A Chinese medicine composition for treating male gouty arthritis and its preparation method and application

Abstract

The present invention belongs to the technical field of Chinese medicine research, and specifically relates to a Chinese medicine composition for treating male gouty arthritis, and a preparation method and application thereof. The Chinese medicine composition of the present invention is composed of the following raw materials in parts by weight: 10-12 parts by weight of holly leaves, 3-5 parts by weight of cicada flower, 5-7.5 parts by weight of Brassica rapa seeds, 0.04 parts by weight of Artemisia selengensis, 20-23 parts by weight of Euonymus fortunei, and 5-6 parts by weight of Sauerkraut seeds. Studies have shown that the Chinese medicine composition of the present invention can improve the degree of joint swelling in the toes of male gouty arthritis Kunming mice, including the degree of joint swelling in the toes of Kunming mice, the inflammation index of the toes of Kunming mice, the antioxidant capacity of Kunming mice, the content of inflammatory mediators in Kunming mice, the main protein and gene expression level in the toes of Kunming mice, and has no effect on the organ index and body weight of the important organs of the male gouty arthritis Kunming mice, indicating that the Chinese medicine composition is effective in treating male gouty arthritis, and has no side effects.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicine research, and particularly relates to a traditional Chinese medicine composition for treating male gouty arthritis, and a preparation method and application thereof. Background Art

[0002] Gout is a common disease that can occur in both men and women (Xu H, Zhang B, Chen Y, et al. Type II collagen facilitates gouty arthritis by regulating MSUcrystallisation and inflammatory cell recruitment[J]. Ann Rheum Dis. 2023Mar;82(3):416-427.). With the increase of social pressure and the existence of bad living habits in men, the number of men suffering from gouty arthritis has increased year by year, which will bring a heavy economic burden to families and society (Abdelnabi M, LeelavawatN, Liao ED, et al. Daptomycin-induced rhabdomyolysis complicated with acutegouty arthritis[J]. Am J Med Sci. 2023 May;365(5):450-456.).

[0003] Men often have bad habits such as smoking, drinking, and irregular living. Combined with mental, social, genetic, and cultural factors, these factors often cause abnormal nucleic acid base metabolism in the body, including abnormal decomposition of guanine and adenine (Yang DH, Chen HC, et al. Early urate-lowering therapy in gouty arthritis with acute flares a double-blind placebo controlled clinical trial[J]. Eur J Med Res. 2023 Jan 6; 28(1): 10.), resulting in many abnormal deposits appearing in places where blood flow is not strong, such as the knee joints, elbow joints, ankle joints, wrist joints, etc., where a large amount of uric acid appears. However, uric acid cannot be transported away quickly with the blood circulation, causing uric acid accumulation (Chen Z, Wang W, Hua Y. Metabolic Regulation of Immune Response and Tissue Remodeling in Gouty Arthritis Review[J]. Crit Rev Eukaryot Gene Expr. 2023; 33(5): 1-16.).

[0004] Studies have shown that humans differ from other animals not only in intellectual development, but also in the lack of certain nucleic acid metabolite processing enzymes. For example, humans cannot metabolize uric acid, which is related to the lack of uricase in humans (Wang Q, Qiu H. Deubiquitinase USP16 induces gouty arthritis via Drp1 dependent mitochondrial fission and NLRP3 inflammasome activation[J]. Arthritis ResTher. 2023 Jul 24; 25(1): 126.). Uricase can metabolize uric acid, the decomposition product of guanine and adenine. Once uricase is lacking, uric acid will accumulate in the body's knee joints, elbow joints, ankle joints, wrist joints, etc., inducing joint pain in the body. This is related to the accumulation of uric acid in the body's knee joints, elbow joints, ankle joints, wrist joints, etc. (Wu Q, Fu C, Lu Z. The risk of myocardial infarction and heart failure in patients with goutyarthritis A systematic review and meta-analysis[J]. Int J Rheum Dis. 2023Mar; 26(3): 415-424.), and effectively reducing the accumulation of uric acid in the body's knee joints, elbow joints, ankle joints, wrist joints, etc., is effective for the treatment of gouty arthritis.

[0005] After a lot of uric acid accumulates in the body's large and small joints (Fu W, Ge M, Li J. Phospholipase A2regulates autophagy in gouty arthritis proteomic and metabolomic studies[J]. J Transl Med. 2023 Apr 17; 21(1): 261.), it often exists in the form of urate in the body's knee joints, elbow joints, ankle joints, wrist joints, etc., causing inflammatory reactions in these joints, including heat, swelling, pain, and redness. In severe cases, deformities such as the knee joints, elbow joints, ankle joints, and wrist joints may also occur, and the patient can only travel with the help of a wheelchair or crutches (Zhu R, Niu Y, Zhou W, et al. Effect of nanoparticles on gouty arthritis a systematic review and meta-analysis[J]. BMC Musculoskelet Disord. 2023 Feb 14; 24(1): 124.).

[0006] In more serious cases, urate is deposited in the kidneys, accompanied by abnormalities in the structure of the renal cortex and medulla, a decrease in the number of glomeruli, tubular obstruction, and tubular degeneration and necrosis (Ge X, Zhang Y, Fang R, et al. Exploring the inhibition mechanism of interleukin-1-beta in gouty arthritis by polygonumcuspidatum using network pharmacology and molecular docking A review[J]. Medicine Baltimore. 2023 Jul 21;102(29):e34396.). The kidneys are unable to perform the functions of concentrating and diluting urine, and the kidneys are also affected in secreting some related hormones and affecting the level of red blood cell metabolism (Li W, Feng H.Lumbarspondylolisthesis associated with gouty arthritis A case report[J]. Asian JSurg. 2023 Jun;46(6):2372-2373.), causing gouty nephropathy and renal failure.

[0007] At present, men often have bad habits such as smoking, drinking, and irregular lifestyle, combined with mental factors, social factors, genetic factors, etc. (Botson JK, Saag K, Peterson J, et al. A Randomized, Placebo-Controlled Study of Methotrexate to Increase Response Rates in Patients with Uncontrolled Gout Receiving Pegloticase Primary Efficacy and Safety Findings[J]. Arthritis Rheumatol. 2023 Feb; 75 (2): 293-304.), which causes uric acid accumulation in the body's knee, elbow, ankle, wrist joints, etc., often inducing gouty arthritis. Therefore, men need to actively change their bad habits of smoking, drinking, and irregular lifestyle (Qin DE, Liang W, Yu Y, et al. Modified Simiaowan prevents and treats gouty arthritis via the Nrf2 / NLRP3 inflammasome signaling pathway[J]. J Ethnopharmacol. 2023 Jul 11;318(Pt A):116906.).

[0008] Studies have shown that eating a lot of fat, coupled with smoking, drinking, and irregular lifestyle, often leads to enzyme activation (Zheng W, Lu P, Jiang D, et al. An ultrasonographic study of gouty arthritis: Synovitis and its relationship to clinical symptoms A retrospective analysis [J]. Health Sci Rep. 2023 Jun 7; 6 (6): e1312.), such as the activation of enzymes related to fatty acid metabolism, abnormal mitochondrial function in cells, and the production of many oxygen free radicals in the body's cells. Oxygen free radicals are a type of factor that promotes the activation of many proteins in the body and can aggravate many diseases (Vaprath Kuniyil AR, Soman D, Kundagol MC, et al. Efficacy of Ayurvedic treatment protocol in gouty arthritis-a clinical study [J]. J Complement Integr Med. 2021 May 11; 20 (1): 278-283.).

[0009] Normally, the metabolism of oxygen free radicals in the body is in a dynamic balance, which can be metabolized by superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX) (Lafforgue A, Lambert C, Dubost JJ, et al. Performance of a diagnostic score for gouty arthritis results from acohort of acute arthritis suspected of being septic[J]. Rheumatol Int. 2023Jan; 43(1): 119-124.), generating malondialdehyde (MDA), which can also activate downstream proteins such as NOD-like receptor protein 3 (NLRP3) (Wang Y. Retraction Note Tripterine ameliorates monosodium uratecrystal-induced gouty arthritis by altering macrophage polarization via themiR-449a / NLRP3 axis[J]. Inflamm Res. 2023 Jan; 72 (1): 3.), promoting the activation of inflammatory pathways and producing a large number of inflammatory mediators, such as the inflammatory mediator interleukin-1β (IL-1β) and the inflammatory mediator tumor necrosis factor-α (TNF-α). These inflammatory mediators interleukin-1β (IL-1β) and inflammatory mediator tumor necrosis factor-α (TNF-α) will promote the aggregation of neutrophils, macrophages, etc. (Zhou Q, Liu L, Sun H, et al. Relief of gouty arthritis in rats by totalsaponins from Dioscorea nipponica Makino through suppression of neutrophilextracellular trap formation via the PI3K / AKT / mTOR axis[J]. Exp Ther Med. 2023Aug 2; 26 (3): 447.), promoting the accumulation of uric acid in the body's knee joints, elbow joints, ankle joints, wrist joints, etc., causing aseptic inflammatory response, and the knee joints, elbow joints, ankle joints, wrist joints, etc. will become hot, swollen, painful, and red (Lien MC, Yeh IH, Lu YC, et al.Plasmonic nanomaterials-based flexible strips for the SERS detection of gout arthritis[J]. Analyst. 2023 Aug 21;148(17):4109-4115.), in severe cases, deformities of the knee, elbow, ankle, wrist and other joints may occur, and further damage to the surrounding tissues of the body may be caused (Kinoshita T,Hashimoto Y,Okano T,et al.Arthroscopic debridement for gouty arthritis of the knee caused by anorexia nervosa A case report[J].J OrthopSci. 2023 Jan;28(1):286-289.), promoting the occurrence and development of gouty arthritis. . Summary of the invention

[0010] The purpose of the present invention is to solve the deficiencies of the prior art and provide a Chinese medicine composition for treating male gouty arthritis and a preparation method and application thereof, specifically adopting the following technical scheme:

[0011] In a first aspect of the present invention, a Chinese medicine composition for treating male gouty arthritis is provided, wherein the raw materials and dosages of the Chinese medicine composition include: 10-12 parts by weight of holly leaves, 3-5 parts by weight of cicada flowers, 5-7.5 parts by weight of Brassica rapa seeds, 0.04 parts by weight of Artemisia suspensa, 20-23 parts by weight of Euonymus fortunei, and 5-6 parts by weight of Sago suspensa seeds. More preferably, the raw materials and dosages of the Chinese medicine composition include: 12 parts by weight of holly leaves, 4.5 parts by weight of cicada flowers, 7.5 parts by weight of Brassica rapa seeds, 0.04 parts by weight of Artemisia suspensa, 22.5 parts by weight of Euonymus fortunei, and 6 parts by weight of Sago suspensa seeds.

[0012] The Chinese medicine composition provided by the present invention comprises six kinds of medicinal materials, among which holly leaves, also known as holly thorn, Chinese magnolia leaf, star anise thorn, are cool in nature, bitter in taste, belong to the main meridians of the liver meridian and kidney meridian of the body, and have the core functions of calming the liver, nourishing yin, benefiting the kidney and clearing away heat; cicada fungus, also known as cicada fungus, cicada pupa grass, or insect flower, etc., are oblong in shape, cold in nature, sweet in taste, belong to the main meridians of the liver meridian of the body, and have the core functions of calming the liver, improving eyesight, removing cataracts and dispelling wind, clearing away rashes and clearing away heat, and calming the nerves and suppressing yang; Brassica rapa seeds, also known as brassica seeds, brassica seeds, rapeseed, etc., are cultivated plants, the seeds are nearly spherical, flat in nature, pungent and sweet in taste, belong to the main meridians of the large intestine meridian and liver meridian of the body, and have the core functions of promoting blood circulation and moistening the intestines, removing blood stasis and dispersing nodules, and relieving constipation and reducing swelling; Artemisia scoparia, also known as Artemisia scoparia, Iron Ox Seven, Iron Hammer, Artemisia selengensis, etc. are perennial herbs, mostly growing on the edge of the forest or in the grass. They are warm in nature, bitter and pungent in taste, and belong to the main meridians of the kidney and liver meridians of the body, with the core functions of promoting blood circulation and removing dampness, relieving pain and dispelling wind, and anti-inflammatory and healing; Fufang Teng, also known as Kaoqiangfeng, Qianjinteng, Huangujin, Climbing Tiger, Yanqingteng, etc., are slightly warm in nature, accompanied by a slightly bitter taste. They belong to the main meridians of the kidney, spleen, and liver meridians of the body, with the core functions of dispelling wind and relaxing muscles, invigorating qi and nourishing the liver, and activating collaterals and nourishing the kidney; Salacia chinensis, also known as Xianli, Pistachio, Solo Fruit, Monkey Chestnut, etc., is a deciduous tree, which can be grown wild or cultivated. It is warm in nature, sweet in taste, and belongs to the main meridians of the stomach and liver meridians of the body, with the core functions of promoting qi and stomach, relieving pain and relieving fullness.

[0013] Men often have abnormal liver qi distribution, which causes wind-toxin accumulation, and wind-toxin accumulation leads to kidney qi deficiency and abnormal metabolism of water and grain essence, accompanied by organ dysfunction. Wind-toxin accumulation will invade blood vessels and joints, causing pain, red face and eyes, and rapid heartbeat when encountering things. Wind-toxin accumulation is prone to occur in people with liver yang hyperactivity syndrome, which often causes palpitations, forgetfulness, dizziness and tinnitus. These symptoms are mostly related to wind-toxin accumulation induced by liver yang hyperactivity syndrome, so it is necessary to dispel wind for men with gouty arthritis.

[0014] When the liver qi is hyperactive, the qi flow of the whole body will be sluggish, the liver's function of storing blood will be abnormal, and the liver cannot effectively play its nourishing role, which will aggravate the symptoms of joint pain. Conversely, smooth qi flow throughout the body will promote the normal function of the liver to store blood. Therefore, qi circulation is necessary for male gouty arthritis.

[0015] In summary, effective treatment of male gouty arthritis in clinical medicine requires calming the liver and activating blood circulation, dispelling wind and promoting qi. The basic treatment principle of this disease focuses on calming the liver and activating blood circulation, dispelling wind and promoting qi, so a Chinese medicine composition that calms the liver and activates blood circulation, dispelling wind and promoting qi can be effectively selected to treat male gouty arthritis.

[0016] In addition, the experimental Chinese medicine composition of the present invention (containing 12.00g of holly leaves, 4.50g of cicada flowers, 7.50g of Brassica rapa seeds, 0.04g of Artemisia scabra, 22.50g of Euonymus fortunei, and 6.00g of Salacia chinensis) has an antioxidant capacity on the toes expressed by SOD and MDA. The study showed that compared with the experimental control group, the SOD content of the toes of Kunming mice (clean grade) in the model control group was significantly reduced, and the MDA was significantly increased, indicating that the model control group had a low ability to effectively scavenge free radicals. After treatment with the Chinese medicine composition (containing 12.00g of holly leaves, 4.50g of cicada fungus, 7.50g of Brassica rapa seeds, 0.04g of Artemisia annua, 22.50g of Euphorbia grandiflora, and 6.00g of Sauvignon ovata), high and medium doses of the Chinese medicine composition (containing 12.00g of holly leaves, 4.50g of cicada fungus, 7.50g of Brassica rapa seeds, 0.04g of Artemisia annua, 22.50g of Euphorbia grandiflora, and 6.00g of Sauvignon ovata) can significantly increase the SOD content in the toes of male gouty arthritis Kunming mice (clean grade), accompanied by a significant reduction in MDA content, indicating that the antioxidant capacity of the high and medium doses of the Chinese medicine composition (containing 12.00g of holly leaves, 4.50g of cicada fungus, 7.50g of Brassica rapa seeds, 0.04g of Artemisia annua, 22.50g of Euphorbia grandiflora, and 6.00g of Sauvignon ovata) is worthy of recognition.

[0017] The levels of IL-1β and TNF-α in the lesions of male gouty arthritis are increased (Qian X, Jiang Y, Luo Y, et al. The Anti-hyperuricemia and Anti-inflammatory Effects of Atractylodes Macrocephala in Hyperuricemia and Gouty Arthritis Rat Models[J]. Comb Chem High Throughput Screen. 2023; 26(5): 950-964.), and the expression of NLRP3 protein and gene is significantly increased (Xue YY, Liu HJ, Sun ZJ, et al. Correction to H19 is involved in the regulation of inflammatory responses in acute gouty arthritis by targeting miR-22-3p[J]. Immunol Res. 2023 Apr; 71(2): 285.). The experimental results of the present invention show that compared with the Kunming mice (clean grade) in the experimental control group, the IL-1β and TNF-α contents in the toes of the male gouty arthritis Kunming mice (clean grade) in the model control group were significantly increased, and the expression of NLRP3 protein and gene was increased, indicating that the aseptic inflammatory response was obvious. After treatment with a high- and medium-dose Chinese herbal composition (containing 12.00g of holly leaves, 4.50g of cicada flowers, 7.50g of Brassica rapa seeds, 0.04g of Artemisia suspensa, 22.50g of Euonymus fortunei, and 6.00g of Sophora japonica), the high- and medium-dose Chinese herbal composition (containing 12.00g of holly leaves, 4.50g of cicada flowers, 7.50g of Brassica rapa seeds, 0.04g of Artemisia suspensa, 22.50g of Sophora japonica, and 6.00g of Sophora japonica) can significantly The IL-1β and TNF-α levels were significantly reduced, accompanied by decreased NLRP3 protein and gene expressions, and aseptic inflammatory response was weakened, indicating that the high and medium doses of the Chinese medicine composition (containing 12.00g of Holly Leaves, 4.50g of Cicadae Concolor, 7.50g of Brassica Fructus, 0.04g of Artemisia Snow onion, 22.50g of Euonymus fortunei, and 6.00g of Sauvignon Blanc) can significantly improve the expression levels of inflammatory mediators and their NLRP3 proteins and genes in the lesions.

[0018] The second aspect of the present invention also provides a traditional Chinese medicine preparation for treating male gouty arthritis, which is prepared by adding the traditional Chinese medicine composition of the present invention to the auxiliary materials required for preparation molding according to the conventional method for preparing pharmaceutical preparations.

[0019] The auxiliary materials include one or more of starch, microcrystalline cellulose, sucrose, lactose, powdered sugar, glucose and vitamin C. The Chinese medicine preparation is a decoction, capsule, pill, granule, tablet or oral solution.

[0020] The third aspect of the present invention also provides a method for preparing the Chinese medicine composition, comprising the following steps:

[0021] (1) Weigh each API according to the weight portion, mix them evenly, add 8-15 times the total weight of the API into distilled water, soak for 2-3 hours, decoct for 2-3 hours, and cool;

[0022] (2) filtering the cooled drug solution in step (1) through gauze to obtain a first filtrate and a filter residue;

[0023] (3) Add the same volume of distilled water as in step (1) to the remaining drug residue in step (2), repeat steps (1) and (2) to obtain a second filtrate, and then combine the first filtrate and the second filtrate and mix them to obtain a water decoction for treating male gouty arthritis.

[0024] Furthermore, the water decoction obtained above is placed in an electric constant temperature air drying oven and dried at 70° C. for 4-5 days to obtain a dry paste of the traditional Chinese medicine composition.

[0025] Furthermore, the dry paste of the traditional Chinese medicine composition obtained above is crushed in a pulverizer, ground, and repeatedly treated with molecular sieves to obtain dry powder of the traditional Chinese medicine composition.

[0026] As a further preferred embodiment, the specific operation of the above-mentioned gauze filtering is to use 2 layers of gauze for filtering, 4 layers of gauze for filtering, 6 layers of gauze for filtering, 8 layers of gauze for filtering, and 10 layers of gauze for filtering in sequence.

[0027] In a fourth aspect of the present invention, the Chinese medicine composition provided by the present invention can be used in the preparation of medicines for treating male gouty arthritis.

[0028] The beneficial effects of the present invention are as follows: the present invention provides a Chinese medicine composition for treating male gouty arthritis, wherein the Chinese medicine composition comprises holly leaves, cicada flower, brassica fruit, Artemisia selengensis, Euonymus fortunei, and Sauerkraut fruit, and has the effects of calming the liver and activating blood circulation, and dispelling wind and promoting qi. The high and medium doses (13.66 g / kg and 6.83 g / kg) of the Chinese medicine composition can improve the experimental indicators of male gouty arthritis Kunming mice (clean grade), including the degree of joint swelling of the toes of Kunming mice (clean grade), the inflammation index of the toes of Kunming mice (clean grade), the antioxidant capacity of Kunming mice (clean grade), the content of inflammatory mediators of Kunming mice (clean grade), and the expression level of main proteins and genes in the toes of Kunming mice (clean grade), while having no effect on the organ index and body weight of important organs of male gouty arthritis Kunming mice (clean grade), indicating that the Chinese medicine composition is effective in treating male gouty arthritis and has no side effects. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 Shown is the effect of the Chinese medicine composition on the swelling degree of the toe joints of the model mice;

[0030] Figure 2 Shown is the effect of the Chinese medicine composition on the inflammation index of the toe joints of the model mice;

[0031] Figure 3 Shown is the effect of the Chinese medicine composition on SOD in the toes of model mice;

[0032] Figure 4 Shown is the effect of the Chinese medicine composition on MDA in the toes of model mice;

[0033] Figure 5 Shown is the effect of the Chinese medicine composition on IL-1β in the toes of model mice;

[0034] Figure 6 Shown is the effect of the Chinese medicine composition on TNF-α in the toes of model mice;

[0035] Figure 7 Shown is the effect of the Chinese medicine composition on NLRP3 protein in the toes of model mice;

[0036] Figure 8 Shown is the effect of the Chinese medicine composition on the NLRP3 gene in the toes of model mice;

[0037] Fig. 9 Shown is the effect of the Chinese medicine composition on the cardiac index and lung index of model mice;

[0038] Fig.10 Shown is the effect of the Chinese medicine composition on the liver index and kidney index of model mice;

[0039] Fig.11Shown is the effect of the Chinese medicine composition on the body weight of model mice. DETAILED DESCRIPTION

[0040] The following will be combined with the embodiments and drawings to clearly and completely describe the concept, specific structure and technical effects of the present invention, so as to fully understand the purpose, scheme and effect of the present invention. It should be noted that the embodiments and features in the embodiments of this application can be combined with each other without conflict.

[0041] Example 1

[0042] Preparation of a Chinese medicine composition for treating male gouty arthritis

[0043] The traditional Chinese medicine composition is composed of the following raw materials by weight: 12.00g of holly leaves, 4.50g of cicada fungus, 7.50g of Brassica rapa seeds, 0.04g of Artemisia selengensis, 22.50g of Euonymus fortunei, and 6.00g of Sauvignon ovata seeds.

[0044] Example 2

[0045] A Chinese medicine composition (water decoction) for treating male gouty arthritis comprises the following steps:

[0046] (1) 12.00 g of holly leaves, 4.50 g of cicada fungus, 7.50 g of Brassica rapa seeds, 0.04 g of Artemisia sibiricum, 22.50 g of Euonymus fortunei and 6.00 g of Sauvignon ovata seeds were uniformly mixed to obtain 52.54 g of a Chinese medicine composition. Ten sets of the Chinese medicine composition were prepared to obtain 525.40 g of the Chinese medicine composition. The mixture of 525.40 g of the medicinal materials was then placed in a 20000 mL large-capacity round-bottom flask, 7000 mL of distilled water was added, and the mixture was soaked for 2.6 h, and then decocted at 100 °C for 2.6 h.

[0047] (2) filtering the cooled drug solution in step (1) through 2 layers of gauze, 4 layers of gauze, 6 layers of gauze, 8 layers of gauze, and 10 layers of gauze, respectively, to obtain a first filtrate and drug residue;

[0048] (3) The remaining drug residue in step (2) is placed in a 20,000 mL large-capacity round-bottom flask, and the same volume of distilled water as in step (1) is added. Steps (1) and (2) are repeated to obtain a second filtrate, and the first filtrate and the second filtrate are combined and mixed to obtain a decoction for treating male gouty arthritis.

[0049] Example 3

[0050] A Chinese medicine composition (dry paste) for treating male gouty arthritis comprises the following steps:

[0051] The decoction obtained in step (3) of the embodiment was placed in a drying oven, set at 65°C, dried for 6.5 days, and weighed and recorded using an electronic balance (PLZOZ-S model) to obtain 125.63g of dry paste of the Chinese medicine composition. The formula for calculating the yield of the dry paste of the Chinese medicine composition is: the yield of the Chinese medicine composition = [dry paste (g) / Chinese medicine composition (g)] × 100%. The yield of the Chinese medicine composition was calculated to be [dry paste (g) / Chinese medicine composition (g)] × 100%, that is, (125.63g / 525.40g) × 100% = 23.91%, that is, 1g of the dry paste of the Chinese medicine composition composed of holly leaves, cicada flowers, Brassica rapa seeds, Artemisia selengensis, Euphorbia fargesii, and Sauerkraut seeds is equivalent to 4.18g of the original medicinal materials of the Chinese medicine composition composed of holly leaves, cicada flowers, Brassica rapa seeds, Artemisia selengensis, Euphorbia fargesii, and Sauerkraut seeds.

[0052] Example 4

[0053] A Chinese medicine composition (dry powder) for treating male gouty arthritis, comprising the following steps:

[0054] The dry paste of the Chinese medicine composition obtained in Example 3 was crushed several times using a pulverizer, carefully ground, and then filtered using a 10-mesh screen, a 20-mesh screen, a 50-mesh screen, a 100-mesh screen, a 150-mesh screen, and a 200-mesh screen in sequence, and weighed and recorded using a PLZOZ-S electronic balance (PLZOZ-S type), to obtain 121.59 g of dry powder of the Chinese medicine composition. The powder yield of the Chinese medicine composition = [dry powder (g) / Chinese medicine composition (g)] × 100%. The powder yield of the traditional Chinese medicine composition was calculated to be [dry powder (g) / traditional Chinese medicine composition (g)] × 100%, that is, (121.59g / 525.40g) × 100% = 23.14%, that is, 1g of dry powder of the traditional Chinese medicine composition composed of holly leaves, cicada flower, Brassica rapa seeds, Artemisia annua, Thunbergia fortunei and Saurus seeds is equivalent to 4.32g of the original medicinal materials of the traditional Chinese medicine composition composed of holly leaves, cicada flower, Brassica rapa seeds, Artemisia annua, Thunbergia fortunei and Saurus seeds.

[0055] The dry powder of the Chinese medicine composition prepared in this example was packaged into bags, 10 g / bag, the last bag was 1.59 g, and there were 13 bags in total. Finally, they were stored in a -80°C refrigerator for later experimental research.

[0056] Example 5

[0057] The efficacy of the Chinese medicine composition of the present invention in treating male gouty arthritis

[0058] 1. Materials

[0059] 1.1 Animals

[0060] Male Kunming mice (clean grade), 105, 12 months old, (29.351±2.633) g, were provided by the Experimental Animal Center of Heilongjiang University of Chinese Medicine (Heilongjiang Province), and the animal qualification certificate number was SCXK (Heilongjiang Province) 2013-004. A dedicated person (professor of the Experimental Animal Center of Heilongjiang University of Chinese Medicine) was responsible for raising 105 Kunming mice (clean grade). The breeding location was the Experimental Animal Center of Heilongjiang University of Chinese Medicine (Heilongjiang Province). The breeding environment was constant humidity, normal temperature, sterile drinking water, sterile feed, sterile bedding, conventional lighting, etc. The 105 Kunming mice (clean grade) were allowed to obtain mouse food and sterile drinking water on their own, and the breeding environment was maintained at 1 / cage. The hair, excrement, mental state, etc. of the Kunming mice (clean grade) were observed at any time.

[0061] 1.2 Drugs

[0062] Allopurinol was purchased from Heilongjiang University of Traditional Chinese Medicine in Chongqing Qingyang Pharmaceutical Co., Ltd., China. Holly leaves, Cicadae, Brassica rapa, Artemisia scabra, Euonymus fortunei, and Sophora japonica were purchased from the First Affiliated Hospital of Heilongjiang University of Traditional Chinese Medicine (Heilongjiang Province). The Chinese medicine composition was identified by professors of Heilongjiang University of Traditional Chinese Medicine and can be used in combination. The dry powder was prepared according to the method of Example 4 and stored in a -80°C refrigerator.

[0063] 1.3 Reagents

[0064] The inflammatory mediator enzyme-linked immunosorbent assay TNF-α and IL-1β kits required for the Chinese herbal composition (Ilex oleifera, Cicadae Concolor, Brassica rapa seeds, Artemisia selengensis, Euonymus fortunei, and Sauerkraut seeds) were purchased from Nanjing Jiancheng Bioengineering Institute (China); the modeling drug sodium urate was purchased from Sigma (USA); the oxygen free radical assessment index enzyme-linked immunosorbent assay MDA and SOD kits were purchased from Nanjing Jiancheng Bioengineering Institute (China); the gene determination β-action and NLRP3 primers were purchased from Santa Cruz Biotechnology (USA); other reagents were domestically produced and analytically pure and provided by a professor from Heilongjiang University of Chinese Medicine (Heilongjiang Province) and an associate professor from Qiqihar Medical College (Heilongjiang Province).

[0065] 1.4 Instruments

[0066] 6100 microplate reader, RT (USA); -20℃ refrigerator, -80℃ refrigerator, Qingdao Haier Electric Co., Ltd. (China); quantitative PCR instrument, Applied Biosystems (USA); high-speed universal grinder, TT30-FW-100, Wenling Linda Machinery Co., Ltd. (China); desktop centrifuge, TGL-16G, Shanghai Anting Scientific Instrument Factory (China); electronic balance, PLZOZ-S, Sartorius (Germany); ultrapure water system, Rsearch UV, Shanghai Hotai Instrument Co., Ltd. (China).

[0067] 2. Methods

[0068] 2.1 Study Groups

[0069] The study requires 105 Kunming mice (clean grade), which are randomly divided into groups according to the counting method of the random number table, with 15 Kunming mice (clean grade) in each group. The Kunming mice (clean grade) are marked with picric acid solution, and the marked parts are the head, trunk, tail, etc. Each group is set to 15, including the experimental control group (15), the surgical control group (15), the model control group (15), the positive control group (15), the high-dose group of the Chinese medicine composition (high-dose group) (15), the medium-dose group of the Chinese medicine composition (medium-dose group) (15), and the low-dose group of the Chinese medicine composition (low-dose group) (15). The hair, excrement, mental state, etc. of each group of Kunming mice (clean grade) need to be observed at any time, and the Kunming mice (clean grade) need to be adapted to feeding for 1 week.

[0070] 2.2 Research model

[0071] Given that urate metabolism has certain rules, Kunming mice (clean grade) need to be treated with two injections (Shao P, Liu H, Xue Y, et al. LncRNA HOTTIP promotes inflammatory response in acute gouty arthritis via miR-101-3p / BRD4 axis[J]. Int J Rheum Dis. 2023 Feb; 26(2): 305-315.). A male gouty arthritis model was prepared according to the reference (Cao Y, Hu Y, Jin XF, et al. Dimethyl fumarate attenuates MSU-induced gouty arthritis by inhibiting NLRP3 inflammasome activation and oxidative stress[J]. Eur Rev Med Pharmacol Sci. 2023 Jan; 27(2): 628-641.). This experiment will select the model control group (15 mice), positive control group (15 mice), high-dose group (15 mice), medium-dose group (15 mice), and low-dose group (15 mice) to prepare the male gouty arthritis model. Experimental process: Prepare sodium urate solution (2.5%) for Kunming mice (clean grade), grab the Kunming mice (clean grade) with the right hand, find the right sole of the foot of the Kunming mice (clean grade), and start injecting 0.05mL of sodium urate solution (2.5%) into the right sole of the foot of the Kunming mice (clean grade) and the right dorsum of the foot of the Kunming mice (clean grade). Observe the bulge of the right sole of the foot of the Kunming mice (clean grade) and the right dorsum of the foot of the Kunming mice (clean grade) at any time. At the same time, 0.05mL of normal saline was injected into the right sole of the foot of the Kunming mice (clean grade) and the right dorsum of the foot of the Kunming mice (clean grade) in the surgical control group (15 mice). In the experimental control group (15 mice), the right sole of the foot of Kunming mice (clean grade) and the right dorsum of the foot of Kunming mice (clean grade) were not injected with sodium urate solution (2.5%) and normal saline.The second modeling process was the same as the first modeling process (Shrateh ON, Jobran AWM, Zaid MA, et al. Unusual location of gouty arthritis with shoulder joint involvement in an elderly male patient a rare case report [J]. Ann Med Surg Lond. 2023 Mar 27; 85 (4): 1158-1161.), and the interval between the two male gouty arthritis models was 7 days, and the total duration of the male gouty arthritis model was 14 days.

[0072] 2.3 Model Evaluation

[0073] After the two male gouty arthritis modeling, the model control group (14 mice), positive control group (14 mice), high-dose group (12 mice), medium-dose group (12 mice), and low-dose group (12 mice) survived, and the toes of Kunming mice (clean grade) were significantly swollen, indicating that the male gouty arthritis model of this experiment was successfully established; no mice died in the surgical control group (15) Kunming mice (clean grade) and the experimental control group (15) Kunming mice (clean grade), and the toes of Kunming mice (clean grade) were not swollen. Finally, the male gouty arthritis model was successfully established, and the number of Kunming mice (clean grade) in each group was model control group (14 mice), positive control group (14 mice), high-dose group (12 mice), medium-dose group (12 mice), and low-dose group (12 mice).

[0074] 2.4 Calculating drug dosage

[0075] This experiment includes a Chinese medicine composition of 12.00g of holly leaves, 4.50g of cicada fungus, 7.50g of Brassica rapa seeds, 0.04g of Artemisia annua, 22.50g of Euphorbia grandiflora, and 6.00g of Sauvignon gras seeds. The safe dose for each portion is 52.54g in total. Calculated according to the body surface area of ​​Kunming mice (clean grade), the safe and effective dose of the Chinese medicine composition containing 12.00g of holly leaves, 4.50g of cicada fungus, 7.50g of Brassica rapa seeds, 0.04g of Artemisia annua, 22.50g of Euphorbia grandiflora, and 6.00g of Sauvignon gras is calculated to be [(52.54g×0.0026) / 20g Kunming mice (clean grade)]×50, i.e. 6.83g / kg. This dose is a medium dose of a Chinese medicine composition including 12.00g of holly leaves, 4.50g of cicada flowers, 7.50g of Brassica rapa seeds, 0.04g of Artemisia scabrae, 22.50g of Euphorbia fargesii, and 6.00g of Sauerkraut seeds. The high dose is 13.66g / kg, and the low dose is 3.42g / kg. The dosage of Kunming mice (clean grade) was calculated with reference to the clinical safety dose of allopurinol (0.026-0.052) g / kg. Based on this, this experiment selected 0.03g / kg of allopurinol for experimental research on a Chinese medicine composition including 12.00g of holly leaves, 4.50g of cicada flowers, 7.50g of Brassica rapa seeds, 0.04g of Artemisia scabrae, 22.50g of Euphorbia fargesii, and 6.00g of Sauerkraut seeds.

[0076] 2.5 Animal Dosing

[0077] In this experiment, the basic status of each group of Kunming mice (clean grade) was observed 4.1 hours after the second modeling of each group of Kunming mice (clean grade), and the drug administration began. The experimental control group Kunming mice (clean grade) were given normal saline (0.3mL) by gavage for 14 days; the surgical control group Kunming mice (clean grade) were given normal saline (0.3mL) by gavage for 14 days; the model control group Kunming mice (clean grade) were given normal saline (0.3mL) by gavage for 14 days; the positive control group was given allopurinol (0.03g / kg) by gavage for 14 days; the high-dose group was given 13.66g / kg of the Chinese medicine composition (equivalent to the original medicinal materials of the Chinese medicine composition, 12.00g of holly leaves, 4.50g of cicada flowers, 7.50g of Brassica rapa, 0.04g of Artemisia selengensis, 22.50g of Euonymus fortunei, 6.00g of Sauerkraut seeds, The mice in the middle-dose group were given 6.83 g / kg of the Chinese medicine composition (equivalent to the original medicinal materials of the Chinese medicine composition, namely, 12.00 g of holly leaves, 4.50 g of cicada fungus, 7.50 g of Brassica rapa seeds, 0.04 g of Artemisia annua, 22.50 g of Euphorbia grandiflora, and 6.00 g of Sauvignon ovata seeds, for a total of 52.54 g) by gavage for 14 days; the mice in the low-dose group were given 3.42 g / kg of the Chinese medicine composition (equivalent to the original medicinal materials of the Chinese medicine composition, namely, 12.00 g of holly leaves, 4.50 g of cicada fungus, 7.50 g of Brassica rapa seeds, 0.04 g of Artemisia annua, 22.50 g of Euphorbia grandiflora, and 6.00 g of Sauvignon ovata seeds, for a total of 52.54 g) by gavage for 14 days. After the intragastric administration of this experiment, the number of surviving Kunming mice (clean grade) in each group included 14 Kunming mice (clean grade) surviving in the experimental control group, surgical control group, model control group, positive control group, high-dose group, medium-dose group, and low-dose group, 14 Kunming mice (clean grade) surviving, 13 Kunming mice (clean grade) surviving, 12 Kunming mice (clean grade) surviving, 11 Kunming mice (clean grade) surviving, 10 Kunming mice (clean grade) surviving, and 11 Kunming mice (clean grade) surviving.

[0078] 2.6 Determination of joint swelling

[0079] This experiment refers to the literature (Zhang Jiping. The effect of Qingre Lishi Xiezhuo Decoction combined with colchicine in the treatment of acute gouty arthritis with damp-heat arthritis and its effects on inflammatory response and blood rheology [J]. Chinese and Foreign Medical Research, 2023, 21 (22): 63-67.) The measured indicators include the degree of joint swelling in the toes (Yang Jun, Yao Jinlong, Liang Shuang, et al. Exploring the mechanism of action of Xuanbi Tongluo paste on acute gouty arthritis based on TLR4 / NF-κB / NLRP3 pathway-mediated synoviocyte pyroptosis [J]. Journal of Traditional Chinese Medicine, 2023, 29 (07): 41-45.). Kunming mice (clean grade) were marked, especially at the knee joint bend. 4.1 hours after the end of intragastric administration, Kunming mice (clean grade) were anesthetized with ether to avoid any unexpected effects of ether on Kunming mice (clean grade). After anesthesia, the toes of Kunming mice (clean grade) were cut off from the knee bend mark, and the volume scale of the toes of Kunming mice (clean grade) in each group was recorded and calculated according to the formula: Kunming mice (clean grade) toe joint swelling (mL) = Kunming mice (clean grade) toe joint volume after the experiment (mL) - Kunming mice (clean grade) toe joint volume before the experiment (mL). Finally, the joint swelling of Kunming mice (clean grade) was evaluated.

[0080] 2.7 Determination of joint inflammation index

[0081] The experimental reference (Wen Xiaofang. Effect of Qingre Lishi Xiezhuo Decoction in acute gouty arthritis of damp-heat blockage type [J]. Medical Theory and Practice, 2023, 36 (16): 2762-2765.) measured indicators, including the inflammation index of the toe joints of Kunming mice (clean grade) with male gouty arthritis. The inflammatory index of Kunming mice (clean grade) includes 5 grades, among which Kunming mice (clean grade) have no swelling at grade 0, 0 points, and clear bone signs; Kunming mice (clean grade) have mild swelling at grade 1, 1-2 points, no bone signs can be seen, and redness and heat appear; Kunming mice (clean grade) have moderate swelling at grade 2, 3-4 points, overall swelling, no bone signs can be seen; Kunming mice (clean grade) have severe swelling at grade 3, 5-6 points, overall swelling, no bone signs can be seen, and obvious swelling of the knee joint; Kunming mice (clean grade) have extremely severe swelling at grade 4, 7-8 points, overall swelling, no bone signs can be seen, obvious swelling of the knee joint, and joint deformity.

[0082] 2.8 Determination of antioxidant capacity

[0083] This experiment refers to the literature (Mohapatra A, Rajendrakumar SK, Chandrasekaran G, et al. Biomineralized Nanoscavenger Abrogates Proinflammatory Macrophage Polarization and Induces Neutrophil Clearance through Reverse Migration during Gouty Arthritis [J]. ACS Appl Mater Interfaces. 2023 Jan 25; 15 (3): 3812-3825.) to determine the antioxidant capacity, and the method used was enzyme-linked immunosorbent assay. In this experiment, Kunming mice (clean grade) were treated with drugs for 4.1 hours, then captured and anesthetized with 10% chloral hydrate. After waiting for the anesthesia effect of Kunming mice (clean grade), the mice (clean grade) were operated on ice, the toes of Kunming mice (clean grade) were collected, the skin of the toes of Kunming mice (clean grade) was removed, the toes of Kunming mice (clean grade) were cut into pieces, the toes of Kunming mice (clean grade) were finely ground, the toes of Kunming mice (clean grade) were homogenized, and the toes of Kunming mice (clean grade) were kept still in a refrigerator (4°C) for 1 hour. The toes of Kunming mice (clean grade) were centrifuged at 3000 rpm in a desktop centrifuge (TGL-16G model) for 20 minutes. The supernatant of the toes of Kunming mice (clean grade) was obtained by pipette, and the SOD and MDA contents of the toes of Kunming mice (clean grade) were determined according to the instructions of the kit.

[0084] 2.9 Determination of inflammatory mediators

[0085] This experiment refers to the literature (Wang W, Kou J, Zhang M, et al. A metabonomic study to explore potential markers of asymptomatic hyperuricemia and acute goutyarthritis. J Orthop Surg Res. 2023 Feb 13; 18 (1): 96.) to determine the content of inflammatory mediators IL-1β and TNF-α, and the method used was enzyme-linked immunosorbent assay. In this experiment, Kunming mice (clean grade) were treated with drugs for 4.1 hours, and then Kunming mice (clean grade) were caught and anesthetized with 10% chloral hydrate. After waiting for the anesthesia effect of Kunming mice (clean grade), Kunming mice (clean grade) were operated on ice, the toes of Kunming mice (clean grade) were collected, the skin of the toes of Kunming mice (clean grade) was removed, the toes of Kunming mice (clean grade) were cut into pieces, the toes of Kunming mice (clean grade) were finely ground, and the toes of Kunming mice (clean grade) were homogenized. (clean grade) toes, place the Kunming mouse (clean grade) toes in a refrigerator (4°C) for 1 hour, centrifuge the Kunming mouse (clean grade) toes at 3000 rpm in a desktop centrifuge (TGL-16G model) for 20 minutes, and obtain the supernatant of the Kunming mouse (clean grade) toes with a pipette to detect the IL-1β and TNF-α protein content of the joint tissue of the toes of Kunming mice (clean grade) in each group, and store the remaining Kunming mouse (clean grade) samples in a -80°C refrigerator for standby use. Determine the absorbance value (OD) at a wavelength of 450nm according to the instructions of the kit to calculate the content of inflammatory mediators IL-1β and TNF-α in the toes of Kunming mice (clean grade).

[0086] 2.10 Determination of NLRP3 protein expression

[0087] In this experiment, Kunming mice (clean grade) were treated with drugs for 4.1 hours, then captured and anesthetized with 10% chloral hydrate. After the anesthesia effect of Kunming mice (clean grade) was good, the Kunming mice (clean grade) were operated on ice, the toes of Kunming mice (clean grade) were collected, the skin of the toes of Kunming mice (clean grade) was removed, and the expression of NLRP3 was detected by immunohistochemistry. The toes of Kunming mice (clean grade) were fixed, embedded, sectioned, dewaxed, dehydrated, repaired, blocked, added with primary antibody working solution (4°C overnight), incubated with HRP-conjugated secondary antibody, counterstained with hematoxylin, observed under an optical microscope, and the mean optical density (MOD) of NLRP3 in the toes of Kunming mice (clean grade) was calculated using Image-Pro Plus 6.0 software to evaluate the expression level of NLRP3.

[0088] 2.11 Determination of NLRP3 gene expression

[0089] In this experiment, quantitative PCR was used to determine the NLRP3 mRNA expression in the joint tissue of the toes of Kunming mice (clean grade) in each group. After 4.1 hours of drug treatment for Kunming mice (clean grade), Kunming mice (clean grade) were captured and injected with 10% chloral hydrate to anesthetize the Kunming mice (clean grade). After waiting for the anesthesia effect of Kunming mice (clean grade), Kunming mice (clean grade) were operated on ice, the toes of Kunming mice (clean grade) were collected, the skin of the toes of Kunming mice (clean grade) was removed, reverse transcription was performed into cDNA of the toes of Kunming mice (clean grade), and primers for NLRP3 mRNA of the toes of Kunming mice (clean grade) were designed. Among them are NLRP3 (Forward: 5'-GAA GTA AGC CGC AGA TAG GGT ATG-3'; Reverse: 3'-CGC TGC ATG GCT GTCACG TGC TAG-5'); β-action (Forward: 5'-GCT CTG ATC CAT CCA TGC ATC CT-3'; Reverse: 3'-CGT TGC ATG CAT TAG CGT TTG GA-5'). For the toes of Kunming mice (clean grade), the experiment needs to add special SYBR Green I fluorescent material to treat the toes of Kunming mice (clean grade). The setting parameters include 50℃ Kunming mice (clean grade) toes treatment for 2 minutes, 95℃ Kunming mice (clean grade) toes treatment for 10 minutes, 95℃ Kunming mice (clean grade) toes treatment for 15 seconds, 60℃ Kunming mice (clean grade) toes treatment for 1 minute. After 40 cycles, the 2-ΔΔCt method is used to calculate the NLRP3 mRNA expression level in the toes of Kunming mice (clean grade).

[0090] 2.12 Determination of organ index

[0091] In this experiment, after 4.1 hours of drug treatment on Kunming mice (clean grade), the Kunming mice (clean grade) were caught and injected with 10% chloral hydrate to anesthetize the Kunming mice (clean grade). After waiting for the anesthesia effect of Kunming mice (clean grade), the Kunming mice (clean grade) were operated on ice, the Kunming mice (clean grade) were killed by decapitation, the Kunming mice (clean grade) were dissected, the organs of Kunming mice (clean grade) were observed, the main organs of Kunming mice (clean grade) including lungs, heart, liver, and kidneys were collected, the main organs of Kunming mice (clean grade) including lungs, heart, liver, and kidneys were weighed, and the organ index of the main organs of Kunming mice (clean grade) including lungs, heart, liver, and kidneys in each group was calculated according to the organ index formula of the main organs of Kunming mice (clean grade) including lungs, heart, liver, and kidneys, and the side effects of the Chinese medicine composition will be evaluated. Calculation formula: organ index (mg / g) of each group of Kunming mice (clean grade) = organ mass (mg) of each group of Kunming mice (clean grade) / body weight (g) of Kunming mice (clean grade).

[0092] 2.13 Determination of mouse body weight

[0093] In this experiment, an electronic balance was used to measure the weight of each group of Kunming mice (clean grade). The measurement schedule was 0d Kunming mice (clean grade), 7d Kunming mice (clean grade), and 14d Kunming mice (clean grade). The weight change patterns of 0d Kunming mice (clean grade), 7d Kunming mice (clean grade), and 14d Kunming mice (clean grade) were analyzed to evaluate the side effects of the Chinese medicine composition.

[0094] 2.14 Statistical analysis

[0095] SPSS 19.0 software was used to analyze the data. ±s indicates that the data of Kunming mice (clean grade) among multiple groups were compared by one-way analysis of variance.

[0096] 3. Results

[0097] 3.1 Effect of Chinese medicine composition on the swelling of toe joints in model mice

[0098] The study showed that compared with the experimental control group of Kunming mice (clean grade), the joint swelling of the toes of male gouty arthritis model Kunming mice (clean grade) in the model control group increased significantly (P < 0.05), the joint swelling of the toes of Kunming mice (clean grade) in the surgical control group did not change significantly (P > 0.05), the joint swelling of the toes of male gouty arthritis model Kunming mice (clean grade) in the positive control group, high-dose group, and medium-dose group decreased significantly (P < 0.05), and the joint swelling of the toes of male gouty arthritis model Kunming mice (clean grade) in the low-dose group did not change significantly (P > 0.05). See Table 1. Figure 1(Note: The horizontal axis is the group, and the vertical axis is the degree of joint swelling (mL)).

[0099] Table 1 Effect of Chinese medicine composition on swelling of toe joints in model mice

[0100]

[0101] Note: Compared with the experimental control group: @ P >0.05, @@ P <0.05; compared with the model control group: & P <0.05, && P >0.05

[0102] 3.2 Effect of Chinese medicine combination on inflammation index of toe joints in model mice

[0103] The study showed that compared with the experimental control group of Kunming mice (clean grade), the inflammation index of the toe joints of the male gouty arthritis model Kunming mice (clean grade) in the model control group was significantly increased ( P＜ 0.05), the inflammation index of the toe joints of Kunming mice (clean grade) in the surgical control group did not change significantly ( P >0.05), the inflammation index of the toe joints of male gouty arthritis model Kunming mice (clean grade) in the positive control group, high-dose group, and medium-dose group was significantly reduced ( P＜ 0.05), the inflammation index of the toe joints of male gouty arthritis model Kunming mice (clean grade) in the low-dose group did not change significantly ( P >0.05). See Table 2. Figure 2 (Note: The horizontal axis is the group and the vertical axis is the joint inflammation index).

[0104] Table 2 Effects of Chinese medicine combination on inflammation index of toe joints in model mice ( ±s)

[0105]

[0106] Note: Compared with the experimental control group: @ P >0.05, @@ P <0.05; compared with the model control group: & P <0.05, && P >0.05

[0107] 3.3 Effect of Chinese herbal medicine combination on SOD in the toes of model mice

[0108] The study showed that compared with the experimental control group of Kunming mice (clean grade), the SOD in the toes of male gouty arthritis model Kunming mice (clean grade) in the model control group was significantly reduced ( P＜ 0.05), and the SOD changes in the toes of Kunming mice (clean grade) in the surgical control group were not obvious ( P >0.05), the SOD in the toes of male gouty arthritis model Kunming mice (clean grade) in the positive control group, high-dose group, and medium-dose group increased significantly ( P＜ 0.05), the SOD changes in the toes of male gouty arthritis model Kunming mice (clean grade) in the low-dose group were not obvious ( P >0.05). See Table 3. Figure 3 (Note: the horizontal axis is the group, and the vertical axis is the specific value of SOD (U / mg) content).

[0109] Table 3 Effects of Chinese medicine composition on SOD in the toes of model mice ( ±s)

[0110]

[0111] Note: Compared with the experimental control group: @ P >0.05, @@ P <0.05; compared with the model control group: & P <0.05, && P >0.05

[0112] 3.4 Effect of Chinese herbal medicine combination on MDA in the toes of model mice

[0113] The study showed that compared with the experimental control group of Kunming mice (clean grade), the MDA in the toes of male gouty arthritis model Kunming mice (clean grade) in the model control group was significantly increased ( P＜ 0.05), the MDA changes in the toes of Kunming mice (clean grade) in the surgical control group were not obvious ( P >0.05), the MDA in the toes of male gouty arthritis model Kunming mice (clean grade) in the positive control group, high-dose group, and medium-dose group was significantly reduced ( P＜ 0.05), the MDA changes in the toes of male gouty arthritis model Kunming mice (clean grade) in the low-dose group were not obvious ( P >0.05). See Table 4. Figure 4 (Note: the horizontal axis is the group; the vertical axis is the specific value of MDA (μmol / g) content).

[0114] Table 4 Effects of Chinese medicine composition on MDA in the toes of model mice ( ±s)

[0115]

[0116] Note: Compared with the experimental control group: @ P >0.05, @@ P <0.05; compared with the model control group: & P <0.05, && P >0.05

[0117] 3.5 Effect of Chinese herbal medicine combination on IL-1β in the toes of model mice

[0118] The study showed that compared with the Kunming mice (clean grade) in the experimental control group, the IL-1β in the toes of the male gouty arthritis model Kunming mice (clean grade) in the model control group was significantly increased ( P＜ 0.05), and the IL-1β in the toes of Kunming mice (clean grade) in the surgical control group did not change significantly ( P >0.05), IL-1β in the toes of male gouty arthritis model Kunming mice (clean grade) in the positive control group, high-dose group, and medium-dose group was significantly reduced ( P＜ 0.05), the IL-1β in the toes of male gouty arthritis model Kunming mice (clean grade) in the low-dose group did not change significantly ( P >0.05). See Table 5. Figure 5 (Note: the horizontal axis is the group; the vertical axis is the specific value of IL-1β (pg / mL) content).

[0119] Table 5 Effects of Chinese medicine composition on IL-1β in the toes of model mice ( ±s)

[0120]

[0121] Note: Compared with the experimental control group: @ P >0.05, @@ P <0.05; compared with the model control group: & P <0.05, && P >0.05

[0122] 3.6 Effect of Chinese medicine combination on TNF-α in the toes of model mice

[0123] The study showed that compared with the Kunming mice (clean grade) in the experimental control group, the TNF-α in the toes of the male gouty arthritis model Kunming mice (clean grade) in the model control group was significantly increased ( P＜ 0.05), and the TNF-α in the toes of Kunming mice (clean grade) in the surgical control group did not change significantly ( P >0.05), TNF-α in the toes of male gouty arthritis model Kunming mice (clean grade) in the positive control group, high-dose group, and medium-dose group was significantly reduced ( P＜ 0.05), the TNF-α in the toes of male gouty arthritis model Kunming mice (clean grade) in the low-dose group did not change significantly ( P >0.05). See Table 6. Figure 6 (Note: The horizontal axis is the group, and the vertical axis is the specific value of TNF-α (pg / mL) content).

[0124] Table 6 Effects of Chinese medicine composition on TNF-α in the toes of model mice ( ±s)

[0125]

[0126] Note: Compared with the experimental control group: @ P >0.05, @@ P <0.05; compared with the model control group: & P <0.05, && P >0.05

[0127] 3.7 Effect of Chinese herbal medicine combination on NLRP3 protein in the toes of model mice

[0128] The study showed that compared with the Kunming mice (clean grade) in the experimental control group, the average optical density of NLRP3 protein in the toes of the male gouty arthritis model Kunming mice (clean grade) in the model control group was significantly increased ( P＜ 0.05), the average optical density of NLRP3 protein in the toes of Kunming mice (clean grade) in the surgical control group did not change significantly ( P >0.05), the average optical density of NLRP3 protein in the toes of male gouty arthritis model Kunming mice (clean grade) in the positive control group, high-dose group, and medium-dose group was significantly reduced ( P＜ 0.05), the average optical density of NLRP3 protein in the toes of male gouty arthritis model Kunming mice (clean grade) in the low-dose group did not change significantly ( P >0.05). See Table 7. Figure 7 (Note: the horizontal axis is the group; the vertical axis is the specific value of NLRP3 (average optical density value)).

[0129] Table 7 Effects of Chinese medicine composition on NLRP3 protein in the toes of model mice ( ±s)

[0130]

[0131] Note: Compared with the experimental control group: @ P >0.05, @@ P <0.05; compared with the model control group: & P <0.05, && P >0.05

[0132] 3.8 Effect of Chinese herbal medicine combination on NLRP3 gene in the toes of model mice

[0133] The study showed that compared with the experimental control group of Kunming mice (clean grade), the relative expression of NLRP3 gene in the toes of male gouty arthritis model Kunming mice (clean grade) in the model control group was significantly increased ( P＜ 0.05), the relative expression of NLRP3 gene in the toes of Kunming mice (clean grade) in the surgical control group did not change significantly ( P >0.05), the relative expression of NLRP3 gene in the toes of male gouty arthritis model Kunming mice (clean grade) in the positive control group, high-dose group, and medium-dose group was significantly reduced ( P＜ 0.05), the relative expression of NLRP3 gene in the toes of male gouty arthritis model Kunming mice (clean grade) in the low-dose group did not change significantly ( P >0.05). See Table 8. Figure 8 (Note: The horizontal axis is the group; the vertical axis is the specific value of NLRP3 (relative expression level)).

[0134] Table 8 Effects of Chinese medicine composition on NLRP3 gene in toes of model mice ( ±s)

[0135]

[0136] Note: Compared with the experimental control group: @ P >0.05, @@ P <0.05; compared with the model control group: & P <0.05, && P >0.05

[0137] 3.9 Effects of Chinese herbal medicine combination on cardiac index and lung index of model mice

[0138] The study showed that compared with the Kunming mice (clean grade) in the experimental control group, the heart index and lung index of the model mice in the model control group did not change significantly ( P >0.05), the cardiac index and lung index of Kunming mice (clean grade) in the surgical control group did not change significantly ( P >0.05), the heart index and lung index of Kunming mice (clean grade) in the positive control group, high-dose group, medium-dose group and low-dose group did not change significantly ( P >0.05). See Table 9. Fig. 9 (Note: the horizontal axis is the group; the vertical axis is the specific values ​​of the cardiac index (mg / g) and the lung index (mg / g)).

[0139] Table 9 Effects of Chinese medicine composition on cardiac index and lung index of model mice ( ±s, n=9)

[0140]

[0141] Note: Compared with the experimental control group: @ P >0.05; compared with the model control group: @@ P >0.05

[0142] 3.10 Effects of Chinese medicine combination on liver index and kidney index of model mice

[0143] The study showed that compared with the Kunming mice (clean grade) in the experimental control group, the liver index and kidney index of the model mice in the model control group did not change significantly ( P >0.05), the liver index and kidney index of Kunming mice (clean grade) in the surgical control group did not change significantly ( P >0.05), the liver index and kidney index of Kunming mice (clean grade) in the positive control group, high-dose group, medium-dose group and low-dose group did not change significantly ( P >0.05). See Table 10. Fig.10 (Note: The horizontal axis is the group; the vertical axis is the specific values ​​of liver index (mg / g) and kidney index (mg / g)).

[0144] Table 10 Effects of Chinese medicine composition on liver index and kidney index of model mice ( ±s, n=9)

[0145]

[0146] Note: Compared with the experimental control group: @ P >0.05; compared with the model control group:@@ P >0.05

[0147] 3.11 Effect of Chinese herbal medicine combination on body weight of model mice

[0148] The study showed that compared with the weight of the experimental control group of 0d Kunming mice (clean grade), 7d Kunming mice (clean grade), and 14d Kunming mice (clean grade), the weight of the model control group and its surgical control group of 0d Kunming mice (clean grade), 7d Kunming mice (clean grade), and 14d Kunming mice (clean grade) did not change significantly ( P >0.05); Compared with the body weight of 0d Kunming mice (clean grade), 7d Kunming mice (clean grade), and 14d Kunming mice (clean grade) in the model control group, the body weight of 0d Kunming mice (clean grade), 7d Kunming mice (clean grade), and 14d Kunming mice (clean grade) in the positive control group, high-dose group, medium-dose group, and low-dose group did not change significantly ( P >0.05). See Table 11. Fig.11 (Note: the horizontal axis is the group; the vertical axis is the body weight at 0d (g), 7d (g), and 14d (g)).

[0149] Table 11 Effects of Chinese medicine composition on body weight of model mice ( ±s, n=9)

[0150]

[0151] Note: Compared with the experimental control group: @ P >0.05; compared with the model control group: @@ P >0.05

[0152] In order to evaluate the side effects of the Chinese medicine composition, this experiment detected the changes in the organ indexes and body weight of the important organs of Kunming mice (clean grade) in each group. The study showed that compared with the experimental control group, the changes in the main organ indexes and body weight of male gouty arthritis Kunming mice (clean grade) in the model control group were not obvious. After treatment with the Chinese medicine composition (12.00g of holly leaves, 4.50g of cicada flowers, 7.50g of Brassica rapa seeds, 0.04g of Artemisia selengensis, 22.50g of Euonymus fortunei, and 6.00g of Sauerkraut seeds), compared with the model control group, the changes in the main organ indexes and body weight of Kunming mice (clean grade) in the positive control group, high-dose group, medium-dose group, and low-dose group were not obvious, indicating that the Chinese medicine composition (12.00g of holly leaves, 4.50g of cicada flowers, 7.50g of Brassica rapa seeds, 0.04g of Artemisia selengensis, 22.50g of Euonymus fortunei, and 6.00g of Sauerkraut seeds) were not obvious. .00g, cicada fungus 4.50g, brassica seeds 7.50g, artemisia annua 0.04g, euonymus fortunei 22.50g, and sarocarpus indica seeds 6.00g) had no effect on the major organ indexes and body weight, indicating that the Chinese medicine composition (holly leaf 12.00g, cicada fungus 4.50g, brassica seeds 7.50g, artemisia annua 0.04g, euonymus fortunei 22.50g, and sarocarpus indica seeds 6.00g) had no side effects.

[0153] The above experiments show that the invention patent discloses a preparation method and application of a Chinese medicine composition that can be used to treat male gouty arthritis. The Chinese medicine composition includes 12.00g of holly leaves, 4.50g of cicada flowers, 7.50g of Brassica rapa seeds, 0.04g of Artemisia suspensa, 22.50g of Euonymus fortunei, and 6.00g of Sauerkraut seeds, which has the effects of calming the liver, activating blood circulation, and dispelling wind and promoting qi. High and medium doses (13.66g / kg, 6.83g / kg) of the Chinese medicine composition can improve the experimental indicators of Kunming mice (clean grade) with gouty arthritis, including the degree of joint swelling in the toes of Kunming mice (clean grade), the joint inflammation index in the toes of Kunming mice (clean grade), the antioxidant capacity of Kunming mice (clean grade), the content of inflammatory mediators in Kunming mice (clean grade), and the expression levels of major proteins and genes in the toes of Kunming mice (clean grade), but have no effect on the organ index and body weight of important organs of Kunming mice (clean grade) with gouty arthritis, indicating that the Chinese medicine composition is effective in treating male gouty arthritis without side effects.

[0154] In summary, the Chinese medicine composition consisting of 6 medicinal materials, including 12.00g of holly leaves, 4.50g of cicada fungus, 7.50g of Brassica rapa seeds, 0.04g of Artemisia annua, 22.50g of Euphorbia fortunei, and 6.00g of Sauvignon granatum, is effective in treating male gouty arthritis without side effects.

[0155] Although the description of the present invention has been quite detailed and specifically describes several described embodiments, it is not intended to be limited to any of these details or embodiments or any particular embodiment, but should be regarded as providing a broad possible interpretation of these claims in view of the prior art by reference to the appended claims, thereby effectively covering the intended scope of the present invention. In addition, the above description of the present invention is based on the embodiments foreseeable by the inventor, and its purpose is to provide a useful description, and those non-substantial changes to the present invention that have not yet been foreseen may still represent equivalent changes to the present invention.

Claims

1. A Chinese medicine composition for treating male gouty arthritis, characterized in that: The raw materials and dosages of the traditional Chinese medicine composition are: 10-12 parts by weight of holly leaves, 3-5 parts by weight of cicada fungus, 5-7.5 parts by weight of Brassica rapa seeds, 0.04 parts by weight of Artemisia annua, 20-23 parts by weight of Euonymus fortunei, and 5-6 parts by weight of Salacia chinensis seeds.

2. The Chinese medicine composition according to claim 1, characterized in that: The raw materials and dosages of the traditional Chinese medicine composition are as follows: 12 parts by weight of holly leaves, 4.5 parts by weight of cicada fungus, 7.5 parts by weight of Brassica rapa seeds, 0.04 parts by weight of Artemisia annua, 22.5 parts by weight of Euonymus fortunei, and 6 parts by weight of Shorea seeds.

3. A Chinese medicine preparation for treating male gouty arthritis, characterized in that: The Chinese medicine composition according to claim 1 or 2 is added with auxiliary materials required for preparation molding, and is prepared according to the conventional method for preparing pharmaceutical preparations.

4. The Chinese medicine preparation according to claim 3, characterized in that: The auxiliary materials include one or more of starch, microcrystalline cellulose, sucrose, lactose, powdered sugar, glucose and vitamin C.

5. The Chinese medicine preparation according to claim 4, characterized in that: The Chinese medicine preparation is a decoction, capsule, pill, granule, tablet or oral liquid.

6. A method for preparing the Chinese medicine composition according to claim 1 or 2, characterized in that: The following steps are involved: (1) Weigh each API according to the weight portion described in claim 1 or 2, mix the APIs evenly, add 8-15 times the total weight of the APIs into distilled water, soak for 2-3 hours, decoct for 2-3 hours, and cool; (2) filtering the cooled drug solution in step (1) through gauze to obtain a first filtrate and a filter residue; (3) Add the same volume of distilled water as in step (1) to the remaining drug residue in step (2), repeat steps (1) and (2) to obtain a second filtrate, and then combine the first filtrate and the second filtrate and mix them to obtain a water decoction for treating male gouty arthritis.

7. The method according to claim 6, characterized in that The obtained water decoction is placed in an electric constant temperature air drying oven and dried at 70° C. for 4-5 days to obtain a dry paste of the traditional Chinese medicine composition.

8. The method according to claim 7, characterized in that The obtained dry paste of the traditional Chinese medicine composition is crushed in a pulverizer, ground, and repeatedly treated with molecular sieves to obtain dry powder of the traditional Chinese medicine composition.

9. The method according to claim 6, characterized in that The specific operation of the gauze filtering is to use 2 layers of gauze for filtering, 4 layers of gauze for filtering, 6 layers of gauze for filtering, 8 layers of gauze for filtering, and 10 layers of gauze for filtering in sequence.

10. Use of the Chinese medicine composition according to claim 1 or 2 in the preparation of a medicament for treating male gouty arthritis.

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