Traditional Chinese medicine bone liquid for inhibiting rheumatism and strengthening bones and preparation method thereof

Abstract

The invention provides traditional Chinese medicine bone liquid for inhibiting rheumatism and strengthening bones and a preparation method thereof, and belongs to the technical field of traditional Chinese medicines. The invention provides traditional Chinese medicine bone liquid for inhibiting rheumatism and strengthening bones. The traditional Chinese medicine bone liquid is prepared from radix angelicae pubescentis, radix clematidis, eucommia ulmoides, rhizoma drynariae, cassia twigs, caulis spatholobi and liquorice. The traditional Chinese medicine bone liquid takes radix angelicae pubescentis and radix clematidis as monarch drugs and has the effects of dispelling wind, eliminating dampness and activating meridians to stop pain; ministerial drugs are eucommia ulmoides and rhizoma drynariae which can tonify liver and kidney and strengthen muscles and bones; cassia twig and suberect spatholobus stem are used as adjuvant drugs for warming meridians and dredging collaterals, and And the guide medicine is liquorice, harmonizes the medicines, and relieves spasm and pain. The traditional Chinese medicine bone liquid disclosed by the invention has both effects of expelling wind-damp and tonifying liver and kidney, not only can dispel wind-damp pathogenic qi, but also can strengthen muscles and bones, is suitable for diseases such as rheumatic arthritis and osteoporosis, and has relatively good comprehensive effects of resisting inflammation, easing pain and promoting bone repair. Different extraction methods are selected according to the characteristics of all the traditional Chinese medicine raw materials, and the effect of treating the rheumatism diseases is enhanced.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicines, and in particular to a traditional Chinese medicine bone fluid for inhibiting rheumatism and strengthening bones and a preparation method thereof. Background Art

[0002] Rheumatic diseases are a type of disease that mainly invades joints, bones, muscles, blood vessels and related soft tissues or connective tissues. Most of them are autoimmune diseases, which are characterized by a long course and high disability rate, seriously affecting the quality of life of patients.

[0003] Currently, the main drugs used in clinical treatment of rheumatic diseases include nonsteroidal anti-inflammatory drugs, glucocorticoids, and immunosuppressants. However, while these drugs can alleviate symptoms to a certain extent, long-term use can cause a number of adverse reactions, including gastrointestinal reactions, liver and kidney damage, and osteoporosis, and they are unable to fundamentally prevent the progression of the disease.

[0004] Traditional Chinese Medicine (TCM) has a long history and unique advantages in treating rheumatic diseases. Many TCM herbs have been shown to possess anti-inflammatory, analgesic, and immunomodulatory properties, effectively improving rheumatic symptoms while minimizing adverse reactions. However, traditional TCM preparations suffer from low extraction rates of active ingredients, poor bioavailability, and insufficient formulation stability, limiting their clinical application and widespread adoption. For example, traditional decoction methods struggle to fully extract the active ingredients in TCM herbs, and some active substances may be destroyed by prolonged high-temperature treatment. Furthermore, simple pulverization and mixing methods result in inefficient absorption and utilization of the drug components in the human body.

[0005] With the development of modern biotechnology and pharmaceutical technology, exploring more efficient and scientific methods for preparing traditional Chinese medicine preparations has become key to improving the efficacy of traditional Chinese medicine in treating rheumatic diseases. Therefore, developing a safe, effective, and technologically advanced traditional Chinese medicine preparation to inhibit rheumatism and strengthen bones is of great practical significance. Summary of the Invention

[0006] The purpose of the present invention is to provide a Chinese medicinal bone liquid for inhibiting rheumatism and strengthening bones and a preparation method thereof. The Chinese medicinal bone liquid provided by the present invention has good anti-inflammatory and bone-strengthening effects and has a good therapeutic effect on treating rheumatic diseases.

[0007] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0008] The invention provides a traditional Chinese medicine bone liquid for inhibiting rheumatism and strengthening bones. The bone liquid comprises the following raw materials in parts by weight: 15-25 parts of angelica dahurica, 12-20 parts of clematis root, 20-30 parts of eucommia bark, 20-35 parts of drynaria rhizome, 10-15 parts of cassia twig, 25-40 parts of millettia reticulata, and 8-12 parts of liquorice.

[0009] The present invention also provides a preparation method of the above-mentioned traditional Chinese medicine bone liquid, comprising the following steps: crushing the angelica dahurica and mixing it with water, inoculating a composite bacterial agent for anaerobic fermentation, and concentrating the fermentation liquid to obtain the angelica dahurica extract; crushing the clematis root and mixing it with an acetic acid solution, ultrasonically treating it, adjusting the pH value, adding a composite enzyme for enzymolysis, and concentrating the enzymatic hydrolysis liquid to obtain the clematis root extract; crushing the eucommia ulmoides, drynaria rhizome, cassia twig and millettia reticulata, mixing them with an ethanol solution, ultrasonically extracting them, filtering them, and concentrating the filtrate to obtain an ethanol extract; crushing the liquorice and mixing it with water, heating it for extraction, filtering it, and concentrating it to obtain the liquorice extract; mixing the angelica dahurica extract, the clematis root extract, the ethanol extract and the liquorice extract, adding auxiliary materials, and homogenizing them to obtain the traditional Chinese medicine bone liquid.

[0010] Preferably, the composite bacterial agent is composed of Bifidobacterium animalis and Clostridium tyrobutyricum; the bacterial count of Bifidobacterium animalis in the composite bacterial agent is 200-500 million CFU / g, and the bacterial count of Clostridium tyrobutyricum is 500-800 million CFU / g; the inoculation amount of the composite bacterial agent is 5‰-10‰ of the weight of Angelica dahurica.

[0011] Preferably, the temperature of the composite bacterial agent fermentation is 35-40°C, the pH value is 6.5-7, and the fermentation time is 48-72 hours.

[0012] Preferably, the volume fraction of the acetic acid solution is 3%-5%, the material-liquid ratio of the ultrasonic treatment is 1:15-20 g / mL, the temperature is 40-50° C., the frequency is 30-40 kHz, and the time is 20-30 min.

[0013] Preferably, the complex enzyme is composed of cellulase and papain in a weight ratio of 2-4:1, and the added amount of the complex enzyme is 3%-8% of the weight of the Clematis chinensis; the enzymatic hydrolysis temperature is 50-55°C, the pH value is 4.5-5.5, and the time is 30-40 minutes.

[0014] Preferably, the volume fraction of the ethanol solution is 70%-80%; the material-liquid ratio of the ultrasonic extraction is 1:10-15 g / mL, the temperature is 45-55° C., the frequency is 25-35 kHz, and the time is 40-50 min.

[0015] Preferably, the mass volume ratio of licorice to water is 1:20-30 g / mL, the extraction temperature is 90-100° C., and the number of extractions is 2-4 times, each time for 30-60 min.

[0016] Preferably, the auxiliary materials include one or more of sweeteners, stabilizers, and antioxidants.

[0017] The present invention also provides the use of the traditional Chinese medicine bone liquid in preparing medicine for treating rheumatism.

[0018] Compared with the prior art, the present invention has the following beneficial effects:

[0019] The present invention provides a Chinese herbal bone liquid for inhibiting rheumatism and strengthening bones, comprising: Angelica dahurica, Clemati radix, Eucommia bark, Drynaria fortunei, Cinnamon twig, Millettia reticulata and Licorice root. The Chinese herbal bone liquid of the present invention uses Angelica dahurica and Clemati radix as the main drugs, which can dispel wind and dampness, dredge meridians and relieve pain; the auxiliary drugs are Eucommia bark and Drynaria fortunei, which can nourish the liver and kidneys and strengthen the tendons and bones; the adjuvant drugs are Cinnamon twig and Millettia reticulata, which can warm the meridians and dredge the meridians, activate blood circulation and dredge meridians; the guiding drug is Licorice root, which can harmonize the various drugs and relieve pain. The present invention selects different extraction methods according to the characteristics of each Chinese herbal raw material, thereby enhancing the effect of treating rheumatic diseases. The Chinese herbal bone liquid of the present invention pays equal attention to dispelling wind and dampness and tonifying the liver and kidneys, can dispel rheumatic evil spirits, and strengthen tendons and bones. It is suitable for diseases such as rheumatic arthritis and osteoporosis, and has a good comprehensive effect of anti-inflammatory, analgesic and bone repair promotion. DETAILED DESCRIPTION

[0020] The invention provides a traditional Chinese medicine bone liquid for inhibiting rheumatism and strengthening bones. The bone liquid comprises the following raw materials in parts by weight: 15-25 parts of angelica dahurica, 12-20 parts of clematis root, 20-30 parts of eucommia bark, 20-35 parts of drynaria rhizome, 10-15 parts of cassia twig, 25-40 parts of millettia reticulata, and 8-12 parts of liquorice.

[0021] In terms of traditional Chinese medicine formula, the main medicine (main medicine, targeting the main symptoms) is Angelica dahurica and Smilax glabra.

[0022] Angelica dahurica is pungent, bitter, and slightly warm. It enters the kidney and bladder meridians. It has the effects of dispelling wind and dampness, relieving numbness and relieving pain. It is an important medicine for treating rheumatic pain, especially rheumatic pain in the lower body.

[0023] Smilax glabra is pungent, salty and warm, and belongs to the bladder meridian. It has the effects of dispelling rheumatism, unblocking meridians and relieving pain. It is especially good at softening bone spurs, relieving joint stiffness, and enhancing the wind-dispelling and dampness-removing effect of Angelica dahurica.

[0024] Angelica dahurica and Clematis chinensis work synergistically as the main medicines, dispelling wind and dampness, unblocking meridians and relieving pain, targeting the core pathogenesis of rheumatic pain.

[0025] The assistant medicines are Eucommia ulmoides and Drynaria fortunei.

[0026] Eucommia ulmoides is sweet and warm, and enters the liver and kidney meridians. It has the effects of nourishing the liver and kidneys, strengthening the tendons and bones, and can improve soreness of the waist and knees and weakness of the tendons and bones caused by rheumatism.

[0027] Drynaria fortunei is bitter and warm, and enters the liver and kidney meridians. It has the effects of nourishing the kidneys and strengthening bones, promoting blood circulation and healing injuries. It can promote bone repair and relieve joint injuries caused by osteoporosis and rheumatism.

[0028] Eucommia ulmoides and Drynaria fortunei work together to assist the main medicine, nourishing the liver and kidneys, strengthening the tendons and bones while eliminating rheumatism, preventing long-term rheumatism from damaging the kidneys, and enhancing bone health.

[0029] Auxiliary drugs (assisting the monarch and ministerial ingredients and treating concurrent symptoms) are cinnamon twig and Millettia reticulata.

[0030] Cinnamon twig is pungent, sweet and warm, and enters the heart, lung and bladder meridians. It has the effects of warming the meridians and dredge the blood vessels, assisting yang and transforming qi, can promote the circulation of qi and blood, enhance the efficacy of anti-rheumatic drugs, and relieve cold and pain in joints.

[0031] Millettia reticulata is bitter, sweet, and warm, and enters the liver and kidney meridians. It has the effects of promoting blood circulation, replenishing blood, relaxing muscles and tendons, and can improve blood stasis and limb numbness caused by rheumatism, and promote blood circulation.

[0032] Cinnamon twig warms and unclogs the meridians, while Millettia reticulata activates blood circulation and unclogs the collaterals. Together, they promote the circulation of Qi and blood, prevent Qi and blood stasis caused by rheumatic obstruction, and enhance the whole prescription's effect of relieving numbness and alleviating pain.

[0033] The guiding drug (to harmonize the medicinal properties and guide the medicine to the meridians) is licorice.

[0034] Licorice is sweet and neutral, and enters the heart, lung, spleen, and stomach meridians. It has the effect of harmonizing various medicines, relieving acute pain, reducing the bias of other medicines, and enhancing the synergistic effect of the whole prescription.

[0035] The Chinese medicine bone liquid of the present invention uses Angelica dahurica and Clemati radix as the main ingredients, which dispel wind and dampness, dredge the meridians and relieve pain; the auxiliary ingredients are Eucommia ulmoides and Drynaria fortunei, which tonify the liver and kidneys and strengthen the tendons and bones; the adjuvant ingredients are cinnamon twig and Millettia reticulata, which warm the meridians and dredge the meridians, promote blood circulation and dredge the meridians; and the guiding ingredient is liquorice, which harmonizes the various ingredients and relieves pain. The Chinese medicine bone liquid of the present invention emphasizes both dispelling wind and rheumatism and tonifying the liver and kidneys, and can not only dispel rheumatic pathogenic qi but also strengthen the tendons and bones. It is suitable for diseases such as rheumatic arthritis and osteoporosis, and has a good comprehensive effect of anti-inflammatory, analgesic, and bone repair promotion.

[0036] The present invention also provides a preparation method of the above-mentioned traditional Chinese medicine bone liquid, comprising the following steps: crushing the angelica dahurica and mixing it with water, inoculating a composite bacterial agent for anaerobic fermentation, and concentrating the fermentation liquid to obtain the angelica dahurica extract; crushing the clematis root and mixing it with an acetic acid solution, ultrasonically treating it, adjusting the pH value, adding a composite enzyme for enzymolysis, and concentrating the enzymatic hydrolysis liquid to obtain the clematis root extract; crushing the eucommia ulmoides, drynaria rhizome, cassia twig and millettia reticulata, mixing them with an ethanol solution, ultrasonically extracting them, filtering them, and concentrating the filtrate to obtain an ethanol extract; crushing the liquorice and mixing it with water, heating it for extraction, filtering it, and concentrating it to obtain the liquorice extract; mixing the angelica dahurica extract, the clematis root extract, the ethanol extract and the liquorice extract, adding auxiliary materials, and homogenizing them to obtain the traditional Chinese medicine bone liquid.

[0037] The composite bacterial agent of the present invention is composed of animal Bifidobacterium and Clostridium tyrobutyricum; the animal Bifidobacterium is purchased from the China Industrial Microorganism Culture Collection Administration Center with a strain number of CICC 24928; the Clostridium tyrobutyricum is Clostridium tyrobutyricum W428, purchased from the China Center for Type Culture Collection with a collection number of CCTCC NO: M 2016090, and disclosed in patent CN105779351 A; the bacterial count of animal Bifidobacterium in the composite bacterial agent is preferably 200-500 million CFU / g, more preferably 300 million CFU / g, and the bacterial count of Clostridium tyrobutyricum is preferably 500-800 million CFU / g, more preferably 600 million CFU / g; the inoculation amount of the composite bacterial agent is 5‰-10‰ of the weight of Angelica dahurica; the fermentation temperature of the composite bacterial agent is preferably 35-40°C, more preferably 37°C, the pH value is preferably 6.5-7, more preferably 6.8, and the fermentation time is preferably 48-72h, more preferably 60h. The specific composite bacterial agent used in the present invention ferments the angelica dahurica, which can transform the components in the angelica dahurica, improve its anti-inflammatory activity, enhance the analgesic and immunomodulatory effects of the angelica dahurica, thereby enhancing the efficacy of rheumatism, and at the same time can also improve bone metabolism, enhance antioxidant activity, and promote bone repair.

[0038] The volume fraction of the acetic acid solution of the present invention is 3%-5%, more preferably 4%, the material-liquid ratio of the ultrasonic treatment is preferably 1:15-20 g / mL, more preferably 1:18 g / mL, the temperature is preferably 40-50°C, more preferably 45°C, the frequency is preferably 30-40 kHz, more preferably 35 kHz, and the time is preferably 20-30 min, more preferably 25 min. The present invention first ultrasonically treats the clematis with an acetic acid solution, which can destroy the cell wall structure, promote the dissolution and release of active ingredients, and promote the enrichment of key components.

[0039] The composite enzyme of the present invention is preferably composed of cellulase and papain in a weight ratio of 2-4:1, more preferably 3:1. The added amount of the composite enzyme is preferably 3%-8% of the weight of the Clematis chinensis, more preferably 5%; the temperature of the enzymatic hydrolysis is preferably 50-55°C, more preferably 52°C, the pH value is preferably 4.5-5.5, more preferably 5, and the time is preferably 30-40min, more preferably 35min. The present invention combines acetic acid ultrasonic pretreatment with composite enzymatic hydrolysis to simultaneously and efficiently extract the key active ingredients in Clematis chinensis, meet the needs of inhibiting joint inflammation and promoting bone regeneration, and meet the therapeutic targets of rheumatic bone disease.

[0040] The volume fraction of the ethanol solution of the present invention is preferably 70%-80%, more preferably 75%; the material-liquid ratio of the ultrasonic extraction is preferably 1:10-15g / mL, more preferably 1:12g / mL, the temperature is preferably 45-55°C, more preferably 50°C, the frequency is preferably 25-35kHz, more preferably 30kHz, and the time is preferably 40-50min, more preferably 45min. Mixed ultrasonic extraction of Eucommia ulmoides, Drynaria fortunei, Cinnamomum cassia twig and Millettia reticulata with ethanol solution can efficiently enrich the active ingredients of the four medicinal materials, improve the anti-rheumatic, analgesic and blood circulation effects, and promote the efficacy of traditional Chinese medicine bone liquid.

[0041] The weight-to-volume ratio of licorice to water of the present invention is preferably 1:20-30 g / mL, more preferably 1:25 g / mL. The extraction temperature is preferably 90-100°C, more preferably 95°C. The number of extractions is preferably 2-4, more preferably 3, and each extraction lasts preferably 30-60 minutes, more preferably 40 minutes. The licorice extract of the present invention can alleviate the potential irritation of medicinal materials such as Angelica dahurica and Clematis chinensis, harmonize the medicinal properties, mask some of the bitterness, and improve oral palatability.

[0042] The auxiliary materials of the present invention preferably include one or more of a sweetener, a stabilizer, and an antioxidant; the sweetener is preferably steviol glycoside, the stabilizer is preferably sucrose fatty acid ester SE-15, and the antioxidant is preferably ascorbic acid.

[0043] The present invention also provides the use of the traditional Chinese medicine bone liquid in preparing medicine for treating rheumatism.

[0044] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions of the present invention. Obviously, the embodiments described are only some of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0045] In the following examples, unless otherwise specified, all methods are conventional.

[0046] Unless otherwise specified, the materials and reagents used in the following examples can be obtained from commercial sources.

[0047] The animal Bifidobacterium involved in the embodiments and comparative examples was purchased from the China Industrial Culture Collection Administration Center for Microorganisms with a strain number of CICC 24928; the Clostridium tyrobutyricum was Clostridium tyrobutyricum W428, purchased from the China Center for Type Culture Collection with a collection number of CCTCC NO: M 2016090, disclosed in patent CN 105779351A; Bifidobacterium longum was purchased from the China Industrial Culture Collection Administration Center for Microorganisms with a strain number of CICC 6205; Clostridium butyricum was purchased from the China Industrial Culture Collection Administration Center for Microorganisms with a strain number of CICC 24854; cellulase was purchased from Anhui Nobet Trading Co., Ltd. with an enzyme activity of 100,000 U / g; papain was purchased from Anhui Nobet Trading Co., Ltd. with an enzyme activity of 100,000 U / g.

[0048] Example 1

[0049] Preparation of Chinese herbal bone fluid

[0050] S1. Weighing

[0051] Weigh 20 parts of Angelica dahurica, 16 parts of Clematidis, 25 parts of Eucommia ulmoides, 28 parts of Drynaria fortunei, 12 parts of Cinnamomum cassia, 30 parts of Millettia repens, 10 parts of Licorice, 0.03 parts of Stevioside, 0.08 parts of Sucrose Fatty Acid Ester SE-15, and 0.02 parts of ascorbic acid respectively;

[0052] 15-25 parts of Angelica dahurica, 12-20 parts of Clematidis, 20-30 parts of Eucommia ulmoides, 20-35 parts of Drynaria fortunei, 10-15 parts of cinnamon twig, 25-40 parts of Millettia reticulata, 8-12 parts of liquorice, 0.01-0.05 parts of steviol glycosides, 0.05-0.15 parts of sucrose fatty acid ester SE-15, 0.01-0.03 parts of ascorbic acid and water.

[0053] S2. Preparation of Angelica dahurica Extract

[0054] The angelica dahurica powder was dried and crushed, passed through a 200-mesh sieve to obtain angelica dahurica powder, mixed with water at a solid-liquid ratio of 1:25 g / mL, sterilized, inoculated with a composite bacterial agent, and anaerobically fermented at 37°C and a pH of 6.8 for 60 h, sterilized, filtered, and the filtrate was concentrated and dried to a water content of 5 wt% to obtain the angelica dahurica extract;

[0055] The composite bacterial agent is composed of Bifidobacterium animalis and Clostridium tyrobutyricum; the bacterial count of Bifidobacterium animalis in the composite bacterial agent is 300 million CFU / g, and the bacterial count of Clostridium tyrobutyricum is 600 million CFU / g; the inoculation amount of the composite bacterial agent is 8‰ of the weight of the Angelica dahurica powder;

[0056] S3. Preparation of Clematis chinensis extract

[0057] The clematis root was crushed and passed through a 150-mesh sieve to obtain clematis root powder, which was mixed with a 4% by volume acetic acid solution at a solid-liquid ratio of 1:18 g / mL, and ultrasonically extracted at 45°C and 35 kHz for 25 minutes. A complex enzyme was added at 5% by weight of the clematis root powder, and enzymatic hydrolysis was performed at 52°C and pH 5 for 35 minutes. After inactivation of the enzyme, the solution was filtered, and the filtrate was concentrated and dried to a water content of 3 wt% to obtain a clematis root extract.

[0058] The complex enzyme consists of cellulase and papain in a weight ratio of 3:1;

[0059] S4. Preparation of ethanol extract

[0060] Eucommia ulmoides, Drynaria fortunei, cinnamon twig and Millettia reticulata were crushed separately, passed through a 300-mesh sieve, mixed, and added with a 75% ethanol solution at a solid-liquid ratio of 1:12 g / mL. Ultrasonic extraction was performed at 50°C and 30 kHz for 45 min, filtered, and the filtrate was concentrated and dried to a water content of 4 wt% to obtain an ethanol extract.

[0061] S5. Preparation of Licorice Extract

[0062] The licorice was dried and crushed, passed through a 100-mesh sieve, and water was added at a solid-liquid ratio of 1:25 g / mL. The mixture was extracted three times at 95°C for 40 min each time. The filtrates were combined, concentrated, and dried to a water content of 3 wt% to obtain a licorice extract.

[0063] S6. Preparation of Chinese medicine bone fluid

[0064] Sucrose fatty acid ester SE-15 and water are mixed in a mass ratio of 1:8, and stirred at 52°C and 1000rpm for 18 minutes to obtain sucrose fatty acid ester SE-15 colloid; Angelica dahurica extract, Clematidis extract, ethanol extract, licorice extract, steviol glycoside, ascorbic acid, sucrose fatty acid ester SE-15 colloid and water (the amount is 5 times the total weight of Angelica dahurica extract, Clematidis extract, ethanol extract, licorice extract, steviol glycoside and ascorbic acid) are mixed, and homogenized at 8000rpm for 15 minutes to obtain the traditional Chinese medicine bone liquid.

[0065] Example 2

[0066] Preparation of Chinese herbal bone fluid

[0067] S1. Weighing

[0068] Weigh 15 parts of Angelica dahurica, 12 parts of Clemati radix, 20 parts of Eucommia ulmoides, 20 parts of Drynaria fortunei, 10 parts of Cinnamomum cassia, 25 parts of Millettia repens, 8 parts of Licorice root, 0.01 parts of Stevioside, 0.05 parts of Sucrose fatty acid ester SE-15, and 0.01 parts of ascorbic acid respectively;

[0069] S2. Preparation of Angelica dahurica Extract

[0070] The angelica dahurica powder was dried and crushed, passed through a 150-mesh sieve to obtain angelica dahurica powder, mixed with water at a solid-liquid ratio of 1:20 g / mL, sterilized, inoculated with a composite bacterial agent, and anaerobically fermented at 35°C and a pH of 6.5 for 72 hours, sterilized, filtered, and the filtrate was concentrated and dried to a water content of 4 wt% to obtain the angelica dahurica extract;

[0071] The composite bacterial agent is composed of Bifidobacterium animalis and Clostridium tyrobutyricum; the bacterial count of Bifidobacterium animalis in the composite bacterial agent is 200 million CFU / g, and the bacterial count of Clostridium tyrobutyricum is 500 million CFU / g; the inoculation amount of the composite bacterial agent is 10‰ of the weight of the Angelica dahurica powder;

[0072] S3. Preparation of Clematis chinensis extract

[0073] The clematis root was crushed and passed through a 100-mesh sieve to obtain clematis root powder, which was mixed with a 3% by volume acetic acid solution at a solid-liquid ratio of 1:15 g / mL, and ultrasonically extracted at 40°C and 30 kHz for 30 minutes. A complex enzyme was added at a concentration of 3% by weight of the clematis root powder, and enzymatic hydrolysis was performed at 50°C and pH 4.5 for 40 minutes. After inactivation of the enzyme, the mixture was filtered, and the filtrate was concentrated and dried to a water content of 2 wt% to obtain a clematis root extract.

[0074] The complex enzyme consists of cellulase and papain in a weight ratio of 2:1;

[0075] S4. Preparation of ethanol extract

[0076] Eucommia ulmoides, Drynaria fortunei, cinnamon twig and Millettia reticulata were crushed separately, passed through a 250-mesh sieve, mixed, and added with a 70% ethanol solution by volume at a solid-liquid ratio of 1:10 g / mL. Ultrasonic extraction was performed at 45°C and 25 kHz for 50 min, filtered, and the filtrate was concentrated and dried to a water content of 5 wt% to obtain an ethanol extract.

[0077] S5. Preparation of Licorice Extract

[0078] The licorice was dried and crushed, passed through an 80-mesh sieve, and water was added at a solid-liquid ratio of 1:20 g / mL. The mixture was extracted at 90° C. for 4 times, each time for 30 min. The filtrates were combined, concentrated, and dried to a water content of 4 wt% to obtain a licorice extract.

[0079] S6. Preparation of Chinese medicine bone fluid

[0080] Sucrose fatty acid ester SE-15 and water are mixed in a mass ratio of 1:7, and stirred at 55°C and 1200rpm for 16 minutes to obtain sucrose fatty acid ester SE-15 colloid; Angelica dahurica extract, Clematidis extract, ethanol extract, licorice extract, steviol glycoside, ascorbic acid, sucrose fatty acid ester SE-15 colloid and water (the amount is 4.5 times the total weight of Angelica dahurica extract, Clematidis extract, ethanol extract, licorice extract, steviol glycoside and ascorbic acid) are mixed, and homogenized at 7000rpm for 20 minutes to obtain the traditional Chinese medicine bone liquid.

[0081] Example 3

[0082] Preparation of Chinese herbal bone fluid

[0083] S1. Weighing

[0084] Weigh 25 parts of Angelica dahurica, 20 parts of Clematidis, 30 parts of Eucommia ulmoides, 35 parts of Drynaria fortunei, 15 parts of Cinnamomum cassia, 40 parts of Millettia repens, 12 parts of Licorice, 0.05 parts of Stevioside, 0.15 parts of Sucrose Fatty Acid Ester SE-15, and 0.03 parts of ascorbic acid respectively;

[0085] S2. Preparation of Angelica dahurica Extract

[0086] The angelica dahurica powder was dried and crushed, passed through a 250-mesh sieve to obtain angelica dahurica powder, mixed with water at a solid-liquid ratio of 1:28 g / mL, sterilized, inoculated with a composite bacterial agent, and anaerobically fermented at 40°C and pH 7 for 48 hours, sterilized, filtered, and the filtrate was concentrated and dried to a water content of 4 wt% to obtain the angelica dahurica extract;

[0087] The composite bacterial agent is composed of Bifidobacterium animalis and Clostridium tyrobutyricum; the bacterial count of Bifidobacterium animalis in the composite bacterial agent is 500 million CFU / g, and the bacterial count of Clostridium tyrobutyricum is 800 million CFU / g; the inoculation amount of the composite bacterial agent is 5‰ of the weight of the Angelica dahurica powder;

[0088] S3. Preparation of Clematis chinensis extract

[0089] The clematis root was crushed and passed through a 200-mesh sieve to obtain clematis root powder, which was mixed with a 5% by volume acetic acid solution at a solid-liquid ratio of 1:20 g / mL, and ultrasonically extracted at 50° C. and 40 kHz for 20 min. A complex enzyme was added at 8% by weight of the clematis root powder, and enzymatic hydrolysis was performed at 55° C. and pH 5.5 for 30 min. After inactivation of the enzyme, the mixture was filtered, and the filtrate was concentrated and dried to a water content of 4 wt% to obtain a clematis root extract.

[0090] The complex enzyme consists of cellulase and papain in a weight ratio of 4:1;

[0091] S4. Preparation of ethanol extract

[0092] Eucommia ulmoides, Drynaria fortunei, cinnamon twig and Millettia reticulata were crushed separately, passed through a 350-mesh sieve, mixed, and added with an 80% ethanol solution by volume at a solid-liquid ratio of 1:15 g / mL. Ultrasonic extraction was performed at 55°C and 35 kHz for 40 min, filtered, and the filtrate was concentrated and dried to a water content of 3 wt% to obtain an ethanol extract.

[0093] S5. Preparation of Licorice Extract

[0094] The licorice was dried and crushed, passed through a 120-mesh sieve, and water was added at a solid-liquid ratio of 1:30 g / mL. The mixture was extracted twice at 100° C. for 60 min each time. The filtrates were combined, concentrated, and dried to a water content of 4 wt % to obtain a licorice extract.

[0095] S6. Preparation of Chinese medicine bone fluid

[0096] Sucrose fatty acid ester SE-15 and water are mixed in a mass ratio of 1:9, and stirred at 55°C and 1200rpm for 15 minutes to obtain sucrose fatty acid ester SE-15 colloid; Angelica dahurica extract, Clematidis extract, ethanol extract, licorice extract, steviol glycoside, ascorbic acid, sucrose fatty acid ester SE-15 colloid and water (the amount is 5.5 times the total weight of Angelica dahurica extract, Clematidis extract, ethanol extract, licorice extract, steviol glycoside and ascorbic acid) are mixed, and homogenized at 9000rpm for 12 minutes to obtain the traditional Chinese medicine bone liquid.

[0097] Comparative Example 1

[0098] This comparative example is the same as the specific implementation method of Example 1, except that "20 parts of Angelica dahurica and 16 parts of Clematidis" in step S1 are replaced by "36 parts of Angelica dahurica", step S3 is deleted, and the Clematidis extract in step S6 is deleted.

[0099] Comparative Example 2

[0100] This comparative example is the same as the specific implementation method of Example 1, except that "20 parts of Angelica dahurica and 16 parts of Clematidis" in step S1 are replaced by "36 parts of Clematidis", step S2 is deleted, and the Angelica dahurica extract in step S6 is deleted.

[0101] Comparative Example 3

[0102] This comparative example is the same as the specific implementation manner of Example 1, except that a different composite bacterial agent is used to prepare the angelica dahurica extract in step S2: the angelica dahurica is dried and crushed, passed through a 200-mesh sieve to obtain angelica dahurica powder, mixed with water at a solid-liquid ratio of 1:25 g / mL, and sterilized. The composite bacterial agent is inoculated and anaerobically fermented at 37°C and a pH value of 6.8 for 60 hours, sterilized, filtered, and the filtrate is concentrated and dried to a water content of 5wt% to obtain the angelica dahurica extract; the composite bacterial agent is composed of Bifidobacterium longum and Clostridium butyricum; the bacterial count of Bifidobacterium longum in the composite bacterial agent is 300 million CFU / g, and the bacterial count of Clostridium butyricum is 600 million CFU / g; the inoculation amount of the composite bacterial agent is 8‰ of the weight of the angelica dahurica powder.

[0103] Comparative Example 4

[0104] The specific implementation of this comparative example is the same as that of Example 1, except that the "acetic acid solution with a volume fraction of 4%" in step S3 is replaced by "water".

[0105] Comparative Example 5

[0106] This comparative example is the same as the specific implementation of Example 1, except that the "ethanol solution with a volume fraction of 75%" in step S4 is replaced by "water".

[0107] Test Example 1

[0108] 190-210 g SD rats were selected and adaptively fed for 7 days (during which they had free access to water and food), and then randomly divided into 11 groups (blank control group, model group, positive control group, and experimental group (Examples 1-3 and Comparative Examples 1-5)), with 10 rats in each group. There were no significant differences in age, weight, health status, etc. among the rats in each group.

[0109] Except for the blank control group, the right hind paws of rats in other groups were disinfected with 75% alcohol and injected with complete Freund's adjuvant at multiple points subcutaneously at a dose of 0.1 mL / rat. The blank control group was injected with an equal volume of normal saline.

[0110] After the modeling was completed, the experimental group was gavaged with the Chinese medicine bone liquid of Examples 1-3 and Comparative Examples 1-5 at a dose of 0.5 mL / kg, the blank control group and the model group were gavaged with an equal volume of normal saline, and the positive control group was gavaged with Compound Rheumatism Ning Tablets (manufacturer Luofushan) at a dose of 300 mg / kg, once a day for 15 consecutive days.

[0111] (1) Determination of the percentage increase in joint swelling

[0112] The thickness of the right hind paw was measured with a vernier caliper before modeling and 15 days after administration, and the percentage increase in paw swelling was calculated. The specific results are shown in Table 1.

[0113] Percentage increase in joint swelling (%) = (right hind paw plantar thickness 15 days after administration - right hind paw plantar thickness before modeling) / right hind paw plantar thickness before modeling × 100%.

[0114] Table 1 Percentage increase in joint swelling in each group (%)

[0115]

[0116]

[0117] As shown in the data of Table 1, the percentage increase in joint swelling in the model group was significantly higher than that in the blank control group, indicating that the animal model was successfully constructed. The percentage increase in joint swelling in the embodiment is lower than that in the positive control group, indicating that the embodiment has a significant therapeutic effect in treating rheumatism. As shown in the data comparison of Example 1 and Comparative Examples 1-2, there is a synergistic effect between the Angelica dahurica extract and the Clematis chinensis extract, and the combination of the two can significantly reduce the percentage increase in joint swelling. As shown in the data comparison of Example 1 and Comparative Examples 3-5, the use of different microorganisms and different solvents to extract Chinese medicine can affect the percentage increase in joint swelling, thereby affecting the effect of treating rheumatism.

[0118] (2) Determination of inflammatory factors

[0119] On the 15th day, 12 hours after administration, the rats were anesthetized, and blood was collected from the abdominal cavity and centrifuged to obtain serum. The levels of inflammatory factors (IL-6, TNFα and PGE2) in the rats of each group were determined by ELISA according to the kit. The specific results are shown in Table 2.

[0120] Table 2 Inflammatory factor levels in each group

[0121]

[0122]

[0123] As shown in Table 2, the content of inflammatory factors in the model group was significantly higher than that in the blank control group, indicating that the rat model was successfully established. The content of inflammatory factors in the embodiment and the positive control group was not much different, indicating that the Chinese medicine bone solution in the embodiment can be used to treat rheumatic diseases. As shown in the data comparison of Example 1 and Comparative Examples 1-2, the compounding of Angelica dahurica extract and Clematis chinensis extract can reduce the inflammatory response caused by complete Freund's adjuvant. As shown in the data comparison of Example 1 and Comparative Examples 3-5, the use of different microorganisms and different solvents to extract Chinese medicine can reduce the inflammatory response.

[0124] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as within the scope of protection of the present invention.

Claims

1. A Chinese medicinal bone liquid for inhibiting rheumatism and strengthening bones, characterized in that: The invention comprises the following raw materials in parts by weight: 15-25 parts of angelica dahurica, 12-20 parts of clematis root, 20-30 parts of eucommia bark, 20-35 parts of drynaria rhizome, 10-15 parts of cassia twig, 25-40 parts of millettia reticulata and 8-12 parts of liquorice.

2. A method for preparing the Chinese medicine bone liquid according to claim 1, characterized in that: The following steps are involved: The angelica dahurica is crushed and mixed with water, and a composite bacterial agent is inoculated for anaerobic fermentation, and the fermentation liquid is concentrated to obtain the angelica dahurica extract; the clematis root is crushed and mixed with an acetic acid solution, and after ultrasonic treatment, the pH value is adjusted, a composite enzyme is added for enzymatic hydrolysis, and the enzymatic hydrolysis liquid is concentrated to obtain the clematis root extract; the eucommia ulmoides, drynaria rhizome, cassia twig and millettia reticulata are crushed, mixed with an ethanol solution, ultrasonically extracted, filtered, and the filtrate is concentrated to obtain an ethanol extract; the licorice is crushed and mixed with water, heated for extraction, filtered, and concentrated to obtain the licorice extract; the angelica dahurica extract, the clematis root extract, the ethanol extract and the licorice extract are mixed, auxiliary materials are added, and homogenized to obtain the traditional Chinese medicine bone liquid.

3. The preparation method according to claim 2, characterized in that The composite bacterial agent is composed of animal Bifidobacterium and Clostridium tyrobutyricum; the bacterial count of animal Bifidobacterium in the composite bacterial agent is 200-500 million CFU / g, and the bacterial count of Clostridium tyrobutyricum is 500-800 million CFU / g; the inoculation amount of the composite bacterial agent is 5‰-10‰ of the weight of Angelica dahurica.

4. The preparation method according to claim 2 or 3, characterized in that The temperature of the composite bacterial agent fermentation is 35-40° C., the pH value is 6.5-7, and the fermentation time is 48-72 hours.

5. The preparation method according to claim 2, characterized in that The volume fraction of the acetic acid solution is 3%-5%, the material-liquid ratio of the ultrasonic treatment is 1:15-20 g / mL, the temperature is 40-50° C., the frequency is 30-40 kHz, and the time is 20-30 min.

6. The preparation method according to claim 2, characterized in that The complex enzyme is composed of cellulase and papain in a weight ratio of 2-4:1, and the added amount of the complex enzyme is 3%-8% of the weight of the clematis. The enzymolysis temperature is 50-55° C., the pH value is 4.5-5.5, and the time is 30-40 minutes.

7. The preparation method according to claim 2, characterized in that The volume fraction of the ethanol solution is 70%-80%; the material-liquid ratio of the ultrasonic extraction is 1:10-15 g / mL, the temperature is 45-55° C., the frequency is 25-35 kHz, and the time is 40-50 min.

8. The preparation method according to claim 2, characterized in that The mass volume ratio of the licorice to water is 1:20-30 g / mL, the extraction temperature is 90-100° C., and the extraction times are 2-4 times, each time for 30-60 minutes.

9. The preparation method according to claim 2, characterized in that The auxiliary materials include one or more of sweeteners, stabilizers, and antioxidants.

10. Use of the traditional Chinese medicine bone liquid according to claim 1 or the traditional Chinese medicine bone liquid obtained by the preparation method according to any one of claims 2 to 9 in preparing a drug for treating rheumatism.

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