CN111375002A - Pharmaceutical composition for treating hyperuricemia and preparation method and application thereof - Google Patents
Pharmaceutical composition for treating hyperuricemia and preparation method and application thereof Download PDFInfo
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- CN111375002A CN111375002A CN202010200055.2A CN202010200055A CN111375002A CN 111375002 A CN111375002 A CN 111375002A CN 202010200055 A CN202010200055 A CN 202010200055A CN 111375002 A CN111375002 A CN 111375002A
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- pharmaceutical composition
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- uric acid
- hyperuricemia
- treating hyperuricemia
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Abstract
The invention discloses a pharmaceutical composition for treating hyperuricemia, which is prepared from the following components: 500 portions of ash bark and 600 portions of clematis root. The pharmaceutical composition for treating hyperuricemia can reduce the concentration of blood uric acid in a human body.
Description
Technical Field
The invention relates to the field of medicines, in particular to a pharmaceutical composition for treating hyperuricemia and a preparation method and application thereof.
Background
Uric acid is a dibasic acid, the pK of whicha1A pK of 5.4a29.8, and is usually present as an anion in humans. Two ways are available in human body to obtain purine to synthesize uric acid, one way is that endogenous purine is metabolized to generate uric acid, the endogenous way is formed by metabolizing intracellular protein purine bases of liver, intestinal tract and other tissues, such as muscle, kidney, vascular endothelium and the like, and 300-400mg uric acid is synthesized every day; one is the generation of uric acid by exogenous purine, and the exogenous way is mainly to take food containing high purine base, and then the food is formed after catabolism in vivo, and about 300mg of uric acid is formed after metabolism every day. Total uric acid production is 1200mg a day for men, and only about 600mg for women. The uric acid is produced by deaminase deamination of adenosine phosphatase (AMP) in human body, dephosphorylation by nucleotidase to form glycosides, and further conversion of glycosides to glycosides by Purine Nucleoside Phosphorylase (PNP)The purine bases hypoxanthine and guanine. Hypoxanthine is then oxidized by Xanthine Oxidase (XO) to form xanthine, and guanine is deaminated by guanine deaminase to form xanthine. Xanthine is again oxidized by xanthine oxidase to form the final product uric acid.
In some mammals, uric acid is metabolized into allantoin, which is readily soluble in water, by uricase, and is excreted dissolved in urine. However, the uricase gene in human and some primates is inactivated, and uric acid in vivo cannot be metabolized into allantoin to be discharged out of the body. The excretion of uric acid in the kidney mainly passes through 4 processes, namely filtration (100%), reabsorption (98-100%), secretion (50%) and reabsorption after secretion (40%), and the transport is completed after the four steps, and only 8% -12% of uric acid is discharged finally. During the excretion of uric acid, the kidney excretes about 2/3, while the digestive tract excretes 1/3, and after uric acid enters the digestive tract, the uric acid is destroyed by escherichia coli enzyme, and the process is the enzymolysis of uric acid. The two most prominent carrier proteins affecting uric acid reabsorption in the kidney are the urate ion transporter 1(URAT1) and the glucose transporter 9 (GLUT-9). URAT1 is a urinary acid anion transporter expressed by SCL22A12 gene, and is located at the brush border side of renal proximal tubular epithelial cells. The immunohistochemical method revealed that URAT1 in the kidney of IgA nephropathy accompanied by hyperuricemia was significantly higher than that in the case of IgA nephropathy without hyperuricemia. It was found that in mice with increased testosterone and in mice with decreased estradiol, the expression of URAT1 was higher and hyperuricemia was more likely to occur than in normal mice. GLUT-9 is a glucose transporter expressed by ALC2A9 gene, plays a major role in uric acid transport in liver and uric acid reabsorption in kidney, is mainly expressed in kidney, and is located in the outer membrane of renal tubular epithelial cell base. Another transporter associated with uric acid reabsorption is the organic anionic protein 4(QAT4), which is located in renal proximal tubular epithelial cells. Primarily involved in the secretion of uric acid is the adenosine triphosphate-binding transporter (ABCG2), a member of the ATP-binding cassette family, located in the apical membrane of the proximal kidney tubule, which is also expressed in small amounts in the intestine.
Normally, uric acid in a human body is about 1200mg, about 600mg is newly generated every day, and 600mg is excreted and kept in a balanced state. However, if too much uric acid is produced in vivo and excretion is not in time or the excretion mechanism of uric acid is degraded, the uric acid in vivo is retained too much, when the blood uric acid concentration is more than 7 mg/dl, the body fluid of a human body is changed into acid, the normal function of human body cells is influenced, and once the uric acid in vivo is too much, the body can not excrete in time, the uric acid is gathered in the body, and the symptom of high uric acid is generated.
Hyperuricemia and metabolic syndrome: the uric acid high-related cardiovascular diseases mainly comprise obesity, hyperglycemia, hypertension and dyslipidemia, and not only increase the occurrence of atherosclerotic vascular diseases, but also increase the risk of type 2 diabetes. Clinical data show that the risk of hypertension increases with increasing blood uric acid, and that every time blood uric acid increases by 60 μmol/L, the risk rate of hypertension increases by 15% -23%, cardiovascular disease mortality increases by 26% in women, 9% in men, 30% in women with ischemic heart disease mortality, 17% in men, and 48% in women with coronary heart disease. Hyperuricemia has a different prognosis and occurrence of coronary heart disease in both men and women, and is likely to be related to estrogen.
High uric acid and gout: gout is a type of inflammatory disease characterized by recurrent attacks due to purine metabolic disorders, and is characterized by red, tender, burning and swollen joints during the attacks. Pathologically gout is defined as inflammation caused by crystalline deposits in joints, tendons and other tissues and uric acid kidney stones. When the pH value is physiological, the binary weak uric acid exists in blood in the form of urate, and when the concentration of the dibasic weak uric acid is too high, the dibasic weak uric acid is precipitated in the form of crystals, and gout is easily complicated when the crystals are precipitated and deposited on joints and the like. In most studies, the uric acid value (SUA) in gouty arthritis is increased, as investigated by VanponeKai et al, and after analysis of relevant biochemical indexes of primary gouty arthritis in Xinjiang, it is found that the levels of blood uric acid (SUA), urine Glucose (GLU) and the like in a gout group are higher than those in a control group, and the difference has statistical significance (P < 0.01). In many studies on gout treatment drugs, it has also been found that SUA increases during molding and decreases after treatment.
Uric acid elevation and nephropathy: studies have shown that uric acid levels in blood are associated with Acute Kidney Injury (AKI), and that the risk of acute kidney injury increases with increased blood uric acid. A large body of clinical data also suggests that HUA is strongly associated with acute kidney injury and Chronic Kidney Disease (CKD). However, epidemiological studies have also shown that there is no link between HUA and chronic kidney disease. Therefore, whether chronic kidney disease is related to HUA is worthy of study.
Therefore, uric acid is high in harm, causes and conditions are complex, allopurinol, febuxostat and the like are generally used as drugs for reducing uric acid at present, but the drugs have obvious side effects, the dosage is not easy to master, gout and the like are induced, and clinically selectable drugs are obviously insufficient.
Disclosure of Invention
In view of the above problems, in one aspect, the present invention provides a pharmaceutical composition for treating hyperuricemia, which can reduce the uric acid concentration in a human body.
The technical scheme is as follows: a pharmaceutical composition for treating hyperuricemia is prepared from the following components:
500 portions and 900 portions of ash bark
400 portions of clematis root and 600 portions of clematis root.
Preferably, the pharmaceutical composition further comprises a diluent and/or a flavoring agent and/or an adhesive as an auxiliary material.
Preferably, the diluent is one or a mixture of sucrose, dextrin, soluble starch, mannitol, β -cyclodextrin and microcrystalline cellulose.
Preferably, the flavoring agent is stevioside and/or aspartame.
Preferably, the binder is water and/or ethanol.
Preferably, the dosage form of the pharmaceutical composition is any one of tablets, granules, capsules, oral liquid and dropping pills.
In one aspect, the invention also provides a method for preparing a pharmaceutical composition for treating hyperuricemia.
The technical scheme is as follows: a method of preparing a pharmaceutical composition for treating hyperuricemia, comprising the steps of:
① collecting cortex Fraxini and radix Clematidis;
② decocting in water, and collecting volatile oil;
③ mixing the decoctions, filtering, and concentrating.
In one aspect, the invention also provides the use of a pharmaceutical composition in the preparation of a medicament for the treatment of hyperuricemia.
The technical scheme is as follows: the application of a pharmaceutical composition in preparing a medicine for treating hyperuricemia is disclosed.
The invention has the beneficial effects that:
the pharmaceutical composition inhibits the expression level of URAT1, thereby inhibiting the reabsorption of uric acid, simultaneously enhancing the expression level of ABCG2, promoting the excretion of uric acid, and further playing the role of reducing blood uric acid, therefore, the pharmaceutical composition can be used as a medicine for reducing uric acid.
Drawings
FIG. 1 is the effect of the pharmaceutical composition of the present invention on the body weight of hyperuricemic mice;
FIG. 2 is the effect of the pharmaceutical composition of the present invention on serum uric acid levels of hyperuricemic mice;
FIG. 3 shows the effect of the pharmaceutical composition of the present invention on renal function of hyperuricemic mouse;
FIG. 4 is the effect of blank groups on renal pathology in hyperuricemic mice;
FIG. 5 is the effect of model groups on renal pathology in hyperuricemic mice;
FIG. 6 is the effect of positive group on kidney pathology in hyperuricemic mice;
FIG. 7 is the effect of low dose group on renal pathology in hyperuricemic mice;
FIG. 8 is the effect of medium dose groups on renal pathology in hyperuricemic mice;
FIG. 9 is the effect of high dose group on renal pathology in hyperuricemic mice;
FIG. 10 is a graph showing the effect of the pharmaceutical composition of the present invention on liver function in hyperuricemic mice;
fig. 11 is the effect of blank groups on liver pathology in hyperuricemic mice (200 ×);
figure 12 is the effect of model group on liver pathology in hyperuricemic mice (200 ×);
figure 13 is the effect of positive groups on liver pathology in hyperuricemic mice (200 ×);
figure 14 is the effect of low dose groups on liver pathology in hyperuricemic mice (200 ×);
figure 15 is the effect of medium dose groups on liver pathology in hyperuricemic mice (200 ×);
figure 16 is the effect of high dose group on liver pathology in hyperuricemic mice (200 ×);
FIG. 17 is the effect of the pharmaceutical composition of the present invention on the body weight of hyperuricemic rats;
FIG. 18 is a graph showing the effect of a pharmaceutical composition of the present invention on serum uric acid levels in hyperuricemic rats;
FIG. 19 shows the effect of the pharmaceutical composition of the present invention on renal function of hyperuricemic rat;
figure 20 is the effect of blank panel on renal pathology in hyperuricemic rats (200 ×);
FIG. 21 is a model set of effects on renal pathology in hyperuricemic rats (200 ×);
figure 22 is the effect of positive groups on hyperuricemic rat renal pathology (200 ×);
figure 23 is the effect of low dose group on renal pathology in hyperuricemic rats (200 ×);
figure 24 is the effect of medium dose groups on renal pathology in hyperuricemic rats (200 ×);
figure 25 is the effect of high dose group on renal pathology in hyperuricemic rats (200 ×);
FIG. 26 is a graph showing the effect of a pharmaceutical composition of the present invention on liver function in hyperuricemic rats;
FIG. 27 is the effect of blank groups on liver pathology in hyperuricemic rats;
FIG. 28 is the effect of model groups on liver pathology in hyperuricemic rats;
FIG. 29 is the effect of positive groups on liver pathology in hyperuricemic rats;
FIG. 30 is the effect of low dose groups on liver pathology in hyperuricemic rats;
FIG. 31 is the effect of medium dose groups on liver pathology in hyperuricemic rats;
FIG. 32 is the effect of high dose groups on liver pathology in hyperuricemic rats;
FIG. 33 is a graph showing the effect of a pharmaceutical composition of the present invention on liver tissue XOD in hyperuricemic rats;
FIG. 34 is a graph showing the effect of the pharmaceutical composition for treating hyperuricemia according to the present invention on the expression of kidney tissue transporter in rat with hyperuricemia;
FIG. 35 is a graph of the effect of blank groups on uric acid rat kidney tissue URAT1 (200 ×);
FIG. 36 is a graph of the effect of model groups on uric acid rat kidney tissue URAT1 (200 ×);
FIG. 37 is the effect of positive group on uric acid rat kidney tissue URAT1 (200 ×);
figure 38 is the effect of low dose group on uric acid rat kidney tissue URAT1 (200 ×);
FIG. 39 is the effect of medium dose groups on uric acid rat kidney tissue URAT1 (200 ×);
FIG. 40 is the effect of high dose group on uric acid rat kidney tissue URAT1 (200 ×);
FIG. 41 is a graph of the effect of blank groups on high uric acid rat kidney tissue GLUT9 (200 ×);
FIG. 42 is a graph of the effect of model group on renal tissue GLUT9 in hyperuricemic rats (200 ×);
FIG. 43 is a graph of the effect of positive groups on high uric acid rat kidney tissue GLUT9 (200 ×);
FIG. 44 is a graph of the effect of low dose groups on GLUT9 in hyperuricemic rat kidney tissue (200 ×);
FIG. 45 is the effect of medium dose group on GLUT9 in hyperuricemic rat kidney tissue (200 ×);
FIG. 46 is the effect of high dose group on GLUT9 in hyperuricemic rat kidney tissue (200 ×);
FIG. 47 is the effect of blank group on renal tissue ABCG2 in hyperuricemic rats (200 ×);
FIG. 48 is a graph of the effect of model group on renal tissue ABCG2 in hyperuricemic rats (200 ×);
figure 49 is the effect of positive group on hyperuricemic rat kidney tissue ABCG2 (200 ×);
figure 50 is the effect of low dose group on renal tissue ABCG2 in hyperuricemic rats (200 ×);
figure 51 is the effect of medium dose group on renal tissue ABCG2 in hyperuricemic rats (200 ×);
figure 52 is the effect of high dose group on renal tissue ABCG2 in hyperuricemic rats (200 ×).
Detailed Description
The invention will be further explained with reference to the drawings.
The examples provided herein are merely to further illustrate the invention and should not be construed as limiting the invention in any way.
It will be apparent to those skilled in the art that the materials and methods of operation used in the present invention are well known in the art, unless otherwise specified, in the following.
Example 1: preparation of medicinal composition granules for treating hyperuricemia
Soaking cortex Fraxini 700kg and radix Clematidis 500kg in water, decocting with 7-12 times of water for 3 times each time, each time for 40-100 min (collecting volatile oil), mixing decoctions, filtering, concentrating to relative density of 1.08-1.12, spray drying to obtain extract powder, sieving, adding dextrin, mixing, adding sweet sugar, and mixing. Adding 92% ethanol, sieving, drying at 40-60 deg.C, sieving, granulating, spraying volatile oil, sealing, moistening for 65-70 min, and packaging to obtain medicinal composition granule for treating hyperuricemia.
Example 2: preparation of pharmaceutical composition oral liquid for treating hyperuricemia
Soaking cortex Fraxini 700kg and radix Clematidis 500kg in water, decocting with 7-12 times of water for 3 times each time, each time for 40-100 min (collecting volatile oil), mixing decoctions, filtering, concentrating to relative density of 1.08-1.12, adding ethanol to ethanol content of 70-80%, standing for 24 hr, filtering, recovering ethanol under reduced pressure, and centrifuging; and adding the collected volatile oil, polysorbate, sodium cyclamate and sorbic acid into the supernatant, uniformly mixing, adding water, filtering, filling and sterilizing to obtain the pharmaceutical composition oral liquid for treating hyperuricemia.
In the following experimental examples 3 to 4:
① Experimental animal
Kunming Mice (KM), 18-22g, male, SPF grade, providing a company with great success at great success.
SD rats, all males, SPF grade, are provided centered on mastery experimental animals ltd.
② Experimental drugs and reagents
Potassium Oxonate (98%), Shanghai Allantin Biochemical technology Co., Ltd. or Shanghai Merlin Biochemical technology Co., Ltd. was prepared into a solution with a concentration of 25mg/ml with physiological saline just before use.
Yeast extract, Beijing Ombo Biotechnology, Inc., was dissolved in hot water to a concentration of 750mg/ml solution just before use.
Allopurinol, Hefeijie pharmaceutical Co., Ltd or Hefeijie pharmaceutical Co., Ltd, was dissolved with hot water to a solution of 2mg/ml immediately before use.
Xanthine Oxidase (XOD) test box, Jiangsu Nanjing institute of bioengineering.
Total protein quantitative test kit (BCA method), Jiangsu Nanjing institute of bioengineering.
Urate anion exchanger (URAT1) antibody: affinity.
Glucose transporter 9(GLUT9) antibody: abcam.
Adenosine triphosphate binding cassette transporter G2(ABCG2) antibody: abcam.
③ Experimental instrument
Fully automatic biochemical analyzers (siemens medical diagnostic products (shanghai) ltd);
microplate reader (MK3 U.S. Thermo);
an ultraviolet visible light photometer (Alpha-1860, a Songjiang high-tech garden in the God river Jing development district of Shanghai city);
electronic balance (HZT-A +200, HZ electronics technologies, Inc., kang, USA).
Microscope (Olympus, Tokyo, Japan, BX53)
Assay of Xanthine Oxidase (XOD) activity in liver tissue: sample pretreatment: accurately weighing the tissue weight (g): adding a 9-fold volume of homogenizing medium (0.9% of normal saline) into the mixture according to the volume (ml) ratio of 1:9, mechanically homogenizing the mixture in an ice-water bath condition to prepare a 10% homogenate, centrifuging the homogenate for 10min at the speed of 2500-3000 r/min, and taking the supernatant for determination.
Immunohistochemistry for the determination of the expression levels of the relevant proteins in the kidney (URAT1, GLUT9, ABCG 2): the tissue slices are hydrated by conventional dewaxing, and the slices are treated by citric acid antigen repairing solution, a peroxidase blocking agent (3% H2O2) is dripped on the tissues, incubated for 15min in a dark place, and washed by PBS for 3 times. Drop primary antibody on the tissue, incubate at 37 ℃ for 45min, after incubation, wash the section with PBS 3 times. Dripping 100ul of solution A (ChemMateTMenvision + HRP) in the two-step anti-rabbit/mouse universal immunohistochemical detection kit on tissues, and incubating for 45min at 37 ℃; after incubation, washing the slices for 3 times by PBS, dripping 100ul of prepared DAB working solution as a color developing agent, and controlling color development under a light mirror; after the color development is finished, the color development is stopped by washing with distilled water. Hematoxylin counterstain, alcohol dehydration of each level, xylene transparency and neutral gum sealing.
And (4) observation: 1 field (non-necrotic and hemorrhagic, non-specifically stained border zone) was randomly selected for each group of tissues using microscope 2-3 and high quality images (2448x 1920pixels) were collected with a CCD. When the image is collected, the consistency of each condition is ensured, namely, the settings of the light source, the aperture size, the white balance, the exposure intensity, the sensitivity, the contrast, the Gamma value and the like are manually adjusted and fixed. Even when the slide is changed, the settings of other components on the microscope, including the magnification of the objective lens, remain unchanged, except for adjusting the focal length and field of view. Then, the objective lens with the magnification of 20 times (the total magnification is 200 times) is used, and each slice randomly captures 10 representative visual fields according to the equidistant sampling principle according to the conditions to acquire images.
Randomly select 5 views to measure the IOD value (absolute value) and divide by the area to give the average IOD value (relative value).
Statistical treatment: all data were taken as mean ± variance
It is shown that,the difference between the two groups was analyzed by t-test of independent samples. The data was processed using GraphPad Prism 5.01(GraphPad Software Inc.), when P<0.05 was considered statistically different.
Example 3 verification of the effect of the pharmaceutical composition for treating hyperuricemia on reducing blood uric acid in mice
The verification method for the effect of the pharmaceutical composition for treating hyperuricemia on reducing the blood uric acid of the mice comprises the following steps:
① modeling and administration method
60 Kunming male mice are purchased, the mice are adapted to one week before the experiment, and are fed with conventional feed during the period, and are freely fed with food and water, and naturally day and night are used for lighting. One week later, 60 mice were randomly divided by body weight into 6 groups of a blank control group, a model control group, an allopurinol group (20mg/kg), a low dose group (2g/kg) of the pharmaceutical composition for the treatment of hyperuricemia of the present invention, a medium dose group (5g/kg) of the pharmaceutical composition for the treatment of hyperuricemia of the present invention, and a high dose group (8g/kg) of the pharmaceutical composition for the treatment of hyperuricemia of the present invention, each group having 10 mice. After grouping, each mouse is marked with picric acid, so that the mice in the group can be distinguished. The mice were weighed at 9:00 a day in the morning, and the administration dose of the mice was calculated based on the body weight, the intragastric volume of the mice was 10ml/kg, and the intraperitoneal injection volume of the mice was 10 ml/kg. Except for the blank group, the other groups were first injected with 7.5g/kg of gastric yeast extract, and then injected with 250mg/kg of oteracil potassium after 10min, while the blank control group was first injected with hot water of the same amount as the gastric yeast extract and then injected with normal saline of the same amount as the gastric yeast extract after 10 min. After 1h of administration, the blank and model groups were given equal amounts of distilled water, and the remaining groups were given the corresponding therapeutic agents, respectively. For 14 consecutive days, during which time the diet was free.
The low-dose group (2g/kg) of the pharmaceutical composition for treating hyperuricemia of the invention refers to the blood-uric-acid-lowering granules prepared in example 1 or the oral liquid low-dose group (2g/kg) prepared in example 2, wherein the blood-uric-acid-lowering granules are 2g granules/kg of mice, and the oral liquid is 2g/kg of active substances in the oral liquid.
The dosage group (5g/kg) of the pharmaceutical composition for treating hyperuricemia of the invention refers to the blood-uric-acid-lowering granules prepared in example 1 or the oral liquid low-dosage group (5g/kg) prepared in example 2, and when the dosage group is the blood-uric-acid-lowering granules, the dosage group is 5g granules/kg of mice, and when the dosage group is the oral liquid, the dosage group refers to 5g/kg of active substances in the oral liquid.
The high-dose group (8g/kg) of the pharmaceutical composition for treating hyperuricemia of the invention refers to the blood-uric-acid-lowering granules prepared in example 1 or the low-dose group (8g/kg) of the oral liquid prepared in example 2, wherein the blood-uric-acid-lowering granules are 8g granules/kg of mice, and the oral liquid is 8g/kg of active substances in the oral liquid.
② pharmacodynamic observation and index detection
And (4) behavioral observation: mice were observed daily for general signs (whether hair was normal, mental status, etc.), and weighed and recorded daily.
And (3) biochemical index determination: after the gavage for 1 hour on day 14, the mouse was subjected to eyeball bleeding, the blood was centrifuged at 3500rpm for 10min, and serum was taken and measured for uric acid level (SUA), creatinine level (SCr), alanine Aminotransferase (ALT), aspartate Aminotransferase (AST) and urea level with a full-automatic biochemical analyzer.
③ histopathological observation of liver and kidney
After the eye was bled, the mice were sacrificed by cervical dislocation and the liver and kidney tissues collected for histopathological analysis:
fixing: liver and kidney tissues were fixed in 10% formalin for 48 h.
And (3) dehydrating: (75%, 85%, 95%, 100%) ethanol was dehydrated stepwise with stirring from time to time during dehydration to achieve sufficient dehydration.
And (3) transparency: xylene was clear (first 50% ethanol + 50% xylene, twice with pure xylene, 1h each).
And (3) infiltration: placing in 50% xylene + 50% powdered paraffin, and opening in an oven at 40 deg.C overnight
Embedding: the tissue was placed in a mold and cooled on a cooling table.
Slicing: firstly, adjusting the thickness of the slice to 5 mu m, cutting off the wax sheet, using a soft brush to lift the cut wax sheet upwards from the knife edge, using tweezers to clamp the wax sheet and drag the wax sheet to the water surface of a spreading instrument, and expanding the wax sheet for about 1-2 min. And vertically inserting the anti-falling glass slide into water to touch the slices, vertically lifting the glass slide attached with the wax sheet, correcting the positions of the slices and numbering, placing the slices in an oven at 60 ℃ for baking for 2 hours, and pasting the slices at 37 ℃ overnight.
HE staining: dewaxing (xylene dewaxing treatment 3 times, 10min each time) → hydration (treatment with 100% ethanol, 95% ethanol, 85% ethanol, 75% ethanol, 3min each time) → distilled water washing (3min) → hematoxylin staining (5-10min) → color separation liquid treatment (5s) → bulk tap water washing treatment (3min) → Li2CO3 treatment (10min) → tap water washing 1min treatment → eosin staining treatment (5min) → tap water washing treatment (1min) → 95% ethanol treatment (1min) → 100% ethanol treatment (1min) → xylene carbolic acid (3:1) treatment (1min) → xylene reprocessing (1min) → finally film sealing with neutral resin.
And (4) observation: using a microscope, 3 fields (non-necrotic and hemorrhagic, non-specifically stained border regions) were randomly selected for each group of tissues and high quality images (2448x 1920pixels) were collected using a CCD. When the image is collected, the consistency of each condition is ensured, namely, the settings of the light source, the aperture size, the white balance, the exposure intensity, the sensitivity, the contrast, the Gamma value and the like are manually adjusted and fixed. Even when the slide is changed, the settings of other components on the microscope, including the magnification of the objective lens, remain unchanged, except for adjusting the focal length and field of view. Then, the objective lens with the magnification of 20 times (the total magnification is 200 times) is used, and each slice randomly captures 10 representative visual fields according to the equidistant sampling principle according to the conditions to acquire images.
④ verification result
A. The influence of the pharmaceutical composition for treating hyperuricemia on the weight of a hyperuricemic mouse
And (3) analyzing the growth state: all 6 mice grew normally, gained consistent weight (see figure 1 and table 1), had better mental status and smooth hair.
TABLE 1 Effect of the pharmaceutical composition for treating hyperuricemia of the present invention on the body weight of mice with hyperuricemia: (
n＝10)
In Table 1, the remaining 5 groups had no significant difference (P >0.05) compared with the blank group
B. The influence of the pharmaceutical composition for treating hyperuricemia on the serum uric acid level of the hyperuricemia mice
Compared with the blank group, the SUA level of the model group is increased and has significant difference (P <0.05), which indicates that the molding is successful. Compared with the model group, the SUA level of the pharmaceutical composition for treating hyperuricemia according to the present invention in the high dose group was significantly reduced (P <0.05), while the SUA level of the pharmaceutical composition for treating hyperuricemia according to the present invention in the low dose group and the pharmaceutical composition for treating hyperuricemia according to the present invention in the dose group was not significantly different from that of the model group (P >0.05), but had a tendency to be reduced. The results are shown in Table 2 and FIG. 2.
TABLE 2 Effect of the pharmaceutical composition for treating hyperuricemia of the present invention on serum uric acid levels in hyperuricemic mice: (
n＝10)
| Uric acid (mu mol/L) | |
| Blank group | 62.48±28.80 |
| Model set | 90.18±17.50* |
| Allopurinol | 5.11±3.80### |
| Low dose group | 74.55±32.47 |
| Middle dose group | 79.88±41.55 |
| High dose group | 65.83±36.21# |
In table 2, # P <0.05 compared to the blank group, # P <0.05, # P <0.001 compared to the model group.
In fig. 2, # P <0.05 compared to the blank group, # P <0.05, # P <0.001 compared to the model group.
C. The influence of the pharmaceutical composition for treating hyperuricemia on the renal function of the hyperuricemia mice
Urea value: compared with a blank group, the Urea level of the model group is obviously increased (p <0.001), which indicates that the model has certain renal injury while establishing a hyperuricemia model. The Urea levels at the NQG low, medium and high doses were all significantly reduced compared to the model group (P < 0.01).
Creatinine value: there was no significant difference in the remaining 5 groups compared to the blank group.
Renal factor: there was no significant difference in the remaining 5 groups compared to the blank group.
The results are shown in Table 3 and FIG. 3.
TABLE 3 Effect of the pharmaceutical composition for treating hyperuricemia of the present invention on renal function levels of hyperuricemic mice: (
n＝10)
| Urea (mmol/L) | Creatinine (mu mol/L) | Kidney coefficient (%) | |
| Blank group | 5.33±0.77 | 32.24±.486 | 1.33±0.05 |
| Model set | 7.53±0.56*** | 34.01±2.46 | 1.48±0.18 |
| Allopurinol | 8.23±3.53 | 35.13±14.63 | 1.27±0.23 |
| Low dose group | 5.53±1.67## | 25.65±10.71 | 1.40±0.12 |
| Middle dose group | 5.80±1.83## | 32.21±8.842 | 1.47±0.18 |
| High dose group | 6.06±1.05## | 31.30±7.785 | 1.36±0.05 |
In table 3, # P <0.001 compared to the blank group, # P <0.01 compared to the model group.
In fig. 3, # P <0.001 compared to the blank group and # P <0.01 compared to the model group.
The pharmaceutical composition for treating hyperuricemia of the invention has the following effects on the kidney pathology of the hyperuricemia mice: HE staining results show that blank mice have clear glomerulus and renal tubule structures, and have no symptoms of inflammatory infiltration, no urate crystallization and the like. Compared with the blank group, the low-dose group, the medium-dose group and the high-dose group of the pharmaceutical composition for treating hyperuricemia have no obvious difference. See fig. 4-9 for comparison. As can be seen from the figure, the remaining 5 groups were not significantly different (P >0.05) compared to the blank group.
D. The effect of the pharmaceutical composition for treating hyperuricemia on the liver function of the hyperuricemia mice
AST and ALT and AST/ALT ratio: compared with the blank group, the NQG low dose group, the medium dose group and the high dose group have no significant difference (P > 0.05). The results are shown in Table 4, FIG. 10.
TABLE 4 Effect of the pharmaceutical composition for treating hyperuricemia of the present invention on hepatic function level of hyperuricemic mouse: (
n＝10)
| AST(U/L) | ALT(U/L) | AST/ALT ratio | |
| Blank group | 144.00±32.89 | 51.50±16.81 | 2.92±0.72 |
| Model set | 154.10±37.6 | 52.00±11.34 | 2.97±0.29 |
| Allopurinol | 137.70±69 | 49.57±10.44 | 2.636±1.06 |
| Low dose group | 137.3±38.03 | 36.25±14.36 | 3.401±0.803 |
| Middle dose group | 129.4±39.67 | 45.00±18.77 | 3.139±0.782 |
| High dose group | 145.70±31.77 | 50.50±9.501 | 2.94±0.70 |
In table 4, the remaining 5 groups were not significantly different (P >0.05) compared to the blank group.
The influence of the pharmaceutical composition for treating hyperuricemia on the liver pathology of the hyperuricemia mice is as follows: the blank group showed glycogen storage, which is a normal physiological phenomenon in mice, and no edema, adiposis, etc. Compared with the blank group, the NQG low dose group, the medium dose group, and the high dose group were not significantly different. The comparison and results are shown in FIGS. 11-16.
Therefore, the pharmaceutical composition for treating hyperuricemia has a certain effect of reducing blood uric acid of mice caused by intraabdominal injection (250mg/kg) of the potassium oxalate in combination with intragastric gavage (7.5g/kg) of the yeast.
Example 4 verification of the effect of the pharmaceutical composition for treating hyperuricemia on reducing blood uric acid in rats
The verification method for the effect of the pharmaceutical composition for treating hyperuricemia on reducing the blood uric acid of the rat comprises the following steps:
① modeling and administration method
36 SD rats were purchased and allowed to acclimate for one week before the experiment, during which the rats were fed with regular diet, freely fed with food and water, and naturally daylit day and night. One week later, 36 rats were randomly divided by body weight into 6 groups of a blank control group, a model group, a positive control group (allopurinol group 10mg/kg), a low dose group (1mg/kg) of the pharmaceutical composition for the treatment of hyperuricemia of the present invention, a medium dose group (3mg/kg) of the pharmaceutical composition for the treatment of hyperuricemia of the present invention, and a high dose group (6mg/kg) of the pharmaceutical composition for the treatment of hyperuricemia of the present invention. After grouping, each rat was marked with picric acid to allow each rat to be distinguished. The rats in the blank group were given free access to normal diet, and the remaining 5 groups were given free access to feed containing 2.5% Potassium Oxonate. Changes in body weight were recorded weekly for 7 weeks. At the end of the fourth week, blood was collected from the tail vein, centrifuged at 3000rpm for 10min, and serum was collected and assayed for uric acid level. Starting from the fifth week, the blank and model groups were gavaged with equal amounts of distilled water and the treatment groups were given the corresponding drugs for 3 consecutive weeks.
The low-dose group (1mg/kg) of the pharmaceutical composition for treating hyperuricemia of the invention refers to the blood uric acid lowering granules prepared in example 1 or the oral liquid low-dose group (1mg/kg) prepared in example 2, and when the granules are blood uric acid lowering granules, the granules are 1mg granules/kg of rats, and when the granules are oral liquid, the content of the effective substances in the oral liquid is 1 mg/kg.
The dosage group (3mg/kg) of the pharmaceutical composition for treating hyperuricemia of the invention refers to the blood uric acid lowering granules prepared in example 1 or the oral liquid low dosage group (3mg/kg) prepared in example 2, and when the dosage group is the blood uric acid lowering granules, the dosage group is 3mg granules/kg of rats, and when the dosage group is the oral liquid, the dosage group refers to the content of the effective substances in the oral liquid, which is 3 mg/kg.
The high-dose group (6mg/kg) of the pharmaceutical composition for treating hyperuricemia of the invention refers to the blood uric acid lowering granules prepared in example 1 or the low-dose group (6mg/kg) of the oral liquid prepared in example 2, wherein the blood uric acid lowering granules are 6mg granules/kg of rats, and the oral liquid is 6mg/kg of active substances in the oral liquid.
② pharmacodynamic observation and index detection
And (4) behavioral observation: rats were observed daily for general signs (whether hair was normal, mental status, etc.), and weighed and recorded weekly.
And (3) biochemical index determination: after 3 weeks of administration of the therapeutic agent, the femoral artery was bled, the blood was centrifuged at 3500r/min for 10min, and serum was taken and assayed for uric acid level (SUA), creatinine level (SCr), alanine Aminotransferase (ALT), aspartate Aminotransferase (AST), and Urea level (Urea) using a full-automatic biochemical analyzer.
③ histopathological observation of liver and kidney
Same as the histopathological observation of liver and kidney in example 3.
④ verification result
A. The influence of the pharmaceutical composition for treating hyperuricemia on the weight of the hyperuricemic rat
And (3) analyzing the growth state: all 6 rats grew normally, gained consistent weight (see fig. 17, table 5), and had better mental status and smooth hair.
TABLE 5 Effect of the pharmaceutical composition for treating hyperuricemia of the present invention on the body weight of hyperuricemic rat
As can be seen from table 5 and fig. 17, there was no significant difference (P >0.05) in the remaining 5 groups compared with the blank group.
B. The effect of the pharmaceutical composition for treating hyperuricemia on the serum uric acid of the hyperuricemic rat
The uric acid number results are shown in Table 6 and FIG. 18. At the end of the fourth week, the uric acid levels of the allopurinol group (positive group), the low dose group, the medium dose group and the high dose group were all significantly increased (P <0.01) compared with the blank group, suggesting that the allopurinol group (positive group), the low dose group, the medium dose group and the high dose group were all successful in establishing a rat hyperuricemia model. At the end of the seventh week, the uric acid levels in the model group were significantly increased (P <0.05) compared with the blank group, and the uric acid levels in the NQG high dose group were significantly decreased (P <0.05) compared with the model group, and the uric acid levels in the NQG medium dose and low dose groups tended to decrease but were not significantly different.
TABLE 6 Effect of the pharmaceutical composition for treating hyperuricemia of the present invention on serum uric acid in hyperuricemic rat: (
n＝6)
| Week 4 uric acid number (μmol/L) | Week 7 uric acid number (μmol/L) | |
| Blank group | 96.44±22.29 | 73.62±20.83 |
| Model set | 197.50±28.92*** | 126.61±36.66* |
| Allopurinol group | 178.50±26.56*** | 58.00±12.74## |
| Low dose group | 158.90±29.19** | 93.30±14.61 |
| Middle dose group | 153.20±27.77** | 84.32±19.5 |
| High dose group | 180.90±29.26*** | 78.18±18.34# |
As can be seen from table 6, # P <0.05, # P <0.01, # P <0.001 compared to the blank group and # P <0.05, # P <0.01 compared to the model group.
As can be seen from fig. 18, P <0.05, # P <0.01, # P <0.001 compared to the blank group, and # P <0.05, # P <0.01 compared to the model group.
C. The influence of the pharmaceutical composition for treating hyperuricemia on the renal function of the hyperuricemic rat
Urea value: compared with the blank group, the urea levels of the model group, the allopurinol group and the NQG low-dose group have no significant difference (P > 0.05); urea was significantly reduced in the NQG medium and high dose groups compared to the blank group (P < 0.01). The results are shown in Table 7, FIG. 19.
Creatinine value: the remaining 5 groups were not significantly different compared to the blank group.
Renal factor: the remaining 5 groups were not significantly different compared to the blank group.
TABLE 7 Effect of the pharmaceutical composition for treating hyperuricemia of the present invention on renal function of hyperuricemic rat: (
n＝6)
In table 7, P <0.01 compared to blank group.
In fig. 19, P <0.01 compared to the blank group.
The pharmaceutical composition for treating hyperuricemia of the invention has the following effects on the kidney pathology of hyperuricemic rats:
blank group: no obvious abnormality is found in the glomerular size, shape, volume, etc. The renal tubular epithelium has no necrosis or desquamation.
Model group: the size, shape, volume and the like of the glomerulus are not obviously abnormal; most rats can see more brown (tan) crystal-like substance precipitation in renal tubules, and have refractivity, and urate crystals are considered; swelling, necrosis and desquamation of part of renal tubular epithelium.
Allopurinol group: glomerular volumes all exhibit varying degrees of shrinkage (considering the potential for atrophy); brown crystals in the renal tubules of the same model group can be seen in the tubular cavities of most rats, and partial swelling and necrosis of the renal tubular epithelium can be seen.
Low dose group: no obvious abnormality is found in the volume, shape, volume and the like of the glomerulus, and brown crystals are found in part of the renal tubules. Swelling and necrosis of renal tubular epithelium are not observed.
The medium dose group: the size, shape and volume of the glomerulus are not obviously abnormal; a small amount of brown crystalline flakes were visible in part of the tubules, and a slight swelling of the renal tubule epithelium was visible.
High dose group: the size, shape, volume and the like of the glomerulus are not obviously abnormal; no crystals were found in the tubules, and a small amount of tubular epithelium was slightly swollen.
Compared to the blank group, the most prominent lesions of the kidney in the model group were urate crystallization with partial swelling of the tubular epithelium and necrotic exfoliation. Compared with the model group, the NQG low-dose kidney lesion is reduced, brown crystals are only contained in part of kidney tubules, the kidney lesion of the NQG medium-dose group is reduced, the brown crystals are not seen in the NQG high-dose group, and the NQG low-dose group, the NQG medium-dose group and the NQG high-dose urate crystals are obviously improved. The results are shown in FIGS. 20-25, where the model set markers are urate crystals.
D. The influence of the pharmaceutical composition for treating hyperuricemia on the liver function of the hyperuricemia rat
AST and ALT: elevated AST and ALT indicate a certain damage to the liver. Compared with the AST level of the blank group, there was no significant difference in AST level of the NQG low dose group, medium dose group and high dose group (P > 0.05). Compared with ALT value of a blank group, the model group, allopurinol group, NQG middle dose group and high dose group are obviously reduced (P is less than 0.05), and liver injury is not suggested. See table 8 and fig. 26.
TABLE 8 Effect of the pharmaceutical composition for treating hyperuricemia of the present invention on liver function of hyperuricemic rat: (
n＝6)
| AST(U/L) | ALT(U/L) | |
| Blank group | 127.40±21.58 | 50.80±6.12 |
| Model set | 142.00±32.64 | 37.50±9.91*** |
| Allopurinol group | 104.08±27.10 | 34.25±2.86*** |
| Low dose group | 100.33±10.54 | 42.33±7.69 |
| Middle dose group | 125.60±21.03 | 39.60±5.67* |
| High dose group | 131.60±19.31 | 34.40±2.93*** |
In table 8, P <0.05, P <0.01, P <0.001 compared to the blank group.
In fig. 26, P <0.05, P <0.01, P <0.001, compared to the blank group.
The pharmaceutical composition for treating hyperuricemia of the invention has the following effects on liver pathology of hyperuricemic rats: edema, fatty lesions, etc. were not observed in the blank group. Compared with the blank group, the pharmaceutical composition for treating hyperuricemia has no significant difference in the low-dose group, the medium-dose group and the high-dose group. The comparison and results are shown in FIGS. 27-32.
E. The pharmaceutical composition for treating hyperuricemia of the invention has the effect on the XOD activity of the liver tissue of the rat with hyperuricemia
Compared with a blank group, the XOD activity of the model group has no significant difference, and the low-dose group, the medium-dose group and the high-dose group of the pharmaceutical composition for treating hyperuricemia have no significant difference. The comparison and results are shown in table 9 and fig. 33.
TABLE 9 Effect of the pharmaceutical composition for treating hyperuricemia of the present invention on liver tissue XOD of hyperuricemic rat: (
n＝6)
| XOD(U/gprot) | |
| Blank group | 29.07±1.35 |
| Model set | 32.46±5.28 |
| Allopurinol group | 26.51±2.67* |
| Low dose group | 29.68±2.73 |
| Middle dose group | 30.29±3.553 |
| High dose group | 29.89±3.83 |
In table 9, P <0.05 compared to blank group.
In fig. 33, P <0.05 compared to blank group.
F. The effect of the pharmaceutical composition for treating hyperuricemia on the expression of the kidney tissue transporter of the rat with hyperuricemia of the invention
Urate anion transporter 1(URAT 1): the mean optical density of the model group was significantly higher (P <0.01) compared to the blank group, indicating an increased level of URAT1 expression. Compared with the model group, the average optical density of the low-dose group, the medium-dose group and the high-dose group of the pharmaceutical composition for treating hyperuricemia is remarkably reduced (P < 0.05). The pharmaceutical composition for treating hyperuricemia can reduce the expression of URAT1, thereby inhibiting the reabsorption of uric acid.
Glucose transporter 9(GLUT 9): the mean optical density of the model group was significantly increased (P <0.01) compared to the blank group, indicating enhanced expression of the model group GLUT 9. Compared with the model group, the mean optical density values of the high-dose group of the pharmaceutical composition for treating hyperuricemia have no significant difference (P >0.05), and the mean optical densities of the medium-dose and low-dose groups have a decreasing trend and have no significant difference (P > 0.05).
Adenosine triphosphate binding cassette transporter (ABCG 2): the mean optical density of the model group was significantly reduced (P <0.001) compared to the blank group, indicating a significant reduction in ABCG2 expression. Compared with the model group, the average optical density values of the low-dose group, the medium-dose group and the high-dose group of the pharmaceutical composition for treating hyperuricemia are obviously increased (P < 0.01). The pharmaceutical composition for treating hyperuricemia can enhance the expression of ABCG2 and increase the excretion of uric acid.
The results are shown in Table 10, FIG. 34, and FIGS. 35-52.
TABLE 10 Effect of the pharmaceutical composition for treating hyperuricemia of the present invention on the expression of renal tissue transporter in rat with hyperuricemia: (
n＝6)
| URAT1 | GLUT-9 | ABCG2 | |
| Blank group | 11.10±8.332 | 118.40±63.24 | 231.8±51.3 |
| Model set | 93.64±48.37** | 666.8±346.90** | 70.61±32.29*** |
| Allopurinol group | 106.60±24.72 | 838.20±45.85 | 50.21±19.32 |
| Low dose group | 30.85±38.92# | 486.80±175.90 | 108.5±60.49## |
| Middle dose group | 4.15±5.022## | 457.7±231.30 | 218.90±47.56### |
| High dose group | 26.06±46.21# | 697.6±103.40 | 179.20±58.49## |
In table 10, # P <0.05, # P <0.01, # P <0.001, compared to the blank group, # P <0.05, # P <0.01, # P <0.001, compared to the model group.
In fig. 34, P <0.05, # P <0.01, # P <0.001, compared to the blank group, # P <0.05, # P <0.01, # P <0.001, compared to the model group.
Therefore, the pharmaceutical composition for treating hyperuricemia has obvious effect of reducing blood uric acid on a hyperuricemia rat model established by potassium oxonate (2.5%) feed feeding, and the mechanism is that the pharmaceutical composition for treating hyperuricemia inhibits the expression level of URAT1, so that the reabsorption of uric acid is inhibited, the expression level of ABCG2 is enhanced, the excretion of uric acid is promoted, and the effect of reducing blood uric acid is exerted.
EXAMPLE 5 clinical trials
In this example, hyperuricemia is defined as hyperuricemia when the blood uric acid concentration is 20% higher than the normal value, i.e. the serum uric acid level of the male patient is more than 504mmol/L, and the serum uric acid level of the female patient is more than 432mmol/L (the blood uric acid concentration is higher than the normal value, the serum uric acid level of the male is not higher than 420mmol/L, and the serum uric acid level of the female is not higher than 360 mmol/L).
45 patients (with unlimited nature and 18-70 years old) were diagnosed with hyperuricemia and subjected to placebo-controlled evaluation randomized double-blind clinical trial (randomized cohort), wherein 30 patients in the drug trial group and 15 patients in the placebo-controlled group were treated.
Both groups of patients were on a normal daily life, diet, and serum uric acid concentrations were measured 1 time every 4 weeks (28 days). Meanwhile, the drug test group was administered with the blood uric acid lowering granules prepared in example 1 plus sodium bicarbonate. The administration method of the granules for reducing blood uric acid prepared in the embodiment 1 comprises 2 bags per day, 1 bag each in the morning and afternoon; the specific usage and dosage of sodium bicarbonate are as follows: 3 times/d, 0.5 g/time, orally, for 12 weeks. Placebo granules (character close to test drug) + sodium bicarbonate tablets, administration methods of placebo granules and sodium bicarbonate tablets, drug test groups were administered to the placebo control group.
During the test period, the blood uric acid level of the patient at different time periods, the liver function index change before and after treatment and the adverse reaction condition are observed. Liver function indices include alanine aminotransferase, aspartate aminotransferase, urea nitrogen, creatinine.
Adverse reactions include liver and kidney dysfunction, gastrointestinal reactions, skin rash, and pruritus. The clinical treatment effect judgment standard is as follows: combining the improvement degree of clinical symptoms of patients, if the serum uric acid level after treatment is reduced by more than or equal to 25 percent compared with that before treatment, or the serum uric acid level is reduced to a normal value range, the effect is obvious; if the serum uric acid level after treatment is reduced by 10 to 25 percent compared with that before treatment, or the serum uric acid level is close to the normal value range, and partial symptoms are effectively improved; it is not effective if the serum uric acid level after treatment is reduced by < 10% compared to that before treatment.
Serum uric acid concentrations were measured and recorded for the test patients at 28 days, 56 days and 74 days after the first dose, respectively, and the results are shown in table 11.
TABLE 11 clinical efficacy observations
As can be seen from table 11, at the end of the 72-day test, the significant efficiency of the drug test group is 63.3%, the effective rate is 26.6%, and the total effective rate of the two groups is 90%; the significant efficiency of the placebo control group is 0%, the effective rate is 13.3%, and the total effective rate of the two groups is 13.3%.
The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (8)
1. The pharmaceutical composition for treating hyperuricemia is characterized by being prepared from the following components:
500 portions and 900 portions of ash bark; and
400 portions of clematis root and 600 portions of clematis root.
2. The pharmaceutical composition for treating hyperuricemia according to claim 1, wherein: the pharmaceutical composition also contains a diluent and/or a flavoring agent and/or an adhesive as auxiliary materials.
3. The pharmaceutical composition for treating hyperuricemia according to claim 2, wherein the diluent is one or a mixture of sucrose, dextrin, soluble starch, mannitol, β -cyclodextrin, microcrystalline cellulose.
4. The pharmaceutical composition for treating hyperuricemia according to claim 2, wherein: the flavoring agent is steviosin and/or aspartame.
5. The pharmaceutical composition for treating hyperuricemia according to claim 2, wherein: the adhesive is water and/or ethanol.
6. The pharmaceutical composition for treating hyperuricemia according to any one of claims 1 to 5, wherein: the dosage form of the pharmaceutical composition is any one of tablets, granules, capsules, oral liquid and dropping pills.
7. A method for preparing a pharmaceutical composition for the treatment of hyperuricemia according to any one of claims 1 to 6, comprising the steps of:
① collecting cortex Fraxini and radix Clematidis;
② decocting in water, and collecting volatile oil;
③ mixing the decoctions, filtering, and concentrating.
8. Use of a pharmaceutical composition according to any one of claims 1 to 6 in the manufacture of a medicament for the treatment of hyperuricemia.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113181377A (en) * | 2021-04-30 | 2021-07-30 | 青岛大学 | Application of ABCG2 protein in preparation of preparation for improving, treating or assisting in treating hyperuricemia with hyperlipidemia and hyperglycemia |
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| CN1695665A (en) * | 2004-11-29 | 2005-11-16 | 成都中汇制药有限公司 | Medication for treating gout |
| CN106266002A (en) * | 2015-05-19 | 2017-01-04 | 天士力制药集团股份有限公司 | A kind of Chinese medicine composition treating hyperuricemia |
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| CN1695665A (en) * | 2004-11-29 | 2005-11-16 | 成都中汇制药有限公司 | Medication for treating gout |
| CN106266002A (en) * | 2015-05-19 | 2017-01-04 | 天士力制药集团股份有限公司 | A kind of Chinese medicine composition treating hyperuricemia |
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| CN113181377A (en) * | 2021-04-30 | 2021-07-30 | 青岛大学 | Application of ABCG2 protein in preparation of preparation for improving, treating or assisting in treating hyperuricemia with hyperlipidemia and hyperglycemia |
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