Traditional Chinese medicine composition for treating rheumatoid arthritis as well as preparation method and application thereof

Abstract

The invention relates to the technical field of traditional Chinese medicine preparations, in particular to a traditional Chinese medicine composition for treating rheumatoid arthritis and a preparation method and application thereof. The traditional Chinese medicine composition is prepared from sargentgloryvine stem, cortex acanthopanacis, caesalpinia cucullata, rhizoma drynariae and garden balsam stem. The preparation method comprises the following steps: extracting; filtering is performed; and concentrating. Animal experiments and clinical tests prove that the traditional Chinese medicine composition provided by the invention has a remarkable treatment effect on rheumatoid arthritis, is good in safety and free of obvious adverse reaction, and has great clinical popularization and application values.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicine preparations, and in particular to a traditional Chinese medicine composition for treating rheumatoid arthritis, and a preparation method and application thereof. Background Art

[0002] Rheumatoid arthritis (RA) is a chronic autoimmune disease caused by a combination of environmental and genetic factors. Its pathological changes are characterized by persistent inflammation of the synovial membrane, pannus formation, and cartilage and bone destruction. RA is characterized by high morbidity, relapse rates, and disability, affecting 0.3% to 1% of the global population. High medical costs and loss of productivity due to joint disability impose a heavy economic burden on society and severely impact patients' quality of life.

[0003] Currently, the drugs used clinically to treat RA include: nonsteroidal anti-inflammatory drugs (NSAIDs) such as ibuprofen and diclofenac sodium; glucocorticoids such as prednisone acetate and dexamethasone acetate; anti-rheumatic drugs such as methotrexate and leflumitol; and biologics such as adalimumab injection and infliximab injection. While these drugs can achieve some efficacy, they still have drawbacks such as significant side effects and high treatment costs. Therefore, there is an urgent need to develop new RA treatments with significant therapeutic effects and minimal side effects.

[0004] To solve the above problems, the present invention provides a traditional Chinese medicine composition obtained by combining Sargentodoxae, Acanthopanax Cortex, Herba Lycopodii, Rhizoma Drynariae and Herba Polygoni Multiflori, aiming to provide a new drug option for the clinical treatment of rheumatoid arthritis and improve the quality of life of patients. Summary of the Invention

[0005] The purpose of the present invention is to provide a traditional Chinese medicine composition for treating rheumatoid arthritis and a preparation method thereof.

[0006] Another object of the present invention is to provide the use of the above-mentioned traditional Chinese medicine composition in the preparation of a medicine for treating rheumatoid arthritis.

[0007] To achieve the above object, the technical solution adopted by the present invention is as follows: The traditional Chinese medicine composition for treating rheumatoid arthritis of the present invention consists of the following components in parts by mass: 20-40 parts of Sargentodon dahurica, 10-20 parts of Cortex Acanthopanacis, 20-40 parts of Herba Cynanchifoliae, 20-40 parts of Drynariae Rhizoma and 10-30 parts of Herba Polygoni Multiflori.

[0008] Preferably, the traditional Chinese medicine composition for treating rheumatoid arthritis of the present invention is composed of the following components in parts by mass: 25-35 parts of Sargentodon grandiflora, 12-18 parts of Acanthopanax acanthopanax, 25-35 parts of Rhizoma Cyperi, 25-35 parts of Rhizoma Drynariae, and 15-25 parts of Herba Polygoni Multiflori.

[0009] Further preferably, the traditional Chinese medicine composition for treating rheumatoid arthritis of the present invention is composed of the following components in parts by mass: 30 parts of Sargentophyllum cuneata, 15 parts of Acanthopanax acanthopanax, 30 parts of Rhizoma Cyperi, 30 parts of Rhizoma Drynariae, and 20 parts of Herba Polygoni Multiflori.

[0010] The preparation method of the Chinese medicine composition for treating rheumatoid arthritis of the present invention comprises the following steps: S1. Extraction: Weigh the five medicinal materials of Sargentodon grandiflorus, Acanthopanax acanthopanax, Radix Angelicae Pubescentis, Rhizoma Drynariae and Herba Polygoni Multiflori according to the formula, add 10-15 times the total weight of the medicinal materials into water, heat to boiling, then simmer for 0.5-1.5 hours; S2, filtration, concentration: Filter the decoction mixed with Chinese medicinal materials obtained in step S1, remove the medicinal residue, take the filtrate, and concentrate it over a slow fire for 0.5-1.5 hours to obtain a decoction; S3. Preparation: The decoction obtained in step S2 is further heated and boiled, and concentrated into a clear paste with a relative density of 1.05-1.15 at 60° C., and pharmaceutically acceptable excipients are added to prepare a pharmaceutically acceptable preparation.

[0011] Preferably, in the method for preparing the traditional Chinese medicine composition for treating rheumatoid arthritis of the present invention, the amount of water added in step S1 is 12 times the total mass of the medicinal materials.

[0012] Preferably, in the method for preparing the traditional Chinese medicine composition for treating rheumatoid arthritis of the present invention, the time for simmering over low heat in step S1 is 1 hour.

[0013] Preferably, in the method for preparing the traditional Chinese medicine composition for treating rheumatoid arthritis of the present invention, the time for slow-heat concentration in step S2 is 1 hour.

[0014] Preferably, in the method for preparing the traditional Chinese medicine composition for treating rheumatoid arthritis of the present invention, the preparation in step S3 includes a solid preparation or a liquid preparation.

[0015] Further preferably, in the method for preparing the traditional Chinese medicine composition for treating rheumatoid arthritis of the present invention, the solid preparations include tablets, capsules, granules, and pills, and the liquid preparations include oral solutions and injections.

[0016] The invention relates to the use of the traditional Chinese medicine composition in preparing medicine for treating rheumatoid arthritis.

[0017] Beneficial effects of the present invention: The present invention provides a traditional Chinese medicine composition for treating rheumatoid arthritis, comprising five medicinal herbs: Sargentodon grandiflora, Acanthopanax acanthopanax, Rhizoma Cyperi, Rhizoma Drynariae, and Herba Polygoni Multiflori. Animal experiments have demonstrated that the present traditional Chinese medicine composition can improve arthritis in a rat model of rheumatoid arthritis (CIA), reduce joint swelling, lower AI scores, decrease secretion of inflammatory factors such as TNF-α, IL-1β, and IL-6, promote IL-10 secretion, regulate the balance of M1 / M2 macrophages, and improve expression of HIF-1α, GPX4, SLC7A11, and GSH proteins, thereby treating RA. Furthermore, clinical trials have demonstrated that the present traditional Chinese medicine composition, combined with conventional Western medicine, can improve the post-treatment ACR20 / 50 / 70 achievement rate and overall efficacy rate (75.76% in the treatment group and 66.67% in the control group) compared to conventional Western medicine treatment for RA with cold-dampness arthritis. This composition significantly improves symptoms of cold-dampness arthritis and enhances patients' quality of life. It is also safe and has no significant adverse reactions, demonstrating its high clinical application value. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 Photos of joint swelling in rats in each group; Figure 2 HE staining results of the joints of rats in each group; Figure 3 Flow cytometry expression of M1 and M2 macrophages in each group; Figure 4 These are the expression bands of HIF-1α, GPX4, SLC7A11, GSH, and β-ACTIN in each group. DETAILED DESCRIPTION

[0019] The following is a detailed description of the technical solution of the present invention in conjunction with specific embodiments. The following embodiments are only for explanation and illustration, and do not constitute a limitation of the technical solution of the present invention.

[0020] Example 1 A traditional Chinese medicine composition for treating rheumatoid arthritis, comprising: 30 g of Sargentodon grandiflora, 15 g of Acanthopanax acanthopanax, 30 g of Radix Glehniae, 30 g of Rhizoma Drynariae, and 20 g of Herba Polygoni Multiflori.

[0021] Example 2 A traditional Chinese medicine composition for treating rheumatoid arthritis, comprising: 20 g of Sargentodon grandiflora, 10 g of Acanthopanax acanthopanax, 20 g of Radix Glehniae, 20 g of Rhizoma Drynariae, and 13 g of Herba Polygoni Multiflori.

[0022] Example 3 A traditional Chinese medicine composition for treating rheumatoid arthritis, comprising: 25 g of Sargentodon grandiflora, 12 g of Acanthopanax acanthopanax, 25 g of Radix Glehniae, 25 g of Rhizoma Drynariae, and 17 g of Herba Polygoni Multiflori.

[0023] Example 4 A traditional Chinese medicine composition for treating rheumatoid arthritis, comprising: 35 g of Sargentodon grandiflorus, 17 g of Acanthopanax acanthopanax, 35 g of Radix Glehniae, 35 g of Rhizoma Drynariae, and 23 g of Herba Polygoni Multiflori.

[0024] Example 5 A traditional Chinese medicine composition for treating rheumatoid arthritis, comprising: 40 g of Sargentodon grandiflora, 20 g of Acanthopanax acanthopanax, 40 g of Radix Glehniae, 40 g of Rhizoma Drynariae, and 27 g of Herba Polygoni Multiflori.

[0025] The Chinese medicine compositions of Examples 1-5 are prepared by any one of the preparation methods of Examples 6-9.

[0026] Example 6 The preparation method of the traditional Chinese medicine composition for treating rheumatoid arthritis comprises the following steps: (1) Extraction: According to the formula composition in Example 1, five medicinal materials, namely, Sargentophyllum grandiflorum, Acanthopanax acanthopanax, Herba Lycopodii, Rhizoma Drynariae and Herba Polygoni Multiflori, were weighed and 1500 mL of water was added. The mixture was heated to boiling and then simmered for 1 h until the volume of the herbal decoction reached 800 mL.

[0027] (2) Filtration and concentration The decoction mixed with Chinese medicinal materials obtained in step (1) was filtered to remove the medicinal residue, and the filtrate was concentrated over a slow fire for 1 h until the volume of the decoction was 300 mL to obtain a decoction.

[0028] Example 7 The preparation method of the traditional Chinese medicine composition for treating rheumatoid arthritis comprises the following steps: (1) Extraction: According to the formula composition in Example 1, five medicinal materials, including Sargentophyllum grandiflorum, Acanthopanax acanthopanax, Radix Angelicae Pubescentis, Rhizoma Drynariae and Herba Polygoni Multiflori, were weighed and added with 1250 mL of water. The mixture was heated to boiling and then simmered for 0.5 h until the volume of the Chinese medicine decoction reached 900 mL.

[0029] (2) Filtration and concentration The decoction mixed with Chinese medicinal materials prepared in step (1) was filtered to remove the medicinal residue, and the filtrate was concentrated over a slow fire for 1.2 h. The decoction was concentrated by heating until the volume of the decoction was 300 mL to obtain a decoction.

[0030] Example 8 The preparation method of the traditional Chinese medicine composition for treating rheumatoid arthritis comprises the following steps: (1) Extraction: According to the formula composition in Example 1, five medicinal materials, namely, Sargentophyllum grandiflorum, Acanthopanax acanthopanax, Herba Lycopodii, Rhizoma Drynariae and Herba Polygoni Multiflori, were weighed and 1800 mL of water was added. The mixture was heated to boiling and then simmered for 1.5 h until the volume of the herbal decoction reached 800 mL.

[0031] (2) Filtration and concentration The decoction mixed with Chinese medicinal materials obtained in step (1) was filtered to remove the medicinal residue, and the filtrate was concentrated over a slow fire for 0.8 h. The decoction was concentrated by heating until the volume of the decoction was 400 mL to obtain a decoction.

[0032] Example 9 The preparation method of the traditional Chinese medicine composition for treating rheumatoid arthritis comprises the following steps: (1) Extraction: According to the formula composition in Example 1, five medicinal materials, namely, Sargentophyllum grandiflorum, Acanthopanax acanthopanax, Herba Lycopodii, Rhizoma Drynariae and Herba Polygoni Multiflori, were weighed and 1500 mL of water was added. The mixture was heated to boiling and then simmered for 1 h until the volume of the herbal decoction reached 800 mL.

[0033] (2) Filtration and concentration The decoction mixed with Chinese medicinal materials obtained in step (1) was filtered to remove the medicinal residue, and the filtrate was concentrated over a slow fire for 1 h until the volume of the decoction was 300 mL to obtain a decoction.

[0034] (3) Preparation The decoction obtained in step S2 is further heated and boiled, concentrated into a clear paste with a relative density of 1.05-1.15 at 60° C., and pharmaceutically acceptable excipients are added to prepare pharmaceutically acceptable preparations, including tablets, capsules, granules, oral liquids, and decoctions.

[0035] Example 10 The Chinese medicine composition prepared by the preparation method of Example 6 is used to treat rheumatoid arthritis. The usage and dosage are as follows: after cooling to room temperature, take 100 mL each after breakfast, lunch, and dinner.

[0036] In order to further verify the reliability of the present invention, the inventors conducted a series of experiments, as follows: 1. Analysis of the formula of Xueteng Qubi Decoction The Xueteng Qubi Decoction, consisting of 30 g of Daxueteng (Snow Vine) root, 15 g of Acanthopanax Cortex, 30 g of Jianxuefei (Snow Fly), 30 g of Drynaria fortunei (Rhizoma Drynariae), and 20 g of Tougucao (Cao Tong Gu Cao), was developed under the guidance of Dong medicine theory. The "Five-in-One" theory is the core of Dong medicine. It states that heaven, earth, air, water, and man form an organic whole. It emphasizes the dialectical relationship between nature, which comprises heaven, earth, air, and water, and man. Dong medicine classifies and names diseases into 12 categories: wind, syndrome, shock, dysentery, sores, sha, and pain. Among these, there are 72 categories of "wind" and "wind-related diseases." Wind-related diseases are caused by wind pathogens or physical weakness, resulting in wind-induced symptoms. Professor Long Yunguang, a renowned traditional Chinese medicine practitioner and Dong medicine scholar, concluded: "The vast majority of conditions referred to as 'wind-related' and 'wind-related diseases' in Dong medicine are related to wind pathogens, with only a few terms having no substantive meaning." Based on the theory of wind-qi diseases and his clinical experience treating RA, Professor Long Yunguang believes that RA is a type of "wind-qi disease." Dong medicine believes that "excess qi or insufficient qi causes illness, and when a person dies, their qi and essence transform into water." A disequilibrium between qi and water leads to illness; the shift from quantitative to qualitative change in qi and water is the pathological process of disease. Therefore, RA is caused by insufficient qi and blood in the body, coupled with the invasion of unhealthy winds during the four seasons, which obstructs the flow of qi in the meridians and internal organs, leading to paralysis. Integrating the Dong medicine principle of "differentiating symptoms, identifying causes, and seeking the underlying cause," Long Yunguang proposed the treatment principle of "dispersing pathogenic winds, harmonizing the internal organs, and supplementing with emotional regulation." Based on the Dong medicine theory of the six properties and six flavors of herbs, the principle for treating RA is that bitter flavors reduce fever, numb (spicy) flavors dispel cold and promote blood circulation, fragrant flavors reduce swelling and relieve pain, and sweet flavors replenish blood and qi.

[0037] The following is an analysis of the Xueteng Qubi Decoction prescription based on the main ingredients, auxiliary ingredients, and supplementary ingredients: (1) Main drug Sargentodoxae, with its bitter and neutral properties, is known for its detoxifying, carbuncle-relieving, blood-activating, pain-relieving, wind-removing, and insecticidal properties. Its bitterness reduces fever, while its cooling properties clear away internal heat and aid in detoxification. Sargentodoxae, with its blood-activating, blood-stasis-removing, and meridian-dredging properties, is also known for its anti-inflammatory and analgesic properties. It is particularly effective for treating joint swelling and pain caused by rheumatoid arthritis (RA).

[0038] Drynaria fortunei, with its bitter flavor and warm nature, is used to tonify the kidneys and bones, heal wounds, and relieve pain. It helps enhance joint repair, reduce damage caused by rheumatoid arthritis, and warms and tonifies cold and dampness, making it suitable for treating joint pain and dysfunction caused by rheumatism, cold, and dampness. Its bitterness has heat-clearing and anti-inflammatory properties.

[0039] (2) Helping with medicine Acanthopanax bark, with a pungent, bitter flavor and warm nature, has the effects of dispelling rheumatism, nourishing the liver and kidneys, and strengthening bones and tendons. It can warm and unclog joints, relieving pain caused by cold and dampness, especially when wind, cold, and dampness block the joints. It can warm and unclog the meridians, dispel cold and dampness, and help the effects of Sargentophyllum scabra and Drynaria fortunei.

[0040] Jianxuefei (Jianxuefei) is pungent and warm in nature, and has the effects of dispelling wind and cold, promoting blood circulation and relaxing tendons, and relieving pain. Jianxuefei helps promote blood circulation, eliminate pain and swelling caused by blood stasis, and enhance the overall effect of the prescription on the joints, further relieving the symptoms of rheumatic joints.

[0041] (3) Prescription Herba pelargonidis (Pelargonidis) is pungent and warm in nature. It has the effects of dispelling wind, removing dampness, relaxing muscles, promoting blood circulation, and relieving pain. By accelerating blood circulation and metabolism, it helps improve blood supply to joints, alleviating swelling and pain caused by inflammation, particularly in the chronic stages of rheumatoid arthritis. Its pungent flavor warms the meridians and promotes smooth blood flow.

[0042] The Xueteng Qubi Decoction formula works together to dispel wind and dampness, activate blood circulation, and dredge the meridians. Its combination, based on the theory of six properties and six flavors, serves a clear therapeutic purpose: the main herbs (Xueteng and Drynaria) focus on reducing fever, promoting blood circulation and removing blood stasis, and promoting menstruation and dispersing blood stasis, helping to alleviate the inflammation and pain of rheumatoid arthritis; the auxiliary herbs (Acanthopanax and Jianxuefei) further enhance the effects of promoting blood circulation and removing blood stasis, eliminating the swelling and pain caused by blood stasis; and the auxiliary herb (Tougucao) supplements the functions of activating meridians and activating meridians, detoxifying, and reducing swelling, further enhancing the efficacy of the entire formula. Xueteng Qubi Decoction dispels wind and dampness, activates blood circulation, and dredges the meridians. When treating rheumatoid arthritis symptoms (such as joint swelling and pain, and limited mobility), this formula can alleviate joint discomfort by comprehensively regulating systemic rheumatic pathology.

[0043] 2. Preparation method of Xueteng Qubi Decoction (1) Extraction Weigh 30 g of Sargentodon grandiflora, 15 g of Acanthopanax acanthopanax, 30 g of Herba Cyperi scabrae, 30 g of Drynaria fortunei, and 20 g of Herba schizonepetae; add 1500 mL of water; heat to boiling; then simmer for 1 h until the volume of the decoction reaches 800–850 mL.

[0044] (2) Filtration Filter the soup mixed with Chinese medicinal materials through a strainer to remove the residue and retain the soup.

[0045] (3) Concentration Transfer the filtered decoction to a decoction pot and concentrate it over low heat for 1 h until the volume reaches 300 mL.

[0046] Xueteng Qubi Decoction Usage and Dosage: Cool to room temperature and take 100 mL each morning, noon, and dinner.

[0047] 3. Animal Experiments to Verify the Efficacy of Xueteng Qubi Decoction 1. Experimental methods (1) Experimental animals Seventy-two female Lewis rats (weighing 200 ± 20 g, clean grade, provided by the Animal Experimental Center of Guizhou University of Traditional Chinese Medicine) were randomly divided into six groups: a blank control group; a model group; low-, medium-, and high-dose Xueteng Qubi Decoction groups; and a HIF-1α inhibitor PX-478 group.

[0048] (2) Animal model preparation The CIA rat model exhibits clinical, histopathological, and immunological characteristics similar to those of humans and is currently an internationally accepted animal model of bone destruction. Type II collagen is dissolved in 0.1 mol / L acetic acid to a concentration of 2 mg / mL. Dissolved with stirring, the solution is refrigerated at 4°C overnight, and then mixed with an equal volume of incomplete Freund's adjuvant to emulsify the solution into a type II collagen emulsion. 0.1 mL of the solution is injected intradermally at multiple points at the base of the rat's tail to induce inflammation, thereby establishing a stable CIA model. A booster immunization is administered once on day 8 using the same method. Model success is assessed using an arthritis scoring system, which measures inflammation greater than 4 points in the joint segment from the toe to the ankle.

[0049] (3) Method of administration Rats with successfully established models were gavage-administered with low, medium, and high doses of Xueteng Qubi Decoction and PX-478, respectively. The blank and model groups were gavage-administered with equal volumes of saline. Starting from the first day of administration, the onset of bilateral hind paw swelling in each group and the time to complete swelling in all model groups (100% swelling time) were recorded. Hind paw volumes were measured using a water volumetric instrument on days 7, 14, 21, and 28. Two hours after gavage on day 28, the rats were sacrificed, blood was collected for serum separation, and the skin of the knee and ankle joints of both hind limbs was removed to obtain synovial and joint tissue for various parameters testing.

[0050] (4) Drug dosage Xueteng Qubi Decoction was prepared by soaking the medicinal materials in 7-fold water for 30 minutes, then boiling for 1 hour. The solution was filtered through gauze, then decocted with 7-fold water for 0.5 hours. The solution was filtered through gauze, and the two solutions were combined and concentrated to a concentration of 5 g / mL. The rat dose was approximately 6.17 times the clinical dose for a 60 kg adult, using a 1:2:4 ratio for low:medium:high doses. The dosing volume was 4 mL. The low-dose group received a concentration of 0.64 g / mL, the medium-dose group received 1.29 g / mL, and the high-dose group received 2.57 g / mL. PX-478 was administered at 10 mg / kg three times a week (Monday, Wednesday, and Friday).

[0051] (5) Index detection 1. Swelling rate and pathological morphology of synovial tissue in rats The right hind ankle joint of the rats was marked, and the volume (mL) below the mark was measured using the isohydration method. Joint swelling was measured before modeling, before drug administration, and on days 7, 14, 21, and 28 of drug administration. The joint swelling rate was calculated as (water volume measured at each dose - water volume measured on day 1) / water volume measured on day 1. Three rats were randomly selected from each experimental group. Synovial and cartilage tissues were obtained and fixed in 10% neutral formalin. After decalcification, dehydration, paraffin infiltration, embedding, and sectioning, H&E staining was performed. Pathological changes in the synovial and cartilage tissues were observed under a light microscope.

[0052] 2. AI Scoring of Rat Arthritis The rats' joint involvement was assessed using a 5-point scoring system. The sum of the joint lesion scores for all four limbs constituted the AI ​​(Intensive Assessment) (AI) for each rat. 0: No arthritis; 1: Mild swelling of the little toe joint; 2: Swelling of the little toe joint and the plantar; 3: Swelling of the paw below the ankle; 4: Swelling of all joints, including the ankle. The AI ​​score was calculated based on the degree of joint swelling and deformity. Higher scores indicate more severe joint inflammation, with the highest AI score for each rat being 16.

[0053] 3.ELISA detection of serum cytokines TNF-α, IL-1β, IL-6, and IL-10 Follow the ELISA kit instructions: Add 50 μL of standard solution of varying concentrations and 50 μL of the sample to be tested to the enzyme-coated plate; add 50 μL of the antibody mixture to each well; shake at 400 rpm for 2 hours at room temperature. Wash the plate three times with 300 μL of 1X wash buffer. Add 100 μL of TMB solution to each well and incubate at 400 rpm for 10 minutes in the dark. Add 100 μL of stop solution to each well and react for 1 minute. Measure the OD at 450 nm to calculate the concentration of each sample.

[0054] 4. FCM detection of CD86 and CD206 expression in tissue cells and calculation of M1 / M2 Synovial membranes were digested with 0.5 g / L collagenase IV for 1 hour and 0.25% trypsin for 30 minutes. Single cells were obtained and blocked with 10% rabbit serum for 30 minutes. The supernatant was discarded after centrifugation. Cells were resuspended in 50 µL of PBS and 10 µL each of PE-CD86 and APC-CD206 antibodies were added. The cells were incubated at 4°C in the dark for 30 minutes. After washing once with PBS, the cells were resuspended in 100 µL of PBS and analyzed by flow cytometry.

[0055] 5. Western blot detection of HIF-1α, SLC7A11, GSH, and GPX4 protein expression in synovial tissue Tissue was homogenized at low temperature, lysed on ice for 40 minutes with protein lysis buffer, and centrifuged at 4°C at 12,000 rpm for 20 minutes. The supernatant was collected. Protein concentration was determined by BCA assay: 20 μg of protein sample was subjected to 10% SDS-PAGE electrophoresis, transferred to a PVDF membrane, and blocked in blocking buffer at 37°C for 1 hour. The membrane was incubated with the primary antibody overnight at 4°C. The membrane was washed and incubated with the secondary antibody on a rocking platform at room temperature for 1.5 hours. The membrane was washed, developer solution was added, and the membrane was exposed and developed using a chemiluminescence imaging system. The image was saved. The relative expression level of each protein was calculated using Image J software.

[0056] (2) Results and Analysis (1) Changes in joint swelling The photos of joint swelling of rats in each group are shown in Figure 2. Figure 1 Results showed that rats in the CIA model group gradually developed symptoms after modeling, exhibiting significant paw redness and swelling. Compared with the CIA model group, joint redness and swelling were significantly reduced in the Xueteng Qubi Decoction and PX-478 groups. Compared with the model group, right hind limb ankle swelling was significantly improved in all treatment groups after 1, 2, 3, and 4 weeks of treatment (P < 0.05). The right hind limb ankle joint drainage volume in each group is shown in Table 1.

[0057] ; (2) HE of synovial tissue The results of HE staining of the joints of rats in each group are as follows Figure 2 Synovial pathology results showed that in the blank control group, the synovial tissue in the joint cavity of rats showed normal nuclear and cytoplasmic structures, with no obvious inflammatory changes or pannus formation. In the CIA model group, a large number of inflammatory cells infiltrated, the synovial tissue proliferated and thickened significantly, showing granulomatous changes, with the appearance of neovascularization and pannus formation, and significant damage to the bone structure. In the Xueteng Qubi Decoction and PX-478 groups, the synovial tissue inflammatory cell infiltration and proliferation were significantly reduced compared with the model group, with very little neovascularization, and no obvious damage to the joint bone and cartilage.

[0058] (3) Joint AI index score The results of the joint AI index scores are shown in Table 2. The results showed that Xueteng Qubi Decoction effectively reduced the AI ​​scores of CIA rats. Compared with the model group, the AI ​​scores of all treatment groups decreased significantly after 4 weeks of treatment (P < 0.05).

[0059] ; (4) Expression of inflammatory factors The expression of inflammatory factors is shown in Table 3. The results showed that after 4 weeks of treatment with Jingxueteng Qubi Decoction, TNF-α, IL-1β, and IL-6 were significantly decreased (P < 0.05), and IL-10 was significantly increased (P < 0.05) compared with the CIA model group.

[0060] ; (5) Expression of M1 / M2 macrophages The expression flow cytometry of M1 and M2 macrophages in each group is shown in Figure 2 Figure 3 The expression of M1 / M2 macrophages in each group is shown in Table 4. The results showed that after treatment with Jingxueteng Qubi Decoction, the M1 / M2 value was significantly decreased compared with the CIA model group (P < 0.05).

[0061] ; (6) HIF-1α, GPX4, SLC7A11, and GSH protein expression The expression bands of HIF-1α, GPX4, SLC7A11, GSH and β-ACTIN in each group are as follows Figure 4 The relative expressions of HIF-1α, GPX4, SLC7A11, and GSH are shown in Table 5. The results showed that after treatment with Jingxueteng Qubi Decoction, the relative expressions of HIF-1α, GPX4, SLC7A11, and GSH were significantly decreased compared with the CIA model group (P < 0.05).

[0062] ; (3) Summary Xueteng Qubi Decoction can improve arthritis in a rat model of rheumatoid arthritis (CIA), reducing joint swelling, lowering AI scores, decreasing the secretion of inflammatory factors such as TNF-α, IL-1β, and IL-6, promoting IL-10 secretion, regulating the balance of M1 / M2 macrophages, and improving the expression of HIF-1α, GPX4, SLC7A11, and GSH proteins, thereby treating RA. In summary, the Dong medicine Xueteng Qubi Decoction can treat the inflammatory model of RA, providing a theoretical basis for its further clinical application.

[0063] IV. Clinical Efficacy Verification (I) Inclusion / Exclusion Criteria (1) Inclusion criteria: 1. Applicants must be between 18 and 75 years old, regardless of gender; 2. Patients who are undergoing treatment for the first time; 3. Patients who meet the 2010 ACR / EULRA diagnostic criteria for RA; 4. No contraindications to methotrexate or hydroxychloroquine; 5. Obtain informed consent; 6. It belongs to the syndrome of cold and dampness obstruction.

[0064] (2) Exclusion criteria: 1. Patients with other autoimmune diseases; 2. Pregnant or breastfeeding women; 3. Patients with a history of cardiovascular and cerebrovascular diseases such as stroke, TIA, MI, HF (NYHA grade II-IV), coronary artery surgery (such as angioplasty, stent implantation, bypass grafting, etc.); 4. Those with a history of gastrointestinal ulcer or gastrointestinal bleeding; 5. Patients with active liver disease or abnormal liver function, with AST, ALT or GGT higher than 1.2 times the upper limit of normal; 6. Abnormal renal function, serum creatinine concentration (sCr) higher than 1.2 times the upper limit of normal value; 7. Abnormal laboratory test 7 days before medication, white blood cell count <4.0×10 9 / L, or platelet count <100×10 9 / L; 8. Patients with malignant tumors or mental illness; 9. Those with a history of alcohol or drug dependence, or those who need to use painkillers daily for a long time for any reason; 10. Participated in other clinical research within 3 months before enrollment; 11. The patient has other conditions that the researchers deem inappropriate for participation in this clinical study.

[0065] (2) Observation indicators (1) Characteristics of joint symptoms The number of swollen joints, number of tender joints and duration of morning stiffness were recorded before and after treatment in the two groups.

[0066] (2) TCM symptom score In accordance with the "Guiding Principles for Clinical Research of New Traditional Chinese Medicines (Trial Implementation)", three primary symptoms were included: joint swelling, joint pain, and morning stiffness; and seven secondary symptoms: joint tenderness, limited flexion and extension, migratory joint pain, joint pain aggravated by cold, heaviness of the limbs, aversion to cold and wind, and fatigue. A four-point scoring system was used for evaluation: primary symptoms were scored as 0 (none), 2 (mild), 4 (moderate), and 6 (severe); secondary symptoms were scored as 0 (none), 1 (mild), 2 (moderate), and 3 (severe). The total score ranged from 0 to 39, with higher scores indicating more severe TCM syndromes.

[0067] (3) Inflammatory indicators Before and after treatment, cubital venous blood was collected from each group on an empty stomach in the morning to detect erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) levels.

[0068] (4) Improvement of disease activity Disease activity was assessed using the Disease Activity Score (DAS-28) for 28 joints. DAS28-ESR = [0.56 × sqrt (T28) + 0.28 × sqrt (SW28) + 0.70 × Ln (ESR)] × 1.08 + 0.16. Note: T28 is the number of tender joints: bilateral proximal interphalangeal joints (10), metacarpophalangeal joints (10), wrist joints (2), elbow joints (2), shoulder joints (2), and knee joints (2), for a total of 28 joints tender or painful during passive motion. SW28 is the number of swollen joints: swollen joint counts are calculated based on whether the above 28 joints are swollen.

[0069] The proportion of patients achieving the ACR response criteria of 20%, 50%, and 70% improvement in disease activity (ACR20 / 50 / 70) was recorded. The ACR response criteria are based on the following seven factors: ① improvement in tenderness count; ② improvement in swollen joint count; ③ patient's assessment of pain; ④ patient's global assessment of disease activity; ⑤ physician's global assessment of disease activity; ⑥ HAO-DI score; and ⑦ improvement in acute phase reactants (ESR, CRP). The percentage improvement for each indicator is calculated as (pre-treatment value - post-treatment value) / pre-treatment value × 100%. ACR20: >20% improvement in all seven criteria; ACR50: >50% improvement in all seven criteria; and ACR70: >70% improvement in all seven criteria.

[0070] (5) TCM syndrome efficacy standards Refer to the "Guiding Principles for Clinical Research of New Traditional Chinese Medicines (Trial Implementation)." Clinical cure: TCM clinical symptoms and signs significantly disappear or essentially disappear, with a >95% reduction in the syndrome score; marked efficacy: TCM clinical symptoms and signs significantly improve, with a ≥70% but <95% reduction in the syndrome score; effective: TCM clinical symptoms and signs improve somewhat, with a ≥30% but <70% reduction in the syndrome score; ineffective: TCM clinical symptoms and signs show no significant improvement or even worsen, with a <30% reduction in the syndrome score. Total effective rate = (pre-treatment score - post-treatment score) / pre-treatment score × 100%.

[0071] (6) Adverse reactions Blood routine tests and liver and kidney function tests were performed at the 1st, 4th, 8th and 12th week after the start of treatment, and adverse reactions during the treatment were recorded.

[0072] (3) Treatment methods The control group received methotrexate tablets (specification: 2.5 mg, Shanghai SPH Sinyi Pharmaceuticals Co., Ltd., approval number: National Medicine Standard H31020644; 10 mg / time, once a week, orally) + folic acid (specification: 2.5 mg, Shanghai SPH Sinyi Pharmaceuticals Co., Ltd., approval number: National Medicine Standard H31020644; 10 mg / time, once a week, orally) + hydroxychloroquine tablets (specification: 0.1 g, Shanghai SPH Sino-Western Pharmaceutical Co., Ltd., approval number: National Medicine Standard H19990263; 0.2 g / time, once a day, orally) for treatment.

[0073] Observation group: methotrexate tablets (10 mg / time, once / week, orally) + folic acid (specification: 2.5 mg, Shanghai SPH Xinyi Pharmaceutical Co., Ltd., approval number: National Medicine Standard H31020644; 10 mg / time, once / week, orally) + hydroxychloroquine tablets (0.2 g / time, once / day, orally) were given Xueteng Qubi Decoction.

[0074] Xueteng Qubi Decoction consists of 30 g of Daxueteng, 15 g of Acanthopanax Cortex, 30 g of Jianxuefei, 30 g of Drynaria Rhizoma, and 20 g of Tougucao. It was prepared by the Chinese medicine pharmacy of the Pharmacy Department of the Second Affiliated Hospital of Guizhou University of Traditional Chinese Medicine. 100 mL was taken three times a day. One month was a course of treatment, and the total treatment was three months.

[0075] (IV) Clinical enrollment Patients with RA, admitted to the outpatient and inpatient Rheumatology Department of the Second Affiliated Hospital of Guizhou University of Traditional Chinese Medicine, were randomly divided into a treatment group and a control group. The treatment group included 33 patients (2 males and 31 females) aged 22 to 72 years (mean, 51.45 ± 10.21) years, with a disease duration of 2 to 18 months (mean, 9.27 ± 3.89) months. The control group included 31 patients (3 males and 28 females) aged 33 to 68 years (mean, 51.10 ± 8.29) years, with a disease duration of 3 to 15 months (mean, 8.26 ± 3.71) months. No statistically significant differences in general characteristics were found between the two groups (P > 0.05), indicating comparability. This study protocol was approved by the Medical Ethics Committee of the Second Affiliated Hospital of Guizhou University of Traditional Chinese Medicine (KY202401212).

[0076] (V) Results and Analysis (1) Comparison of joint swelling, joint tenderness, and morning stiffness before and after treatment The results of the comparison of the number of swollen joints, number of tender joints, and duration of morning stiffness before and after treatment in each group are shown in Table 6. The results show that after treatment, the number of swollen joints, number of tender joints, and duration of morning stiffness in the treatment group were all reduced compared with those before treatment (P < 0.05); after treatment, the number of swollen joints, number of tender joints, and duration of morning stiffness in the control group were all reduced (P < 0.05); after treatment, the number of tender joints and duration of morning stiffness in the treatment group were both lower than those in the control group.

[0077] ; (2) Comparison of TCM symptom scores, ESR, and CRP before and after treatment The results of the comparison of TCM symptom scores, ESR, and CRP before and after treatment in each group are shown in Table 7. The results show that after treatment, the TCM symptom scores, ESR, and CRP in the treatment group were lower than those before treatment (P<0.05); after treatment, the TCM symptom scores, ESR, and CRP in the control group were lower (P<0.05); after treatment, the TCM symptom scores, ESR, and CRP in the treatment group were lower than those in the control group (P<0.05).

[0078] ; (3) Comparison of DAS28, ACR20, ACR50 and ACR70 before and after treatment The comparison results of DAS28, ACR20, ACR50, and ACR70 before and after treatment in each group are shown in Table 8. The results show that after treatment, DAS28 in the treatment group was lower than that in the control group (P<0.05); after treatment, DAS28 in the treatment group was lower than that in the control group (P<0.05); after treatment, DAS28 in the treatment group was lower than that in the control group (P<0.05); after treatment, ACR20, ACR50, and ACR70 in the treatment group were all higher than those in the control group (P<0.05).

[0079] ; (4) Comparison of TCM syndrome efficacy The results of the comparison of TCM symptom efficacy are shown in Table 9. The results show that the total effective rate of the treatment group was 75.76%, and the total effective rate of the control group was 66.67%. The TCM symptom efficacy of the treatment group was better than that of the control group (P < 0.05).

[0080] ; (5) Adverse reactions During the treatment, 1 case of nausea and 1 case of mild abdominal distension occurred in the treatment group, with an adverse reaction rate of 6.06%. 2 cases of upper respiratory tract infection and nausea occurred in the control group, with an adverse reaction rate of 6.45%. There was no statistically significant difference in the incidence of adverse reactions between the two groups (P>0.05).

[0081] (VI) Summary Compared with conventional Western medicine treatment, Xueteng Qubi Decoction combined with conventional Western medicine for the treatment of RA with cold-dampness arthralgia syndrome can increase the ACR20 / 50 / 70 achievement rate after treatment, improve the symptoms of cold-dampness arthralgia syndrome, and generally improve the quality of life of patients. It has good safety and no obvious adverse reactions, and has great clinical promotion and application value.

[0082] Although the present invention has been described in detail above using general descriptions, specific embodiments, and experiments, it will be apparent to those skilled in the art that modifications and improvements may be made based on the present invention. Therefore, such modifications and improvements, which do not depart from the spirit of the present invention, are intended to be within the scope of protection claimed herein.

Claims

1. A Chinese medicine composition for treating rheumatoid arthritis, characterized in that: The traditional Chinese medicine composition is composed of the following components in parts by mass: 20-40 parts of Sargentodon grandiflora, 10-20 parts of Acanthopanax acanthopanax, 20-40 parts of Rhizoma Cyperi, 20-40 parts of Rhizoma Drynariae, and 10-30 parts of Herba Polygoni Multiflori.

2. The Chinese medicine composition for treating rheumatoid arthritis according to claim 1, characterized in that The traditional Chinese medicine composition consists of the following components in parts by mass: 25-35 parts of Sargentodon grandiflora, 12-18 parts of Cortex Acanthopanacis, 25-35 parts of Radix Sanguisorbae, 25-35 parts of Rhizoma Drynariae, and 15-25 parts of Herba Scutellariae.

3. The Chinese medicine composition for treating rheumatoid arthritis according to claim 2, characterized in that: The traditional Chinese medicine composition consists of the following components in parts by mass: 30 parts of Sargentodon grandiflorus, 15 parts of Acanthopanax acanthopanax, 30 parts of Rhizoma Cyperi, 30 parts of Rhizoma Drynariae, and 20 parts of Herba Polygoni Multiflori.

4. The method for preparing a Chinese medicine composition for treating rheumatoid arthritis according to any one of claims 1 to 3, wherein: The preparation method comprises the following steps: S1. Extraction: Weigh the five medicinal herbs of Sargentodon grandiflorus, Acanthopanax acanthopanax, Radix Angelicae Pubescentis, Rhizoma Drynariae, and Herba Polygoni Multiflori according to the formula, add 10-15 times the total weight of the herbs in water, heat to boiling, then simmer for 0.5-1.5 h. S2, filtration, concentration: Filter the decoction mixed with Chinese medicinal materials obtained in step S1 to remove the medicinal residue, take the filtrate, and concentrate it over a slow fire for 0.5-1.5 hours to obtain a decoction; S3. Preparation: The decoction obtained in step S2 is further heated and boiled, and concentrated into a clear paste with a relative density of 1.05-1.15 at 60° C., and pharmaceutically acceptable excipients are added to prepare a pharmaceutically acceptable preparation.

5. The preparation method according to claim 4, characterized in that The amount of water added in step S1 is 12 times the total mass of the medicinal materials.

6. The preparation method according to claim 4, characterized in that The simmering time in step S1 is 1 h.

7. The preparation method according to claim 4, characterized in that The time for slow fire concentration in step S2 is 1 h.

8. The preparation method according to claim 4, characterized in that The preparation in step S3 includes a solid preparation or a liquid preparation.

9. The preparation method according to claim 8, characterized in that The solid preparations include tablets, capsules, granules, and pills, and the liquid preparations include oral solutions and injections.

10. Use of the traditional Chinese medicine composition according to any one of claims 1 to 3 in the preparation of a medicament for treating rheumatoid arthritis.

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