CN116350728B - Traditional Chinese medicine composition for reducing uric acid and/or resisting inflammation and relieving pain and application thereof - Google Patents

Traditional Chinese medicine composition for reducing uric acid and/or resisting inflammation and relieving pain and application thereof Download PDF

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CN116350728B
CN116350728B CN202310460391.4A CN202310460391A CN116350728B CN 116350728 B CN116350728 B CN 116350728B CN 202310460391 A CN202310460391 A CN 202310460391A CN 116350728 B CN116350728 B CN 116350728B
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chinese medicine
traditional chinese
medicine composition
uric acid
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CN116350728A (en
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林昌松
刘清平
徐强
刘敏莹
刘小宝
雷旭杰
陈淑琳
韩玉凤
刘明岭
张明英
刘丽娟
柯雪红
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First Affiliated Hospital of Guangzhou University of Chinese Medicine
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Abstract

The invention provides a traditional Chinese medicine composition for reducing uric acid and/or resisting inflammation and relieving pain and application thereof, wherein the active ingredients of the traditional Chinese medicine composition are prepared from the following raw materials in parts by weight: 15-60 parts of glabrous greenbrier rhizome, 6-12 parts of ash bark, 2-5 parts of coptis chinensis, 3-12 parts of amur corktree bark, 5-12 parts of twotooth achyranthes root, 5-10 parts of beautiful sweetgum fruit, 6-12 parts of lily, 9-30 parts of astragalus root and 9-15 parts of giant knotweed. The traditional Chinese medicine composition can be used for preparing medicines for treating gouty arthritis and/or hyperuricemia. The traditional Chinese medicine composition is provided for the pathogenesis of gout damp-heat phlegm turbidity and blood stasis, and has the effects of clearing heat and promoting diuresis, discharging turbidity and removing blood stasis and dredging joints. Pharmacodynamic experiments show that the traditional Chinese medicine composition has good uric acid reducing and anti-inflammatory analgesic effects on SD rat models of gouty arthritis and hyperuricemia, and the traditional Chinese medicine composition has good curative effects, and has more remarkable anti-inflammatory analgesic effects while reducing uric acid.

Description

Traditional Chinese medicine composition for reducing uric acid and/or resisting inflammation and relieving pain and application thereof
Technical Field
The invention belongs to the technical field of traditional Chinese medicines, and particularly relates to a traditional Chinese medicine composition for reducing uric acid and/or resisting inflammation and relieving pain and application thereof.
Background
Gouty arthritis is recurrent joint inflammation caused by elevated serum uric acid level and sodium urate crystal deposition on tissues such as joint capsules or tendons, and is characterized by red, swelling, heat and pain of single joints or multiple joints, severe patients cannot walk, the pain degree reaches the peak within 24 hours of onset, colchicine can be rapidly relieved after taking, and ankle joints and first metatarsophalangeal joints are the most frequently onset parts of gouty arthritis. Hyperuricemia is a direct cause of gouty arthritis, and high purine diet or high alcohol and fructose intake easily causes purine metabolic disorder, excessive uric acid generation or excretion disorder, thereby causing elevated blood uric acid concentration and urate deposition, and gouty arthritis is produced under the stimulation of severe exercise, diet, wound and other causes.
With the improvement of living conditions, the incidence rate of gouty arthritis and hyperuricemia in China rises year by year, and the incidence population has a tendency of younger. Gouty arthritis can increase the risks of diabetes, hyperlipidemia and cardiovascular and cerebrovascular diseases, and the urates can be deposited on various parts of a human body, can cause joint deformity and joint movement disorder when deposited on joints, can cause kidney stones when deposited on kidneys, and can cause kidney function injury and even kidney failure when serious patients can greatly influence the life quality of patients.
The current gouty arthritis treatment drugs mainly comprise glucocorticoids, non-steroidal anti-inflammatory drugs, colchicine and uric acid lowering drugs, wherein the non-steroidal anti-inflammatory drugs comprise naproxen, diclofenac sodium, ibuprofen, celecoxib, meloxicam, indomethacin and the like, and the uric acid lowering drugs comprise allopurinol, febuxostat, tribromone, probenecid and polyethylene glycol uricase. Glucocorticoids, non-steroidal anti-inflammatory drugs, colchicine are often used in the acute onset of gout, whereas uric acid lowering drugs are mostly used in the intermittent gout stage and hyperuricemia. However, glucocorticoids may increase the risk of infection or cause the rise of blood sugar in diabetics, nonsteroidal anti-inflammatory drugs may damage the kidney, cardiovascular and gastrointestinal functions of patients, and long-term use of uric acid lowering drugs may cause impaired liver and kidney functions and cardiovascular diseases, and some patients have drug resistance to the drugs and poor uric acid level control.
The traditional Chinese medicine has the characteristics of low toxic and side effect, good curative effect and difficult drug resistance, and various traditional Chinese medicine compositions or traditional Chinese medicine preparations for treating gout or hyperuricemia exist at present, for example, the Chinese patent application of publication No. CN112022966A (12 months 4 days of publication No. 2020) discloses a traditional Chinese medicine composition for preventing, relieving and/or treating hyperuricemia. However, the gouty arthritis patient is in the alternate process of the acute attack stage and the remission stage for a long time, so that the medicine capable of treating the anti-inflammatory pain in the acute attack stage and the uric acid in the remission stage of gout or hyperuricemia can reduce the dosage, and can also reduce the toxic and side effects of the medicine and the economic burden of the patient. However, there is currently no drug with both uric acid lowering and anti-inflammatory analgesic effects, and patient needs are difficult to meet.
Disclosure of Invention
Based on the above, the invention aims to provide a traditional Chinese medicine composition for reducing uric acid and/or resisting inflammation and relieving pain and application thereof. The traditional Chinese medicine composition has the effects of reducing uric acid and resisting inflammation and relieving pain, and can be used for simultaneously treating gouty arthritis and hyperuricemia.
In order to achieve the above purpose, the present invention adopts the following technical scheme.
The traditional Chinese medicine composition for reducing uric acid and/or resisting inflammation and relieving pain is prepared from the following raw materials in parts by weight: 15-60 parts of glabrous greenbrier rhizome, 6-12 parts of ash bark, 2-5 parts of coptis chinensis, 3-12 parts of amur corktree bark, 5-12 parts of twotooth achyranthes root, 5-10 parts of beautiful sweetgum fruit, 6-12 parts of lily, 9-30 parts of astragalus root and 9-15 parts of giant knotweed.
In some preferred embodiments, the active ingredients of the traditional Chinese medicine composition are prepared from the following raw materials in parts by weight: 30-60 parts of glabrous greenbrier rhizome, 8-12 parts of ash bark, 2-5 parts of coptis chinensis, 5-12 parts of amur corktree bark, 5-12 parts of twotooth achyranthes root, 5-10 parts of beautiful sweetgum fruit, 6-12 parts of lily, 15-25 parts of astragalus root and 9-15 parts of giant knotweed.
More preferably, the active ingredients of the traditional Chinese medicine composition are prepared from the following raw materials in parts by weight: 60 parts of glabrous greenbrier rhizome, 10 parts of ash bark, 5 parts of coptis chinensis, 10 parts of amur corktree bark, 10 parts of twotooth achyranthes root, 10 parts of beautiful sweetgum fruit, 10 parts of lily, 20 parts of astragalus root, and 15 parts of giant knotweed.
The invention also provides a preparation method of the traditional Chinese medicine composition, which comprises the following steps: all the raw materials are weighed according to the proportion, and extracted by using an extraction reagent to obtain the traditional Chinese medicine composition.
In some embodiments, the extraction reagent is water.
In some embodiments, the preparation method comprises the steps of: all the raw materials are weighed according to the proportion, soaked in water, decocted and extracted for 1 to 3 times, each time for 0.5 to 1.5 hours, and the decoction is combined.
The invention also provides application of the traditional Chinese medicine composition or the traditional Chinese medicine composition prepared by the preparation method in preparation of medicines for treating gouty arthritis and/or hyperuricemia.
The invention also provides a medicine for treating gouty arthritis and/or hyperuricemia, which comprises the traditional Chinese medicine composition or the traditional Chinese medicine composition prepared by the preparation method and pharmaceutically acceptable auxiliary materials.
In some embodiments, the medicament is in an oral dosage form.
In some preferred embodiments, the medicament is a decoction, a powder, a capsule, a tablet, a honeyed pill, a water-honeyed pill, a watered pill, a concentrated pill, a paste pill, a wax pill, a granule, an oral liquid, or a drop pill.
The invention obtains a traditional Chinese medicine composition with uric acid reducing, anti-inflammatory and analgesic effects through optimization, and the preparation raw materials of the traditional Chinese medicine composition comprise glabrous greenbrier rhizome, ash bark, coptis chinensis, amur corktree bark, twotooth achyranthes root, beautiful sweetgum fruit, lily, astragalus root and giant knotweed rhizome in proper dosage, and the compatibility principle is as follows: rhizoma smilacis glabrae dehumidifies and detoxifies, relieves the pain of the joints, and is reused as a monarch drug; coptis chinensis, phellodendron bark, cortex fraxini, polygonum cuspidatum and fructus forsythiae Lu Tongqing are used as ministerial drugs for removing dampness-heat in the triple energizer, dispersing blood stasis and relieving pain; achyranthes root, lily and astragalus root, radix astragali have the effects of strengthening spleen, tonifying kidney and tonifying lung and are adjuvant drugs. The 9 traditional Chinese medicines are matched with each other to supplement each other, and have the effects of discharging turbidity, removing dampness, removing blood stasis and relieving pain. Pharmacodynamic experiments show that the traditional Chinese medicine composition has good uric acid reducing and anti-inflammatory analgesic effects on SD rat models of gouty arthritis and hyperuricemia, and the traditional Chinese medicine composition has good curative effects, has more remarkable anti-inflammatory analgesic effects while reducing uric acid, and has statistical significance on differences.
Drawings
FIG. 1 is a graph showing the effect of the Chinese medicinal composition on the inflammatory index of right ankle joint of SD rat with gouty arthritis.
Detailed Description
In order that the invention may be understood more fully, a more particular description of the invention will be rendered by reference to specific embodiments thereof which are illustrated in the appended claims. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. These embodiments are provided so that this disclosure will be thorough and complete. It should be understood that the experimental methods in the following examples, in which specific conditions are not noted, are generally performed under conventional conditions or under conditions suggested by the manufacturer. The various reagents commonly used in the examples are all commercially available products.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. The term "and/or" as used herein includes any and all combinations of one or more of the associated listed items.
Gout and hyperuricemia are caused by improper diet and weak intrinsic constitution of patients. The patient overeating thick paste flavor, spleen and stomach weakness does not transport and transform food, food stagnates to become water-damp, and damp gradually transforms into heat, which is phlegm for a long time. The internal generation of phlegm-heat and damp turbidity blocks the triple energizer, the lung lobes do not transport water and food grains, the spleen and stomach do not transport water and moisture, and the kidneys do not transport urine, so that food metabolites such as uric acid are accumulated in the body and cannot be normally discharged out of the body, and hyperuricemia is formed; the accumulation of phlegm-heat locally blocks the channels and collaterals, and pain is caused by obstruction of collaterals, so the joint is red, swollen, hot and painful, gouty arthritis, long-term accumulation of phlegm-turbidity, obstruction of blood vessels, blood stasis and phlegm-turbidity cementation are caused, and the joint is tophus for a long time.
The traditional Chinese medicine composition for gout and hyperuricemia provided by the invention comprises the following components: smilax glabra is sweet in nature, light in taste, flat, and capable of returning to liver and stomach channels, removing dampness, expelling turbidity, removing toxin, promoting joint function, helping spleen and stomach transport and transform water dampness, and is reused as a monarch drug; coptis root has bitter and cold nature and enters heart, spleen, stomach, liver, gall bladder and large intestine meridians to clear damp-heat in upper and middle energizer; cortex Fraxini has bitter, astringent and cold properties, and can enter liver, gallbladder and large intestine channels to clear damp-heat in middle-jiao and lower-jiao; huang Baixing the Chinese medicinal composition has the effects of bitter and cold taste, invigorating kidney and bladder meridian, clearing away damp-heat, and helping the lower-jiao kidney to open and close; polygonum cuspidatum has slightly bitter and slightly cold taste, enters liver, gall and lung meridians, and can promote diuresis, clear heat, dissipate blood stasis and relieve pain; the fructus liquidambaris is bitter in property, flat, enters liver and kidney meridians, and can dispel wind, activate collaterals and promote diuresis; the five flavors are combined as ministerial drugs to assist the monarch drugs to clear damp-heat in the triple energizer. Achyranthes root has bitter and sweet taste, is sour and flat, and has the functions of invigorating liver and kidney, strengthening tendons and bones, promoting urination, eliminating dampness and removing blood stasis; lily is sweet and cold in nature, returns to heart and lung channels, can moisten and nourish joints, and prevents monarch and minister drugs from damaging yin due to excessive dryness; astragalus root has sweet and slightly warm taste, enters lung and spleen meridian, and can replenish qi, invigorate spleen, invigorate lung, assist upper and middle-jiao to deliver and transport water; the three medicines are combined together, and the assistant and assistant medicines invigorate the spleen, benefit the kidney and tonify the lung, restore the function of lung, spleen and kidney triple energizer to distribute body fluid and remove damp turbidity. Simultaneously, the fructus liquidambaris, the achyranthes root, the polygonum cuspidatum and the like have the effects of activating blood, dredging collaterals and relieving pain, so that blood stasis is removed, and joints are facilitated to relieve pain. The whole formula has the effects of expelling turbidity, removing dampness, removing blood stasis and relieving pain, and is suitable for arthralgia syndrome due to damp-heat accumulation, gout, hyperuricemia and other diseases.
The subject of administration of the Chinese medicinal composition and/or the medicament of the present invention is a mammal, preferably a human, in need thereof.
According to the total mass of the traditional Chinese medicine composition, when the traditional Chinese medicine composition or the medicine is applied to a person in need thereof, the daily dosage is 1.0-2.8 g/kg.
Pharmaceutically acceptable excipients of the present invention include, but are not limited to, (1) diluents such as starches, powdered sugar, dextrins, lactose, pregelatinized starches, microcrystalline fibers, inorganic calcium salts (e.g., calcium sulfate, dibasic calcium phosphate, medicinal calcium carbonate, etc.), mannitol, vegetable oils, polyethylene glycols, etc.; (2) Binders such as distilled water, ethanol, starch slurry, sodium carboxymethyl cellulose, hydroxypropyl cellulose, methyl cellulose, and ethyl cellulose, hydroxypropyl methylcellulose, and the like; (3) Disintegrants such as dry starch, sodium carboxymethyl starch, low-substituted hydroxypropylcellulose, crosslinked polyvinylpyrrolidone, crosslinked sodium carboxymethyl cellulose, etc.; (4) Lubricants such as magnesium stearate, silica gel micropowder, talc, hydrogenated vegetable oil, polyethylene glycols, magnesium lauryl sulfate, etc.; (5) solvents such as water, ethanol solutions, and the like.
In the present invention, the "parts" or "parts by weight" refers to the relative mass ratio between the related substances, and does not represent a specific mass number of a specific substance. Depending on the production, clinical practice, 1 part or 1 part by weight may be 1g, 10g, 500g, 1kg, and any other mass number.
The following description is made with reference to specific embodiments.
Example 1
The embodiment provides a water decoction for reducing uric acid and/or resisting inflammation and relieving pain, which comprises the following raw material medicines in parts by weight:
60g of glabrous greenbrier rhizome, 10g of ash bark, 5g of coptis chinensis, 10g of amur corktree bark, 10g of twotooth achyranthes root, 10g of beautiful sweetgum fruit, 10g of lily, 20g of astragalus root and 15g of giant knotweed.
The specific preparation method comprises the following steps:
1) Removing impurities from rhizoma Smilacis Glabrae, cortex Fraxini, coptidis rhizoma, cortex Phellodendri, achyranthis radix, fructus Lipuidambaris, bulbus Lilii, radix astragali, and rhizoma Polygoni Cuspidati, and cleaning to obtain a raw material mixture;
2) Soaking the raw material mixture in the step 1) in 6-10 times of purified water for 1.5 hours, decocting with fast fire, decocting slowly for 1 hour, pouring out the decoction, adding 8 times of purified water, decocting slowly for 45 minutes, pouring out the decoction, mixing the two decoctions, and stirring uniformly.
The decoction is administered once a day, and is divided into two times a day, and is administered in the morning and evening.
Example 2
This example investigated the pharmacodynamic test of the traditional Chinese medicine composition (hereinafter referred to as a turbidity-relieving pain-relieving prescription) of the invention on the SD rat hyperuricemia model caused by hypoxanthine and potassium oxazinate.
1. Experimental materials
1.1 laboratory animals
36 SPF-grade 6-week male SD rats were provided by the first affiliated hospital laboratory animal center, university of Guangzhou traditional Chinese medicine, 200+ -20 g body weight, eligibility number: no.44829700007976, experimental license number: SYXK (Guangdong) 2018-0092. The strain is fed to an SPF-grade animal experiment center of a first affiliated hospital of Guangzhou university of Chinese medicine, the room temperature is maintained at (23+/-2 ℃), the humidity is maintained at (55% +/-5), the light and shade are alternately changed for 12 hours, and the strain is adaptively fed for 7 days.
1.2 Experimental drugs
1) A Chinese medicinal composition comprises: 60g of glabrous greenbrier rhizome, 10g of ash bark, 5g of coptis chinensis, 10g of amur corktree bark, 10g of twotooth achyranthes root, 10g of beautiful sweetgum fruit, 10g of lily, 20g of astragalus root and 15g of giant knotweed rhizome are decocted according to a clinical traditional Chinese medicine decoction method. The first decoction is fully soaked in deionized water with the mass volume ratio of 10 times, then the mixture is boiled with strong fire and then decocted with slow fire for 1h, the water decoction is collected, and the obtained water decoction is filtered by gauze and a 200-mesh screen to obtain the water decoction of the 1 st time. The second decoction is carried out by decocting the medicine with deionized water with the mass of 8 times of medicine for 45min, filtering again and collecting filtrate, and mixing the two water decoctions uniformly. Concentrating under reduced pressure by rotary evaporator, controlling water bath temperature at 70deg.C, concentrating the water decoction to obtain water decoction of Chinese medicinal composition.
2) Allopurinol tablet: produced by Shanghai friendship universal pharmaceutical industry stock company, [ approval mark ] national drug standard character H31020334; grinding into fine powder, adding appropriate amount of pure water, and making into allopurinol solution.
1.3 Experimental reagents
1) Potassium oxazinate: sigma, cat No.: 156124, lot number: BCCG6675.
2) Hypoxanthine: sigma, cat No.: h9377, lot number: SLCG4177.
2. Experimental method
2.1 modeling and administration
The 36 SD rats with SPF level were randomly divided into a normal group, a model group, a low dose group of the Chinese medicinal composition, a medium dose group of the Chinese medicinal composition, a high dose group of the Chinese medicinal composition, and an allopurinol group, each group being 6. In addition to the normal group, each group was given 500mg/kg of oral hypoxanthine per day for intragastric administration, and 100mg/kg of potassium oxazinate was intraperitoneally injected for a total of 7 days. On the 8 th day of molding, each dosage group of the traditional Chinese medicine composition and allopurinol are respectively administered with traditional Chinese medicine composition decoction (6.60 g/kg, 13.19g/kg, 26.38 g/kg) and allopurinol solution (10 mg/kg) after half an hour of molding, and the normal group and the model group are filled with distilled water, and the gastric lavage volume is 10ml/kg. All doses of the modeling agent, drug and distilled water were converted according to the body surface area of the rats, and each administration group was given 1 time per day by oral gavage and intraperitoneal injection for a total of 21 days.
2.2 serum and urine sample collection
Serum samples were collected 3h after the 3h post-treatment on day 21 of modeling. After the experiment is completed, the abdominal cavity is anesthetized with chloral hydrate (350 mg/kg), the anesthetized rat is fixed on a homemade operation plate, abdominal hair is cut off, after conventional sterilization, the abdomen is opened, the abdominal aorta is moved to collect blood, 5ml of blood is collected by a red vacuum tube (non-anticoagulation), high-speed centrifugation is carried out at 3500rpm for 10min, and the obtained blood supernatant is stored in a refrigerator at-80 ℃.
2.3 Biochemical index detection
Using a fully automated biochemical analyzer (Roche)8000 Serum samples stored in-80 ℃ refrigerator were tested for uric acid (SUA), creatinine (SCr), urea nitrogen (BUN), glutamic-oxaloacetic transaminase (AST), glutamic-pyruvic transaminase (ALT).
2.4 statistical methods
Statistical analysis was performed using SPSS 26.0. The results of each group of data were expressed as Mean ± standard deviation (Mean ± SD), and the significance of the difference between the two groups was checked using independent sample T-test. * P is less than 0.05, and the difference has statistical significance; ** p is less than 0.01, and the difference has obvious statistical significance.
3. Results
The effect results of the traditional Chinese medicine composition on hyperuricemia SD rat blood uric acid and liver and kidney functions are shown in table 1:
TABLE 1
Note that: in comparison with the normal group, ## P<0.01, # P<0.05; in comparison with the set of models, ** P<0.01, * P<0.05。
the results in Table 1 show that the SUA level of SD rats in the model group is significantly higher than that in the normal group on the 21 st day of model formation, the difference has statistical significance (P < 0.05), and the model group SD rats are successfully model formation, meanwhile, the SCr level of SD rats in the model group is significantly increased (P < 0.05) compared with that in the normal group, and the BUN average value is increased, so that the kidney function of SD rats can be influenced by hyperuricemia, and the kidney function of the rats is damaged. After 14 days of positive control (allopurinol tablet) and the high, medium and low dose treatments of the turbid expelling and pain relieving prescription, the SUA level of SD rats is obviously reduced, and compared with a model group, the high dose group (P < 0.05) and the low dose group (P < 0.05) have statistical differences, so that the turbid expelling and pain relieving prescription is indicated to reduce the SUA level of the SD rats with hyperuricemia. Compared with the model group, the SCr, BUN, ALT, AST level of the SD rats in the high and medium dose groups is reduced, the SCr, BUN, ALT level of the SD rats in the low dose group is not obviously different from that of the SD rats in the normal group, the AST level is reduced compared with the model group, the turbidity discharging pain relieving formula is indicated to have no damage to liver and kidney functions, and the renal function injury caused by hyperuricemia can be reversed, and the hepatotoxicity can be prevented.
The effect of the traditional Chinese medicine composition on the weight of the hyperuricemia SD rat is shown in table 2:
TABLE 2
Table 2 (subsequent)
Note that: in comparison with the normal group, ## P<0.01, # P<0.05; in comparison with the set of models, ** P<0.01, * P<0.05。
the results in Table 2 show that SD rats in the low dose group of the turbidity-relieving pain-relieving prescription have significantly increased weight (P < 0.05) in the 1 st balance compared with the normal group, and significantly reduced weight (P < 0.05) in the 10 th and 16 th balances compared with the normal group, after the administration treatment from the 8 th day to the 21 st day of molding, the weight levels of SD rats in the low dose group, the medium and high dose groups are not significantly different from the normal group, and the weight levels of rats in the medium dose group and the low dose group are higher than those in the model group, so that the turbidity-relieving pain-relieving prescription has no influence on the general nutrition state of rats.
Example 3
The present example studied the pharmacodynamic test of the traditional Chinese medicine composition (hereinafter referred to as a turbidity-relieving pain-relieving prescription) of the present invention on sodium urate-induced SD rat gouty arthritis model.
1. Experimental materials
1.1 laboratory animals
48 SPF-grade 6-week male SD rats were provided by the first affiliated hospital laboratory animal center, university of Guangzhou traditional Chinese medicine, 200+ -20 g body weight, eligibility number: no.44829700009906, experimental license number: SYXK (Guangdong) 2018-0092. This was fed to the SPF class animal center of the first affiliated Hospital, guangzhou university of Chinese medicine under the same conditions as in example 2, item 1.1.
1.2 Experimental drugs
1) A Chinese medicinal composition comprises: the preparation of the water decoction of the traditional Chinese medicine composition is the same as 1.2.1) in the example 2.
2) Colchicine tablet: produced by Xishuangbanna pharmaceutical industry limited liability company, [ approved literature ] national drug standard H53021369; grinding into fine powder, adding appropriate amount of pure water, and making into allopurinol solution.
1.3 Experimental reagents
1) Sodium urate: sigma, cat No.: u2875, lot number: BCCH3341.
2) IL-1 beta enzyme-linked immunosorbent assay kit: jiang Lai organism, cat No.: JL20884, lot number: 032716003208840411.IL-6 ELISA kit: jiang Lai organism, cat No.: JL20896, lot number: 032716003208960411. TNF-alpha ELISA kit: jiang Lai organism, cat No.: JL13202, lot number: 032716003132020411.
2. experimental method
2.1 modeling and administration
2.1.1 grouping and administration
The 48 SD rats with SPF level were randomly divided into a normal group, a model group, a low-dose group of the Chinese medicinal composition, a medium-dose group of the Chinese medicinal composition, a high-dose group of the Chinese medicinal composition, and a colchicine group, 8 of which were each. The stomach of all rats is irrigated, and each dosage group of the traditional Chinese medicine composition and colchicine are respectively administrated with traditional Chinese medicine composition decoction (6.60 g/kg, 13.19g/kg, 26.38 g/kg) and colchicine solution (0.3 mg/kg) for oral administration, and distilled water is orally administrated to the normal group and the model group, the stomach irrigation volume is 10ml/kg, and the stomach is irrigated continuously for 7 days once a day.
2.1.2 establishment of acute gouty arthritis model
On day 5 of gavage, the rats of the other groups, except the normal group, replicate the acute gouty arthritis model, and 0.2ml of physiological saline is injected into the ankle joint on the right rear side of the normal group. And (5) continuing to irrigate the stomach for administration according to groups for 2 hours after molding.
The molding method comprises the following steps: the method comprises the steps of selecting a puncture part at the rear of the outer side of a right ankle joint of an SD male rat, bending the puncture part and carrying out local disinfection, enabling an upward needle inserting direction to form an included angle of 30-45 degrees with tibia, injecting 0.2ml of sodium urate crystal solution with the concentration of 25mg/ml into a joint cavity of the rat by using a 4.5-gauge needle, and constructing an animal model of acute gouty arthritis by taking the opposite side bulge of a joint capsule as an injection standard.
Molding success standard: after sodium urate crystals are injected for 2 hours, compared with a normal group, the gouty arthritis model rat has the advantages that the red and swollen skin temperature of the right ankle joint is increased, the osseous mark is disappeared, the color of the sole is dark red purple, walking is slow, feet are curled, and even the hind limbs are overturned, the feet are in a trekking gait and the modeling is successful.
2.2 serum sample collection
After 2h of gastric lavage on day 7, 1% pentobarbital is injected into the abdominal cavity for anesthesia, the anesthetic dose is 6ml/kg, 5ml blood is drawn through abdominal aorta, the abdominal aorta is placed in a centrifuge after standing for 30min, the centrifuge is centrifuged for 15min at 3500r/min, upper serum is sucked, and the sub-package is kept at-20 ℃ for standby. Rats were sacrificed after blood collection.
2.3 rat joint inflammation index and ankle swelling degree measurement
All rats were subjected to measurements of joint inflammation index and ankle swelling degree before (0 h) and after (2 h, 6h, 12h, 24h, 48 h) molding.
Index grading of joint inflammation: 0. normal; 1. erythema, mild swelling, and visible bony signs of joint skin; 2. obvious redness and swelling of the joint, disappearance of the bony marks, but swelling is limited to the joint parts; 3. the limb is swollen outside the joint.
Ankle swelling degree: before molding, a clear straight line is drawn at the position 0.5mm below the ankle joint of the rat by using a marker pen, the joint diameter of the same part of different time nodes before molding and after inducing inflammation is measured by using a vernier caliper for 3 times, an average value is obtained, and the difference value between the average value and the joint diameter measured for 0h is used as the swelling degree.
2.4 serum index detection
The concentrations of IL-1. Beta., IL-6 and TNF-. Alpha.in rat serum were measured by ELISA according to the kit instructions.
2.5 statistical methods
Statistical analysis was performed using SPSS 26.0. The results of the data of the metrology data were expressed as Mean ± standard deviation (Mean ± SD), and the significance of the differences between the two groups was checked using independent sample T-test. The data results of the count data were expressed using frequency (N) and the significance of the differences between the two groups was checked using the Wilcoxon Mann-Whitney test. * P is less than 0.05, and the difference has statistical significance; ** p is less than 0.01, and the difference has obvious statistical significance.
3. Results
The effect of the Chinese medicinal composition on the inflammatory index of right ankle of SD rat with gouty arthritis (N) results are shown in Table 3:
TABLE 3 Table 3
Table 3 (subsequent)
Note that: in comparison with the normal group, ## P<0.01; in comparison with the set of models, * P<0.05。
in the molding process, 1 rat in the blank group and the low-dose group of the traditional Chinese medicine composition dies after anesthesia, and the rest rats are not abnormal. The results in table 3 show that the right ankle inflammation index of the model group SD rats at 2, 6, 12, 24, 48 hours after molding was significantly increased (P < 0.01) compared to the normal group rats, suggesting that molding was successful. Fig. 1 shows that compared with the model group, average right ankle joint inflammation index of the rats in the middle and low dose groups is reduced after 6 hours and 12 hours after molding, and average joint inflammation index of the rats in the high and low dose groups is continuously reduced after 12 hours, 24 hours and 48 hours after molding, which indicates that the middle and low dose groups can effectively reduce joint inflammation index of the SD rat with gouty arthritis.
The effect of the traditional Chinese medicine composition on the ankle joint swelling degree of SD rat with gouty arthritis is shown in Table 4:
TABLE 4 Table 4
Note that: in comparison with the normal group, ## P<0.01, # P<0.05; in comparison with the set of models, ** P<0.01, * P<0.05。
the results in table 4 show that model group SD rats had significantly increased ankle swelling levels (P < 0.05) at 2 hours post-molding compared to the normal group, and significantly increased ankle swelling levels (P < 0.01) at 6, 12, 24, 48 hours post-molding, suggesting successful molding. Compared with a model group, the maximum ankle joint swelling degree of rats in a high-dose group, a medium-dose group and a low-dose group is obviously reduced, the average joint swelling degree of rats in a high-dose group is obviously reduced after 6 hours (P < 0.05) and 48 hours (P < 0.01) after molding compared with the model group, the average joint swelling degree of rats in a medium-dose group is obviously reduced after 48 hours (P < 0.05) after molding compared with the model group, and the method for prompting the turbidity relief and pain relief can effectively reduce the joint swelling of SD rats with gouty arthritis.
The effect results of the traditional Chinese medicine composition on the serum index of SD rat with gouty arthritis are shown in Table 5:
TABLE 5
Note that: in comparison with the normal group, ## P<0.01, # P<0.05; in comparison with the set of models, ** P<0.01, * P<0.05; in comparison with the colchicine group, △△ P<0.01, P<0.05。
the results in Table 5 show that the serum inflammatory factors IL-1 beta, IL-6, TNF-alpha levels were significantly elevated 48h after molding (P < 0.01) in SD rats in the model group compared to the normal group, suggesting that gouty arthritis may lead to joint inflammation in SD rats. Compared with a model group, the high, medium and low dose groups SD rats have obviously reduced IL-1 beta, IL-6 and TNF-alpha levels (P < 0.01), and are dose dependent, which shows that the high dose groups SD rats have no statistical difference (P > 0.05) between the TNF-alpha level and the normal groups SD rats, and the high dose groups SD rats have obviously reduced arthritis factors of gouty arthritis SD rats and reduced arthritis, which shows that the high dose groups SD rats have normal TNF-alpha level induced by sodium urate crystals and good anti-inflammatory effect. Compared with colchicine group (positive control drug), the IL-6 (P < 0.05) and TNF-alpha (P < 0.01) levels of the high-dose group rats of the turbidity-relieving pain-relieving prescription are obviously reduced, the TNF-alpha (P < 0.01) levels of the high-dose group rats are obviously reduced, and the average IL-6, TNF-alpha level of the low-dose group rats, the medium-dose group rats and the high-dose group rats of the turbidity-relieving pain-relieving prescription and the average IL-1 beta level of the medium-dose group rats and the high-dose group rats are reduced compared with the colchicine group, so that the efficacy of the turbidity-relieving pain-relieving prescription in reducing inflammatory factors of the SD rats with gouty arthritis is suggested to be better than that of colchicine.
In conclusion, the traditional Chinese medicine composition has good uric acid reducing, anti-inflammatory and analgesic effects on SD rat models of gouty arthritis and hyperuricemia.
The foregoing examples illustrate only a few embodiments of the invention and are described in detail herein without thereby limiting the scope of the invention. It should be noted that it will be apparent to those skilled in the art that several variations and modifications can be made without departing from the spirit of the invention, which are all within the scope of the invention. Accordingly, the scope of protection of the present invention is to be determined by the appended claims.

Claims (9)

1. The traditional Chinese medicine composition for reducing uric acid and/or treating gouty arthritis is characterized by comprising the following active ingredients in parts by weight: 15-60 parts of glabrous greenbrier rhizome, 6-12 parts of ash bark, 2-5 parts of coptis chinensis, 3-12 parts of amur corktree bark, 5-12 parts of twotooth achyranthes root, 5-10 parts of beautiful sweetgum fruit, 6-12 parts of lily, 9-30 parts of astragalus root and 9-15 parts of giant knotweed.
2. The uric acid lowering and/or gouty arthritis treating traditional Chinese medicine composition according to claim 1, wherein the active ingredients of the traditional Chinese medicine composition are prepared from the following raw materials in parts by weight: 30-60 parts of glabrous greenbrier rhizome, 8-12 parts of ash bark, 2-5 parts of coptis chinensis, 5-12 parts of amur corktree bark, 5-12 parts of twotooth achyranthes root, 5-10 parts of beautiful sweetgum fruit, 6-12 parts of lily, 15-25 parts of astragalus root and 9-15 parts of giant knotweed.
3. The uric acid lowering and/or gouty arthritis treating traditional Chinese medicine composition according to claim 2, wherein the active ingredients of the traditional Chinese medicine composition are prepared from the following raw materials in parts by weight: 60 parts of glabrous greenbrier rhizome, 10 parts of ash bark, 5 parts of coptis chinensis, 10 parts of amur corktree bark, 10 parts of twotooth achyranthes root, 10 parts of beautiful sweetgum fruit, 10 parts of lily, 20 parts of astragalus root, and 15 parts of giant knotweed.
4. A method for preparing a Chinese medicinal composition according to any one of claims 1 to 3, comprising the steps of: weighing all the raw materials according to a proportion, and extracting by using an extracting reagent to obtain the traditional Chinese medicine composition; the extraction reagent is water.
5. The method of manufacturing as claimed in claim 4, comprising the steps of: all the raw materials are weighed according to the proportion, soaked in water, decocted and extracted for 1 to 3 times, each time for 0.5 to 1.5 hours, and the decoction is combined.
6. Use of a traditional Chinese medicine composition according to any one of claims 1-3 or a traditional Chinese medicine composition obtained by a preparation method according to any one of claims 4-5 in the preparation of a medicament for treating gouty arthritis and/or hyperuricemia.
7. A medicament for treating gouty arthritis and/or hyperuricemia, characterized in that the medicament comprises the traditional Chinese medicine composition according to any one of claims 1-3 or the traditional Chinese medicine composition prepared by the preparation method according to any one of claims 4-6 and pharmaceutically acceptable auxiliary materials.
8. The medicament of claim 7, wherein the medicament is in an oral dosage form.
9. The medicament according to claim 8, wherein the medicament is a decoction, a powder, a capsule, a tablet, a honeyed pill, a water honeyed pill, a watered pill, a concentrated pill, a paste pill, a wax pill, a granule, an oral liquid, or a drop pill.
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* Cited by examiner, † Cited by third party
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双黄痛风胶囊抗痛风性关节炎作用研究;张白嘉等;中国药房;-;第19卷(第36期);2816-2817 *

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