A traditional Chinese medicine compound for treating gouty arthritis

Abstract

The application provides a traditional Chinese medicine compound for treating gouty arthritis, and relates to the technical field of traditional Chinese medicine pharmacy.The traditional Chinese medicine compound takes chicory and kidney tea as monarch drugs, takes anemarrhena as a minister drug, takes mulberry branch, oil pine knot, sophora fruit, celery seed, cassia twig and white peony root as auxiliary drugs, and takes rattan fig as a minister drug, and the whole prescription uses cold and warm drugs together, uses attack and tonification together, and considers both symptoms and root causes, and jointly plays the effects of clearing heat and reducing dampness, dredging collaterals and relieving pain, and invigorating spleen and eliminating turbidity.The application evaluates the curative effect of the traditional Chinese medicine compound on gouty arthritis by constructing a gouty arthritis cell model and a model, and the experimental results show that the traditional Chinese medicine compound reduces the expression level of inflammatory mediators in joint tissues, improves the pathological condition of joint tissues, achieves good reduction of the joint swelling degree of GA model rats, improves the joint dysfunction condition, and has a significant therapeutic effect on gouty arthritis rats.The traditional Chinese medicine compound is expected to be further researched and applied as a new drug for treating gouty arthritis.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine manufacturing technology, specifically to a traditional Chinese medicine compound for treating gouty arthritis. Background Technology

[0002] Gouty arthritis (GA) is an inflammatory disease caused by long-term purine metabolism disorders or uric acid excretion disorders, leading to elevated uric acid levels in the blood and subsequent deposition of urate crystals in the joint cavity and surrounding tissues. This deposition is the basis for clinical symptoms such as joint redness, swelling, severe pain, and even deformity. The disease generally affects the first articular joint (R joint) and is usually unilateral. With the continuous development of my country's social economy and the gradual increase in people's income levels, dietary structures have changed accordingly, resulting in an incidence rate of gout in my country between 1% and 3%, showing a year-on-year upward trend. Western medicine generally uses three main classes of drugs: non-steroidal anti-inflammatory drugs (NSAIDs), colchicine, and glucocorticoids. These drugs are effective in reducing inflammation and pain and slowing disease progression. However, it is important to note that these drugs often have many adverse reactions, are not suitable for all patients, and are prone to recurrence. In traditional Chinese medicine theory, gouty arthritis is classified under various conditions such as Bi syndrome, turbid blood stasis Bi syndrome, arthralgia, and wandering wind syndrome. The pathogenesis of gouty arthritis is largely related to the invasion of cold and dampness, leading to dysfunction of kidney qi transformation, disordered water metabolism, and retention of dampness in the body. Long-term dampness accumulation in the body disrupts qi circulation, causing phlegm and blood stasis to obstruct the meridians of the limbs. Combined with a diet high in fatty and sweet foods, this generates internal damp-heat, which invades the internal organs, tendons, and joints, resulting in joint pain. The combination of internal and external pathogenic factors leads to this disease, causing pain and swelling in the limbs and joints, and potentially causing limb deformities. Traditional Chinese medicine has used herbal medicine to treat gout for over two thousand years. Many Chinese herbs and their active ingredients have advantages in treating gouty arthritis, including multi-target effects, low toxicity, and significant efficacy. Against this background, this invention aims to provide a traditional Chinese medicine compound for treating gouty arthritis, evaluate its therapeutic effect, and explore its anti-inflammatory and uric acid-lowering pharmacological effects, providing new ideas and directions for the treatment of gouty arthritis. Summary of the Invention

[0003] (a) Technical problems to be solved

[0004] In view of the shortcomings of the existing technology, the present invention provides a traditional Chinese medicine compound for the treatment of gouty arthritis.

[0005] (II) Technical Solution

[0006] To achieve the above objectives, the present invention provides the following technical solution:

[0007] In a first aspect, the present invention provides a traditional Chinese medicine compound for treating gouty arthritis, the traditional Chinese medicine compound being composed of the following traditional Chinese medicinal materials in parts by weight: 5-22 parts of cinnamon twig, 5-20 parts of white peony root, 5-22 parts of anemarrhena rhizome, 5-20 parts of chicory, 5-22 parts of kidney tea, 5-20 parts of sophora flower bud, 5-22 parts of mulberry twig, 5-20 parts of pine resin, 5-20 parts of celery seed, and 5-25 parts of five-finger peach.

[0008] Preferably, the above-mentioned traditional Chinese medicine compound is composed of the following traditional Chinese medicinal materials in parts by weight: 8-18 parts of cinnamon twig, 5-15 parts of white peony root, 8-18 parts of anemarrhena rhizome, 5-15 parts of chicory, 8-18 parts of kidney tea, 5-15 parts of sophora flower bud, 8-18 parts of mulberry twig, 5-15 parts of pine resin, 5-15 parts of celery seed, and 10-20 parts of five-finger peach.

[0009] Preferably, the above-mentioned traditional Chinese medicine compound is composed of the following parts by weight of Chinese medicinal materials: 12 parts of cinnamon twig, 9 parts of white peony root, 12 parts of anemarrhena rhizome, 9 parts of chicory, 12 parts of kidney tea, 9 parts of sophora flower bud, 12 parts of mulberry twig, 9 parts of pine resin, 10 parts of celery seed, and 15 parts of five-finger peach.

[0010] Secondly, the present invention provides the application of the above-mentioned traditional Chinese medicine compound in the preparation of a drug for treating gouty arthritis, wherein the drug is made from the above-mentioned parts by weight of traditional Chinese medicinal materials as raw materials, supplemented with pharmaceutically acceptable excipients, and prepared into any one of pills, powders, granules, oral liquids, ointments, tablets, powders, and decoctions.

[0011] Cinnamon twig: The dried tender twig of cinnamon (Cinnamomum cassia), a plant in the Lauraceae family; pungent and hot in taste; enters the lung, heart, and bladder meridians; has the effects of inducing sweating and relieving exterior syndromes, warming and unblocking the meridians, and warming and assisting yang qi; used for colds due to wind-cold, various pain syndromes caused by cold stagnation and blood stasis, phlegm retention, water retention syndrome, and palpitations.

[0012] White peony root: the dried root of Paeonia lactiflora Pall., a plant in the Ranunculaceae family; bitter and sour in taste, slightly cold in nature, and enters the liver and spleen meridians; it has the effects of nourishing blood and regulating menstruation, astringing yin and stopping sweating, softening the liver and relieving pain, and calming liver yang. It is often used for blood deficiency and chlorosis, irregular menstruation, spontaneous sweating, night sweats, hypochondriac pain, abdominal pain, limb spasms, headache and dizziness.

[0013] Anemarrhena asphodeloides: the dried rhizome of Anemarrhena asphodeloides, a monocotyledonous plant belonging to the Liliaceae family; bitter, cold, and enters the lung, stomach, and kidney meridians; it has the effects of clearing heat and purging fire, nourishing yin and moistening dryness; it is used for symptoms such as fever and thirst, dry cough due to lung heat, bone steaming fever, internal heat and thirst, and constipation due to intestinal dryness.

[0014] Chicory: The dried aerial parts or roots of *Cichori-μm glandulosum* Boiss. et Huet or *Cichorium intybus* L., belonging to the Asteraceae family; slightly bitter and salty in taste, cool in nature; enters the spleen, liver, and bladder meridians; has the effects of clearing the liver and gallbladder, strengthening the stomach and promoting digestion, and promoting diuresis and reducing swelling; used for damp-heat jaundice, stomach pain with poor appetite, edema with scanty urine.

[0015] Clerodendranthus spicatus: A perennial herbaceous plant of the family Lamiaceae and the genus Clerodendranthus spicatus, also known as Orthosiphon aristatus; it is sweet, slightly bitter, and cool in nature. It is used for clearing heat, promoting diuresis, and expelling stones. It is mainly used to treat acute and chronic nephritis, cystitis, urinary tract stones, gallstones, and rheumatoid arthritis.

[0016] Flos Sophorae Immaturus: The dried flower buds of Sophora japonica, a plant of the Leguminosae family; it has the effects of cooling blood and stopping bleeding, clearing the liver and purging fire; it is used for hematochezia, hemorrhoids, dysentery, metrorrhagia and metrostaxis, hematemesis, epistaxis, red eyes due to liver heat, headache and dizziness.

[0017] Ramulus Mori: The dried tender branches of Morus alba L., a plant of the Moraceae family; it is bitter in taste and neutral in nature, and belongs to the Liver Meridian. It has the effects of dispelling wind-dampness and promoting joint movement, and is mainly used to treat rheumatic arthralgia and soreness and numbness of the shoulder and arm joints.

[0018] Oleum Pini Nodi: The dried tuberous nodes or branched nodes of Pinus tabuliformis or other plants of the same genus in the family Pinaceae, which belongs to traditional Chinese medicine for dispelling wind-dampness and promoting blood circulation to remove stasis. Its nature and flavor are bitter, pungent, and warm, and it belongs to the Liver and Kidney Meridians. It has the effects of expelling wind and dampness, dredging collaterals and relieving pain, etc., and is commonly used to treat wind-cold-damp arthralgia, joint pain, traumatic injury and other diseases.

[0019] Celery Seed: The dried mature seeds of Apium graveolens, a plant of the Umbelliferae family. It is flat in nature and sweet in taste, and belongs to the Liver and Stomach Meridians. It has the effects of diuretic and detumescence, suppressing the hyperactive liver-yang, expelling wind and dampness, etc., and is commonly used to regulate edema, hypertension and rheumatic arthralgia.

[0020] Radix Fici Hirta: The roots of Ficus simplicissima Lour., a plant of the Moraceae family; it is sweet in taste and neutral in nature, and enters the Spleen, Lung, and Liver Meridians; it has the effects of strengthening the spleen and tonifying the lung, promoting qi and activating dampness, and relaxing muscles and activating collaterals. It is mainly used to treat spleen deficiency with edema, anorexia and weakness, pulmonary tuberculosis cough, night sweats, leukorrhea, lack of lactation after childbirth, rheumatic arthralgia, edema, liver cirrhosis ascites, hepatitis, and traumatic injury.

[0021] (III) Beneficial effects

[0022] The traditional Chinese medicine compound of the present invention is formulated to clear heat and dredge collaterals, strengthen the spleen and promote diuresis for dampness, and is used for the syndrome of damp-heat obstruction in gouty arthritis. In the formula, chicory and Clerodendranthus spicatus are used as the monarch drugs. Both of them are cold in nature and mainly enter the liver, kidney and bladder meridians. They are good at clearing and promoting damp-heat, diuresing and discharging turbidity, aiming to eliminate and promote the excretion of uric acid from the source, directly hitting the core of the accumulation of gouty turbid toxin. Anemarrhena asphodeloides is used as the ministerial drug, taking its functions of clearing heat and purging fire, nourishing yin and moistening dryness. It can not only enhance the power of clearing damp-heat in the lower jiao, but also prevent the monarch drugs from injuring yin due to diuresis, embodying the meaning of "treating arthralgia and protecting yin". Among the adjuvant drugs, mulberry branches and pine cone joints dispel wind and dampness, relax muscles and activate collaterals to specifically treat joint pain; sophora japonica and celery seeds clear heat and cool blood, and assist in reducing turbidity; cinnamon twig warms and dredges the meridians, prevents cold from stagnating, and also helps to activate collaterals; white peony nourishes the blood and soothes the liver, alleviates pain by relaxing spasms, and combines with cinnamon twig to harmonize nutrient qi and defensive qi. The guiding drug Ficus hirta Vahl can strengthen the spleen and tonify the lung, promote qi and remove dampness. It can not only support healthy qi, strengthen the spleen to cut off the source of damp-turbidity, but also relax muscles and activate collaterals, leading all the drugs to reach the extremities. The whole formula combines cold and warm herbs, combines attacking and tonifying, takes into account both the root and the branch, and together achieves the effects of clearing heat and promoting diuresis, dredging collaterals and relieving pain, strengthening the spleen and discharging turbidity.

[0023] The present invention successfully constructed a cell model of gouty arthritis using MSU, and evaluated the efficacy of the traditional Chinese medicine compound of the present invention in the treatment of gouty arthritis. The experimental results showed that the traditional Chinese medicine compound of the present invention reduced the expression levels of inflammatory mediators in joint tissues, improved the pathological conditions of joint tissues, achieved better reduction of the joint swelling degree in GA model rats, improved their joint dysfunction, and had a significant therapeutic effect on gouty arthritis rats. The traditional Chinese medicine compound of the present invention is expected to be further studied and applied as a new drug for the treatment of gouty arthritis. Description of the Drawings

[0024] Figure 1 Results of the tests on the contents of inflammatory mediators IL-β and TNF-α and cell viability in the supernatant of cells in each group; note, ****P<0.0001.

[0025] Figure 2 Results of the scores of joint swelling degree and joint disorder index in each group of rats after successful modeling and at 6h, 12h, 24h, 48h, 72h after administration; note, at the same measurement time point, compared with the blank group, **P<0.01; compared with the model group, # P<0.05, ## P<0.01; compared with the low-dose traditional Chinese medicine group, & P<0.05, && P<0.01.

[0026] Figure 3 Pathological results of the ankle joint synovial tissue in each group of rats.

[0027] Figure 4The levels of inflammatory mediators IL-6, TNF-α, and IL-β in the synovial tissue of the ankle joint of rats in each group were measured. Note: Compared with the control group, **P<0.01; compared with the model group, ## P<0.01. Detailed Implementation

[0028] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions of the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0029] Example 1

[0030] A traditional Chinese medicine compound for treating gouty arthritis, the compound consisting of the following medicinal materials in parts by weight: 5 parts cinnamon twig, 5 parts white peony root, 5 parts anemarrhena rhizome, 5 parts chicory, 5 parts kidney tea, 5 parts sophora flower bud, 5 parts mulberry twig, 5 parts pine resin, 5 parts celery seed, and 5 parts five-finger peach.

[0031] The above-mentioned traditional Chinese medicine compound is used to prepare a drug for treating gouty arthritis. The drug is made from the above-mentioned proportions of traditional Chinese medicinal materials by weight, supplemented with pharmaceutically acceptable excipients, and prepared into any one of the following: pills, powders, granules, oral liquids, ointments, tablets, powders, and decoctions.

[0032] Example 2

[0033] The difference between this embodiment and Embodiment 1 is that the traditional Chinese medicine compound for treating gouty arthritis is composed of the following parts by weight of Chinese medicinal materials: 22 parts of cinnamon twig, 20 parts of white peony root, 22 parts of anemarrhena rhizome, 20 parts of chicory, 22 parts of kidney tea, 20 parts of sophora flower bud, 22 parts of mulberry twig, 20 parts of pine resin, 20 parts of celery seed, and 25 parts of five-finger peach.

[0034] Example 3

[0035] The difference between this embodiment and Embodiment 1 is that the traditional Chinese medicine compound for treating gouty arthritis is composed of the following parts by weight of Chinese medicinal materials: 20 parts of cinnamon twig, 16 parts of white peony root, 20 parts of anemarrhena rhizome, 16 parts of chicory, 20 parts of kidney tea, 18 parts of sophora flower bud, 20 parts of mulberry twig, 16 parts of pine resin, 16 parts of celery seed, and 22 parts of five-finger peach.

[0036] Example 4

[0037] The difference between this embodiment and Embodiment 1 is that the traditional Chinese medicine compound for treating gouty arthritis is composed of the following parts by weight of Chinese medicinal materials: 7 parts of cinnamon twig, 5 parts of white peony root, 7 parts of anemarrhena rhizome, 6 parts of chicory, 6 parts of kidney tea, 5 parts of sophora flower bud, 6 parts of mulberry twig, 5 parts of pine resin, 6 parts of celery seed, and 9 parts of five-finger peach.

[0038] Example 5

[0039] The difference between this embodiment and Embodiment 1 is that the traditional Chinese medicine compound for treating gouty arthritis is composed of the following parts by weight of Chinese medicinal materials: 8 parts of cinnamon twig, 5 parts of white peony root, 8 parts of anemarrhena rhizome, 5 parts of chicory, 8 parts of kidney tea, 5 parts of sophora flower bud, 8 parts of mulberry twig, 5 parts of pine resin, 5 parts of celery seed, and 10 parts of five-finger peach.

[0040] Example 6

[0041] The difference between this embodiment and Embodiment 1 is that the traditional Chinese medicine compound for treating gouty arthritis is composed of the following parts by weight of Chinese medicinal materials: 18 parts of cinnamon twig, 15 parts of white peony root, 18 parts of anemarrhena rhizome, 15 parts of chicory, 18 parts of kidney tea, 15 parts of sophora flower bud, 18 parts of mulberry twig, 15 parts of pine resin, 15 parts of celery seed, and 20 parts of five-finger peach.

[0042] Example 7

[0043] The difference between this embodiment and Embodiment 1 is that the traditional Chinese medicine compound for treating gouty arthritis is composed of the following parts by weight of Chinese medicinal materials: 10 parts of cinnamon twig, 8 parts of white peony root, 12 parts of anemarrhena rhizome, 8 parts of chicory, 14 parts of kidney tea, 10 parts of sophora flower bud, 14 parts of mulberry twig, 9 parts of pine resin, 12 parts of celery seed, and 14 parts of five-finger peach.

[0044] Example 8

[0045] The difference between this embodiment and Embodiment 1 is that the traditional Chinese medicine compound for treating gouty arthritis is composed of the following parts by weight of Chinese medicinal materials: 14 parts of cinnamon twig, 12 parts of white peony root, 16 parts of anemarrhena rhizome, 10 parts of chicory, 14 parts of kidney tea, 10 parts of sophora flower bud, 16 parts of mulberry twig, 12 parts of pine resin, 12 parts of celery seed, and 18 parts of five-finger peach.

[0046] Example 9

[0047] The difference between this embodiment and Embodiment 1 is that the traditional Chinese medicine compound for treating gouty arthritis is composed of the following parts by weight of Chinese medicinal materials: 12 parts of cinnamon twig, 9 parts of white peony root, 12 parts of anemarrhena rhizome, 9 parts of chicory, 12 parts of kidney tea, 9 parts of sophora flower bud, 12 parts of mulberry twig, 9 parts of pine resin, 10 parts of celery seed, and 15 parts of five-finger peach.

[0048] Experimental Example 1

[0049] The formulation screening of traditional Chinese medicine compound of this invention

[0050] 1 Experimental Drugs

[0051] Drug 1: Cinnamon twig 12 parts, white peony root 9 parts, anemarrhena rhizome 12 parts, chicory 9 parts, kidney tea 12 parts, sophora flower bud 9 parts, mulberry twig 12 parts, pine resin 9 parts, celery seed 10 parts, five-finger peach 15 parts

[0052] Drug 2: Cinnamon twig 12 parts, white peony root 9 parts, anemarrhena rhizome 12 parts, sophora flower bud 9 parts, mulberry twig 12 parts, pine resin 9 parts, celery seed 10 parts

[0053] Drug 3: Cinnamon twig 12 parts, Anemarrhena rhizome 12 parts, Chicory 9 parts, Kidney tea 12 parts, Celery seed 10 parts, Five-finger peach 15 parts

[0054] Drug 4: White peony root 9 parts, chicory 9 parts, kidney tea 12 parts, sophora japonica flower bud 9 parts, mulberry twig 12 parts, pine resin 9 parts, five-finger peach 15 parts

[0055] Drug 5: Cinnamon twig 9 parts, white peony root 9 parts, anemarrhena rhizome 12 parts, ephedra 6 parts, prepared aconite root 6 parts, atractylodes rhizome 9 parts, saposhnikovia root 9 parts, fresh ginger 15 parts, licorice root 6 parts (classic formula: Cinnamon Twig, Peony Root and Anemarrhena Rhizome Decoction).

[0056] For each 1g portion, weigh the required Chinese medicinal materials for drugs 1-5 according to the above-mentioned weight proportions. After washing, soak them in 10 times the amount of 75% ethanol for 6 hours, heat to 60℃ and sonicate for 3 hours. After filtration, obtain filtrate 1. Then, continue to soak the residue in 8 times the amount of 75% ethanol at 60℃ and sonicate for 2 hours, filter, and repeat twice to obtain filtrate 2 and filtrate 3. Combine the three filtrates and concentrate under reduced pressure. The concentrate is then freeze-dried under vacuum to obtain drugs 1-5. Each drug should be dissolved in dimethyl sulfoxide and diluted with culture medium to a suitable drug concentration before use, ensuring that the DMSO content in the culture medium is less than 0.1% at the time of administration.

[0057] 2. Establishment and grouping of cell models for gouty arthritis

[0058] Human monocytes (THP-1) were revived and passaged in DMEM medium containing 1% penicillin-streptomycin and 10% fetal bovine serum, and then cultured at 37°C in a 5% CO2 incubator. Cells in the logarithmic growth phase were harvested and cultured at 10⁻⁶ cells per ... 5 Cells were seeded at a density of 100 cells / mL in 96-well plates and divided into 7 groups: normal group (NC group), model group (M group), and experimental groups 1-5 (S1-S5 groups). The normal group was given serum-free DMEM medium, the model group was given serum-free DMEN medium containing 0.05 μg / mL monosodium urate (MSU), and the experimental groups 1-5 were given serum-free DMEN medium containing 0.05 μg / mL MSU + 50 μg / mL drug 1-5. Each group was set up with 6 replicates, and the cells in each group were treated as described above for 24 h.

[0059] 3. ELISA detection of inflammatory mediator expression in cell supernatant

[0060] After THP-1 cells were grouped and treated, the cell culture supernatant of each group was collected in sterile tubes, centrifuged at 300×g for 20 min, and the supernatant was collected. The IL-β and TNF-α in the cell supernatant were measured according to the ELISA kit instructions.

[0061] 4. Cell viability assay

[0062] Human monocytes (THP-1) were revived and passaged in DMEM medium containing 1% penicillin-streptomycin and 10% fetal bovine serum, and then cultured at 37°C in a 5% CO2 incubator. Cells in the logarithmic growth phase were harvested and cultured at 10⁻⁶ cells per ... 5 Cells were seeded at a density of [number] cells / mL in 96-well plates and divided into a control group and experimental groups 1-5. Cells in experimental groups 1-5 were treated with serum-free DMEN medium containing 50 μg / mL of the corresponding drug. Each group had six replicates, and cells were cultured overnight as described above. After overnight culture, MTT solution was added to each well for 3-4 hours of incubation, followed by removal of the supernatant. Cell viability was calculated by measuring absorbance (A) at 490 nm. Cell viability = (Adrug group / Acontrol group) × 100%.

[0063] 5 Statistical Methods

[0064] Data were processed and analyzed using Prism 8.0.2 software. Data are expressed as mean ± standard deviation. Independent samples t-test was used for comparisons between groups, and one-way ANOVA was used for comparisons among multiple groups. P < 0.05 was considered statistically significant.

[0065] 6 Results

[0066] This experiment used MSU to construct a GA cell model and evaluated the application potential of different traditional Chinese medicine compound formulas in the treatment of GA by detecting the expression of inflammatory mediators IL-β and TNF-α. The results are as follows: Figure 1 As shown, compared with the blank group, the levels of inflammatory mediators IL-β and TNF-α in the cell supernatant of the model group were significantly increased (P<0.0001), indicating that the GA cell model was successfully constructed. Compared with the model group, the levels of inflammatory mediators IL-β and TNF-α in the cell supernatant of each treatment group were significantly decreased (P<0.0001); among them, the other herbal compound drugs (S2-S5) were more effective in significantly reducing the levels of inflammatory mediators IL-β and TNF-α in the cell supernatant of the GA model compared with the herbal compound (S1) of this invention (P<0.0001). The above results indicate that the various components of the herbal compound of this invention complement each other, synergistically enhance each other, and jointly achieve the best technical effect of inhibiting the expression of inflammatory mediators in GA model cells.

[0067] Experimental Example 2

[0068] Evaluation of the efficacy of the traditional Chinese medicine compound of this invention on rats with acute gouty arthritis.

[0069] 1. Materials and Methods

[0070] 1.1 Laboratory Animals

[0071] Sixty SPF-grade male SD rats, weighing 200-220g, were selected and acclimatized for one week in a standard animal laboratory with an ambient temperature of 20-25℃, relative humidity of 45%-55%, and alternating light and dark for 12 hours before the experiment.

[0072] 1.2 Experimental Drugs

[0073] The compound extract of traditional Chinese medicine in this invention consists of: 12 parts cinnamon twig, 9 parts white peony root, 12 parts anemarrhena rhizome, 9 parts chicory, 12 parts kidney tea, 9 parts sophora flower bud, 12 parts mulberry twig, 9 parts pine resin, 10 parts celery seed, and 15 parts five-finger peach.

[0074] Control drug 1: Cinnamon twig 9 parts, white peony root 9 parts, anemarrhena rhizome 12 parts, ephedra 6 parts, prepared aconite root 6 parts, atractylodes rhizome 9 parts, saposhnikovia root 9 parts, fresh ginger 15 parts, licorice root 6 parts.

[0075] For each part by weight of 1g, the required Chinese medicinal materials for the compound Chinese medicine of the present invention and the control drug 1 are weighed according to the above-mentioned parts by weight. After washing, they are soaked in 10 times the amount of 75% ethanol for 6 hours, heated to 60°C and sonicated for 3 hours. After filtration, filtrate 1 is obtained. Then, the residue is soaked in 8 times the amount of 75% ethanol at 60°C and sonicated for 2 hours. After filtration, the process is repeated twice to obtain filtrate 2 and filtrate 3. The three filtrates are combined and concentrated under reduced pressure. The concentrate is then freeze-dried under vacuum to obtain the extract of the compound Chinese medicine of the present invention and the control drug 1.

[0076] Comparative drug 2: Etoricoxib tablets (National Drug Approval Number HJ20181027).

[0077] 1.3 Animal Model Construction and Grouping

[0078] Rats were randomly divided into four groups: a blank control group (NC group), a model group (M group), a low-dose traditional Chinese medicine group (Low group, 5 g / kg of the compound extract of the traditional Chinese medicine of this invention), a high-dose traditional Chinese medicine group (High group, 10 g / kg of the compound extract of the traditional Chinese medicine of this invention), a control group 1 (DZ-1 group, 5 g / kg of control drug 1), and a control group 2 (DZ-2 group, etoricoxib 5.4 mg / kg), with 10 rats in each group. Except for the blank control group, the rats in the other groups were intraperitoneally injected with 3% potassium oxonate solution at a dose of 1 mL / 100 g twice daily for one week. Then, 30 μL of 50 mg / mL MSU suspension (prepared with 0.9% sodium chloride solution) was injected into the medial Achilles tendon joint cavity of the right ankle of the rats using an injection needle. The rats in the blank control group were injected intraperitoneally and into the right ankle joint cavity with an equal volume of 0.9% sodium chloride solution. After successful modeling, the drug administration groups were given the corresponding drugs by gavage, while the model group and the blank control group were given an equal volume of distilled water. Each group was administered the drugs continuously for 3 days.

[0079] 1.4 Observation Indicators

[0080] 1.4.1 Measurement of bare joint swelling in rats

[0081] The diameter of the right ankle joint of rats was measured with vernier calipers before modeling, after successful modeling (recorded as 0h), and at 6h, 12h, 24h, 48h, and 72h after drug administration, and the measurements were marked. The ankle joint swelling degree was calculated according to the formula: Right ankle joint swelling degree (%) = (Ankle joint diameter of rats at each time point after modeling - Ankle joint diameter of rats before modeling) / Ankle joint diameter of rats before modeling × 100%.

[0082] 1.4.2 Joint Dysfunction Index

[0083] After successful gouty arthritis modeling, and at 6h, 12h, 24h, and 48h after drug administration, the joint dysfunction index scores were observed and recorded. The scoring criteria were as follows: 0 points: normal walking with both feet evenly touching the ground; 1 point: feet lightly touching the ground with mild limping; 2 points: feet flexed with toes touching the ground with obvious limping; 3 points: feet completely off the ground with a tripedal gait.

[0084] 1.4.2 Histopathological observation of rat ankle joint synovial tissue

[0085] After the last administration, anesthesia was administered via injection of 1% sodium pentobarbital. Synovial tissue from the ankle joint was harvested and fixed by immersion in 0.4% paraformaldehyde. Following dehydration, paraffin embedding, and sectioning, pathological sections were prepared, stained with hematoxylin and eosin (HE), and observed under an optical microscope for histopathological morphological changes.

[0086] 1.4.3 ELISA measurement of inflammatory factor levels in rat ankle synovial tissue

[0087] After the last administration, the animals were anesthetized by injection of 1% sodium pentobarbital. The synovial tissue of the ankle joint was then collected and tested according to the instructions of the ELISA kit to determine the levels of IL-6, TNF-α, and IL-β in the synovial tissue of the ankle joint of each group of animals.

[0088] 1.5 Statistical Analysis

[0089] Data were processed and analyzed using Prism 8.0.2 software. Data are expressed as mean ± standard deviation. Independent samples t-test was used for comparisons between groups, and one-way ANOVA was used for comparisons among multiple groups. P < 0.05 was considered statistically significant.

[0090] 2 Results

[0091] 2.1 Degree of joint swelling

[0092] At 0 h, compared with the control group, the joint swelling of rats in each model group was significantly increased (P<0.01), indicating that the rat model of gouty arthritis was successfully established. At 6 h, 12 h, 24 h, 48 h, and 72 h after administration, compared with the model group, the joint swelling of rats in each administration group was significantly reduced (P<0.01). Among them, at 24 h, 48 h, and 72 h after administration, the joint swelling of rats in the low-dose traditional Chinese medicine group was significantly lower than that in control group 1 (P<0.05, P<0.01), while there was no statistically significant difference in joint swelling between the low-dose traditional Chinese medicine group and control group 2 (P>0.05). At each time point, the degree of joint swelling of rats in the high-dose traditional Chinese medicine group was significantly lower than that in the low-dose traditional Chinese medicine group and control groups 1-2. The above results indicate that the traditional Chinese medicine compound of the present invention can effectively reduce the degree of joint swelling in rats with gouty arthritis, and its effect is comparable to that of commonly used Western medicines, and is significantly better than Guizhi Shaoyao Zhimu Decoction, and has a certain dose-effect relationship. Figure 2 As shown.

[0093] 2.2 Joint dysfunction score

[0094] At 0 h, compared with the control group, the joint dysfunction scores of rats in each model group were significantly increased (P<0.01), indicating that the rat model of gouty arthritis was successfully established. At 6 h, 12 h, 24 h, 48 h, and 72 h after drug administration, compared with the model group, the joint dysfunction scores of rats in each drug administration group were significantly reduced (P<0.05, P<0.01). Among them, at each time point, the joint dysfunction scores of rats in the low-dose traditional Chinese medicine group were significantly lower than those in control group 1 and comparable to those in control group 2, while the joint dysfunction scores of rats in the high-dose traditional Chinese medicine group were significantly lower than those in the low-dose traditional Chinese medicine group and control groups 1-2. The above results indicate that the traditional Chinese medicine compound of this invention can effectively improve the degree of joint dysfunction in rats with gouty arthritis, and its effect is comparable to that of commonly used Western medicines, and significantly better than Guizhi Shaoyao Zhimu Decoction, and has a certain dose-effect relationship.Figure 2 As shown.

[0095] 2.3 Histopathological observation of ankle joint synovial tissue and expression of inflammatory mediators

[0096] The pathological conditions of the ankle joint synovial tissue in each group of rats are as follows: Figure 3 As shown, the synovial tissue structure in the control group was clear, with no inflammatory cell infiltration or synovial hyperplasia; compared with the control group, the synovial tissue structure in the model group was disordered, with a large number of inflammatory cells infiltrating. Compared with the model group, the synovial tissue structure of rats in each drug-treated group was significantly improved, and the inflammatory cell infiltration was reduced to varying degrees.

[0097] Compared with the control group, the levels of inflammatory mediators IL-6, TNF-α, and IL-β in the synovial tissue of the ankle joint of rats in the model group were significantly increased (P<0.01). Compared with the model group, the levels of inflammatory mediators IL-6, TNF-α, and IL-β in the synovial tissue of the ankle joint of rats in each treatment group were significantly decreased (P<0.01). Among them, the levels of inflammatory mediators IL-6, TNF-α, and IL-β in the synovial tissue of the ankle joint of rats in the low-dose traditional Chinese medicine group were significantly lower than those in control group 1 and comparable to those in control group 2. The levels of inflammatory mediators IL-6, TNF-α, and IL-β in the synovial tissue of the ankle joint of rats in the high-dose traditional Chinese medicine group were significantly lower than those in the low-dose traditional Chinese medicine group and control groups 1-2. These results indicate that the traditional Chinese medicine compound of this invention can effectively reduce the expression level of inflammatory mediators in the synovial tissue of the ankle joint of rats with gouty arthritis. Its effect is comparable to that of commonly used Western medicines and is significantly better than that of Guizhi Shaoyao Zhimu Decoction, and it has a certain dose-effect relationship. Figure 4 As shown.

[0098] 3. Conclusion

[0099] This experiment successfully established a rat model of gouty arthritis using MSU (methylformamide). The efficacy of the traditional Chinese medicine compound of this invention in treating gouty arthritis was evaluated. The results show that the traditional Chinese medicine compound of this invention significantly reduces joint swelling and improves joint dysfunction in GA model rats by lowering the expression levels of inflammatory mediators in joint tissues and improving the pathological condition of joint tissues. This demonstrates a significant therapeutic effect on gouty arthritis in rats. The traditional Chinese medicine compound of this invention holds promise for further research and application as a novel drug for treating gouty arthritis.

[0100] The above embodiments are only used to illustrate the technical solutions of the present invention, and are not intended to limit it. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A traditional Chinese medicine compound for treating gouty arthritis, characterized in that, The traditional Chinese medicine compound is made from the following medicinal materials in parts by weight: 5-22 parts cinnamon twig, 5-20 parts white peony root, 5-22 parts anemarrhena rhizome, 5-20 parts chicory, 5-22 parts kidney tea, 5-20 parts sophora flower bud, 5-22 parts mulberry twig, 5-20 parts pine resin, 5-20 parts celery seed, and 5-25 parts five-finger peach. Weigh each medicinal material according to the above parts by weight, wash them, soak them in 10 times the amount of 75% ethanol for 6 hours, heat to 60℃ and sonicate for 3 hours, filter to obtain filtrate 1, then continue to soak the residue in 8 times the amount of 75% ethanol at 60℃ and sonicate for 2 hours, filter, repeat twice to obtain filtrate 2 and filtrate 3, combine the three filtrates and concentrate under reduced pressure, freeze dry the concentrate under vacuum to obtain the traditional Chinese medicine compound extract for the treatment of gouty arthritis.

2. The traditional Chinese medicine compound for treating gouty arthritis as described in claim 1, characterized in that, The herbal compound is made from the following parts by weight of Chinese medicinal materials: 8-18 parts of cinnamon twig, 5-15 parts of white peony root, 8-18 parts of anemarrhena rhizome, 5-15 parts of chicory, 8-18 parts of kidney tea, 5-15 parts of sophora flower bud, 8-18 parts of mulberry twig, 5-15 parts of pine resin, 5-15 parts of celery seed, and 10-20 parts of five-finger peach.

3. A traditional Chinese medicine compound for treating gouty arthritis as described in claim 1, characterized in that, The traditional Chinese medicine compound is made from the following Chinese medicinal materials in parts by weight: 12 parts cinnamon twig, 9 parts white peony root, 12 parts anemarrhena rhizome, 9 parts chicory, 12 parts kidney tea, 9 parts sophora flower bud, 12 parts mulberry twig, 9 parts pine resin, 10 parts celery seed, and 15 parts five-finger peach.

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