CN116549552A - Traditional Chinese medicine composition for treating osteoarthritis and preparation method thereof - Google Patents
Traditional Chinese medicine composition for treating osteoarthritis and preparation method thereof Download PDFInfo
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Abstract
The invention provides a traditional Chinese medicine composition for treating osteoarthritis and a preparation method thereof, wherein the traditional Chinese medicine composition comprises the following components in parts by mass: 50 parts of Chinese angelica, 30 parts of white mulberry root-bark, 16 parts of ground beetle, 8 parts of pseudo-ginseng, 16 parts of dahurian angelica root and 3 parts of dragon's blood. Animal experiment and research results show that the traditional Chinese medicine composition provided by the invention has the effect of treating osteoarthritis diseases.
Description
Technical Field
The invention belongs to the technical field of medicines, and in particular relates to a traditional Chinese medicine composition for treating osteoarthritis and a preparation method thereof.
Background
Osteoarthritis (OA) is a endemic disease worldwide. It is a major cause of disability in the elderly and causes pain, loss of function and reduced quality of life (QOL). Knee osteoarthritis (KneeOsteoarthritis, KOA) is a chronic degenerative disease characterized by the formation of osteophytes and loss of cartilage on the surface of the knee, clinically characterized by pain, stiffness, limited movement of the knee, and in severe cases, can lead to swelling and deformity of the knee. KOA belongs to the category of "arthralgia syndrome" and "bone arthralgia" in traditional Chinese medicine. The Chinese medicine has knowledge of arthralgia syndrome, and is recorded in the yellow emperor's internal passage as early as "the disease is in bone, the bone weight is not the same, and the bone marrow ache and the cold qi is the bone arthralgia. Along with the promotion of social development degree and population aging, the probability of osteoarthritis of the population over 50 years old in China is about 5%, the incidence rate of the population over 60 years old can reach 20%, and the proportion of KOA patients can reach 78.5% at the highest. The incidence of KOA has increased significantly and has tended to be younger, and thus it is imperative to find more scientific and effective treatments for KOA.
The research on the pathogenesis of KOA is more and complex, the basic principle is that the adverse factors of balance destruction between anabolic and catabolic functions of articular cartilage are increased, wherein MMP13 is considered as a key enzyme for rational destruction of cartilage, plays an important role in skeletal development and skeletal reconstruction, is a main enzyme for targeting cartilage degradation, and can degrade not only type II collagen in cartilage, but also proteoglycan, type IV collagen, type IX collagen and the like in cartilage. The activation of catabolic axis plays an important role, mainly through the regulation of growth factors, hormones, cytokines and transcription factors on the transcription level, and researches show that the levels of TNF-alpha, IL-8 and IL-18 in serum are positively correlated with MMP13, and researches show that IL-1 beta can induce the expression of MMP13 in chondrocytes. These results all indicate that MMP13 may play an important role in the progression of OA, and as its research is advanced, more and more related molecular pathways and targets are discovered, which lays a foundation for the study of the pathogenesis of OA.
At present, the clinical treatment methods are various, and the traditional Chinese medicine treatment mainly comprises the oral administration, acupuncture, massage, external application of the traditional Chinese medicine and the like, and the western medicine treatment comprises the medicine treatment, the medicine injection of the joint cavity, the joint replacement, the rehabilitation therapy and the like. The oral anti-inflammatory analgesic drug is mainly used for treatment, but has great side effects, especially intestinal side effects, and is difficult for patients to take and endure for a long time. The cartilage protecting medicine, such as glucosamine and hyaluronic acid, only temporarily relieves the illness state, has single effect, long administration time and undefined curative effect. The surgical treatment is mainly used for severe patients, the surgical wound is large, the long-term curative effect is undefined, and the surgical cost is high, so that the patients are not accepted.
Compared with western medicine, the traditional Chinese medicine for treating knee osteoarthritis has the advantages of small side effect, low price and the like, wherein the traditional Chinese medicine for external application has unique curative effect after history verification in centuries. The Chinese herbal medicine composition has the advantages of convenient and flexible use, convenient taking and wearing, pure natural Chinese herbal medicine preparation, safety, no side effect, administration through skin permeation effect on meridian points, no oral gastrointestinal absorption, no toxic or side effect on internal organs such as liver, kidney, spleen, stomach and the like, and suitability for long-term use. And unlike the plaster which is tightly stuck on the skin, the daily flower has the opportunity to relieve illness state within 30-60 minutes, is not allergic and does not irritate the skin, thus being not limited by seasons and being suitable for all seasons.
Disclosure of Invention
The invention aims to provide a traditional Chinese medicine composition for treating osteoarthritis related diseases, which fully exerts the advantages of traditional Chinese medicine therapy and provides an effective method for preventing and/or treating osteoarthritis related diseases.
In order to achieve the above purpose, the present invention provides the following technical solutions:
in a first aspect, the invention provides a traditional Chinese medicine composition for treating osteoarthritis, which comprises the following components in parts by mass: 50 parts of Chinese angelica, 30 parts of white mulberry root-bark, 16 parts of ground beetle, 8 parts of pseudo-ginseng, 16 parts of dahurian angelica root and 3 parts of dragon's blood.
Further, the traditional Chinese medicine composition also comprises a pharmaceutically acceptable carrier.
Further, the Chinese medicinal composition is used for treating or improving osteoarthritis-related symptoms.
In the above aspect, the osteoarthritis-related symptoms include, but are not limited to: chronic progressive polyarthritis, articular cartilage injury, hyperosteogeny, inflammatory cell joint infiltration, synovitis.
More preferably, treating or ameliorating an osteoarthritis-related symptom comprises: relieving pain caused by osteoarthritis, and/or relieving inflammation caused by osteoarthritis, and/or reducing cartilage destruction caused by osteoarthritis.
In a second aspect, the invention further provides a formulation for the treatment of osteoarthritis comprising a Chinese medicinal composition as described above.
Preferably, the preparation comprises an application, a paste, a gel, a decoction, a granule, a tablet, a capsule, an oral liquid, a mixture, a syrup or a pill.
In a third aspect, the present invention also provides the use of a Chinese medicinal composition as described above for the manufacture of a medicament for the treatment of an osteoarthritis-related disorder.
In a fourth aspect, the present invention also provides a method for preparing the above-described Chinese medicinal composition, comprising mixing the following ingredients in proportion to prepare the composition: 50 parts of Chinese angelica, 30 parts of white mulberry root-bark, 16 parts of ground beetle, 8 parts of pseudo-ginseng, 16 parts of dahurian angelica root and 3 parts of dragon's blood.
Compared with the prior art, the invention has the beneficial effects that: the invention provides a traditional Chinese medicine composition, which has the effect of treating osteoarthritis, effectively relieves joint pain and joint inflammation caused by the osteoarthritis, and reduces cartilage damage caused by the osteoarthritis. The invention fully plays the advantages of the traditional medicine, references and develops the traditional medicine, provides scientific basis for developing the medicine for treating the osteoarthritis by taking the traditional medicine recipe as the raw material, and has great application value.
Drawings
Fig. 1 is a graph showing the trend of cold pain threshold change over a month for different group administrations, p < 0.001 (n=10) compared to the ACLT model group;
FIG. 2 is a graph of red and green staining of different groupings of knee joints;
fig. 3 is a graph of OARSI scores for solid green staining of different groupings of knee safranin, p <0.05 (n=3) compared to ACLT model group;
fig. 4 is a graph of immunohistochemical staining of different groupings of knee MMP 13.
Detailed Description
The invention discloses a traditional Chinese medicine composition with an effect of treating osteoarthritis and application thereof, wherein the traditional Chinese medicine compound is prepared from the following raw materials in proportion: 50g of angelica sinensis, 30g of cortex mori radicis, 16g of ground beetle, 8g of pseudo-ginseng, 16g of radix angelicae, and 3g of dragon's blood. The traditional Chinese medicine composition with the effect of treating osteoarthritis is prepared based on a hundred-year clinical curative effect on the basis of a family ancestral bone-setting prescription. The inventor selects the raw material compatibility formula according to the traditional Chinese medicine theory and the pathogenesis of osteoarthritis, and the proportion of each component is scientific and reasonable. Animal experiment and research results show that the externally applied traditional Chinese medicine provided by the invention has the effect of treating osteoarthritis diseases, especially knee osteoarthritis.
The invention provides a product for preventing and/or treating osteoarthritis, which is prepared from the following raw materials in parts by weight: 50g of angelica sinensis, 30g of white mulberry root-bark, 16g of ground beetle, 8g of pseudo-ginseng, 16g of angelica dahurica and 3g of dragon's blood.
The product with the effect of preventing and/or treating osteoarthritis comprises a medicine or a traditional Chinese medicine composition.
Furthermore, the product with the effect of preventing and/or treating osteoarthritis is also added with pharmaceutically acceptable auxiliary materials to prepare a pharmaceutically acceptable preparation; the pharmaceutically acceptable preparation comprises application, ointment, gel, decoction, granule, tablet, capsule, oral liquid, mixture, syrup or pill, etc.
The above-mentioned products are useful for preventing and/or treating osteoarthritis-related disorders, including chronic progressive polyarthritis caused by osteoarthritis, articular cartilage damage, hyperosteogeny, inflammatory cell joints, synovitis, and the like.
The treatment effect is as follows:
(1) Relieving pain caused by osteoarthritis; and/or
(2) Reducing inflammation caused by osteoarthritis; and/or
(3) Reduce cartilage damage caused by osteoarthritis.
The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
In the description of the present invention, it should be noted that the directions or positional relationships indicated by the terms "upper", "lower", "inner", "outer", "front", "rear", "both ends", "one end", "the other end", etc. are based on the directions or positional relationships shown in the drawings, are merely for convenience of describing the present invention and simplifying the description, and do not indicate or imply that the devices or elements referred to must have a specific direction, be configured and operated in the specific direction, and thus should not be construed as limiting the present invention. Furthermore, the terms "first," "second," and the like, are used for descriptive purposes only and are not to be construed as indicating or implying relative importance.
The experimental methods used in the following examples are all conventional methods unless otherwise specified; reagents, biological materials, etc. used in the examples described below are commercially available unless otherwise specified.
The reagents and biological materials used in the following examples were derived from:
animal health clean grade male C57BL/6 mice: jiangsu Huazhuang Xinnuo pharmaceutical technology Co., ltd;
celecoxib: qingdao ocean pharmaceutical Co., ltd.
Example 1:
40C 57BL/6 male mice (20+/-2 g) are randomly divided into 4 groups, and 10 mice in each group are respectively a sham operation control group, a model group, a positive drug control group (celecoxib) and a drug group of the invention, wherein the mice are respectively treated by a blank group, a model group and a drug administration group as follows:
control group: 10 sham-treated C57BL/6 male mice were randomized and given double distilled water with a lavage volume of 0.2mL.
Model group: 10 ACLT model mice were randomly picked and given double distilled water with a lavage volume of 0.2mL.
Positive drug group: 10 ACLT model mice are randomly taken, celecoxib is converted into the mouse administration dose (37 mg/kg) according to the clinical administration dose of human, and the mice are administrated 1 time a day, and the stomach irrigation volume is 0.2mL.
The medicine group of the invention comprises: 10 ACLT model mice were randomly picked up, and the drug was uniformly applied to the knee joints of the mice twice daily.
The method for establishing the ACLT model mice is as follows:
taking 40 healthy clean C57BL/6 male mice, adaptively feeding the mice for one week, injecting 5% chloral hydrate into abdominal cavity, anesthetizing the mice at a dose of 0.2ml each, taking right hind limbs of the mice, shaving the mice with a radius of 1cm around knee joints, disinfecting the iodophor surface after the skin is completely exposed, longitudinally cutting the skin on the knee joint surface by a surgical knife, stripping connective tissue on the knee joint surface by a surgical forceps after the knee joint is exposed, cutting the knee joint ligament, exposing the knee joint cavity, debridement, picking and separating the anterior cruciate ligament by a surgical needle, cutting the anterior cruciate ligament by a small surgical scissors under direct vision (the joint cavity is opened by a prosthetic surgical group, separating the anterior cruciate ligament but not cutting, and other operations are consistent), and then sequentially suturing the knee ligament and the skin on the knee joint surface of the mice. After the operation, the mice were sterilized on the wound surface with iodophor and returned to the cage, and the administration was started after one month of normal feeding.
Example 2:
in this example, cold pain test was performed on mice in the administration period to examine the effect of the drug of the present invention on the pain sensitivity of mice, and the cold pain test procedure is as follows:
mice were evaluated for cold pain threshold using cold plate experiments: and (3) placing the mice on a cold plate with a preset temperature value of 4 ℃, recording the time when the mice perform the actions of licking, lifting legs, backing, jumping and the like, keeping the time of the mice on the cold plate for at most 60s, and stopping the experiment immediately if the time exceeds 60s and the mice still do not react. Three replicates were required for each cold plate experiment, the time interval between each replicate was not less than 10min, and the average of the time recorded from the three replicates was taken as the cold pain threshold for the mice. The results of the test are shown in FIG. 1, starting from the day before administration and measuring every 7 days until the end of the administration.
As can be seen from fig. 1, from day 0 of administration, mice that have undergone modeling treatment begin to develop disease, and the detection results of cold pain show that the cold pain threshold of mice in the model group and the administration group is lower than that of mice in the control group (sham operation group), and that the cold pain threshold of mice in the ACLT model group gradually decreases with the development of arthritis inflammation, indicating rapid increase in pain sensitivity and exacerbation of inflammation pain. Compared with the sensitivity of ACLT mice to pain, the administration group gradually relieves the pain of the joints of the limbs of the mice, the cold pain threshold of the mice gradually rises, and the mice are obviously improved at weeks 1 and 2, and compared with the model control group, the mice have obvious difference (P < 0.0001). This demonstrates that the external application of the drug of the invention can significantly alleviate joint symptoms of ACLT model mice.
In this example, the right hind limb of the mice was also harvested 35 days after the above administration, fixed with formalin, and stained for safranin and fast green, as follows:
(1) Drawing materials: storing the ankle joints of the mice in a 4% paraformaldehyde solution;
(2) Dehydrating: placing the tissue into a dehydration box, placing the dehydration box into a full-automatic dehydrator for dehydration, and dehydrating by using ethanol with concentration gradient;
(3) Embedding: after the tissue has been dehydrated, it is completely immersed in melted paraffin. Paraffin wax was placed in an embedding machine for embedding and then frozen at-20 ℃. After trimming the paraffin blocks, preserving at 4 ℃;
(4) Slicing and pasting: cutting the wax block into 5-8 mu m thick pieces, and then attaching the wax block to a glass slide;
(5) Dehydrating: respectively dehydrating with xylene and alcohol with different concentrations, and washing with distilled water for several times;
(6) Dewaxing and dyeing: dyeing the dehydrated tissue with 0.5% fast green for 20min, slightly washing with 1% acetic acid for 3 times, and washing with running water for 3s;0.5% safranin is dripped on the surface of a sample, and the temperature is 37 ℃ for 5min;
(7) And (3) removing the water sealing piece: dehydrating with xylene and alcohol with different concentrations, placing neutral resin on a cover glass, sealing, and air drying;
(8) And (5) microscopic examination and photographing: and observing and photographing under a microscope.
Sections after safranin fast green staining was completed, each section was scored by the observer according to the principles of OARSI scoring in table 1.
TABLE 1OARSI scoring items
| Symptom manifestation | Score value |
| Cartilage is free from abrasion | 0 |
| The cartilage surface has abnormal safranin deficiency but no structural change | 0.5 |
| Cartilage surface fibrous destruction with small fissures but no loss of cartilage | 1 |
| Cartilage surface fibrous destruction is 1 severe and cartilage loss occurs | 2 |
| Cartilage wear is less than one quarter and breaks down to subchondral bone | 3 |
| Cartilage wear is less than one half and breaks down to subchondral bone | 4 |
| Cartilage wears out more than half and breaks down to subchondral bone | 5 |
Fig. 2 shows the red and green fixation staining of knee joint in different groups, wherein the cartilage matrix of the prosthetic operation group is uniformly stained, the wet line structure is complete, the distribution of each layer is clear, the cartilage cells of the model group are arranged in a disordered manner, a large number of necrotic cartilage cells exist, the cartilage matrix is shallowly stained, the cartilage surface is seriously damaged, the wet line structure is damaged, and part of fiber cracks are visible, so that the success of molding is demonstrated. After the positive medicine is given, the cartilage surface layer is not obviously thinned, each layer is more distinct than the model component, and the wet line structure is basically complete. After the medicine is administered, the cartilage matrix is dyed uniformly compared with the model group, and the wet line structure is slightly damaged, so that the medicine can improve the cartilage damage of the mice with knee osteoarthritis to a certain extent.
Fig. 3 is OARSI scores on safranin and fast green staining of knee joints in different groups, with significant differences (P < 0.01) between control and model groups, indicating successful modeling. The drug of the invention was significantly different (P < 0.05) from the model group, indicating a decrease in knee osteoarthritis index.
In this example, the right hind limb of the mice was also harvested 35 days after the administration and fixed with formalin, and immunohistochemical staining was performed on the mice, as follows:
(1) Dewaxing: dewaxing the slices by using xylene I, xylene II, xylene III, 100% alcohol I, 100% alcohol II, 95% alcohol and 85% alcohol, and washing the slices by using distilled water for a plurality of times;
(2) Antigen retrieval: adding 3% hydrogen peroxide. Washing with distilled water for several times, adding antigen retrieval liquid, and washing with distilled water for several times again; closing: adding 5% serum, and sealing for 30min;
(3) Adding an antibody: adding corresponding primary antibody, and standing at 4 ℃ overnight;
(4) Adding a secondary antibody: washing for several times by PBS, adding corresponding secondary antibodies, and incubating for 2 hours at room temperature; adding a color-developing agent: washing for several times by PBS, and adding a color developing agent;
(5) Counterstaining: washing with distilled water for several times, staining with hematoxylin, differentiating, returning to blue, and washing with running water;
(6) And (3) removing the water sealing piece: dehydrating with xylene and alcohol with different concentrations, placing neutral resin on a cover glass, sealing, and air drying;
(7) And (5) microscopic examination and photographing: photographs were taken under observation under a microscope and analyzed with ImageJ.
FIG. 4 shows immunohistochemical staining of knee joint in different groups, and shows that positive expression rate of MMP-13 in cartilage tissue of mice in model group is increased compared with that in sham operation group, and the positive expression rate is mainly concentrated in the region with serious degeneration of cartilage surface layer, and the staining is dark brown. The distribution is uneven. After the positive medicine is treated, a small amount of MMP-13 positive particles can be seen, and can be seen in the surface layer and the shallow layer, and the dyeing is shallower than that of a model group. After the medicine is given, MMP-13 positive particles are basically distributed uniformly, and the dyeing is shallower than that of a model group. The above results demonstrate that the inventive drug can reduce the expression of knee osteoarthritis chondrocyte MMP 13.
The present invention is not described in detail in the present application, and is well known to those skilled in the art.
Finally, what is to be described is: the above embodiments are only for illustrating the technical solution of the present invention and not for limiting the same, and although the present invention has been described in detail with reference to the examples, it should be understood by those skilled in the art that modifications and equivalents may be made thereto without departing from the spirit and scope of the technical solution of the present invention, which is intended to be covered by the scope of the claims of the present invention.
Claims (9)
1. The traditional Chinese medicine composition for treating osteoarthritis is characterized by comprising the following components in parts by mass: 50 parts of Chinese angelica, 30 parts of white mulberry root-bark, 16 parts of ground beetle, 8 parts of pseudo-ginseng, 16 parts of dahurian angelica root and 3 parts of dragon's blood.
2. The traditional Chinese medicine composition for treating osteoarthritis as claimed in claim 1, further comprising a pharmaceutically acceptable carrier.
3. The traditional Chinese medicine composition for treating osteoarthritis as claimed in claim 1, which is used for treating or improving osteoarthritis-related symptoms.
4. A traditional Chinese medicinal composition for the treatment of osteoarthritis as claimed in claim 3, wherein said symptoms include, but are not limited to: chronic progressive polyarthritis, articular cartilage injury, hyperosteogeny, inflammatory cell joint infiltration, synovitis.
5. The traditional Chinese medicine composition for treating osteoarthritis as claimed in claim 3, wherein treating or improving osteoarthritis-related symptoms comprises: relieving pain caused by osteoarthritis, and/or relieving inflammation caused by osteoarthritis, and/or reducing cartilage destruction caused by osteoarthritis.
6. A formulation for use in the treatment of osteoarthritis, comprising a Chinese medicinal composition according to any one of claims 1 to 5.
7. The formulation for treating osteoarthritis as claimed in claim 6, wherein the formulation comprises an application, a paste, a gel, a decoction, a granule, a tablet, a capsule, an oral liquid, a mixture, a syrup or a pill.
8. Use of a traditional Chinese medicine composition according to any one of claims 1 to 5 for the manufacture of a medicament for the treatment of an osteoarthritis-related disorder.
9. The method for preparing a Chinese medicinal composition according to any one of claims 1 to 5, comprising mixing the following ingredients in proportion to prepare a composition: 50 parts of Chinese angelica, 30 parts of white mulberry root-bark, 16 parts of ground beetle, 8 parts of pseudo-ginseng, 16 parts of dahurian angelica root and 3 parts of dragon's blood.
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