Application of rhizoma smilacis glabrae extract in preparation of product for treating hyperuricemia

Abstract

The invention discloses application of a rhizoma smilacis glabrae extract in preparation of a product for treating hyperuricemia. The content of tannin in the rhizoma smilacis glabrae extract is greater than 20%, or the rhizoma smilacis glabrae extract also contains astilbin composition components. Through systematic research and animal experiments, it is found that the rhizoma smilacis glabrae extract containing the tannin or the composition containing the tannin and astilbin has the effect of treating hyperuricemia caused by oteracil potassium and adenine and hyperuricemia induced by fructose in a multi-way and multi-target mode; the compound is effective to multiple targets including xanthine oxidase, adenosine deaminase, urate transporter and the like.

Description

Technical Field

[0001] The present invention relates to the technical field of smilax glabra extracts, and in particular to an application of the smilax glabra extract in preparing a product for treating hyperuricemia. Background Art

[0002] Hyperuricemia (HUA) refers specifically to serum uric acid levels exceeding the normal physiological threshold (>420 μmol / L). The pathological mechanism of this condition stems from an imbalance in uric acid metabolism, including hypersynthesis or impaired excretion. This disruption of homeostasis can trigger HUA, increasing the burden on the liver and kidneys, leading to inflammatory responses and organ damage, and resulting in a series of clinical complications, such as gout (specific deposition of urate crystals in synovial tissue), uric acid nephropathy (renal interstitial fibrosis), cardiovascular disease (hypertension), and metabolic syndrome, accompanied by organ damage.

[0003] Uric acid, the end product of purine metabolism, is primarily synthesized in the liver, skeletal muscle, and gastrointestinal mucosa. Xanthine oxidase (XOD) and adenosine deaminase (ADA) are key regulatory enzymes. Uric acid is eliminated through bidirectional regulation of reabsorption and secretion by transporters in the renal excretion system. A high-purine diet is closely associated with HUA. Recent studies have found that fructose, when metabolized in the liver, consumes significant amounts of adenosine triphosphate (ATP). The resulting adenosine monophosphate (AMP) is further degraded by ADA into hypoxanthine, which is ultimately converted into uric acid, leading to HUA. Therefore, a high-fructose diet is considered a significant contributor to the high incidence and younger age of HUA. A high-fructose, high-purine, and high-fat diet can also significantly impact the balance of the intestinal microbiome, further exacerbating HUA.

[0004] Currently, chemical drugs for treating hyperuricemia include uricase XOD inhibitors (allopurinol, febuxostat, and topiroxetine), uricosuric agents (probenecid, sulfinpyrazone, and benzbromarone), urate oxidase analogs, and probiotics (lactic acid bacteria). However, these drugs have varying degrees of adverse reactions and poor long-term medication compliance. Discovering safe and effective drugs for treating HUA from traditional Chinese medicine has significant clinical value.

[0005] Smilax glabra and its compounds are frequently used in traditional medicine to treat HUA. Extensive research has been conducted on its chemical constituents and pharmacological effects, suggesting its efficacy in warming the middle and strengthening the spleen, promoting diuresis and reducing swelling, and providing tranquilizing and sedative effects. Modern pharmacological studies have shown that Smilax glabra exhibits significant pharmacological activities in uric acid-lowering, anti-tumor, antioxidant, anti-inflammatory, analgesic, and antibacterial properties. Literature reports indicate that Smilax glabra contains over 200 chemical constituents, including flavonoids, phenols, phenolic acids, organic acids, and phenylpropanoids. Among these, astilbin has been shown to exhibit excellent therapeutic effects against HUA induced by a high-purine diet. However, due to the complex etiology of HUA, further research is needed to explore the efficacy of Smilax glabra and the composition of its active ingredients. Summary of the Invention

[0006] In response to the deficiencies in the prior art, the present invention provides an application of a Smilax glabra extract in the preparation of a product for treating hyperuricemia. The extract comprising Smilax glabra tannins or a composition comprising Smilax glabra tannins and astilbin is obtained by extracting Smilax glabra. The extract has a therapeutic effect on hyperuricemia caused by potassium oxonate and adenine, as well as fructose-induced hyperuricemia, and has no toxic side effects.

[0007] In order to solve the above technical problems, the present invention provides an application of a Smilax glabra extract in preparing a product for treating hyperuricemia, wherein the Smilax glabra extract includes Smilax glabra tannins; or includes a composition of tannins and astilbin.

[0008] Furthermore, the smilax glabra tannin is a proanthocyanidin formed by polymerization of catechin and epicatechin, and the mass content of the smilax glabra tannin in the smilax glabra extract is greater than 20%.

[0009] Furthermore, the astilbin composition includes one or more of neoastilbin, astilbin, neoisoastilbin, isoastilbin, and chrysanthemoside, wherein the mass content of astilbin in the Smilax glabra extract is greater than 15%.

[0010] Furthermore, the preparation method of the Smilax glabra extract comprises the following steps:

[0011] S1. Extract the Rhizoma Smilacis Glabrae by heating and refluxing with 40%-75% ethanol, filter, and concentrate to obtain an extract;

[0012] S2. The extract is adsorbed on a macroporous resin column, eluted with 30%-70% ethanol, concentrated under reduced pressure, and dried to obtain Smilax glabra extract A.

[0013] Furthermore, the Smilax glabra extract A is prepared into a pH 6-8 solution, extracted with ethyl acetate, and the aqueous layer after extraction is adjusted to pH 2-4, concentrated under reduced pressure, and dried to obtain Smilax glabra extract B.

[0014] Furthermore, the Smilax glabra extract reduces uric acid production by inhibiting xanthine oxidase.

[0015] Furthermore, the Smilax glabra extract reduces uric acid production by inhibiting the expression of adenosine deaminase.

[0016] Furthermore, the Smilax glabra extract improves the pathological state of liver and kidney tissues by reducing serum uric acid, creatinine, urea nitrogen and kidney index.

[0017] Furthermore, the Smilax glabra extract improves the body's inflammatory response by inhibiting the release of serum inflammatory factors TNF-α, IL-6, IL-1β, IL-17A and the formation of NETs (neutrophil granulocyte nets).

[0018] Furthermore, the Smilax glabra extract reduces uric acid production by inhibiting the expression of ADA (adenosine dehydrogenase).

[0019] Furthermore, the Smilax glabra extract promotes uric acid excretion by inhibiting the expression of uric acid transporters GLUT9 and URAT1 and promoting the expression of uric acid transporters ABCG2 and OAT1.

[0020] Furthermore, the product is a medicine, a health product or a food.

[0021] Furthermore, the hyperuricemia is hyperuricemia caused by potassium oxonate and adenine or fructose-induced hyperuricemia.

[0022] Beneficial effects of the present invention:

[0023] Through systematic research and animal experiments, the present invention found that the tannin component (TFL-5) in the Smilax glabra extract can be used alone to resist HUA and has an inhibitory effect on XOD; astilbin can also be used alone to resist HUA, but is ineffective against the XOD enzyme; the Smilax glabra extract (TFL-3) composed of tannins and astilbin has the efficacy of both components, and also shows multiple effects such as regulating blood lipids and anti-inflammatory, with multi-pathway and multi-target effects in treating hyperuricemia caused by potassium oxonate and adenine, as well as fructose-induced hyperuricemia, without obvious toxic and side effects. On this basis, the preparation technology and quality control method of the Smilax glabra extract were established through research to ensure that the content of the active ingredients of tannins and astilbin in the extract is greater than that of conventional water extracts, thereby ensuring the beneficial effect of the Smilax glabra extract in reducing HUA. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] In order to more clearly illustrate the technical solution of the present invention, the following briefly introduces the drawings required for use in the embodiments. Obviously, the drawings described below are merely embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without creative work.

[0025] Figure 1 Figures AF are HPLC results of the Smilax glabra extracts TFL-1 to TFL-5 and astilbin of the present invention, respectively; in the figure, peak positions 1-5 are neoastilbin, astilbin, neoisoastilbin, isoastilbin, and chrysanthemum glycosides, and P1 is 5-O-caffeoylshikimic acid;

[0026] Figure 2 The present invention relates to the effects of the Smilax glabra extract on the levels of serum uric acid, uric acid in urine, urea nitrogen, creatinine, and AST / ALT in rats with hyperuricemia induced by adenine and potassium oxonate (n=5);

[0027] Figure 3 The present invention relates to the effects of the Smilax glabra extract on organ indexes in rats with hyperuricemia induced by adenine and potassium oxonate;

[0028] Figure 4 The present invention is to investigate the effect of the Smilax glabra extract on the blood routine of rats with hyperuricemia induced by adenine and potassium oxonate;

[0029] Figure 5 The present invention relates to the effect of the Smilax glabra extract on blood lipids in rats with hyperuricemia induced by adenine and potassium oxonate;

[0030] Figure 6 The present invention is about the effect of the Smilax glabra extract on NETs in the serum of rats with hyperuricemia induced by adenine and potassium oxonate;

[0031] Figure 7 The present invention relates to the effect of the Smilax glabra extract on TNF-α, IL-1β, IL-6 and IL-17A in rats with hyperuricemia induced by adenine and potassium oxonate;

[0032] Figure 8 The present invention relates to the effect of the Smilax glabra extract on the liver XOD of rats with hyperuricemia induced by adenine and potassium oxonate;

[0033] Figure 9 The present invention is to investigate the effect of the Smilax glabra extract on the expression of GLUT9, ABCG2 and OAT1 mRNA in the kidneys of rats with hyperuricemia induced by adenine and potassium oxonate;

[0034] Figure 10 The present invention relates to the effect of the Smilax glabra extract on the levels of urea nitrogen, creatinine, AST / ALT, and uric acid in urine of fructose-induced hyperuricemia rats;

[0035] Figure 11 The effect of the Smilax glabra extract of the present invention on organ indexes in fructose-induced hyperuricemia rats;

[0036] Figure 12 The effect of the Smilax glabra extract of the present invention on the blood routine of fructose-induced hyperuricemia rats;

[0037] Figure 13 The present invention relates to the effect of the Smilax glabra extract on blood lipids in fructose-induced hyperuricemia rats;

[0038] Figure 14 The present invention is about the effect of the Smilax glabra extract on NETs in fructose-induced hyperuricemia rats;

[0039] Figure 15 The present invention relates to the effect of the Smilax glabra extract on TNF-α, IL-1β, IL-6 and IL-17A in rats with hyperuricemia induced by fructose;

[0040] Figure 16 The present invention is to investigate the effect of the Smilax glabra extract on FA and ADA in fructose-induced hyperuricemia rats;

[0041] Figure 17 The effect of the Smilax glabra extract of the present invention on URAT1, GLUT9, ABCG2 and OAT1 in fructose-induced hyperuricemia rats;

[0042] In the accompanying drawings, compared with the blank control group, # P<0.05, ## P<0.01, ### P<0.001, #### P<0.0001; compared with the model group, * P<0.05, ** P<0.01, *** P<0.001, **** P<0.0001. DETAILED DESCRIPTION

[0043] The following will provide a clear and complete description of the technical solutions of the present invention in conjunction with specific embodiments of the present invention. Obviously, the embodiments described are only some embodiments of the present invention, not all embodiments. All other embodiments obtained by ordinary technicians in this field based on the embodiments of the present invention without making any creative efforts shall fall within the scope of protection of the present invention.

[0044] Example 1

[0045] The present embodiment relates to a method for preparing a polyphenol extract of Smilax glabra, comprising the following steps: weighing 1 kg of Smilax glabra slices, adding 50% ethanol at a material-liquid ratio of 1:10, heating and reflux extraction twice, each time for 1 hour, combining the extracts, concentrating to 500 mL, loading onto an AB-8 macroporous resin (BV = 3.5 L) chromatographic column, eluting with deionized water (5 BV), 50% ethanol (3 BV), and 95% ethanol (2 BV) in sequence, collecting the 50% ethanol elution fraction, and concentrating under reduced pressure to dryness to obtain a polyphenol extract TFL-3 (17.7 g) of Smilax glabra.

[0046] Example 2

[0047] This embodiment relates to a method for preparing a polyphenol extract of Smilax glabra, comprising the following steps: taking 13 g of the TFL-3 extract obtained in Example 1, adding 4.0 L of water, adjusting the pH to 6-8 with 0.1 mol / L NaOH solution, extracting three times with 4.0 L of ethyl acetate, combining the ethyl acetate extracts, and concentrating under reduced pressure to dryness to obtain a polyphenol extract TFL-4 (7.2 g). The aqueous layer solution after extraction was adjusted to a pH of 2-4 with 0.1 mol / L HCl solution, and concentrating under reduced pressure to dryness to obtain a polyphenol extract TFL-5 (6.3 g).

[0048] Example 3

[0049] The present embodiment relates to a preparation method of a polyphenol extract of Smilax glabra, comprising the following steps: weighing 1 kg of Smilax glabra slices, adding water at a material-liquid ratio of 1:10, heating and refluxing, extracting twice, each time for 1 hour, combining the extracts, concentrating to 1.0 L, taking 500 mL, concentrating, and drying to obtain Smilax glabra polyphenol extract TFL-1 (9.3 g). The remaining 500 mL was taken, filtered, and loaded onto an AB-8 macroporous resin (BV=1.5 L) chromatographic column, eluted with deionized water (5 BV), 50% ethanol (3 BV), and 95% ethanol (2 BV) in sequence, collecting the 50% ethanol elution portion, and concentrating under reduced pressure to dryness to obtain Smilax glabra polyphenol extract TFL-2 (8.2 g).

[0050] Test Case

[0051] The content of astilbin and total flavonoid glycosides (the sum of the peak area contents of peaks 1-5, calculated based on the astilbin reference substance) was determined by HPLC. The chromatographic column was C 18 -AR-Ⅱ (4.6ID×250mm), detection wavelength 291nm, flow rate 1mL / min, column temperature 25℃, injection volume 10μL; mobile phase A (0.1% acetic acid aqueous solution) and B (acetonitrile) gradient elution, B% 19 (0min)-20 (10min)-30 (30min)-40 (35min)-19 (45min), Figure 1AF are HPLC analysis of the Smilax glabra extracts TFL-1 to TFL-5 and astilbin of the present invention, respectively. The total polyphenol and tannin contents of the Smilax glabra extract were determined (based on the gallic acid reference substance content) with reference to the 2020 edition of the Pharmacopoeia, Part 4, General Chapter 2202, "Determination of Tannin Content." The results are shown in Table 1.

[0052] Among the various Smilax glabra extracts, TFL-3, prepared by 50% ethanol extraction-macroporous resin refining process, contains 47% total flavonoids and 37% tannins, of which the content of astilbin is as high as 25.8%. TFL-4 mainly contains 64% total flavonoids, as well as other polyphenols such as catechins and epicatechin. TFL-5 mainly contains 29% tannins. After further liquid chromatography-mass spectrometry analysis, it was found that it is mainly a proanthocyanidin polymerized by catechins and epicatechins, and also contains 5-O-caffeoylshikimic acid (P1). As a control, the extracts of TFL-1 and TFL-2 prepared by water extraction process also contain a combination of tannins (17.5%) and astilbin (3.2%), but the content is significantly lower than that of TFL-3.

[0053] Table 1 Determination of tannin and flavonoid contents in Smilax glabra polyphenol extract samples

[0054] extract TFL-1 TFL-2 TFL-3 TFL-4 TFL-5 Neoastilbin% 0.85 5.43 7.30 10.89 0 Astilbin% 0.51 3.20 25.81 33.47 0 Neoisoastilbin% 0.22 1.79 7.32 11.23 0 Isoflavin% 0.19 1.41 4.92 5.06 0 Huangqi glycoside% 0.02 0.18 2.03 3.13 0 Total flavonoid glycosides% 1.81 12.01 47.39 63.77 0 Total polyphenols% 1.38 41.2 56.0 57.6 34.1 Tannin% 1.06 17.5 37.1 32.7 29.3

[0055] Application Example 1: Application of Smilax glabra extract in hyperuricemia induced by adenine and potassium oxamate

[0056] Adenine and potassium oxonate-induced hyperuricemia rat model was established, and allopurinol (10 mg / kg), TFL-1 (600 mg / kg), TFL-2 (100 mg / kg), TFL-3 (100 mg / kg), TFL-4 (25 mg / kg) and TFL-5 (75 mg / kg) were administered by gavage for 14 consecutive days. The allopurinol group was used as the positive group, and a blank group and a model group were given distilled water to study the effects of extracts from different parts of Smilax glabra on hyperuricemia.

[0057] The results of the preliminary efficacy evaluation experiments are shown in Table 2. Compared with the blank control group, serum uric acid (SUA) levels in both the model group and the drug-treated group were significantly increased after modeling (P < 0.001), indicating that the hyperuricemia rat model was successfully established. After drug administration, serum uric acid levels in the TFL-1, TFL-2, TFL-3, and positive groups were significantly decreased (P < 0.0001), indicating that each drug-treated group had a significant therapeutic effect on hyperuricemia. In terms of dosage, the TFL-2 and TFL-3 groups had high levels of active ingredients, with an effect being seen at 100 mg / kg. Combined analysis of Tables 1 and 2 shows that the TFL-3 group had higher levels of tannins and total flavonoids than the TFL-2 group, demonstrating a better HUA-lowering effect.

[0058] Table 2 Changes in serum uric acid in hyperuricemic rats (n=5)

[0059]

[0060] Further efficacy evaluation results are shown in Table 3. Benzbromarone (10 mg / kg) was used as the active agent in this experiment. Following administration, serum uric acid levels significantly decreased in the TFL-3, TFL-4, and TFL-5 groups, as well as the active group (P < 0.0001), indicating that all treatment groups demonstrated significant therapeutic effects on hyperuricemia. Compared with the TFL-5 group, the TFL-4 group showed a more significant decrease in serum uric acid levels (P < 0.01). The TFL-3 group also demonstrated a significant uric acid-lowering effect (P < 0.05).

[0061] Table 3 Changes in serum uric acid in hyperuricemic rats (n=5)

[0062]

[0063] Compared with the blank group, the levels of creatinine and urea nitrogen in rats in each group after modeling were significantly increased (P<0.0001), and the uric acid in urine was decreased (P<0.0001); compared with the model group, the levels of creatinine and urea nitrogen in rats in the TFL-3, TFL-4, TFL-5 and positive groups were significantly decreased (P<0.0001), and the uric acid in urine of the TFL-3 and TFL-4 groups was increased (P<0.05). Compared with the blank group, although AST / ALT increased in the model group, it decreased in the drug-treated group. Figure 2 shown.

[0064] Compared with the blank control group, the kidney index of the rats in the model group was significantly increased (P<0.0001), and compared with the model group, the kidney index of the drug-treated groups decreased (P<0.01). Compared with the blank group, the liver index of the rats in the model group decreased, and compared with the model group, the liver index of the drug-treated groups increased, but there was no statistical difference. Compared with the blank group, the spleen index of the rats in the model group was significantly increased (P<0.001), and compared with the model group, the spleen index of the drug-treated groups decreased, and TFL-3 was more significant (P<0.05). Figure 3 shown.

[0065] Compared with the blank control group, the lymphocyte ratio of the model group was significantly decreased (P<0.001), and compared with the model group, the lymphocyte ratio of the TFL-3 group was significantly increased (P<0.01); compared with the blank group, the monocyte ratio of the model group was significantly decreased (P<0.001), and compared with the model group, the monocyte ratio of the TFL-4 group was significantly increased (P<0.05); compared with the blank group, the neutrophil ratio of the model group was significantly increased (P<0.0001), and compared with the model group, the neutrophil ratio of the TFL-4 group, TFL-5 group and positive group was significantly decreased (P<0.001), although the neutrophil ratio of the TFL-3 group was decreased, but there was no significant difference; compared with the blank group, the total platelet count of the model group was significantly increased (P<0.0001), and compared with the model group, the total platelet count of the TFL-3 group, TFL-4 group, TFL-5 group and positive group was significantly decreased (P<0.01). Figure 4 Neutrophils are the primary component of white blood cells, responsible for defending against bacterial infection and participating in inflammatory responses. Changes in their number and ratio are often used to diagnose infectious and inflammatory diseases. Neutrophil count, the ratio of neutrophil to lymphocyte count, provides a more comprehensive picture of the body's immune status.

[0066] Compared with the blank group, the triglyceride level in the model group was significantly increased (P<0.0001), indicating that lipid metabolism may be abnormal. The TFL-3 group had a significant effect in reducing triglyceride (P<0.01), while the total cholesterol, low-density lipoprotein, and high-density lipoprotein levels in the other groups were not statistically significant. Figure 5 shown.

[0067] Compared with the blank group, the NETs concentration in the model group was significantly increased (P<0.0001), and compared with the model group, the NETs concentration in the TFL-3, TFL-4, TFL-5 and positive groups was significantly decreased (P<0.0001), indicating that Smilax glabra has an inhibitory effect on NETs. Figure 6 shown.

[0068] Compared with the blank control group, the serum TNF-α, IL-1β, IL-6, and IL-17A levels of the rats in the model group were significantly increased (P<0.0001). Compared with the model group, the serum TNF-α, IL-1β, IL-6, and IL-17A levels of the rats in the TFL-4, TFL-5, and positive groups were significantly decreased (P<0.0001). The TNF-α (P<0.01), IL-1β (P<0.001), IL-6 (P<0.0001), and IL-17A (P<0.001) levels in the TFL-3 group were also decreased. This indicates that the Smilax glabra groups may improve the inflammatory response of rats through the IL-17 pathway. Figure 7 shown.

[0069] Compared with the blank group, the XOD activity in the liver of the model group increased (P<0.0001); compared with the model group, the XOD activity in the TFL-3 group, TFL-5 group and positive group decreased (P<0.0001), while the TFL-4 group decreased but there was no significant change. Figure 8 shown.

[0070] Compared with the blank group, the expression of reabsorption protein GLUT9 in the model group increased (P<0.01), and the expression of secretory proteins ABCG2 and OAT1 decreased (P<0.05). Compared with the model group, the expression of GLUT9 in the TFL-4 group decreased most significantly (P<0.0001). Compared with the model group, the expression of ABCG2 in the TFL-3, TFL-4, and TFL-5 groups increased (P<0.05). Compared with the model group, the expression of OAT1 in the drug-treated group increased, but there was no statistical difference. Figure 9 shown.

[0071] Application Example 2: Application of Smilax glabra extract in fructose-induced hyperuricemia

[0072] A fructose-induced hyperuricemia rat model was established. Allopurinol (10 mg / kg), TFL-3 (50 mg / kg), TFL-4 (25 mg / kg), TFL-5 (25 mg / kg) and astilbin (25 mg / kg) were administered orally for 4 consecutive weeks. The allopurinol group was used as the positive group, and a blank group and a model group were given distilled water to study the effects of extracts from different parts of Smilax glabra on hyperuricemia.

[0073] Four weeks after modeling, except for the blank group, the uric acid concentration in the serum of rats in all groups increased significantly (P<0.0001); after the start of administration, the TFL-3 (P<0.001), TFL-4 (P<0.0001), TFL-5 (P<0.001), and Astilbin groups (P<0.0001) all decreased significantly, indicating that each Smilax glabra administration group can reduce its uric acid level, as shown in Table 4.

[0074] Table 4 Changes in serum uric acid in hyperuricemic rats (n=5)

[0075] Group 0 weeks 4 weeks 5 weeks 6 weeks 7 weeks 8 weeks Blank group 74.5±19.6 98.4±7.4 83.6±23.4 94.9±16.7 92.0±30.8 88.4±8.2 Model Group 82.9±16.6 <![CDATA[173.9±32.8 ## ]]> <![CDATA[180.9±30.6 ### ]]> <![CDATA[167.7±22.9 ## ]]> <![CDATA[187.1±12.1 ### ]]> <![CDATA[194.0±29.5 ### ]]> TFL-3 76.8±13.2 <![CDATA[165.5±36.7 ## ]]> 157.8±49.1 <![CDATA[124.0±38.9 * ]]> <![CDATA[95.1±38.6 *** ]]> <![CDATA[112.4±15.1 *** ]]> TFL-4 78.1±10.0 <![CDATA[165.6±46.2 ## ]]> <![CDATA[131.6±39.5 * ]]> <![CDATA[132.8±35.5 * ]]> <![CDATA[116.2±53.3 ** ]]> <![CDATA[98.1±8.7 *** ]]> TFL-5 71.8±17.5 <![CDATA[173.2±45.2 ## ]]> 162.1±28.0 <![CDATA[131.8±15.3 * ]]> <![CDATA[121.2±26.3 ** ]]> <![CDATA[113.8±15.6 *** ]]> Astilbin 86.8±13.8 <![CDATA[159.9±21.1 # ]]> 147.4±11.3 139.6±39.2 <![CDATA[122.5±28.1 ** ]]> <![CDATA[101.1±14.1 *** ]]> Positive group 88.4±20.4 <![CDATA[224.6±70.0 ### ]]> <![CDATA[131.7±11.9 * ]]> <![CDATA[79.0±19.4 *** ]]> <![CDATA[71.4±20.0 *** ]]> <![CDATA[60.2±1.1 *** ]]>

[0076] Compared with the blank control group, the creatinine (P<0.001), AST / ALT (P<0.05), and uric acid in urine (P<0.001) of the model group were all increased. Compared with the model group, the creatinine of rats in the TFL-4 (P<0.01) and Astilbin groups (P<0.05) groups was significantly reduced; the AST / ALT of the TFL-3 group was decreased (P<0.05), indicating that its liver function was improved. Compared with the model group, the TFL-3, TFL-5 and Astilbin groups were all decreased (P<0.0001). Figure 10 shown.

[0077] Compared with the blank control group, the renal index of the rats in the model group was significantly increased (P<0.01). Compared with the model group, the renal index of the TFL-4 group and the Astilbin group decreased (P<0.05). Compared with the blank group, the liver index of the rats in the model group increased (P<0.0001). Compared with the model group, the liver index of the rats in the treatment group decreased, but there was no statistical difference. Compared with the blank group, the spleen index of the rats in the model group did not change significantly, indicating that no obvious damage to the spleen was caused. Figure 11 shown.

[0078] Compared with the blank control group, the white blood cell ratio in the model group was significantly increased (P<0.01), and compared with the model group, the white blood cell ratio in the TFL-3 group was significantly decreased (P<0.05); compared with the blank group, the neutrophil ratio in the model group was significantly increased (P<0.05), and compared with the model group, the neutrophil ratio in the TFL-3 group and the TFL-5 group was significantly decreased (P<0.05); compared with the blank group, the total platelet count in the model group was significantly increased (P<0.05), and compared with the model group, the total platelet count in the TFL-4 group (P<0.05) and the Astilbin group (P<0.01) was significantly decreased. Figure 12 shown.

[0079] Compared with the blank group, the triglyceride level in the model group was significantly increased (P<0.001), indicating that lipid metabolism may be abnormal. The TFL-3 group had a significant effect in reducing triglyceride (P<0.01). The low-density lipoprotein cholesterol in the model group was significantly increased (P<0.0), and the TFL-4 group had a significant effect in reducing low-density lipoprotein cholesterol (P<0.01). There was no statistically significant difference in total cholesterol and high-density lipoprotein cholesterol. Figure 13 shown.

[0080] Compared with the blank group, the NETs concentration in the model group was significantly increased (P<0.0001). Compared with the model group, the NETs concentration in the TFL-3, TFL-5, Astilbin groups and the positive group was significantly decreased (P<0.05), and the TFL-4 group also decreased (P<0.001), indicating that each drug-treated group had an inhibitory effect on NETs. Figure 14 shown.

[0081] Compared with the blank control group, the serum TNF-α, IL-1β, IL-6, and IL-17A levels of the rats in the model group were significantly increased (P<0.0001). Compared with the model group, the serum TNF-α, IL-1β, and IL-6 levels of the rats in the TFL-3, TFL-5, and Astilbin groups were significantly decreased (P<0.0001). The TNF-α (P<0.001), IL-1β (P<0.01), IL-6 (P<0.001), and IL-17A (P<0.001) levels of the TFL-4 group were also decreased. This indicates that the Smilax glabra groups may improve the inflammatory response of rats through the IL-17 pathway. Figure 15 shown.

[0082] Compared with the blank group, the activities of FA and ADA in the liver of the model group increased (P<0.0001). Compared with the model group, the ADA activities of the TFL-3, TFL-4, TFL-5, and positive groups decreased significantly (P<0.0001), and the Astilbin group also decreased (P<0.01). Among them, only the TFL-4 group (P<0.01) and the positive group (P<0.0001) decreased the FA activity. Figure 16 shown.

[0083] Compared with the blank group, the expressions of GLUT9 (P<0.0001) and URAT1 (P<0.001) increased in the model group, while the expressions of ABCG2 and OAT1 decreased (P<0.0001). Compared with the model group, the expressions of GLUT9 decreased in the TFL-3, TFL-4, and TFL-5 groups (P<0.001), and significantly decreased in the astilbin group (P<0.0001). The expressions of URAT1 decreased in the TFL-3 group (P<0.01), TFL-4 group (P<0.05), TFL-5 group (P<0.01), and astilbin group (P<0.05). Compared with the model group, the expression of ABCG2 (P<0.01) in the TFL-3, TFL-4, and Astilbin groups was significantly increased, and the expression of OAT1 (P<0.05) in the TFL-3, TFL-4, and Astilbin groups was significantly increased. The experimental results showed that each drug-treated group had a significant regulatory effect on the gene expression of the model group, including reducing the reabsorption of GLUT9 and URAT1 transporters and promoting the secretion of ABCG2 and OAT1 proteins. These two pathways regulate the level of uric acid in the body, such as Figure 17 shown.

[0084] In summary, TFL-3, TFL-4, TFL-5, and astilbin reduced serum uric acid, urinary uric acid, creatinine, and renal index in rats with hyperuricemia, inhibited the release of serum inflammatory factors TNF-α, IL-6, IL-1β, and IL-17A, and the formation of NETs, ​​inhibited the expression of adenosine dehydrogenase, reduced uric acid production, inhibited the expression of uric acid transporters GLUT9 and URAT1, and promoted the expression of uric acid transporters ABCG2 and OAT1. In addition, the TFL-3 group significantly reduced triglycerides, and the TFL-4 and TFL-5 groups reduced the neutrophil ratio. Therefore, astilbin is effective in the fructose model of HUA but has no effect on XOD. Smilax glabra polyphenols and their combination with astilbin are effective in the fructose model of HUA and have an anti-XOD effect.

[0085] The present invention has been described in detail above with reference to specific embodiments and exemplary examples. However, these descriptions should not be construed as limiting the present invention. Those skilled in the art will appreciate that various equivalent substitutions, modifications, or improvements may be made to the technical solutions and implementations of the present invention without departing from the spirit and scope of the present invention, all of which fall within the scope of the present invention. The scope of protection of the present invention shall be determined by the appended claims.

Claims

1. An application of a Smilax glabra extract in preparing a product for treating hyperuricemia, characterized in that: The smilax glabra extract comprises smilax glabra tannin, or comprises a combination of tannin and astilbin.

2. The use according to claim 1, characterized in that The smilax glabra tannin is a proanthocyanidin formed by polymerization of catechin and epicatechin. The mass content of the smilax glabra tannin in the smilax glabra extract is greater than 20%.

3. The use according to claim 1, characterized in that The astilbin composition comprises one or more of neoastilbin, astilbin, neoisoastilbin, isoastilbin and chrysanthemoside, wherein the mass content of astilbin in the Smilax glabra extract is greater than 15%.

4. The use according to claim 1, wherein The preparation method of the Smilax glabra extract comprises the following steps: S1. Extract the Rhizoma Smilacis Glabrae by heating and refluxing with 40%-75% ethanol, filter, and concentrate to obtain an extract; S2. The extract is adsorbed on a macroporous resin column, eluted with 30%-70% ethanol, concentrated under reduced pressure, and dried to obtain Smilax glabra extract A.

5. The use according to claim 4, characterized in that The Smilax glabra extract A is prepared into a pH 6-8 solution, extracted with ethyl acetate, and the aqueous layer after extraction is adjusted to pH 2-4, concentrated under reduced pressure, and dried to obtain the Smilax glabra extract B.

6. The use according to claim 1, wherein The Smilax glabra extract reduces uric acid production by inhibiting xanthine oxidase.

7. The use according to claim 1, wherein The Smilax glabra extract reduces uric acid production by inhibiting the expression of adenosine deaminase.

8. The use according to claim 1, wherein The smilax glabra extract promotes uric acid excretion by inhibiting the expression of uric acid transporters GLUT9 and URAT1 and promoting the expression of uric acid transporters ABCG2 and OAT1.

9. The use according to claim 1, wherein The product is a medicine, health product or food.

10. The use according to claim 1, wherein The hyperuricemia is hyperuricemia caused by potassium oxonate and adenine or fructose-induced hyperuricemia.

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