Application of small-leaf broadleaf holly leaf in reducing uric acid

Abstract

The invention belongs to the technical field of traditional Chinese medicine application, and particularly relates to application of small-leaf broadleaf holly leaves in reducing uric acid. The small-leaf broadleaf holly leaf can significantly inhibit the activity of xanthine oxidase in serum and liver so as to reduce the generation of uric acid and increase the uric acid content in urine so as to promote uric acid excretion, so that the level of uric acid in serum of an individual with hyperuricemia is reduced, and the effect of treating hyperuricemia and gout is achieved. As a traditional medicinal tea dual-purpose beverage, the small-leaf broadleaf holly leaf disclosed by the invention provides a safe and effective natural medicine for treating hyperuricemia and gout due to the efficient uric acid reducing effect of the small-leaf broadleaf holly leaf.

Description

Technical Field

[0001] The invention belongs to the technical field of application of traditional Chinese medicines, and particularly relates to the application of small-leaf kudingcha in reducing uric acid. Background Art

[0002] Hyperuricemia (HUA) is a common metabolic disease, and its incidence has been gradually increasing in recent years. Under normal circumstances, the body's uric acid production and excretion are in a dynamic equilibrium. However, when uric acid is produced excessively or excreted less, it will lead to increased uric acid levels in the blood, causing hyperuricemia. According to the "Chinese Multidisciplinary Expert Consensus on the Diagnosis and Treatment of Hyperuricemia-Related Diseases (2023 Edition)" (Chinese Journal of Practical Internal Medicine 2023, 43(06): 461-480.), serum uric acid (SUA) levels exceeding the saturation concentration (420 μmol / L) can induce gout attacks. Aging, obesity, high-purine diet, excessive alcohol intake, insulin resistance, hypertension and excessive medication are the main causes of hyperuricemia. Hyperuricemia is also associated with the occurrence of diabetes, hypertension, cardiovascular disease and kidney disease.

[0003] At present, drugs for the treatment of hyperuricemia / gout are mainly divided into the following categories. The first is drugs that inhibit uric acid production, such as allopurinol and febuxostat. Allopurinol reduces the production of uric acid by inhibiting xanthine oxidase (XOD); febuxostat has a highly selective inhibitory effect on xanthine oxidase and has a significant effect on lowering uric acid. The second is drugs that promote uric acid excretion, such as benzbromarone. It increases uric acid excretion by inhibiting the reabsorption of uric acid by the renal tubules. However, since the treatment of hyperuricemia or gout requires long-term use of uric acid-lowering drugs, the side effects of existing drugs limit their widespread and long-term use. Long-term use of the above-mentioned chemical drugs may cause hyponatremia, hypokalemia, and hypocalcemia, and also have obvious side effects on the liver and kidneys. Therefore, the development of uric acid-lowering drugs with small toxic side effects that can be used for a long time and have both the effects of promoting uric acid excretion and inhibiting XOD has become an important research and development direction for the prevention and treatment of hyperuricemia.

[0004] Plant medicine is an important natural treasure trove for finding and developing drugs for treating hyperuricemia and gout. As a traditional medicinal tea, small-leaf Kudingcha is a relatively outstanding variety among non-Camelliateas. It has a slightly bitter smell and a slightly cold nature. It has the effects of clearing away heat and detoxifying, reducing swelling and relieving pain, resisting rheumatism, scalding, and treating headaches, furuncles, and arthritis. Small-leaf Kudingcha mainly contains a large amount of terpenoids, phenylpropanoids, and flavonoid compounds. It is also rich in volatile oils, organic acids, and polysaccharides. Modern pharmacological studies have shown that it has multiple effects such as improving the cardiovascular system, lowering blood sugar and lipids, antibacterial and anti-inflammatory, antioxidant, cough and expectorant, weight loss and liver protection, and is safe and non-toxic ([1] Yi Fan, Peng Yong, Xu Lijia, et al. Research progress of small-leaf Kudingcha [J]. Modern Chinese Medicine, 2013, 15(10): 906-912.). However, it is not clear whether small-leaf Kudingcha can play a role in lowering uric acid and there are few related research reports.

[0005] Patent CN201910134870.0 discloses a Kudingcha preparation with anti-gout effect, its preparation method and application, and points out that Kudingcha extract oral liquid can inhibit uric acid in hyperuricemia mice (the inhibition rate of low to high dose groups is between 25.57-47.97%), but does not show strong uric acid lowering activity. The small-leaf Kudingcha of the present invention has an inhibition rate of 94.5% on uric acid at high doses, showing very strong uric acid lowering activity.

[0006] Therefore, the present invention aims to provide an application of small-leaf Kudingcha tea which has obvious innovative applications and can be used to reduce blood uric acid levels. Summary of the invention

[0007] To solve the above problems, the present invention proposes the application of small leaf Kudingcha in reducing uric acid. The present invention has found through research that small leaf Kudingcha can reduce the uric acid content in the serum of individuals with hyperuricemia, and has good application prospects in anti-hyperuricemia.

[0008] The technical solution adopted by the present invention is as follows:

[0009] The application of small-leaf Kuding tea in lowering uric acid is specifically used to prepare pharmaceutical preparations for preventing and treating hyperuricemia and gout.

[0010] The application of the small-leaf Kudingcha tea in reducing uric acid can also be used to prepare food that assists in reducing uric acid.

[0011] The small-leaf Kudingcha can be Ligustrum robustum (Roxb.) Bl., Ligustrum henryi Hemsl., Ligustrum pricei Hayata or Ligustrums inense var. myrianthum of the genus Ligustrum of the family Oleaceae.

[0012] The small leaf Kudingcha tea can be a single leaf of small leaf Kudingcha tea, powder made from small leaf Kudingcha tea leaves, an extract of small leaf Kudingcha tea leaves, or an effective part of the extract of small leaf Kudingcha tea leaves.

[0013] The extract of small leaf Kudingcha tea is any one of a water extract, an alcohol extract, and an alcohol-water mixed extract of small leaf Kudingcha tea.

[0014] The small-leaf Kudingcha water extract is obtained by using water as a solvent and adopting any one or both of the extraction methods of decoction and reflux.

[0015] The small leaf Kudingcha alcohol extract is an ethanol or methanol extract of small leaf Kudingcha.

[0016] The small leaf Kudingcha alcohol-water mixed extract is any one or both of a small leaf Kudingcha aqueous ethanol extract and an aqueous methanol extract.

[0017] The use of the small-leaf Kuding tea in the preparation of a pharmaceutical preparation for preventing and treating hyperuricemia and gout is characterized in that the pharmaceutical preparation is prepared by combining small-leaf Kuding tea or a traditional Chinese medicine composition containing small-leaf Kuding tea with pharmaceutically acceptable excipients to prepare the desired preparation.

[0018] The use of the small-leaf Kuding tea in preparing food that assists in reducing uric acid is characterized in that the food is a small-leaf Kuding tea raw material for human consumption or drinking or a finished product containing the small-leaf Kuding tea.

[0019] The beneficial effects of the present invention are as follows: the small-leaf Kuding tea of ​​the present invention is a dual-purpose beverage of medicinal tea, has high safety, can be well applied to the field of regulating uric acid, treating hyperuricemia-related diseases or for preparing drugs for treating, preventing, alleviating and / or relieving uric acid-related diseases, as well as foods for regulating uric acid, and provides a new, safe and effective natural medicine for treating hyperuricemia and gout. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 .The effect of small-leaf Kudingcha on serum uric acid in hyperuricemia mice;

[0021] Figure 2.The results of the effect of small-leaf Kudingcha on xanthine oxidase (XOD) in the serum and liver of hyperuricemia mice;

[0022] Figure 3 .The effect of small-leaf Kudingcha on uric acid in the urine of hyperuricemia mice;

[0023] Figure 4 .The effect of small-leaf Kudingcha on serum creatinine and urea nitrogen levels in hyperuricemia mice;

[0024] Figure 5 .The results of the effect of small-leaf Kudingcha on organ coefficients in hyperuricemia mice. DETAILED DESCRIPTION

[0025] In order to explain the present invention more clearly, the present invention is further described below in conjunction with preferred embodiments. It should be understood by those skilled in the art that the following specific description is illustrative rather than restrictive, and should not be used to limit the scope of protection of the present invention.

[0026] Preparation of water extract of small leaf kudingcha:

[0027] Weigh a certain amount of small-leaf Kudingcha tea and put it into a round-bottom flask, add 10 times the amount of water and soak for 30 minutes, boil for 1 hour, filter, then add the same volume of water and boil for 1 hour, filter, combine the two decoctions, concentrate, vacuum dry, and crush to obtain the small-leaf Kudingcha tea water extract, with an extraction rate of 36.72%.

[0028] Preparation of ethanol extract of small leaf Kudingcha:

[0029] Weigh a certain amount of small-leaf Kudingcha tea into a round-bottom flask, add 10 times the amount of ethanol, heat and reflux to extract for 1 hour, filter, add the same volume of ethanol to the residue again, extract for 1 hour, filter, combine the two filtrates, concentrate to an extract, vacuum dry, and crush to obtain small-leaf Kudingcha tea ethanol extract with an extraction rate of 17.85%.

[0030] Preparation of 50% ethanol extract of Kudingcha microphylla:

[0031] Weigh a certain amount of small-leaf Kudingcha tea and put it into a round-bottom flask, add 10 times the amount of 50% ethanol-water mixed solvent, heat and reflux to extract for 1 hour, filter, add the same volume of 50% ethanol-water mixed solvent to the residue again, extract for 1 hour, filter, combine the two filtrates, concentrate to an extract, vacuum dry, and crush to obtain a 50% ethanol extract of small-leaf Kudingcha tea with an extraction rate of 25.14%.

[0032] Dosage setting basis of Kudingcha microphylla extract:

[0033] According to the "Chinese Materia Medica", the usage and dosage of Kuding Tea is 3-9g of medicinal materials. The low, medium and high doses were selected as 2.25g, 4.5g and 9g respectively. The human dose was converted into the mouse dose. Finally, the low, medium and high doses of small-leaf Kuding Tea water extract were converted through extraction rate to obtain 125.31mg / kg, 250.63mg / kg and 501.25mg / kg respectively.

[0034] Example 1 Therapeutic effect of water extract of small leaf kudingcha on hyperuricemia mice

[0035] 1. Experimental Animals

[0036] SPF male Kunming mice, weighing 20 ± 2 g, were purchased from Changsha Tianqin Biotechnology Co., Ltd. (License No.: SCXK (Xiang) 2019-0014). All mice were adaptively fed and raised at 23-27 ° C and 50-70% humidity for one week, with free access to water and food. All animal experiments followed the protocol approved by the Ethics Committee of Guizhou Medical University (No.: 201800246).

[0037] 2. Effect of Kudingcha xiaoye on serum uric acid in hyperuricemia mice

[0038] 2.1 Establishment of hyperuricemia mouse model

[0039] 70 male Kunming mice were randomly divided into normal control group, model group, allopurinol group (positive control, 10 mg / kg), benzbromarone group (positive control, 10 mg / kg) and low, medium and high dose groups of water extract of small leaf Kuding tea (125.31, 250.63, 501.25 mg / kg), with 10 mice in each group. Except for the normal control group, the other groups were gavaged with hypoxanthine (PO) 500 mg / kg and intraperitoneally injected with potassium oxonate (HX) 100 mg / kg at 9 o'clock every day for 7 consecutive days. The mice in the normal control group were gavaged and intraperitoneally injected with the same volume of normal saline every day. Starting from the third day of the experiment, the drug group was gavaged with the corresponding drug 1 hour after the modeling, while the normal control group and the model group were gavaged with the same volume of normal saline. Starting from the third day of the experiment, the drug group was gavaged with the corresponding drug 1 hour after the modeling, while the normal control group and the model group were gavaged with the same volume of normal saline. On the sixth day of the experiment, one hour after the administration of the corresponding drug, the mice were placed in metabolic cages according to the groups, and the 24-hour urine of the mice was collected. On the seventh day of the experiment, one hour after the administration, the mice were anesthetized, blood was collected from the orbits, and then they were killed and whole blood samples were collected. After coagulation for 2 hours at room temperature, the blood was centrifuged (3000rpm, 10min), and the resulting serum was used for subsequent detection and analysis of uric acid, XOD and other kits.

[0040] 2.2 Experimental Results

[0041] Figure 1 .Effect of small-leaf Kudingcha on serum uric acid in hyperuricemia mice ( n=10). Compared with the normal group, # P<0.05, ## P<0.01, ### P<0.001; compared with the model group, * P<0.05, ** P<0.01, *** P<0.001.

[0042] The results are as follows Figure 1 As shown in the figure, compared with the normal control group, the blood uric acid content of mice in the model group was significantly increased (P < 0.001), indicating that the hyperuricemia mouse model was successfully established. Compared with the model group, the blood uric acid content of mice in each dose group of small leaf Kudingcha, allopurinol group, and benzbromarone group was significantly reduced, indicating that small leaf Kudingcha has a significant effect of lowering uric acid (P < 0.001).

[0043] 3. Effect of small-leaf Kudingcha on serum and liver xanthine oxidase (XOD) in hyperuricemia mice

[0044] 3.1 Experimental methods

[0045] After the serum was naturally precipitated for 2 hours, it was centrifuged at 3000 rpm / min for 10 minutes at 4°C, and the supernatant was used to determine XOD. In addition, 80 mg of liver tissue from each group of mice was accurately weighed, 0.72 ml of normal saline was added, and a 10% homogenate was prepared in an ice water bath, centrifuged at 3000 rpm / min for 10 minutes, and the supernatant was taken to detect XOD activity according to the instructions of the XOD kit.

[0046] 3.2 Experimental Results

[0047] Figure 2 .Effects of small-leaf Kudingcha on XOD in serum and liver of hyperuricemia mice ( n=10) (Figure A is serum XOD, Figure B is liver XOD). Compared with the normal group, # P<0.05, ## P<0.01, ### P<0.001; compared with the model group, * P<0.05, ** P<0.01, *** P<0.001.

[0048] The results are as follows Figure 2As shown in the results, compared with the normal group, the serum and liver XOD activities of the mice in the model group were significantly increased (P < 0.001); compared with the model group, different doses of small-leaf kudingcha could significantly reduce the XOD activities in the serum and liver of the hyperuricemia mice (P < 0.01 or P < 0.001) in a dose-dependent manner, indicating that small-leaf kudingcha can reduce the production of uric acid by inhibiting the XOD activity in mice to achieve the effect of lowering uric acid levels.

[0049] 4. Effect of Micro-leaf Kudingcha on Uric Acid in Hyperuricemic Mice

[0050] 4.1 Experimental methods

[0051] The collected mouse urine was used to detect the uric acid level according to the instructions of the uric acid kit.

[0052] 4.2 Experimental Results

[0053] Figure 3 .Effect of small-leaf Kudingcha on uric acid in the urine of hyperuricemic mice ( n=10). Compared with the normal group, # P<0.05, ## P<0.01, ### P<0.001; compared with the model group, * P<0.05, ** P<0.01, *** P<0.001.

[0054] The results are as follows Figure 3 As shown, compared with the blank control group, the urine uric acid level of mice in the hyperuricemia group was significantly decreased (P < 0.001); compared with the model group, the urine uric acid level in the low, medium and high dose groups of benzbromarone and small-leaf kudingcha was significantly increased (P < 0.05 or P < 0.001), indicating that each dose of benzbromarone and small-leaf kudingcha can significantly promote the excretion of uric acid in the body, thereby reducing the blood uric acid level.

[0055] Effects of 5-leaf Kudingcha on serum creatinine and urea nitrogen levels in hyperuricemia mice

[0056] 5.1 Experimental Methods

[0057] The serum was allowed to settle naturally for 2 h, then centrifuged at 3000 rpm / min for 10 min at 4°C, and the supernatant was used to determine creatinine and urea nitrogen.

[0058] 5.2 Experimental Results

[0059] Figure 4 .Effects of small-leaf Kudingcha on serum creatinine and urea nitrogen levels in hyperuricemia mice ( n=10) (Figure A is serum creatinine, Figure B is blood urea nitrogen). Compared with the normal group, # P<0.05, ## P<0.01, ### P<0.001; compared with the model group, * P<0.05, ** P<0.01, *** P<0.001.

[0060] The results are as follows Figure 4 As shown in the figure, compared with the normal control group, the serum creatinine and urea nitrogen levels of the mice in the model group were significantly increased (P < 0.05), indicating that hyperuricemia has damaged the renal function of the mice. Compared with the model group, the serum creatinine and urea nitrogen levels of the mice in the allopurinol group were significantly increased (P < 0.001 or P < 0.01), indicating that allopurinol significantly deteriorated the renal function of the hyperuricemia mice, but the serum urea nitrogen content of the mice in the small leaf Kudingcha dose groups showed a downward trend, and the blood creatinine content of the small leaf Kudingcha medium and high dose groups was significantly reduced (P < 0.05). The above results show that small leaf Kudingcha has a significant improvement on the renal function of hyperuricemia mice.

[0061] 6. Effect of Micro-leaf Kudingcha on Organ Coefficients in Hyperuricemia Mice

[0062] 6.1 Experimental methods

[0063] After the experimental method of Example 1 was completed, the weight, liver, kidney and spleen of the mice were weighed and calculated according to the formula: organ coefficient = organ mass / mouse weight*100% to investigate the effect of small leaf kudingcha on the organ index of hyperuricemia mice.

[0064] 6.2 Experimental Results

[0065] Figure 5 .Effects of small-leaf Kudingcha on organ coefficients in hyperuricemia mice ( n=10) (Figure A is body weight, Figure B is kidney coefficient, Figure C is liver coefficient, and Figure D is spleen coefficient). Compared with the normal group, # P<0.05, ## P<0.01, ### P<0.001; compared with the model group, * P<0.05, ** P<0.01, *** P<0.001.

[0066] The results are as follows Figure 5As shown, compared with the normal control group, the body weight of the mice in the model group was significantly decreased on the 5th and 7th days (P < 0.05), indicating that hyperuricemia produced toxic side effects on the mice; compared with the model group, the body weight of the mice in the allopurinol group was significantly decreased on the 5th and 7th days after administration (P < 0.01 or P < 0.001), indicating that the toxic side effects of allopurinol inhibited the natural growth of the body weight of the mice, while there was no significant change in the body weight of the mice in each dose group of small-leaf Kuding Tea, indicating that small-leaf Kuding Tea had no toxic side effects on mice.

[0067] Compared with the normal group, the liver, kidney and spleen coefficients of the mice in the model group were increased (the kidney coefficient was P < 0.001), indicating that hyperuricemia damaged the liver, kidney and spleen of the mice, but only the kidney damage was more significant; compared with the model group, the kidney coefficients of each dose group of small-leaf Kuding Tea were reduced, among which the medium and high doses were more significant (P < 0.05), indicating that the medium and high dose groups of small-leaf Kuding Tea significantly improved the kidney damage of mice; the liver coefficients of each dose group of small-leaf Kuding Tea were reduced, among which the high dose group was more significant (P < 0.05); compared with the blank group, there was no significant change in the spleen coefficients of the model group and each dose group of small-leaf Kuding Tea.

[0068] The above activity test results show that small-leaf Kuding tea can reduce the production of uric acid by inhibiting the activity of xanthine oxidase in serum and liver, and promote the excretion of uric acid by increasing the content of uric acid in urine, and finally reduce the content of uric acid in the serum of hyperuricemia model mice. The results of serum urea nitrogen, creatinine and organ indexes all show that small-leaf Kuding tea is safe and has no side effects. In summary, small-leaf Kuding tea can effectively reduce the blood uric acid of hyperuricemia individuals by inhibiting uric acid production and promoting uric acid excretion, and can thus be used to prepare drugs or foods for the treatment of hyperuricemia-related diseases.

[0069] Example 2 Preparation of solid beverage

[0070] 1. Raw material preparation: Select high-quality small-leaf Kuding tea leaves, wash and dry the tea leaves to remove impurities and moisture. Prepare sweeteners (xylitol), fillers (maltodextrin), etc.

[0071] 2. Extraction of active ingredients: Weigh a certain amount of small-leaf Kudingcha tea, add 10 times the amount of water, boil for 1 hour, filter, then add the same volume of water and boil for 1 hour, filter, combine the two decoctions, concentrate, vacuum dry, crush and pass through an 80-mesh sieve to obtain the small-leaf Kudingcha tea water extract.

[0072] 3. Mixing and bagging: Mix the raw materials according to 2g of water extract of small-leaf kuding tea, 5.2g of filler, and 0.8g of sweetener, making a total of 8g, mix them evenly and bag them. When eating, add 150ml of hot water to brew, stir evenly and dissolve.

[0073] Example 3 Preparation of tablets

[0074] 1. Raw material preparation: Select high-quality small-leaf Kuding tea leaves, wash and dry the tea leaves to remove impurities and moisture.

[0075] 2. Extraction of effective ingredients: Weigh a certain amount of small-leaf Kudingcha, add 10 times the amount of water and boil for 1 hour, filter, then add the same volume of water and boil for 1 hour, filter, combine the two decoctions to obtain the aqueous extract of the drug of the present invention.

[0076] 3. Preparation of soft material: Mix the concentrated extract with appropriate amount of filler (starch), binder (starch slurry), disintegrant (sodium carboxymethyl starch) and other auxiliary materials.

[0077] 4. Granulation: The soft material is squeezed through a screen by an extrusion granulator to form granules, and the obtained granules are placed in an oven for drying to remove moisture from the granules.

[0078] 5. Tablet pressing: Add the dried granules into the tablet press, adjust the parameters of the tablet press, and perform tablet pressing operation.

[0079] Finally, it should be pointed out that the above embodiments are only representative examples of the present invention. Obviously, the technical solution of the present invention is not limited to the above embodiments, and there are many variations. All variations that can be directly derived or associated with the content disclosed by ordinary technicians in this field should be considered as the protection scope of the present invention.

Claims

1. The application of small leaf Kudingcha in reducing uric acid, characterized in that: The small-leaf Kudingcha tea is used for preparing a medicinal preparation for preventing and treating hyperuricemia and gout.

2. The use according to claim 1, characterized in that The pharmaceutical preparation is prepared by combining small-leaf Kudingcha or a Chinese medicinal composition containing small-leaf Kudingcha with pharmaceutically acceptable auxiliary materials to prepare the desired preparation.

3. The application of small leaf Kudingcha in reducing uric acid, characterized in that: The small-leaf Kudingcha tea is used for preparing food for assisting in reducing uric acid.

4. The use according to claim 1 or 2, characterized in that: The small-leaf Kudingcha can be Ligustrum robusta, Ligustrum lucidum, Ligustrum pedunculata or Ligustrum lucidum.

5. The use according to claim 1 or 2, characterized in that: The small leaf Kudingcha tea can be a single leaf of small leaf Kudingcha tea, powder made from small leaf Kudingcha tea leaves, an extract of small leaf Kudingcha tea leaves, or an effective part of the extract of small leaf Kudingcha tea leaves.

6. The use according to claim 5, characterized in that The extract of small leaf Kudingcha tea is any one of a water extract, an alcohol extract, and an alcohol-water mixed extract of small leaf Kudingcha tea.

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