Traditional Chinese medicine composition for treating rheumatic arthritis as well as preparation method and application of traditional Chinese medicine composition

Abstract

The invention discloses a traditional Chinese medicine composition for treating rheumatic arthritis and a preparation method and application thereof.Acanthopanax senticosus and schisandra chinensis are compatible to form the traditional Chinese medicine composition, the Acanthopanax senticosus contains multiple active ingredients such as eleutheroside and the like, has a remarkable anti-inflammatory effect and can inhibit release of inflammatory mediators, and therefore the arthritis reaction is relieved; schisandra chinensis also contains anti-inflammatory components such as schisandrin, and the schisandra chinensis and schisandrin are combined for use, so that the anti-inflammatory effect is enhanced through a synergistic effect, infiltration of inflammatory cells in joint parts is reduced, and joint swelling and pain are relieved.

Description

Technical Field

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

[0002] At present, there are two methods for treating rheumatoid arthritis: traditional Chinese medicine and Western medicine. The treatment method mainly based on Western medicine includes: using non-steroidal anti-inflammatory drugs such as aspirin enteric-coated tablets, ibuprofen capsules, etc., which can inhibit the synthesis of inflammatory factors; or using glucocorticoid drugs such as prednisone acetate tablets, dexamethasone acetate tablets, etc., which can reduce the release of inflammatory factors and thus relieve the discomfort caused by rheumatoid arthritis. However, Western medicine has great side effects and is not suitable for long-term use.

[0003] Acanthopanax and Schisandra chinensis have been widely used as medicines in Chinese medicine for a long time. Acanthopanax has the effects of "tonifying the middle and benefiting the essence, strengthening the tendons and bones, and strengthening the will". Taking it for a long time can "lighten the body and resist aging". When combined with other medicines, it can also "improve diet, strengthen the energy, and not forget things". Schisandra chinensis has the effects of astringing, strengthening qi and promoting body fluid, and tonifying the kidney and calming the mind. There are related Chinese medicine patents reporting that a single medicinal material is used to treat rheumatoid arthritis, such as CN102058677B, a Chinese medicine composition for treating rheumatoid arthritis: the Chinese medicine The composition is composed of the following raw materials in weight ratio, wherein: 3-15% of Angelica dahurica, 5-20% of Acanthopanax senticosus, 5-25% of Caulis Sinomenii, 5-20% of Caulis Piperis Kadzus, 3-15% of Notopterygium wilfordii, 5-25% of Sargentodoxae, 5-20% of Polygonum cuspidatum, 6-30% of Sargentodoxae, and 8-25% of Cibotium barometz. It has the functions of dispelling wind and dampness, relieving pain, and is used for treating rheumatoid arthritis and the like. However, in the preparation process, the raw materials need to be soaked overnight after being added, and then distilled and extracted for 3-7 hours, which takes a long time, has low production efficiency, and is not conducive to industrialization. There is no research report on the combination of Acanthopanax senticosus and Schisandra chinensis for treating rheumatoid arthritis.

[0004] Moreover, some conventional methods for extracting traditional Chinese medicine, such as fluid extract preparation, ethanol reflux or water extraction and alcohol precipitation, cannot obtain the ideal content of effective ingredients when extracting Acanthopanax and Schisandra chinensis, which increases the cost and waste of resources. Summary of the invention

[0005] The main purpose of the present invention is to provide a Chinese medicine composition for treating rheumatoid arthritis and its preparation method and application, aiming to solve the technical problems of the existing Chinese medicine composition for treating rheumatoid arthritis, such as long preparation process and low transfer rate of effective ingredients.

[0006] In order to achieve the above object, the first aspect of the present invention provides a method for preparing a Chinese medicine composition for treating rheumatoid arthritis, comprising the following steps:

[0007] S1: Take Acanthopanax senticosus and Schisandra chinensis and add them to ethanol for reflux extraction. After filtering the extract, concentrate the filtrate, add water to dilute the concentrate, filter again, and take the supernatant for later use;

[0008] S2: adsorbing the supernatant with a first resin, washing with water until neutral, discarding the washing liquid, eluting the resin with a sodium hydroxide solution to obtain an eluent A, then washing with water until neutral, adding ethanol for elution, and obtaining an eluent B;

[0009] S3: The eluent A is adjusted to neutral with a pH adjuster, centrifuged and filtered, the supernatant is adsorbed by a second resin, washed with water until it is salt-free, and then eluted with ethanol, the eluent is collected and concentrated, and then vacuum dried to obtain a dry paste A;

[0010] S4: vacuum concentrating the eluate B, and vacuum drying to obtain a dry paste B;

[0011] S5: Post-processing the dry paste A and the dry paste B to prepare various dosage forms of Chinese medicine compositions for treating rheumatoid arthritis.

[0012] Furthermore, the step S1 comprises: adding 6 to 8 times the total amount of ethanol of the Acanthopanax and Schisandra chinensis, refluxing extraction at 80 to 100° C. for 1 to 3 hours, and repeating 1 to 3 times; vacuum concentrating the filtrate at 40 to 80° C. to a relative density of 1.15 to 1.25 (60° C.) to obtain the concentrated solution; diluting the concentrated solution with 2 to 4 times the amount of water, filtering, and obtaining a supernatant.

[0013] Further, the step S2 comprises: adsorbing the supernatant with the first resin, first adding 1 to 3 times the amount of water to wash until neutral, discarding the water washing liquid, then eluting the first resin with 1 to 3 times the amount of 0.1% sodium hydroxide solution to obtain eluent A, then washing with water until neutral, adding 5 to 8 times the amount of 50% to 70% ethanol to elute, and obtaining eluent B;

[0014] The step S3 comprises: adjusting the eluent A to neutrality with hydrochloric acid, adsorbing the supernatant with a second resin, washing with water until it is salt-free, eluting with 4 to 6 times 50% to 70% ethanol, collecting and concentrating the eluent A', and vacuum drying to obtain a dry paste A.

[0015] Furthermore, the volume percentage concentration of the ethanol is 50% to 70%.

[0016] Furthermore, the preparation process of the dry paste includes: vacuum concentrating the eluent A' at 40-80°C to a relative density of 1.20-1.30 (60°C) to obtain a concentrated solution, and vacuum drying the concentrated solution at a temperature of 50-70°C to a moisture content of 3% to obtain the dry paste A; vacuum concentrating the eluent B at 40-80°C to a relative density of 1.20-1.30 (60°C) to obtain a concentrated solution, and vacuum drying the concentrated solution at a temperature of 50-70°C to a moisture content of 3% to obtain a dry paste B.

[0017] Furthermore, the first resin is a D101 macroporous resin with a diameter-to-height ratio of 1:2 to 1:4; and the second resin is a polyamide resin with a diameter-to-height ratio of 1:2 to 1:4.

[0018] Furthermore, the weight ratio of the Acanthopanax senticosus to the Schisandra chinensis is 1:3 to 3:1.

[0019] The second aspect of the present invention provides a Chinese medicine composition for treating rheumatoid arthritis, which is prepared by any one of the above preparation methods.

[0020] Furthermore, the Chinese medicine composition is in the form of granules, capsules or tablets.

[0021] Furthermore, the preparation process of the granules includes: mixing dry pastes A and B, grinding and sieving, adding 1 to 3 times the amount of dextrin to the obtained dry powder and mixing evenly, then adding 50% to 90% ethanol to granulate, the addition amount is 20% to 40%, and drying to obtain the granules;

[0022] The preparation process of the capsule comprises: mixing dry paste A and B, grinding and sieving, adding 1 to 6 times the amount of starch to the obtained dry powder and mixing evenly, then adding 50% to 90% ethanol to granulate, the addition amount is 15% to 30%, drying, mixing and filling in capsules to obtain the obtained capsule;

[0023] The preparation process of the tablet comprises: mixing dry paste A and B, grinding and sieving, adding 4 to 7 times amount of starch to the obtained dry powder and mixing evenly, then adding 50% to 90% ethanol to granulate, the added amount is 15% to 35%, drying, mixing and tableting to obtain the tablet.

[0024] The third aspect of the present invention provides a use of a Chinese medicine composition in preparing a drug for treating rheumatoid arthritis.

[0025] Beneficial effects:

[0026] The invention discloses a Chinese medicine composition for treating rheumatoid arthritis, and a preparation method and application thereof. The Chinese medicine composition is prepared by combining Acanthopanax senticosus and Schisandra chinensis. Acanthopanax senticosus contains multiple active ingredients, such as eleutheroside, which has a significant anti-inflammatory effect and can inhibit the release of inflammatory mediators, thereby alleviating joint inflammatory reactions; Schisandra chinensis also contains ingredients with anti-inflammatory effects, such as schisandrin. The two are used in combination to enhance the anti-inflammatory effect through synergistic effect, reduce the infiltration of inflammatory cells in joints, and relieve joint swelling and pain.

[0027] The preparation method of the Chinese medicine composition of the present invention adopts the first resin and the second resin for adsorption, which can specifically combine the effective ingredients in the Chinese medicine extract; multiple elution processes are successively used, first washing with water to neutrality to remove part of the impurities, and then eluting with sodium hydroxide solution and ethanol respectively to obtain different eluates, and the staged elution method can more finely separate the effective ingredients of Acanthopanax senticosus and the effective ingredients of Schisandra chinensis, and the obtained eluate is further concentrated, thereby improving the purity of the dry paste and the transfer rate of the effective ingredients. At the same time, in the preparation method of the present application, the raw materials do not need to be soaked overnight before distillation, which shortens the extraction cycle and improves the production efficiency.

[0028] The Chinese medicine composition of the present invention effectively regulates the production of various cytokines by inhibiting the expression of chemokines and adhesion factors, as well as the activity of key enzymes in the development of inflammation, thereby affecting the aggregation, adhesion and infiltration of leukocytes to the inflammatory site, thereby achieving the effect of treating rheumatoid arthritis. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 Figure 2 is the change of paw volume of CIA rats in each group;

[0030] Figure 2 This is the rheumatoid arthritis score of CIA rats in each group;

[0031] Figure 3 The expression of serum inflammatory factors in CIA rats in each group;

[0032] Figure 4 are the superoxide dismutase and catalase activities of CIA rats in each group;

[0033] Figure 5 Glutathione peroxidase activity of CIA rats in each group.

[0034] The realization of the purpose, functional features and advantages of the present invention will be further explained in conjunction with embodiments and with reference to the accompanying drawings. DETAILED DESCRIPTION

[0035] The experimental methods in the following examples of the present invention, where no specific conditions are specified, are usually carried out under conventional conditions or under conditions recommended by the manufacturers. The various commonly used chemical reagents used in the examples are all commercially available products.

[0036] Unless otherwise defined, all technical and scientific terms used in the present invention have the same meaning as those commonly understood by those skilled in the art of the present invention. The terms used in the specification of the present invention are only for the purpose of describing specific embodiments and are not intended to limit the present invention.

[0037] The terms "including" and "having" and any variations thereof of the present invention are intended to cover non-exclusive inclusions. For example, a process, method, device, product or equipment comprising a series of steps is not limited to the listed steps or modules, but may optionally include steps not listed, or may optionally include other steps inherent to these processes, methods, products or equipment.

[0038] In order to make the purpose, technical scheme and advantages of the present invention clearer, the present invention is further described in detail below in conjunction with specific embodiments. It should be understood that these descriptions are exemplary only and are not intended to limit the scope of the present invention. In addition, in the following description, the description of known structures and technologies is omitted to avoid unnecessary confusion of the concept of the present invention.

[0039] The present invention is further described in the following examples, but the examples are not intended to limit the scope of protection of the present invention.

[0040] One embodiment of the present invention provides a method for preparing a Chinese medicine composition for treating rheumatoid arthritis, comprising the following steps:

[0041] S1: Take Acanthopanax senticosus and Schisandra chinensis and add them to ethanol for reflux extraction. After filtering the extract, concentrate the filtrate, add water to dilute the concentrate, filter again, and take the supernatant for later use;

[0042] S2: adsorbing the supernatant with a first resin, washing with water until neutral, discarding the washing liquid, eluting the resin with a sodium hydroxide solution to obtain an eluent A, then washing with water until neutral, adding ethanol for elution, and obtaining an eluent B;

[0043] S3: The eluent A is adjusted to neutral with a pH adjuster, centrifuged and filtered, the supernatant is adsorbed by a second resin, washed with water until it is salt-free, and then eluted with ethanol, the eluent is collected and concentrated, and then vacuum dried to obtain a dry paste A;

[0044] S4: vacuum concentrating the eluate B, and vacuum drying to obtain a dry paste B;

[0045] S5: Post-processing the dry paste A and the dry paste B to prepare various dosage forms of Chinese medicine compositions for treating rheumatoid arthritis.

[0046] In the above embodiment, Acanthopanax and Schisandra chinensis are combined into a traditional Chinese medicine composition. Acanthopanax contains multiple active ingredients, such as eleutheroside, which has significant anti-inflammatory effect and can inhibit the release of inflammatory mediators, thereby alleviating joint inflammation reaction; Schisandra chinensis also contains ingredients with anti-inflammatory effect such as schisandrin. The combination of the two can enhance the anti-inflammatory effect through synergistic effect, reduce the infiltration of inflammatory cells in the joints, and relieve joint swelling and pain.

[0047] In the above embodiment, the first resin and the second resin are used for adsorption, which can specifically bind the effective ingredients in the Chinese medicine extract; multiple elution processes are used successively, first washing with water to neutrality to remove some impurities, and then eluting with sodium hydroxide solution and ethanol respectively to obtain different eluates. The staged elution method can more finely separate the effective ingredients of Acanthopanax senticosus and the effective ingredients of Schisandra chinensis, and the obtained eluate is further concentrated to improve the purity of the dry paste.

[0048] In the above embodiments, the Chinese medicine composition effectively regulates the production of various cytokines by inhibiting the expression of chemokines and adhesion factors, as well as the activity of key enzymes in the development of inflammation, thereby affecting the aggregation, adhesion and infiltration of leukocytes to the inflammatory site, thereby achieving the effect of treating rheumatoid arthritis.

[0049] In one embodiment, the step S1 comprises: adding 6 to 8 times the total amount of ethanol of the Acanthopanax and Schisandra chinensis, refluxing and extracting at 80 to 100° C. for 1 to 3 hours, and repeating 1 to 3 times; vacuum concentrating the filtrate at 40 to 80° C. to a relative density of 1.15 to 1.25 (60° C.) to obtain the concentrated solution; diluting the concentrated solution with 2 to 4 times the amount of water, filtering, and obtaining a supernatant.

[0050] In one embodiment, the step S2 comprises: adsorbing the supernatant with the first resin, first adding 1 to 3 times the amount of water to wash until neutral, discarding the water washing liquid, then eluting the first resin with 1 to 3 times the amount of a sodium hydroxide solution with a mass percentage concentration of 0.1% to obtain an eluent A, then washing with water until neutral, adding 5 to 8 times the amount of ethanol with a volume percentage concentration of 50% to 70% to elute, and obtaining an eluent B;

[0051] The step S3 comprises: adjusting the eluent A to neutrality with hydrochloric acid, adsorbing the supernatant with a second resin, washing with water until it is salt-free, eluting with 4 to 6 times the volume percentage of 50% to 70% ethanol, collecting and concentrating the eluent A', and vacuum drying to obtain a dry paste A.

[0052] In one embodiment, the preparation process of the dry paste includes: vacuum concentrating the eluent A' or the eluent B at 40-80°C to a relative density of 1.20-1.30 (60°C) to obtain a concentrated solution, and vacuum drying the concentrated solution at a temperature of 50-70°C to a moisture content of 3% to obtain the dry paste A or dry paste B.

[0053] In one embodiment, the first resin is a D101 macroporous resin with an aspect ratio of 1:2 to 1:4, and the second resin is a polyamide resin with an aspect ratio of 1:2 to 1:4.

[0054] In one embodiment, the weight ratio of the Acanthopanax senticosus to the Schisandra chinensis is 1:3 to 3:1.

[0055] In one embodiment, the dosage form of the Chinese medicine composition includes: granules, capsules and tablets.

[0056] In one embodiment, the preparation process of the granules includes: mixing dry pastes A and B, grinding and sieving, adding 1 to 3 times the amount of dextrin to the obtained dry powder and mixing evenly, then adding 50% to 90% ethanol to granulate, the addition amount is 20% to 40%, and drying to obtain;

[0057] The preparation process of the capsule comprises: mixing dry paste A and B, grinding and sieving, adding 1 to 6 times the amount of starch to the obtained dry powder and mixing evenly, then adding 50% to 90% ethanol to granulate, the addition amount is 15% to 30%, drying, mixing and filling in capsules to obtain the obtained capsule;

[0058] The preparation process of the tablet comprises: mixing dry paste A and B, grinding and sieving, adding 4 to 7 times amount of starch to the obtained dry powder and mixing evenly, then adding 50% to 90% ethanol to granulate, the added amount is 15% to 35%, drying, mixing and tableting to obtain the tablet.

[0059] The specific embodiments are as follows:

[0060] Example 1

[0061] S1: Take 750g of Acanthopanax senticosus and 250g of Schisandra chinensis, add them to 8000g of 65% ethanol by volume, extract them under reflux for 3 times at 80-100°C, filter the extract to obtain a filtrate, concentrate the filtrate under vacuum at 40-80°C to a relative density of 1.25 (60°C), dilute the concentrate with 4 times the amount of water, let it stand, centrifuge and filter, and take the supernatant for later use;

[0062] S2: The supernatant is adsorbed by D101 resin with a diameter-to-height ratio of 1:2, and firstly 3 times the amount of water as the supernatant is added to wash until neutral, the washing liquid is discarded, and then 3 times the amount of sodium hydroxide solution with a mass percentage concentration of 0.1% is used to elute the resin to obtain eluent A, and then washed with water until neutral, and 8 times the amount of ethanol with a volume percentage concentration of 65% is added to elute to obtain eluent B;

[0063] S3: The pH of the eluent A is adjusted to neutral with hydrochloric acid, centrifuged and filtered, the supernatant is adsorbed by a polyamide resin with a diameter-to-height ratio of 1:4, washed with water until salt-free, and then eluted with 6 times the volume of the supernatant and 70% ethanol, the eluent is collected and concentrated to a relative density of 1.20 (60°C), and then vacuum dried at 60°C to a moisture content of 3%, to obtain a dry paste A;

[0064] S4: vacuum concentrating the eluent B at 40-80°C to a relative density of 1.25 (60°C), and vacuum drying at 60°C to a moisture content of 3%, to obtain a dry paste B;

[0065] S5: After the dry paste A and the dry paste B are crushed, 1 times the amount of dextrin is added to the obtained dry powder and mixed evenly, and then 20% of ethanol with a volume percentage concentration of 85% is added for granulation, and the mixture is dried and mixed to obtain a granular Chinese medicine composition for treating rheumatoid arthritis.

[0066] Example 2

[0067] S1: Take 500g of Acanthopanax senticosus and 500g of Schisandra chinensis, add them to 8000g of 65% ethanol by volume, extract them under reflux for 3 times at 80-100°C, filter the extract to obtain a filtrate, concentrate the filtrate under vacuum at 40-80°C to a relative density of 1.25 (60°C), add 4 times the amount of water to dilute the concentrate, let it stand, centrifuge and filter, and take the supernatant for later use;

[0068] S2: The supernatant is adsorbed by D101 resin with a diameter-to-height ratio of 1:2, and firstly 3 times the amount of water as the supernatant is added to wash until neutral, the washing liquid is discarded, and then the resin is eluted with 3 times the amount of sodium hydroxide solution as the supernatant to obtain eluent A, and then washed with water until neutral, and 8 times the amount of 65% by volume ethanol is added to elute to obtain eluent B;

[0069] S3: The pH of the eluent A is adjusted to neutral with hydrochloric acid, centrifuged and filtered, the supernatant is adsorbed by a polyamide resin with a diameter-to-height ratio of 1:4, washed with water until salt-free, and then eluted with 6 times the volume of the supernatant and 70% ethanol, the eluent is collected and concentrated to a relative density of 1.20 (60°C), and then vacuum dried at 60°C to a moisture content of 3%, to obtain a dry paste A;

[0070] S4: vacuum concentrating the eluent B at 40-80°C to a relative density of 1.25 (60°C), and vacuum drying at 60°C to a moisture content of 3%, to obtain a dry paste B;

[0071] S5: After the dry paste A and the dry paste B are crushed, 1 times the amount of dextrin is added to the obtained dry powder and mixed evenly, and then 85% by volume ethanol is added to granulate, the added amount is 20%, dried, and the total mixing is to obtain a granular Chinese medicine composition for treating rheumatoid arthritis.

[0072] Example 3

[0073] S1: Take 250g of Acanthopanax senticosus and 750g of Schisandra chinensis and add them to 8000g of 65% ethanol by volume, extract them under reflux for 3 times at 80-100°C, filter the extract to obtain a filtrate, concentrate the filtrate under vacuum at 40-80°C to a relative density of 1.25 (60°C), add 4 times the amount of water to dilute the concentrate, let it stand, centrifuge and filter, and take the supernatant for later use;

[0074] S2: The supernatant is adsorbed by D101 resin with a diameter-to-height ratio of 1:2, and water is added 3 times as much as the supernatant to wash until neutral, and the washing liquid is discarded, and then the resin is eluted with a sodium hydroxide solution 3 times as much as the supernatant to obtain an eluent A, and then washed with water until neutral, and ethanol with a volume percentage concentration of 65% is added 8 times as much as the supernatant to elute, to obtain an eluent B;

[0075] S3: The pH of the eluent A is adjusted to neutral with hydrochloric acid, centrifuged and filtered, the supernatant is adsorbed by a polyamide resin with a diameter-to-height ratio of 1:4, washed with water until salt-free, and then eluted with 6 times the volume of the supernatant and 70% ethanol, the eluent is collected and concentrated to a relative density of 1.20 (60°C), and then vacuum dried at 60°C to a moisture content of 3%, to obtain a dry paste A;

[0076] S4: vacuum concentrating the eluent B at 40-80°C to a relative density of 1.25 (60°C), and vacuum drying at 60°C to a moisture content of 3%, to obtain a dry paste B;

[0077] S5: After the dry paste A and the dry paste B are crushed, 1 times the amount of dextrin is added to the obtained dry powder and mixed evenly, and then 85% by volume ethanol is added to granulate, the added amount is 20%, dried, and the total mixing is to obtain a granular Chinese medicine composition for treating rheumatoid arthritis.

[0078] Comparative Example 1 (fluid extract preparation method)

[0079] Extraction: Grind 750g of Acanthopanax senticosus and 250g of Schisandra chinensis into coarse powder, add 3000g of 75% ethanol (3 times the total amount of Acanthopanax senticosus and Schisandra chinensis) and stir evenly, moisten and seal for 7 hours to allow it to fully expand, and put the expanded coarse powder into the percolation barrel with a filling height of 2 / 3 of the percolation barrel, flatten it, and slowly add 75% ethanol (volume percentage concentration) until the air between the powder particles is discharged and a proper amount of ethanol flows out, close the discharge valve, cover the filter cylinder, and soak it. Soak for 24 hours. After the soaking reaches the specified time, open the material valve for percolation, add 75% ethanol with a volume percentage concentration of 5 times the total amount of Acanthopanax and Schisandra while percolating, control the percolation rate to 3 ml / min, collect the filtrate, and mix it with the ethanol flowing out when exhausting the air. Concentrate the mixture of the filtrate and ethanol in vacuum at 60°C to a relative density of 1.30 (60°C) to obtain a concentrated solution, and vacuum dry the concentrated solution at 60°C to a moisture content of 3% to obtain a dry paste for standby use.

[0080] Preparation: Crush the dry paste through a 120-mesh sieve to obtain dry powder, add 3 times the amount of dextrin and mix evenly, then add 80% by volume ethanol to granulate, the added amount is 32%, dry, and mix to obtain the sample.

[0081] Comparative Example 2 (Ethanol Reflux Method)

[0082] Extraction: Add 750g of Acanthopanax and 250g of Schisandra chinensis into 70% ethanol (volume percentage concentration is 10 times the total amount of Acanthopanax and Schisandra chinensis), reflux extraction at 80-100°C for 1.5h, extract 3 times, filter after the extraction to obtain a filtrate, vacuum concentrate the filtrate at 60°C to a relative density of 1.30 (60°C) to obtain a concentrated solution, vacuum dry the concentrated solution at 60°C to a moisture content of 3%, obtain a dry paste, and set aside.

[0083] Preparation: Crush the dry paste through a 120-mesh sieve to obtain dry powder, add 2 times the amount of starch and mix evenly, then add 85% by volume ethanol to granulate, the added amount is 23%, dry, mix, and fill in No. 1 capsules to obtain the sample.

[0084] Comparative Example 3 (Water Extraction and Alcohol Precipitation Method)

[0085] Extraction: Add 750g of Acanthopanax senticosus and 250g of Schisandra chinensis to 10 times the total amount of water, and reflux extract at 80-100°C for 1.5h. Extract 3 times. Filter after extraction to obtain filtrate. The filtrate is vacuum concentrated at 60°C to 3g of medicinal material per milliliter. After cooling, 95% ethanol by volume is slowly added under stirring until the alcohol content is more than 70%. Then, seal and refrigerate for 12h, filter, and concentrate the filtrate to a relative density of 1.30 (60°C) to obtain a concentrated solution. The concentrated solution is vacuum dried at 60°C to a moisture content of 3% to obtain a dry paste for standby use.

[0086] Preparation: Crush the dry paste through a 120-mesh sieve to obtain dry powder, add 5 times the amount of starch and mix evenly, then add 85% by volume ethanol to granulate, the addition amount is 23%, dry, mix, and tablet to obtain the sample.

[0087] The Chinese medicine compositions prepared in Examples 1 to 3 and the samples prepared in Comparative Examples 1 to 3 were tested, and the dry paste yield and the effective ingredient content were as shown in Table 1:

[0088] Table 1:

[0089]

[0090] Referring to Table 1, the average transfer rate of eleutheroside E, the main active ingredient of Acanthopanax senticosus, is about 90%, and the average transfer rate of schisandrin A, the main active ingredient of Schisandra chinensis, is as high as 88%. It can be seen that the preparation method of the present invention can effectively adsorb active ingredients, remove impurities, and concentrate essence compared with Comparative Examples 1 to 3.

[0091] The therapeutic effect of the Chinese medicine composition of the present invention on rats with rheumatoid arthritis is shown in the following efficacy test:

[0092] (1) Grouping: 90 rats were divided into a blank group, a model group, and a positive group according to the random number table method. The Chinese medicine composition obtained in Example 1 was divided into a high-dose group, a medium-dose group, and a low-dose group (the doses were 1.0 g / kg, 0.5 g / kg, and 0.25 g / kg, respectively), namely, a high-dose group, a medium-dose group, and a low-dose group, for a total of 6 groups, with 15 rats in each group.

[0093] (2) Construction of collagen-induced arthritis (CIA) model: Except for the blank group, bovine type II collagen (2 mg / mL) was dissolved in acetic acid (0.5 mol / L) overnight under sterile conditions at 4°C in all other groups, and then fully emulsified with an equal volume of incomplete Freund's adjuvant (10 mL) to prepare bovine type II collagen emulsion. After the rats were anesthetized intraperitoneally with 10% sodium pentobarbital (0.3 mL / 100 g), 0.1 mL of the emulsion was subcutaneously injected into the plantar surface of the rats' feet every day for 21 days. Successful modeling criteria: Inflammatory reactions in the ankles and toes of rats were manifested as redness and swelling of the paws, and the growth range of the paws was >0.8 cm.

[0094] (3) Dosage regimen: Starting from the first day after successful modeling, the blank group and the model group were gavaged with normal saline, the positive group was gavaged with methotrexate (at a dose of 1 mg / kg), and the high-dose, medium-dose, and low-dose groups were gavaged with 1.0 g / kg, 0.5 g / kg, and 0.25 g / kg of the Chinese medicine composition, respectively. Each group was gavaged once a day for 28 days.

[0095] (4) Index detection: Rats were anesthetized and killed, and whole blood was collected and centrifuged to obtain serum. At the same time, liver tissue was collected and stored at low temperature for later use.

[0096] 4.1 The changes in the paw volume of CIA rats were observed. The paw swelling was estimated weekly using a limb swelling volume meter based on the volume of the right hind paw to evaluate the effect of the Chinese medicine composition on the paw volume of CIA rats.

[0097] Refer to Table 2 and Figure 1 From the changes in the paw volume (mL) of CIA rats, it can be seen that compared with the model group, the positive group and the drug-medium and drug-high groups can significantly alleviate the swelling of the rat paws and are close to the state of normal rats.

[0098] Table 2:

[0099]

[0100] 4.2 Rheumatoid arthritis score: The severity of rheumatoid arthritis in rats was assessed every 2 days using a 5-point scoring scale. The scoring criteria are: 4 points: swelling from the ankle joint to the entire paw; 3 points: swelling from the toes to the ankle joint; 2 points: swelling of the toes and toe joints; 1 point: erythema or swelling of the toe joints; 0 points: normal. The arthritis score of each rat is the sum of the four limbs. The rheumatoid arthritis scores of CIA rats are shown in Tables 3 and Figure 2 .according to Figure 2 As shown in Table 3, all CIA rats did not develop arthritis symptoms until the 15th day. Figure 1 It can be seen that compared with the model group, both the administration composition-medium and the administration group-high can improve the symptoms of CIA model arthritis, and the effect of the administration group-high in improving CIA arthritis is similar to that of the positive drug methotrexate.

[0101] Table 3:

[0102]

[0103]

[0104] 4.3 The levels of serum IL-6, IL-1β and TNF-α in rats of each group were detected by ELISA method to evaluate the effect of the Chinese medicine composition of the present invention on the immune cytokines of CIA rats.

[0105] like Figure 3 As shown in Table 4, compared with the blank group, **P<0.01; compared with the model group, ##P<0.01; comparison between different dose groups, &&P<0.01, the model group had significant differences compared with the blank group, indicating that the CIA rat model was successfully established. Compared with the model group, the positive group and different dose groups can improve the inflammatory response of CIA rats, with significant differences. In addition, there are significant differences between the high, medium and low dose groups, showing obvious dose dependence. The larger the dose, the more obvious the improvement of the inflammatory response of CIA rats. By reducing the expression of proinflammatory factors and reducing the permeability of microvessels, the occurrence of redness, swelling, heat and pain in arthritis is alleviated.

[0106] Table 4:

[0107] Grouping IL-1β (pg / mL) TNF-ɑ(pg / mL) IL-6 (pg / mL) Blank Group 14.56 98.39 35.88 Model Group 58.22 158.15 63.88 Positive group 18.49 105.25 35.15 Drug group-low 45.74 137.22 55.22 Drug group-medium 33.86 128.08 47.89 Drug group-high 20.16 111.22 38.99

[0108] 4.4 Prepare liver tissue homogenate and operate according to the instructions of the kit to determine the activities of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px). The results are shown in Table 1. Figure 4 And Table 5, among which, compared with the blank group, **P<0.01; compared with the model group, ##P<0.01; comparison between different dose groups, &&P<0.01.

[0109] Oxidative stress is an important cause of rheumatoid arthritis (RA) and one of the pathological consequences of RA. Oxidative stress refers to the increase in the body's lipid peroxidation reaction, which in turn causes an increase in cytokine secretion and aggravates the inflammatory response of the joints. Superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) are antioxidant enzymes that can remove reactive oxygen species in the body and protect cells from oxidative stress damage. The effect of the Chinese medicine composition on the oxidative stress response of CIA rats is shown in Figure 2. Figures 4-5As shown in Table 5, the activities of SOD, CAT and GSH-Px enzymes in CIA model rats were reduced, indicating that CIA rats had oxidative stress damage. Compared with the model group, the positive group and different doses of the Chinese medicine composition could significantly increase the activities of SOD, CAT and GSH-Px enzymes, and had the effect of regulating oxidative stress damage in CIA rats.

[0110] Table 5:

[0111] Grouping SOD(u / mg) CAT(u / mg) GSH (u / mL) Blank Group 35.08 71.06 850 Model Group 11.89 25.76 435 Positive group 33.88 66.97 821 Drug group-low 19.08 41.22 635 Drug group-medium 25.87 53.67 725 Drug group-high 31.97 63.42 815

[0112] The above description is only a preferred embodiment of the present invention, and does not limit the patent scope of the present invention. Any equivalent structure or equivalent process transformation made by using the contents of the present invention specification and drawings, or directly or indirectly applied in other related technical fields, are also included in the patent protection scope of the present invention.

Claims

1. A method for preparing a Chinese medicine composition for treating rheumatoid arthritis, characterized in that: The following steps are involved: S1: Take Acanthopanax senticosus and Schisandra chinensis and add them to ethanol for reflux extraction. After filtering the extract, concentrate the filtrate, add water to dilute the concentrate, filter again, and take the supernatant for later use; S2: adsorbing the supernatant with a first resin, washing with water until neutral, discarding the washing liquid, eluting the resin with a sodium hydroxide solution to obtain an eluent A, then washing with water until neutral, adding ethanol for elution, and obtaining an eluent B; S3: The eluent A is adjusted to neutral with a pH adjuster, centrifuged and filtered, the supernatant is adsorbed by a second resin, washed with water until it is salt-free, and then eluted with ethanol, the eluent is collected and concentrated, and then vacuum dried to obtain a dry paste A; S4: vacuum concentrating the eluate B, and vacuum drying to obtain a dry paste B; S5: Post-processing the dry paste A and the dry paste B to prepare a traditional Chinese medicine composition for treating rheumatoid arthritis.

2. The method for preparing the Chinese medicine composition for treating rheumatoid arthritis according to claim 1, characterized in that: The step S1 comprises: adding 6 to 8 times the total amount of ethanol of Acanthopanax senticosus and Schisandra chinensis, refluxing and extracting at 80 to 100° C. for 1 to 3 hours, and repeating 1 to 3 times; vacuum concentrating the filtrate at 40 to 80° C. to a relative density of 1.15 to 1.25 (60° C.) to obtain the concentrated solution; diluting the concentrated solution with 2 to 4 times the amount of water, filtering, and obtaining a supernatant.

3. The method for preparing the Chinese medicine composition for treating rheumatoid arthritis according to claim 1, characterized in that: The step S2 comprises: adsorbing the supernatant with the first resin, first adding 1 to 3 times the amount of water to wash until neutral, discarding the water washing liquid, then eluting the first resin with 1 to 3 times the amount of 0.1% sodium hydroxide solution to obtain eluent A, then washing with water until neutral, adding 5 to 8 times the amount of ethanol to elute, and obtaining eluent B; The step S3 comprises: adjusting the eluent A to neutrality with hydrochloric acid, adsorbing the supernatant with a second resin, washing with water until it is salt-free, eluting with 4 to 6 times ethanol, collecting and concentrating the eluent A', and vacuum drying to obtain a dry paste A.

4. The method for preparing the Chinese medicine composition for treating rheumatoid arthritis according to claim 3, characterized in that: The volume percentage concentration of the ethanol is 50% to 70%.

5. The method for preparing the Chinese medicine composition for treating rheumatoid arthritis according to claim 3, characterized in that: The preparation process of the dry paste includes: vacuum concentrating the eluent A' at 40-80°C to a relative density of 1.20-1.30 (60°C) to obtain a concentrated solution, and vacuum drying the concentrated solution at a temperature of 50-70°C to a moisture content of 3% to obtain the dry paste A; vacuum concentrating the eluent B at 40-80°C to a relative density of 1.20-1.30 (60°C) to obtain a concentrated solution, and vacuum drying the concentrated solution at a temperature of 50-70°C to a moisture content of 3% to obtain a dry paste B.

6. The method for preparing the Chinese medicine composition for treating rheumatoid arthritis according to claim 1, characterized in that: The first resin is a D101 macroporous resin with a diameter-to-height ratio of 1:2 to 1:4, and the second resin is a polyamide resin with a diameter-to-height ratio of 1:2 to 1:

4.

7. The method for preparing the Chinese medicine composition for treating rheumatoid arthritis according to claim 1, characterized in that: The weight ratio of the Acanthopanax senticosus to the Schisandra chinensis is 1:3 to 3:

1.

8. A Chinese medicine composition for treating rheumatoid arthritis, characterized in that: The method is prepared by any one of claims 1 to 7.

9. The Chinese medicine composition for treating rheumatoid arthritis according to claim 8, characterized in that: The traditional Chinese medicine composition is in the form of granules, capsules or tablets.

10. Use of the Chinese medicine composition according to claim 8 in preparing a medicine for treating rheumatoid arthritis.

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