lowering effect surpass that of chlorogenic acid monomers. It can be inferred that sugarcane polyphenol extract has the synergistic mechanism advantage of "multi-component-multi-target-multi-pathway".

[0142] Determination of Xanthine Oxidoreductase (XOR) Activity in Rat Liver Tissue

[0143] Part of the rat liver tissue was taken to prepare a tissue homogenate, and the supernatant was collected after centrifugation. The XOR content in the rat liver tissue was detected using a kit (enzyme colorimetric method).

[0144] Hypoxanthine is converted into uric acid during metabolism in the liver, in which xanthine oxidoreductase (XOR) plays a key role. Figure 6 The data obtained can show that:

[0145] Compared with the blank group (Normal), the XOR expression level in the model group (Model, 140.5±1.21μmol / L) increased by 77.66%. This is because XOR catalyzes the conversion of hypoxanthine to uric acid in the liver, which in turn leads to an increase in the serum uric acid level in the model group rats.

[0146] Compared with the model group, the XOR level in the sugarcane polyphenol intervention group decreased significantly. Among them, the low-dose sugarcane polyphenol group (SPL, 12.10±1.29μmol / L) decreased by 13.84%, the medium-dose sugarcane polyphenol group (SPM, 11.04±0.56μmol / L) decreased by 21.40%, and the high-dose sugarcane polyphenol group (SPH, 10.40±0.98μmol / L) decreased by 25.93%. These results indicate that sugarcane polyphenols have good XOR inhibitory ability; the positive control chlorogenic acid group (CGA, 9.80±1.13μmol / L) decreased by 30.27% compared with the model group, which further proves that chlorogenic acid has a good ability to reduce liver XOR.

[0147] Renal excretion also plays a vital role in blood uric acid levels. The expression of uric acid transporters in rat kidneys was tested. Figure 7 、 Figure 8 and Figure 9 As shown:

[0148] pass Figure 7 It can be seen that compared with the blank group (Normal), the expression level of URAT1, a transporter responsible for uric acid reabsorption, in the model group (Model) was significantly increased (p < 0.05); while the sugarcane polyphenol intervention group effectively slowed down the expression of the reabsorption transporter URAT1, thereby reducing the reabsorption of uric acid from urine to serum.

[0149] ATP binding cassette superfamily Gmember 2 (ABCG2) is an important transporter protein that regulates uric acid excretion. Studies have shown that the loss of ABCG2 function leads to the blockage of urate secretion in the human body, thereby increasing uric acid levels. Figure 8 It can be seen that after nutritional intervention in the sugarcane polyphenol intervention group (SPL, SPM, SPH), the expression level of the secretory transporter ABCG2 was increased, thereby promoting the secretion of uric acid from cells to the lumen.

[0150] The uric acid transporter GLUT9 protein can transport uric acid from cells to the blood, leading to an increase in the uric acid level in the blood. Figure 9As can be seen, the sugarcane polyphenol intervention groups (SPL, SPM, and SPH) can downregulate the elevated expression of GLUT9 in rats with high uric acid levels, and this downregulation effect increases with increasing sugarcane polyphenol dosage. Compared with the positive control chlorogenic acid (CGA) group, the SPH group significantly downregulated GLUT9 expression (P < 0.05) at the same polyphenol content, demonstrating the beneficial GLUT9 regulatory effects of sugarcane polyphenols. Furthermore, the unmodified control group showed no significant differences in the regulation of the three transporters compared to the blank group.

[0151] Determination of liver function and kidney function indicators in rats:

[0152] After the rat plasma was placed at room temperature for 1 hour, it was centrifuged at 3500 rpm for 10 minutes to obtain serum. The levels of blood urea nitrogen (BUN), creatinine (Cr), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) in the rat serum were measured using commercial kits. The results are as follows:

[0153] The kidneys are an important site for uric acid excretion. Decreased renal function will affect uric acid excretion in the body. Creatinine (CRE) and urea nitrogen (BUN) are important indicators for evaluating renal function. Both are filtered by the glomeruli and excreted by the renal tubules. The higher the BUN and CRE levels in the serum, the more severe the renal function damage.

[0154] pass Figure 10 It can be seen that compared with the blank group (Normal), the BUN level of the model group (Model, 11.12±0.92μmol / L) was significantly increased, 1.67 times that of the blank group. The sugarcane polyphenol intervention groups (SPL, SPM, SPH) and the positive control chlorogenic acid group (CGA) were able to significantly reduce serum BUN levels (p < 0.05). By comparing SPL, SPM, and SPH, it can be seen that sugarcane polyphenol extract has a certain dose effect. By comparing SPH and CGA, it can be seen that when the polyphenol content is the same, the effect of SPH is significantly better than CGA.

[0155] pass Figure 11 It can be seen that compared with the blank group (Normal), the CRE level in the model group (Model, 67.47±5.15μmol / L) was significantly increased, 2.09 times that of the blank group. The sugarcane polyphenol intervention groups (SPL, SPM, SPH) and the positive control chlorogenic acid group (CGA) were able to significantly reduce the serum CRE level (p < 0.05). By comparing SPL, SPM and SPH, it can be seen that the sugarcane polyphenol extract has a certain dose effect. By comparing SPH and CGA, it can be seen that when the polyphenol content is the same, the effect of SPH is significantly better than CGA.

[0156] In summary, both the sugarcane polyphenols intervention group and the positive control chlorogenic acid group were able to reduce the expression levels of CRE and BUN in serum, and the sugarcane polyphenols intervention group also showed a dose effect. A comparison of SPH and CGA showed that, given the same polyphenol content, SPH was significantly more effective than CGA. This also suggests that the abundant polyphenols in SP have a significant synergistic effect with each other, and their combined action can better alleviate renal damage in hyperuricemia.

[0157] The glomerulus, a key structural unit in the kidney's excretion function, is a crucial component of the kidney's excretion system. Glomerular filtration rate (GFR) is a core indicator for assessing the kidney's filtration and excretion functions. It can typically be accurately calculated by measuring serum creatinine levels, reflecting the body's filtration efficiency and excretion capacity.

[0158] like Figure 12 As shown in the study, under the pathological condition of hyperuricemia, the glomerular filtration rate of experimental rats showed a significant downward trend, directly reflecting the damage of hyperuricemia to the renal filtration function. However, under the intervention measures of each treatment group, the glomerular filtration rate of the rats showed varying degrees of recovery. In particular, the glomerular filtration rate of rats in the SPH treatment group recovered significantly, reaching a level that was insignificantly different from that of the blank control group. This fully demonstrates the effectiveness of SPH treatment in improving renal function damage caused by hyperuricemia.

[0159] High levels of uric acid accumulation in the body can damage multiple organ systems. Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) are commonly used clinical indicators for liver function assessment. These two enzymes are primarily present in the cytoplasm of hepatocytes. When hepatocytes are damaged, the integrity of the cell membrane is disrupted, and large amounts of ALT and AST are released into the blood, leading to abnormally elevated serum levels of these two enzymes.

[0160] like Figure 13 and Figure 14As shown in the results of a study, compared with the blank control group, serum ALT and AST levels in model rats induced by hypoxanthine (HX) and potassium oxalate (PO) were significantly elevated (P < 0.05). This result clearly demonstrates that hyperuricemia causes severe liver damage in rats. Both the sugarcane polyphenol intervention groups (SPL, SPM, and SPH) and the positive control chlorogenic acid (CGA) group reduced serum ALT and AST levels to some extent, effectively alleviating liver damage in rats. The SPH (sugarcane polyphenol) group showed the most significant therapeutic effect. Compared with the model group, ALT levels in the SPH group decreased by 20.26% (P < 0.05) and AST levels decreased by 18.14% (P < 0.05). This result not only fully demonstrates the excellent efficacy of SPH in protecting liver function and alleviating liver damage caused by hyperuricemia, but also demonstrates that, given the same polyphenol content, SPH is significantly more effective than CGA, indicating a significant synergistic effect between the abundant polyphenols in SP.

[0161] Uric acid (UA) is the end product of purine metabolism in nucleoproteins and nucleic acids, and most of it is excreted through the kidneys. Its excretion process is complex: after filtration by the glomeruli, UA undergoes secretion, reabsorption, and resecretion in the renal tubules, ultimately being excreted in the urine. To assess the extent of damage caused by hyperuricemia, H&E staining of kidney sections was performed to observe pathological changes. The specific method is as follows:

[0162] After fixation in paraformaldehyde for 48 hours, rat kidneys and ileum were dehydrated through graded ethanol (70% to absolute ethanol), cleared with xylene, and then impregnated with paraffin before embedding into paraffin blocks. The paraffin blocks were sectioned, flattened, mounted on glass slides, and baked to enhance adhesion. Paraffin sections were stained with hematoxylin and erythrin, and then mounted. Histological structure was observed under an inverted microscope.

[0163] Here are the results:

[0164] like Figure 15As shown, the nephron structures of the rats in the blank and sample control groups were intact and compact, with neatly arranged renal tubules and intact lumens. There was no glomerular proliferation or degeneration, indicating that the kidneys of both groups were in a normal physiological state. However, the kidneys of the rats in the model group showed severe morphological damage. Numerous tubular epithelial cells in the model group atrophied and shed, the brush border disappeared, and the tubular lumen became significantly dilated and irregular in shape. Glomeruli also showed signs of sclerosis, atrophy, and even necrosis. Inflammatory cell infiltration was also present in the renal tissue. This indicates that the model group rats developed acute tubular necrosis 16 days after HX+PO treatment, leading to renal inflammation. Compared with the model group, the pathological changes in the rats' kidneys were significantly improved after the nutritional intervention with sugarcane polyphenols (SP) and chlorogenic acid (CGA). Necrosis of tubular epithelial cells and tubular dilation were alleviated, the severity of glomerular lesions was reduced, and inflammatory cell infiltration was significantly reduced. What is particularly noteworthy is that the renal pathological sections of the SPH group and the CGA group were almost the same as those of the blank group, which fully demonstrated that SP and CGA have significant effects in the treatment of kidney damage caused by hyperuricemia, and can effectively protect the kidneys and reduce the damage of hyperuricemia to the kidneys.

[0165] When uric acid is overproduced in the body, high concentrations of uric acid in the blood, after being filtered through the glomeruli, exceed the reabsorption capacity of the proximal renal tubules. Consequently, large amounts of uric acid remain in the renal tubules. When the accumulation reaches a certain level, it triggers a series of inflammatory responses in the kidneys. By measuring the expression of proinflammatory cytokines (IL-1β, IL-6, and TNF-α), the effects of SP and CGA on renal inflammation in rats were determined.

[0166] The specific method is as follows:

[0167] Partial renal tissue was collected from rats to prepare tissue homogenates, which were then centrifuged and the supernatant collected. Enzyme-linked immunosorbent assays were used to measure the levels of interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor (TNF-α) in renal tissues. Results were corrected for protein concentration in each sample.

[0168] Here are the results:

[0169] like Figure 16 、 Figure 17 and Figure 18As shown, the pro-inflammatory factors in the sample control group were not significantly different from those in the blank group. However, compared with the blank group, the levels of inflammatory factors IL-1β, IL-6, and TNF-α in the kidneys of hyperuricemia rats in the model group were significantly increased by 164.16%, 72.47%, and 66.64%, respectively (P<0.05), indicating that hyperuricemia caused the occurrence of renal inflammatory reactions in rats. In the SP and CGA groups, the expression levels of inflammatory factors in the kidneys of rats decreased to varying degrees. Among them, the SPM, SPH, and CGA groups all showed good inflammatory factor clearance effects, and the inflammatory factor levels were not significantly different from those in the blank group, indicating that SP can alleviate renal inflammation caused by hyperuricemia to a certain extent.

[0170] The present invention further detects the expression levels of p-PI3K / PI3K, p-AKT / AKT and p-NF-κB / NF-κB pathways in kidney. Figure 19 、 Figure 20 and Figure 21 As can be seen, the ratios of p-PI3K / PI3K, p-AKT / AKT, and p-NF-κB / NF-κB in the model group were significantly higher than those in the normal group, strongly indicating activation of these signaling pathways. In contrast, phosphorylation levels of these pathway proteins decreased in the SPL, SPM, SPH, and CGA groups. The SPH group showed the most significant effect. This suggests that SP exhibits significant synergistic effects among its polyphenol monomers, effectively inhibiting the activation of the p-PI3K / PI3K, p-AKT / AKT, and p-NF-κB / NF-κB pathways, thereby suppressing renal inflammation in rats.

[0171] It is noteworthy that the method of western blot analysis of the present invention is:

[0172] Prepare an appropriate amount of rat tissue, wash with 0.01M PBS, and add 5 volumes of RIPA lysis buffer (containing 1% PMSF and 2% phosphatase inhibitors) to prepare a tissue homogenate. Lyse on ice for 30 minutes. Centrifuge the tissue homogenate at 12,000 rpm at 4°C for 10 minutes. Remove the supernatant, determine and equalize the protein concentration, add 5x loading buffer, and denature. Proteins in the samples were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to PVDF membranes. The membranes were blocked with 5% skim milk at room temperature for 1 h and then incubated with primary antibodies against URAT1 (1:1500), ABCG2 (1:1000), GLUT9 (1:1000), PI3K (1:1000), p-PI3K (1:1000), AKT (1:1000), p-AKT (1:1000), NFKB (1:3000), p-NFKB (1:1000), and GAPDH (1:1000) overnight at 4°C. After incubation, the PVDF membranes were washed with TBST. Secondary antibodies were then applied, and the blots were detected using ECL luminescence. Protein expression was quantified by measuring grayscale values using Image J software, and the expression of the target protein in each group was normalized to the internal control GAPDH.

[0173] 10. Statistical analysis

[0174] All data are expressed as mean ± standard deviation (SD). Data were analyzed using one-way analysis of variance and Duncan's test (SPSS 23.0). P < 0.05 was considered significant. Graphs were created using Graphpad Pism 9.5 software. For the same indicator, different letters between groups indicate significant differences.

[0175] From the above experimental results, we can see that:

[0176] 1. Sugarcane polyphenol extract exhibited excellent uric acid-lowering activity both in vitro and in vivo, significantly outperforming apple polyphenols, citrus polyphenols, and chlorogenic acid at equivalent phenolic content. Sugarcane polyphenols effectively reduced serum uric acid levels in hyperuricemic rats in a dose-dependent manner, with the SPH group (19.07 μg SP / g BW / d) demonstrating a particularly significant nutritional intervention effect.

[0177] 2. Sugarcane polyphenols can exert their effects through dual pathways: on the one hand, they can inhibit the activity of xanthine oxidase (XO), a key enzyme in the production of uric acid, thereby reducing the excessive production of uric acid; on the other hand, sugarcane polyphenols can also regulate the expression of uric acid transporters (such as URAT1, ABCG2, and GLUT9) in the kidneys, promoting the excretion of uric acid from the blood into the urine.

[0178] 3. After nutritional intervention with sugarcane polyphenols, the impaired liver and kidney function of rats with hyperuricemia was significantly restored. Furthermore, while lowering serum uric acid levels, sugarcane polyphenols also effectively reduced the expression of inflammatory factors by regulating the PI3K / AKT / NF-κB signaling pathway, thereby alleviating the kidney inflammation and resulting damage caused by hyperuricemia.

[0179] 4. Chlorogenic acid, as a characteristic component of sugarcane polyphenols, participates in its uric acid-lowering activity in vivo to a certain extent. However, under the same phenol content conditions, by comparing the SPH group and the CGA group, it can be seen that the effect of chlorogenic acid alone on lowering the serum uric acid level in rats is not as good as that of sugarcane polyphenols as a whole. This shows that the synergistic effect between the polyphenol monomers of sugarcane polyphenols has a "multi-component-multi-target-multi-pathway" synergistic mechanism, which can improve the uric acid-lowering effect and can play a better role in drugs or health products for gout, chronic gout, liver damage and kidney damage caused by hyperuricemia.

[0180] It is worth noting that the SP mentioned in this invention refers to sugarcane polyphenols. The SP group and sugarcane polyphenol intervention group mentioned in this invention both refer to experimental groups containing sugarcane polyphenols, including SPH, SPM, and SPL. The chlorogenic acid group mentioned in this invention is the CGA group.

[0181] In the above embodiments, the description of each embodiment has its own focus. For parts that are not described in detail in a certain embodiment, reference can be made to the relevant descriptions of other embodiments.

[0182] The above description is merely a preferred embodiment of the present invention and does not constitute any form of limitation to the present invention. Although the present invention has been disclosed as a preferred embodiment as above, it is not intended to limit the present invention. Any technician familiar with this profession can make some changes or modifications to equivalent embodiments of the technical contents disclosed above without departing from the scope of the technical solution of the present invention. However, any simple modifications, equivalent changes and modifications made to the above embodiments based on the technical essence of the present invention without departing from the content of the technical solution of the present invention are still within the scope of the technical solution of the present invention.

Claims

1. A use of a sugarcane polyphenol extract in preparing a drug for preventing and / or treating hyperuricemia, characterized in that: The sugarcane polyphenol extract is used in medicines or health products for gout, chronic gout, liver damage and kidney damage caused by hyperuricemia; the sugarcane polyphenol extract specifically includes: chlorogenic acid, p-coumaric acid, vanillin, syringic acid, caffeic acid, isorhamnetin, vitexin, tangerin, diosmin, naringenin, quercetin, rutin, isorhamnetin, catechin gallate, wheat flavonoids, biochanin A, umbelliferone, 5,4'-dihydroxy-3,3'-dimethoxy 6,7-methylenedioxyflavone 4'-O-glucuronide, apigenin 7-O-glucuronide, rough ragweed, isoquercetin, psoralen, (-)-epigallocatechin, eriodictyol, proanthocyanidin trimer C1, 7,3',4'-trihydroxyflavone, 6"-O-malonyl glycitin, theaflavin 3'-O-gallate, myricetin, pinorexin, peonidin, delphinidin 3-O-(6"-acetyl-galactoside).

2. The use according to claim 1, characterized in that In terms of mass percentage, the sugarcane polyphenol extract includes: chlorogenic acid 51.14%, p-coumaric acid 9.46%, vanillin 5.38%, syringic acid 3.69%, caffeic acid 3.23%, isorhamnetin 1.10%, vitexin 0.88%, flavonoids 0.74%, diosmin 0.65%, naringenin 0.58%, quercetin 0.57%, rutin 0.54%, isorhamnetin 0.52%, catechin gallate 0.46%, wheat flavonoids 0.43%, biochanin A 0.34%, umbelliferone 0.27%, 5,4'-dihydroxy-3,3'-dimethoxy-6,7-methylenedioxyflavone 4'-O-glucose Glucuronide 0.19%, apigenin 7-O-glucuronide 0.16%, ragweed 0.15%, isoquercetin 0.12%, psoralen 0.11%, (-)-epigallocatechin 0.10%, eriodictyol 0.06%, proanthocyanidin trimer C1 0.05%, 7,3',4'-trihydroxyflavone 0.04%, 6"-O-malonyl glycitin 0.04%, theaflavin 3'-O-gallate 0.02%, myricetin 0.02%, pinoresin 0.01%, peonyin 0.01%, delphinidin 3-O-(6"-acetyl-galactoside) 0.003%, and the rest are active side effects.

3. The use according to claim 2, characterized in that The polyphenol content of the sugarcane polyphenol extract is 17.92-19.18 mg GAE / g.

4. The use according to claim 3, characterized in that The preparation method of the sugarcane polyphenol extract specifically comprises: 1) At room temperature, weigh 100 mL of Mackey's terminal molasses, add water, mix, and then stir for 10 to 15 minutes to obtain a mixture A; the mixture A is stored at 26 to 28° C.; the volume of the mixture A is 200 mL; the Brix value of the mixture A is 48; and the pH value of the mixture A is 5.4 to 5.6; 2) stirring the mixture A at 26-28° C. and slowly adding ethanol while continuously stirring during the addition of ethanol to obtain a mixture B; the ethanol concentration of the mixture B is 83% v / v; 3) Continue stirring the mixture B until it becomes turbid and a black gel-like precipitate appears, then centrifuge at 4000 rpm and collect the supernatant to obtain a crude product; the volume of the crude product is 880 mL; 4) heating the crude product in a water bath at 45° C. until no ethanol is contained in the crude product, thereby obtaining an extract; the extract has a Brix value of 64 to 65; the extract is dark or yellow in color and does not contain any particulate matter.

5. The use according to claim 4, characterized in that In step 1) and step 4), the Brix content is tested by taking 1 mL of the sample, diluting it with 1 mL of water, mixing it thoroughly, and then placing a drop of the sample on an Ella refractometer to test the Brix content.

6. A composition for preventing and / or treating hyperuricemia, gout, chronic gout, liver damage and kidney damage caused by hyperuricemia, characterized in that: The composition includes a sugarcane polyphenol extract; the sugarcane polyphenol extract specifically includes: chlorogenic acid, p-coumaric acid, vanillin, syringic acid, caffeic acid, isorhamnetin, vitexin, scutellarin, diosmin, naringenin, quercetin, rutin, isorhamnetin, catechin gallate, wheat flavonoids, biochanin A, umbelliferone, 5,4'-dihydroxy-3,3'-dimethoxy-6,7-methylenedioxyflavone 4' -O-glucuronide, apigenin 7-O-glucuronide, rough ragweed, isoquercetin, psoralen, (-)-epigallocatechin, eriodictyol, proanthocyanidin trimer C1, 7,3',4'-trihydroxyflavone, 6"-O-malonyl glycitin, theaflavin 3'-O-gallate, myricetin, pinorexin, peonidin, delphinidin 3-O-(6"-acetyl-galactoside).

7. The use according to claim 6, characterized in that In terms of mass percentage, the sugarcane polyphenol extract includes: chlorogenic acid 51.14%, p-coumaric acid 9.46%, vanillin 5.38%, syringic acid 3.69%, caffeic acid 3.23%, isorhamnetin 1.10%, vitexin 0.88%, flavonoids 0.74%, diosmin 0.65%, naringenin 0.58%, quercetin 0.57%, rutin 0.54%, isorhamnetin 0.52%, catechin gallate 0.46%, wheat flavonoids 0.43%, biochanin A 0.34%, umbelliferone 0.27%, 5,4'-dihydroxy-3,3'-dimethoxy-6,7-methylenedioxyflavone 4'-O-glucose Glucuronide 0.19%, apigenin 7-O-glucuronide 0.16%, ragweed 0.15%, isoquercetin 0.12%, psoralen 0.11%, (-)-epigallocatechin 0.10%, eriodictyol 0.06%, proanthocyanidin trimer C1 0.05%, 7,3',4'-trihydroxyflavone 0.04%, 6"-O-malonyl glycitin 0.04%, theaflavin 3'-O-gallate 0.02%, myricetin 0.02%, pinoresin 0.01%, peonyin 0.01%, delphinidin 3-O-(6"-acetyl-galactoside) 0.003%, and the rest are active side effects.

8. The composition according to claim 7, characterized in that The polyphenol content of the sugarcane polyphenol extract is 17.92-19.18 mg GAE / g.

9. The composition according to claim 8, characterized in that The preparation method of the sugarcane polyphenol extract specifically comprises: 1) At room temperature, weigh 100 mL of Mackey's terminal molasses, add water, mix, and then stir for 10 to 15 minutes to obtain a mixture A; the mixture A is stored at 26 to 28° C.; the volume of the mixture A is 200 mL; the Brix value of the mixture A is 48; and the pH value of the mixture A is 5.4 to 5.6; 2) stirring the mixture A at 26-28° C. and slowly adding ethanol while continuously stirring during the addition of ethanol to obtain a mixture B; the ethanol concentration of the mixture B is 83% v / v; 3) Continue stirring the mixture B until it becomes turbid and a black gel-like precipitate appears, then centrifuge at 4000 rpm and collect the supernatant to obtain a crude product; the volume of the crude product is 880 mL; 4) heating the crude product in a water bath at 45° C. until no ethanol is contained in the crude product, thereby obtaining an extract; the extract has a Brix value of 64 to 65; the extract is dark or yellow in color and does not contain any particulate matter.

10. The composition according to claim 9, characterized in that In step 1) and step 4), the Brix content is tested by taking 1 mL of the sample, diluting it with 1 mL of water, mixing it thoroughly, and then placing a drop of the sample on an Ella refractometer to test the Brix content.

Images