Traditional Chinese medicine composition for treating early rheumatoid arthritis and preparation method thereof

Abstract

The invention provides a traditional Chinese medicine composition for treating early rheumatoid arthritis and a preparation method thereof, and the traditional Chinese medicine composition is prepared from the following raw materials in parts by weight: 8-12 parts of radix bupleuri, 8-12 parts of cassia twig, 8-12 parts of radix stephaniae tetrandrae, 15-25 parts of radix clematidis, 10-20 parts of rhizoma corydalis, 5-15 parts of radix scutellariae, 8-16 parts of radix angelicae, 15-25 parts of radix paeoniae alba and 3-8 parts of liquorice. The traditional Chinese medicine composition has the beneficial effects that the traditional Chinese medicine composition for treating early rheumatoid arthritis is a pure traditional Chinese medicine prescription, is good in clinical curative effect, can promote recovery of joint meridians and collaterals, is small in toxic and side effects, is wide in medicine source, is low in cost, and is suitable for popularization and application. The traditional Chinese medicine composition is of great significance in further improving the clinical curative effect of traditional Chinese medicine on early rheumatoid arthritis and preventing joint damage, deformity or dysfunction, and is suitable for clinical popularization and application.

Description

Technical Field

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

[0002] Rheumatoid arthritis (RA) can cause synovial damage and joint destruction in the early stage of the disease. 70% of patients will experience irreversible joint destruction, deformity or loss of function within 2 years after the onset. Moreover, there is currently no specific treatment for this disease. In the early stage, slow-acting anti-rheumatic drugs, non-steroidal anti-inflammatory drugs, glucocorticoids, etc. are usually selected for treatment, but the curative effect is limited and the side effects are obvious. Therefore, strengthening the treatment of early RA is an urgent problem to be solved in clinical practice.

[0003] At present, traditional Chinese medicine compositions are widely used in the preparation of drugs for treating rheumatoid arthritis. However, there is not enough research on the combination treatment of traditional Chinese medicine for early rheumatoid arthritis. Summary of the Invention

[0004] The present application provides a traditional Chinese medicine composition for treating early rheumatoid arthritis and a preparation method thereof, which have good curative effect, high efficiency, short course of treatment, small toxic and side effects and low cost.

[0005] According to some embodiments of the traditional Chinese medicine composition for treating early rheumatoid arthritis of the present application, the traditional Chinese medicine composition comprises the following raw materials in parts by weight: 8 - 12 parts of Bupleuri Radix, 8 - 12 parts of Ramulus Cinnamomi, 8 - 12 parts of Stephaniae Tetrandrae Radix, 15 - 25 parts of Clematidis Radix, 10 - 20 parts of Corydalis Rhizoma, 5 - 15 parts of Scutellariae Radix, 8 - 16 parts of Angelicae Dahuricae Radix, 15 - 25 parts of Paeoniae Radix Alba, and 3 - 8 parts of Glycyrrhizae Radix.

[0006] The traditional Chinese medicine composition for treating early rheumatoid arthritis of the present application can reduce RA disease-related inflammatory indexes such as ESR and CRP, inhibit the abnormal proliferation of RA synovial fibroblasts, reduce the expression of pro-inflammatory factor IL-6, improve synovial inflammation, and improve symptoms such as the number of joint tenderness, swelling number and morning stiffness time of patients, and there is no obvious adverse phenomenon after long-term application.

[0007] According to some embodiments of the traditional Chinese medicine composition for treating early rheumatoid arthritis of the present application, the traditional Chinese medicine composition comprises the following raw materials in parts by weight: 9 - 11 parts of Bupleuri Radix, 9 - 11 parts of Ramulus Cinnamomi, 9 - 11 parts of Stephaniae Tetrandrae Radix, 18 - 22 parts of Clematidis Radix, 12 - 18 parts of Corydalis Rhizoma, 8 - 12 parts of Scutellariae Radix, 10 - 14 parts of Angelicae Dahuricae Radix, 18 - 22 parts of Paeoniae Radix Alba, and 4 - 6 parts of Glycyrrhizae Radix.

[0008] According to some embodiments of the traditional Chinese medicine composition for treating early rheumatoid arthritis described in the present application, the traditional Chinese medicine composition comprises raw materials in the following parts by weight: 10 parts of Bupleuri Radix, 10 parts of Cinnamomi Ramulus, 10 parts of Stephaniae Tetrandrae Radix, 20 parts of Clematidis Radix, 15 parts of Corydalis Rhizoma, 10 parts of Scutellariae Radix, 12 parts of Angelicae Dahuricae Radix, 20 parts of Paeoniae Radix Alba, and 5 parts of Glycyrrhizae Radix.

[0009] Bupleuri Radix is pungent, bitter, and slightly cold in nature. It belongs to the liver, gallbladder, and lung meridians. It has the effects of dispelling wind-dampness and relieving pain, and promoting the flow of blood vessels. Clinically, it is commonly used for symptoms such as common cold with fever, alternating chills and fever, distending pain in the chest and hypochondrium, irregular menstruation, uterine prolapse, rectal prolapse, etc.

[0010] Cinnamomi Ramulus is pungent, sweet, and warm in nature. It belongs to the heart, lung, and bladder meridians. It has the effects of inducing sweating to relieve the exterior syndrome, warming the channels to promote blood circulation, assisting yang to transform qi, and calming the adverse qi to descend. Clinically, it is commonly used for symptoms such as wind-cold cold, cold pain in the epigastrium and abdomen, blood-cold amenorrhea, arthralgia pain, phlegm retention, edema, palpitation, and rushing of qi.

[0011] Stephaniae Tetrandrae Radix is bitter and cold in nature. It belongs to the bladder and lung meridians. It has the effects of expelling wind and relieving pain, and promoting diuresis to reduce edema. Clinically, it is commonly used for symptoms such as wind-dampness arthralgia, edema and beriberi, dysuria, eczema and sores and toxins.

[0012] Clematidis Radix is pungent, salty, and warm in nature. It belongs to the bladder meridian. It has the effects of dispelling wind-dampness and dredging the channels. Clinically, it is commonly used for symptoms such as wind-dampness arthralgia, limb numbness, muscle contracture, and difficulty in flexion and extension.

[0013] Corydalis Rhizoma is pungent, bitter, and warm in nature. It belongs to the liver and spleen meridians. It has the effects of promoting blood circulation, promoting qi movement, and relieving pain. Clinically, it is commonly used for symptoms such as pain in the chest and hypochondrium, epigastrium and abdomen, angina pectoris, amenorrhea and dysmenorrhea, stasis after childbirth, and swelling and pain due to falls and bruises.

[0014] Scutellariae Radix is bitter and cold in nature. It belongs to the lung, gallbladder, spleen, large intestine, and small intestine meridians. It has the effects of clearing heat and drying dampness, purging fire and detoxifying, stopping bleeding, and preventing miscarriage. Clinically, it is commonly used for symptoms such as damp-warm syndrome, summerheat-dampness syndrome, chest distress and nausea, damp-heat stuffiness and fullness, diarrhea and dysentery, jaundice, cough due to lung heat, high fever and restlessness, hematemesis and epistaxis due to blood heat, carbuncles and sores and toxins, and threatened abortion.

[0015] Paeoniae Radix Alba is bitter, sour, and slightly cold in nature. It belongs to the liver and spleen meridians. It has the effects of nourishing blood and regulating menstruation, astringing yin and stopping sweating, soothing the liver and relieving pain, and suppressing the hyperactive liver yang. Clinically, it is commonly used for symptoms such as sallow complexion due to blood deficiency, irregular menstruation, spontaneous sweating, night sweating, hypochondriac pain, abdominal pain, spasmodic pain in the limbs, headache and dizziness.

[0016] Angelicae Dahuricae Radix is pungent and warm in nature. It belongs to the stomach, large intestine, and lung meridians. It has the effects of inducing sweating to dispel cold, expelling wind and relieving pain, promoting nasal passage, drying dampness and stopping leukorrhea, and expelling pus and promoting tissue regeneration. Clinically, it is commonly used for symptoms such as headache due to common cold, pain in the supraorbital bone, nasal congestion and runny nose, allergic rhinitis, nasal sinusitis, toothache, leukorrhea, and swelling and pain of sores and ulcers.

[0017] Licorice root, sweet in flavor and neutral in nature. It acts on the heart, lung, spleen, and stomach meridians. It has the effects of invigorating the spleen and replenishing qi, clearing heat and detoxifying, resolving phlegm and relieving cough, alleviating spasm and pain, and coordinating various medicinal herbs. Clinically, it is commonly used for symptoms such as weakness of the spleen and stomach, lassitude and fatigue, palpitations and shortness of breath, excessive phlegm and cough, spasmodic pain in the epigastric and abdominal regions and the limbs, carbuncles and sores, and relieving the toxicity and potency of drugs.

[0018] In the second aspect of the present application, there is provided a method for preparing the traditional Chinese medicine composition for treating early rheumatoid arthritis described in the first aspect of the present application, comprising the following steps: (1) Decoct and extract Bupleurum chinense, Cinnamon twig, Stephania tetrandra, Clematis chinensis, Corydalis yanhusuo, Scutellaria baicalensis, Dahurian angelica root, White peony root, and Licorice root with water respectively, combine the extraction liquids, filter, and concentrate the filtrate; (2) Mix and stir the concentrate obtained in step (1) with ethanol, let it stand, take the supernatant, and concentrate to obtain a clear extract.

[0019] In some embodiments of the preparation method according to the present application, in step (1), the decocting temperature is 60 - 100 °C, such as 60 °C, 65 °C, 70 °C, 80 °C, 88 °C, 90 °C, 95 °C, 100 °C, etc., and the decocting time is 30 - 60 min, such as 30 min, 35 min, 40 min, 43 min, 48 min, 53 min, 56 min, 60 min, etc.

[0020] In some embodiments of the preparation method according to the present application, in step (2), the mass concentration of the ethanol is 40% - 60%, such as 40%, 46%, 50%, 53%, 60%, etc.

[0021] In some embodiments of the preparation method according to the present application, in step (2), the mass ratio of the concentrate to the ethanol is 1:(8 - 12), such as 1:8, 1:9, 1:10, 1:12, etc.

[0022] In some embodiments of the preparation method according to the present application, in step (2), the viscosity of the clear extract is 1.10 - 1.15 Pa·s.

[0023] In some embodiments of the preparation method according to the present application, the preparation method further comprises mixing the clear extract, sucrose, and dextrin, making soft materials, granulating, and drying to obtain the traditional Chinese medicine composition for treating early rheumatoid arthritis.

[0024] In some embodiments of the preparation method according to the present application, the drying temperature is 40 - 60 °C, such as 40 °C, 50 °C, 55 °C, 60 °C, etc., and the drying time is 1 - 3 h, such as 1 h, 2 h, 3 h, etc.

[0025] The beneficial effects of this application include: The traditional Chinese medicine composition for treating early rheumatoid arthritis in this application uses Bupleuri Radix and Ramulus Cinnamomi as the monarch drugs, which can warm and dredge the meridians, benefit the joints, activate blood circulation, dispel wind-dampness; Stephaniae Tetrandrae Radix is most proficient in expelling wind and dampness; Clematidis Radix is warm in nature, pungent and salty in taste, and is fierce and good at moving, passing through the twelve meridians, which can strengthen the effect of expelling wind-dampness and dredging the meridians; Corydalis Rhizoma is warm in nature, pungent and bitter in taste, promoting blood circulation and regulating qi to relieve pain, and can "regulate qi stagnation in blood and blood stasis in qi, so it can specifically treat various pains in the whole body", and it is the ministerial drug; Scutellariae Radix clears the pathogenic heat in Shaoyang. When combined with Bupleuri Radix, it clears the stagnant heat and promotes the smooth flow of the pivot mechanism. Bupleuri Radix is ascending and dispersing, and when combined with the descending and purging of Scutellariae Radix, there will be no harm of ascending yang and robbing yin. When the healthy qi is strong, the pathogen will have no chance to turn inward and can be directly resolved from the outside, and it is the adjuvant drug; Paeoniae Radix Alba enters the blood aspect, can harmonize the blood vessels and relieve spasm and pain; Angelicae Dahuricae Radix is warm in nature and pungent in taste, and its functions are to disperse cold and expel wind, and promote qi and blood circulation, and it is the adjuvant drug; The dosage of Glycyrrhizae Radix should be light. When Paeoniae Radix Alba is combined with Glycyrrhizae Radix, it is the Paeoniae-Glycyrrhizae Decoction. Zhang Zhongjing used it to relieve spasm and pain, and it is the guiding drug. The whole formula together exerts the effects of expelling wind-dampness and dredging collaterals to relieve pain, and its treatment method is in line with the basic pathogenesis of the wind-damp obstruction syndrome in early RA.

[0026] The traditional Chinese medicine composition for treating early rheumatoid arthritis in this application is a pure traditional Chinese medicine prescription, which has good clinical efficacy, can promote the recovery of joint meridians, has small toxic and side effects. In addition, its medicinal sources are wide and the cost is low, which is of great significance for further improving the clinical efficacy of traditional Chinese medicine in treating early rheumatoid arthritis, preventing joint destruction, deformity or loss of function, and is suitable for clinical promotion and application. Brief Description of the Drawings

[0027] Figure 1 It is an analysis chart of the average level of the inflammatory index ESR of the patient after taking the decoction of the traditional Chinese medicine composition in Example 1 of this application for 4 weeks; Figure 2 It is an analysis chart of the integral of traditional Chinese medicine symptoms of the patient after taking the decoction of the traditional Chinese medicine composition in Example 1 of this application for 4 weeks; Figure 3 It is a microscopic cell morphology chart of the influence of the traditional Chinese medicine composition in this application on human rheumatoid arthritis cells; Figure 4 It is a chart of the influence of the traditional Chinese medicine composition in this application on the proliferation of human rheumatoid arthritis cells; Figure 5 It is a chart of the influence of the traditional Chinese medicine composition in this application on the protein blotting of IL-6 in human rheumatoid arthritis cells; Figure 6 It is a chart of the influence of the traditional Chinese medicine composition in this application on the protein expression of IL-6 in human rheumatoid arthritis cells; Figure 7 It is a chart of the influence of the traditional Chinese medicine composition in this application on the mRNA expression of IL-6 in human rheumatoid arthritis cells. Detailed Description of the Invention

[0028] Embodiments of the present invention will be described in detail below. Examples of the embodiments are shown in the accompanying drawings. The embodiments described below with reference to the accompanying drawings are exemplary and are intended to explain the present invention, and should not be construed as limiting the present invention.

[0029] In the present invention, the terms "one embodiment", "some embodiments", "example", "specific example", or "some examples", etc. mean that the specific features, structures, materials, or characteristics described in connection with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials, or characteristics described can be combined in any one or more embodiments or examples in a suitable manner. In addition, without contradiction, those skilled in the art can combine and combine the different embodiments or examples described in this specification and the features of different embodiments or examples.

[0030] Example 1 A traditional Chinese medicine composition for treating early rheumatoid arthritis, comprising the following raw materials in parts by weight: 8 g of Bupleuri Radix, 8 g of Ramulus Cinnamomi, 9 g of Stephaniae Tetrandrae Radix, 15 g of Clematidis Radix, 12 g of Corydalis Rhizoma, 8 g of Scutellariae Radix, 10 g of Angelicae Dahuricae Radix, 18 g of Paeoniae Radix Alba, and 5 g of Glycyrrhizae Radix.

[0031] The preparation method of the traditional Chinese medicine composition for treating early rheumatoid arthritis according to Example 1 comprises the following steps: (1) Decoct Bupleuri Radix, Ramulus Cinnamomi, Stephaniae Tetrandrae Radix, Clematidis Radix, Corydalis Rhizoma, Scutellariae Radix, Angelicae Dahuricae Radix, Paeoniae Radix Alba, and Glycyrrhizae Radix separately with water at 80 °C for 40 min for extraction, combine the extraction solutions, filter, and concentrate the filtrate; (2) Mix and stir the concentrate obtained in step (1) with ethanol having a mass concentration of 45%, wherein the mass ratio of the concentrate to the ethanol is 1:10, let stand, take the supernatant, and concentrate to obtain a clear paste with a viscosity of 1.10 Pa·s; (3) Mix the clear paste obtained in step (2) with sucrose and dextrin, make soft materials, granulate, and dry at 50 °C for 2 h to obtain the traditional Chinese medicine composition for treating early rheumatoid arthritis.

[0032] Example 2 A traditional Chinese medicine composition for treating early rheumatoid arthritis, comprising the following raw materials in parts by weight: 10 g of Bupleuri Radix, 10 g of Ramulus Cinnamomi, 10 g of Stephaniae Tetrandrae Radix, 20 g of Clematidis Radix, 15 g of Corydalis Rhizoma, 8 g of Scutellariae Radix, 13 g of Angelicae Dahuricae Radix, 20 g of Paeoniae Radix Alba, and 5 g of Glycyrrhizae Radix.

[0033] The preparation method of the traditional Chinese medicine composition for treating early rheumatoid arthritis according to Example 2 comprises the following steps: (1) Decoct Bupleuri Radix, Ramulus Cinnamomi, Stephaniae Tetrandrae Radix, Clematidis Radix, Corydalis Rhizoma, Scutellariae Radix, Angelicae Dahuricae Radix, Paeoniae Radix Alba and Glycyrrhizae Radix separately with water at 80 °C for 40 min for extraction, combine the extraction solutions, filter, and concentrate the filtrate; (2) Mix and stir the concentrate obtained in step (1) with ethanol having a mass concentration of 45%, wherein the mass ratio of the concentrate to the ethanol is 1:10, let it stand and take the supernatant, and concentrate to obtain a clear extract with a viscosity of 1.10 Pa·s; (3) Mix the clear extract obtained in step (2) with sucrose and dextrin, prepare soft materials, granulate, and dry at 50 °C for 2 h to obtain the traditional Chinese medicine composition for treating early rheumatoid arthritis.

[0034] Example 3 A traditional Chinese medicine composition for treating early rheumatoid arthritis, comprising the following raw materials in parts by weight: 12 g of Bupleuri Radix, 12 g of Ramulus Cinnamomi, 12 g of Stephaniae Tetrandrae Radix, 25 g of Clematidis Radix, 20 g of Corydalis Rhizoma, 15 g of Scutellariae Radix, 16 g of Angelicae Dahuricae Radix, 25 g of Paeoniae Radix Alba, and 8 g of Glycyrrhizae Radix.

[0035] The preparation method of the traditional Chinese medicine composition for treating early rheumatoid arthritis according to Example 3 comprises the following steps: (1) Decoct Bupleuri Radix, Ramulus Cinnamomi, Stephaniae Tetrandrae Radix, Clematidis Radix, Corydalis Rhizoma, Scutellariae Radix, Angelicae Dahuricae Radix, Paeoniae Radix Alba and Glycyrrhizae Radix separately with water at 80 °C for 40 min for extraction, combine the extraction solutions, filter, and concentrate the filtrate; (2) Mix and stir the concentrate obtained in step (1) with ethanol having a mass concentration of 45%, wherein the mass ratio of the concentrate to the ethanol is 1:10, let it stand and take the supernatant, and concentrate to obtain a clear extract with a viscosity of 1.10 Pa·s; (3) Mix the clear extract obtained in step (2) with sucrose and dextrin, prepare soft materials, granulate, and dry at 50 °C for 2 h to obtain the traditional Chinese medicine composition for treating early rheumatoid arthritis.

[0036] Example 4 A traditional Chinese medicine composition for treating early rheumatoid arthritis, comprising the following raw materials in parts by weight: 10 g of Bupleuri Radix, 10 g of Ramulus Cinnamomi, 10 g of Stephaniae Tetrandrae Radix, 20 g of Clematidis Radix, 12 g of Corydalis Rhizoma, 10 g of Scutellariae Radix, 12 g of Angelicae Dahuricae Radix, 18 g of Paeoniae Radix Alba, and 6 g of Glycyrrhizae Radix.

[0037] The preparation method of the traditional Chinese medicine composition for treating early rheumatoid arthritis according to Example 4 comprises the following steps: (1) Decoct Bupleuri Radix, Ramulus Cinnamomi, Stephaniae Tetrandrae Radix, Clematidis Radix, Corydalis Rhizoma, Scutellariae Radix, Angelicae Dahuricae Radix, Paeoniae Radix Alba and Glycyrrhizae Radix separately with water at 80 °C for 40 min for extraction, combine the extraction solutions, filter, and concentrate the filtrate; (2) Mix and stir the concentrate obtained in step (1) with ethanol having a mass concentration of 45%, wherein the mass ratio of the concentrate to the ethanol is 1:10. Let it stand and take the supernatant, and concentrate to obtain a clear extract with a viscosity of 1.10 Pa·s; (3) Mix the clear extract obtained in step (2) with sucrose and dextrin, make soft materials, granulate, and dry at 50 °C for 2 h to obtain the traditional Chinese medicine composition for treating early rheumatoid arthritis.

[0038] Comparative Example 1 The difference between Comparative Example 1 and Example 1 is only that: the traditional Chinese medicine composition for treating early rheumatoid arthritis in Comparative Example 1 does not contain the raw materials Corydalis yanhusuo, Paeonia lactiflora, Clematis chinensis, and Glycyrrhiza uralensis.

[0039] Comparative Example 2 The difference between Comparative Example 2 and Example 1 is only that: the traditional Chinese medicine composition for treating early rheumatoid arthritis in Comparative Example 2 does not contain the raw materials Bupleurum chinense, Cinnamomum cassia, Angelica dahurica, Stephania tetrandra, and Scutellaria baicalensis.

[0040] Comparative Example 3 The difference between Comparative Example 3 and Example 1 is only that: the traditional Chinese medicine composition for treating early rheumatoid arthritis in Comparative Example 3 does not contain the raw material Bupleurum chinense, uses Gentiana macrophylla instead of Stephania tetrandra, and Tripterygium wilfordii instead of Angelica dahurica.

[0041] Comparative Example 4 The difference between Comparative Example 4 and Example 1 is only that: the traditional Chinese medicine composition for treating early rheumatoid arthritis in Comparative Example 4 does not contain the raw material Scutellaria baicalensis, uses Sarcandra glabra instead of Angelica dahurica, and Gentiana macrophylla instead of Stephania tetrandra.

[0042] Comparative Example 5 The difference between Comparative Example 5 and Example 1 is only that: the traditional Chinese medicine composition for treating early rheumatoid arthritis in Comparative Example 5 does not contain the raw material Clematis chinensis, and uses Smilax glabra instead of Stephania tetrandra.

[0043] Comparative Example 6 The difference between Comparative Example 6 and Example 1 is only that: the traditional Chinese medicine composition for treating early rheumatoid arthritis in Comparative Example 6 uses Sarcandra glabra instead of Stephania tetrandra, and adds 5 g of Sinomenium acutum.

[0044] Comparative Example 7 The difference between Comparative Example 7 and Example 1 is only that: the traditional Chinese medicine composition for treating early rheumatoid arthritis in Comparative Example 7 uses Tripterygium wilfordii instead of Scutellaria baicalensis.

[0045] Comparative Example 8 The difference between Comparative Example 8 and Example 1 is only that: the traditional Chinese medicine composition for treating early rheumatoid arthritis in Comparative Example 8 uses Smilax glabra instead of Angelica dahurica.

[0046] Comparative Example 9 The difference between Comparative Example 9 and Example 1 is only that: the traditional Chinese medicine composition for treating early rheumatoid arthritis described in Comparative Example 9 does not contain the raw material Angelica dahurica.

[0047] Efficacy study of the traditional Chinese medicine composition for treating early rheumatoid arthritis described in Example 1 of this application and the traditional Chinese medicine compositions for treating early rheumatoid arthritis described in Comparative Examples 1-9.

[0048] 1. Research and test method of the traditional Chinese medicine composition for treating early rheumatoid arthritis in eliminating inflammation and improving symptoms: Randomly select 20 patients with early rheumatoid arthritis, select the traditional Chinese medicine composition for treating early rheumatoid arthritis described in Example 1, use water (the amount of water used for decoction is 10 times the weight of the traditional Chinese medicine composition) to decoct at 80 °C for 40 min, extract and filter 400 ml, take one dose per day, divide it into 2 times, warm and take it in the morning and evening. The average levels of the inflammatory index ESR and the TCM symptom scores of the patients before and after taking the medicine for 4 weeks are respectively as Figure 1 and Figure 2 shown; From Figure 1 and Figure 2 it can be seen that: after taking the decoction of the traditional Chinese medicine composition described in Example 1 of this application for 4 weeks, the ESR level and the TCM symptom scores of the patients decreased significantly (***P<0.001), that is, the traditional Chinese medicine composition described in this application has good clinical efficacy in treating early rheumatoid arthritis.

[0049] Note: Clinical symptom scoring standard: Record the changes in clinical symptoms of patients with early rheumatoid arthritis before and after clinical treatment, and use the scoring method to record according to the degree: none - 0 points, mild - 1 point, moderate - 2 points, severe - 3 points.

[0050] Evaluation of TCM symptom scores: Before and after treatment, refer to the "Guiding Principles for Clinical Research of New Chinese Medicines", and quantify the TCM syndromes according to mild, moderate and severe. Efficacy determination: Clinical cure: TCM clinical symptoms and signs disappear or basically disappear, and the syndrome score is reduced by ≥95%; Marked effect: TCM clinical symptoms and signs are significantly improved, and the syndrome score is reduced by ≥70% and less than 95%; Effective: TCM clinical symptoms and signs are all improved, and the syndrome score is reduced by ≥30% and less than 70%; Ineffective: TCM clinical symptoms and signs are not significantly improved, or even aggravated, and the syndrome score is reduced by less than 30%. The improvement rate of TCM syndrome score = (score before treatment - score after treatment) / score before treatment × 100%.

[0051] 2. Similarity screening: Identify the medical records with a coincidence degree of less than 0.7 between the traditional Chinese medicine compositions in Comparative Examples 1-9 and the traditional Chinese medicine composition of the present invention, and classify them into Group A; the medical records with a coincidence degree of ≥0.7 with the traditional Chinese medicine composition of the present invention are classified into Group B, and the syndrome efficacy of the two groups is evaluated. The classification of Group A and Group B is shown in Table 1.

[0052] Randomly select 250 patients with early rheumatoid arthritis and divide them into 10 groups on average. The traditional Chinese medicine composition for treating early rheumatoid arthritis described in Example 1 of the present application and the traditional Chinese medicine compositions for treating early rheumatoid arthritis described in Comparative Examples 1-9 are respectively decocted with water (the amount of water used for decoction is 10 times the weight of the traditional Chinese medicine composition) at a temperature of 80°C for 40 minutes, filtered to obtain 400 ml, and the 10 groups of patients with early rheumatoid arthritis respectively take the decoctions of the traditional Chinese medicine compositions described in Example 1 and Comparative Examples 1-9. One dose per day, taken in two divided doses, warm taken in the morning and evening. The treatment results before and after taking the medicine for 4 weeks are shown in Table 1.

[0053] Table 1 It can be seen from Table 1 that: 150 medical records with a coincidence degree of ≥0.7 with the traditional Chinese medicine composition of the present invention have a relatively high clinical effective rate. In addition, the medical records with a coincidence degree of ≥0.95 have the highest clinical effective rate and obvious effective rate, that is, the traditional Chinese medicine composition with a relatively high coincidence degree with the present application is an optimized prescription for treating early rheumatoid arthritis, with significant curative effect in clinic and research value.

[0054] 3. Research on the inhibitory effect of the traditional Chinese medicine composition for treating early rheumatoid arthritis on the proliferation of human rheumatoid arthritis synovial fibroblasts (MH7A cells)

Preparation of medicated serum

[0055] According to the surface area conversion method for humans and animals, the dosage of CBD was 4 times the clinical dosage for humans. Rats in the blank serum group were given an equal amount of distilled water by gavage. In Example 1, 32 g / kg was given by gavage (gavage with the decoction of the traditional Chinese medicine composition described in Example 1 of the present application, and the decoction method refers to the above text). In Comparative Example 1, 14.8 g / kg was given by gavage, and in Comparative Example 2, 17.2 g / kg was given by gavage. Gavage administration was continuously carried out for 7 days. The methotrexate group was given methotrexate tablets at 2 mg / kg by gavage, once every 3 days for 25 consecutive days. 1 h after the last gavage, the rats were anesthetized, and blood was taken from the abdominal aorta under sterile conditions. After standing for 1 h, it was centrifuged at 3000 rpm for 10 min at 4°C, and the supernatant was taken and placed in another EP tube. The collected serum was inactivated at 56°C for 30 min, filtered and sterilized, and then stored in a -80°C refrigerator for standby.

[0056] The cells were divided into a blank group, a model group (10 ng / ml TNF-α), a blank serum group (10 ng / ml TNF-α + NS drug-containing serum), an Example 1 group (10 ng / ml TNF-α + Example 1 drug-containing serum), a Comparative Example 1 group (10 ng / ml TNF-α + Comparative Example 1 drug-containing serum), a Comparative Example 2 group (10 ng / ml TNF-α + Comparative Example 2 drug-containing serum), and a methotrexate group (10 ng / ml TNF-α + MTX drug-containing serum). Test method: MTT method was used to detect the viability of MH7A cells: Human RA-FLS (MH7A cells) (BFN60805399, Qingqi (Shanghai) Biotechnology Co., Ltd.), and the cells were cultured in DMEM medium containing 10% fetal bovine serum and placed in a constant temperature incubator at 37°C with 5% CO2. Take normally cultured cells and seed them in a 96-well plate at 2×10 4 Cells / mL. Add 100 μL of cell suspension to each well. After culturing for 24 h, add the medicinal liquids of different groups to the wells, 180 μL per well, and continue to culture for 24 and 48 h. Add 20 μL of MTT solution to the 96-well plate, incubate for 4 h, discard the supernatant, add 150 μL of DMSO to each well to dissolve the purple crystals, avoid light, gently shake on a shaker for 10 min, and measure the absorbance value (OD) at a wavelength of 490 nm.

[0057] Cell viability (%) = (OD value of sample well - OD value of blank well) / (OD value of normal well - OD value of blank well) × 100%.

[0058] Morphological diagrams of human rheumatoid arthritis cells in Example 1, Comparative Example 1, Comparative Example 2, and the methotrexate control group are as Figure 3 shown.

[0059] The proliferation of human rheumatoid arthritis cells in Example 1, Comparative Example 1, Comparative Example 2, and the methotrexate control group is as Figure 4 shown.

[0060] From Figure 3 and Figure 4 it can be seen that: The traditional Chinese medicine compositions described in Example 1, Comparative Example 1, and Comparative Example 2 can all inhibit the proliferation of MH7A cells induced by TNF-a, increase the reduction or death of adherent cell numbers, and reduce cell-cell connections. The MTT results show that the methotrexate control group significantly inhibits the proliferation of MH7A cells. Example 1, Comparative Example 1, and Comparative Example 2 all inhibit the proliferation of MH7A cells. Among them, the inhibitory effect of the Example 1 group on MH7A cells is the best. Compared with the Example 1 group, the inhibitory effect of the Comparative Example 1 group is weaker, and there is no significant difference between the Comparative Example 1 and Comparative Example 2 groups.

[0061] 4. Research on the traditional Chinese medicine composition for treating early rheumatoid arthritis in reducing the expression of pro-inflammatory factor IL-6 (1) Test method: ① Western blot detection of IL-6 protein expression in MH7A cells: Take normally cultured cells and seed them in a 6-well plate at 2×10 5 Cells / mL, with 2 mL of cell suspension in each well. Incubate for 24 h, and then add the medicinal liquids of different groups (prepared according to the medicinal liquid preparation method in the above 3 test methods) into the wells and continue to incubate for 24 h. Prepare the corresponding volume of lysis buffer at a ratio of RIPA lysis buffer:PMSF:phosphatase inhibitor = 100:1:2 at 4°C, centrifuge at 12,000 rpm for 15 min, and take the supernatant. Use a BCA kit to measure the concentration, calculate the protein concentration, and adjust the samples to the same concentration. The detection of IL-6 protein blotting and expression in MH7A cells is respectively as Figure 5 and Figure 6 shown.

[0062] (2) Quantitative PCR detection of IL-6 mRNA expression in MH7A cells: Use TRIzol reagent (103104, Invitrogen) to isolate total RNA from the isolated MH7A cells. Use Abm Master Mix (G490, Abm) samples for reverse transcription to generate the first-strand complementary DNA. Use SYBR Premix Ex Taq (MasterMix-R, Abm) for quantitative real-time polymerase chain reaction (PCR) on ABI7500, and the results are as Figure 7 shown.

[0063] From Figures 5 - 7It can be seen that the methotrexate control group significantly inhibited the expression of IL-6 protein and mRNA in MH7A cells induced by TNF-α. Examples 1, Comparative Example 1 and Comparative Example 2 all inhibited the expression of IL-6 protein and mRNA in MH7A cells induced by TNF-α. Among Examples 1, Comparative Example 1 and Comparative Example 2, the inhibitory effect of the group of Example 1 on the expression of IL-6 mRNA in MH7A cells induced by TNF-α was the best; there was no significant difference between the groups of Comparative Example 1 and Comparative Example 2. It shows that the traditional Chinese medicine composition for treating early rheumatoid arthritis has significant advantages in reducing the expression of the pro-inflammatory factor IL-6.

[0064] Although the above embodiments have been shown and described, it can be understood that the above embodiments are exemplary and should not be construed as limiting the present invention. Any changes, modifications, substitutions and variations made by those of ordinary skill in the art to the above embodiments are within the protection scope of the present invention.

Claims

1. A traditional Chinese medicine composition for treating early rheumatoid arthritis, characterized in that, It comprises the following raw materials in parts by weight: 8 - 12 parts of Bupleuri Radix, 8 - 12 parts of Ramulus Cinnamomi, 8 - 12 parts of Stephaniae Tetrandrae Radix, 15 - 25 parts of Clematidis Radix, 10 - 20 parts of Corydalis Rhizoma, 5 - 15 parts of Scutellariae Radix, 8 - 16 parts of Angelicae Dahuricae Radix, 15 - 25 parts of Paeoniae Radix Alba, and 3 - 8 parts of Glycyrrhizae Radix et Rhizoma.

2. The traditional Chinese medicine composition for treating early rheumatoid arthritis according to claim 1, wherein, It comprises the following raw materials in parts by weight: 9 - 11 parts of Bupleuri Radix, 9 - 11 parts of Ramulus Cinnamomi, 9 - 11 parts of Stephaniae Tetrandrae Radix, 18 - 22 parts of Clematidis Radix, 12 - 18 parts of Corydalis Rhizoma, 8 - 12 parts of Scutellariae Radix, 10 - 14 parts of Angelicae Dahuricae Radix, 18 - 22 parts of Paeoniae Radix Alba, and 4 - 6 parts of Glycyrrhizae Radix et Rhizoma.

3. The traditional Chinese medicine composition for treating early rheumatoid arthritis according to claim 1, wherein, It comprises the following raw materials in parts by weight: 10 parts of Bupleuri Radix, 10 parts of Ramulus Cinnamomi, 10 parts of Stephaniae Tetrandrae Radix, 20 parts of Clematidis Radix, 15 parts of Corydalis Rhizoma, 10 parts of Scutellariae Radix, 12 parts of Angelicae Dahuricae Radix, 20 parts of Paeoniae Radix Alba, and 5 parts of Glycyrrhizae Radix et Rhizoma.

4. The preparation method of the traditional Chinese medicine composition for treating early rheumatoid arthritis according to any one of claims 1-3, characterized in that, It comprises the following steps: (1) Decoct and extract Bupleuri Radix, Ramulus Cinnamomi, Stephaniae Tetrandrae Radix, Clematidis Radix, Corydalis Rhizoma, Scutellariae Radix, Angelicae Dahuricae Radix, Paeoniae Radix Alba, and Glycyrrhizae Radix et Rhizoma with water respectively, combine the extraction solutions, filter, and concentrate the filtrate; (2) Mix and stir the concentrate obtained in step (1) with ethanol, let it stand and take the supernatant, and concentrate to obtain a clear extract.

5. The preparation method of the traditional Chinese medicine composition for treating early rheumatoid arthritis according to claim 4, characterized in that, In step (1), the decocting temperature is 60 - 100 °C, and the decocting time is 30 - 60 min.

6. The preparation method of the traditional Chinese medicine composition for treating early rheumatoid arthritis according to claim 4, characterized in that, In step (2), the mass concentration of the ethanol is 40% - 60%.

7. The preparation method of the traditional Chinese medicine composition for treating early rheumatoid arthritis according to claim 4, wherein In step (2), the mass ratio of the concentrate to the ethanol is 1:(8 - 12).

8. The preparation method of the traditional Chinese medicine composition for treating early rheumatoid arthritis according to claim 4, characterized in that, In step (2), the viscosity of the clear extract is 1.10 - 1.15 Pa·s.

9. The preparation method of the traditional Chinese medicine composition for treating early rheumatoid arthritis according to claim 4, characterized in that, The preparation method further comprises mixing the clear extract, sucrose, and dextrin, making soft materials, granulating, and drying to obtain the traditional Chinese medicine composition for treating early rheumatoid arthritis.

10. The preparation method of the traditional Chinese medicine composition for treating early rheumatoid arthritis according to claim 9, characterized in that, The drying temperature is 40 - 60 °C, and the drying time is 1 - 3 h.

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