Traditional Chinese medicine preparation for treating early-stage and middle-stage knee osteoarthritis

Abstract

The invention discloses a traditional Chinese medicine preparation for treating early and middle knee osteoarthritis, which comprises the following components in terms of one dose: 20g of purslane speedwell herb, 20g of glabrous sarcandra herb, 15g of epimedium, 15g of ligusticum wallichii, 15g of eucommia ulmoides, 15g of vinegar-processed corydalis tuber, 20g of bighead atractylodes rhizome, 12g of codonopsis pilosula, 20g of angelica sinensis and 15g of radix achyranthis bidentatae. The preparation method comprises the following steps: putting the components into a container, adding a proper amount of cold water, soaking the medicinal materials in the water for 30 minutes to 1 hour, and enabling the water to be 2-3cm higher than the medicinal materials; putting the soaked medicinal materials and soaking water into a decocting container, adding a small amount of water, boiling with big fire, then turning to soft fire, decocting for 20-30 minutes while keeping a slight boiling state, pouring out the liquid medicine after the first decoction, adding a proper amount of cold water again, boiling with big fire until the water immerses the dregs and is 1-2cm higher than the dregs, pouring out the liquid medicine after the first decoction, pouring out the liquid medicine after the second decoction, adding a proper amount of cold water until the water is 1-2cm higher than the dregs, boiling with big fire until the water is 1-2cm higher than the dregs, and pouring out the liquid medicine after the second decoction. Then decocting for 20-30 minutes with soft fire; and respectively pouring out the liquid medicine decocted for the first time and the liquid medicine decocted for the second time, and combining together. The traditional Chinese medicine is used for treating early-stage and middle-stage knee osteoarthritis.

Description

Technical Field

[0001] The invention belongs to the technical field of ethnic medicine, and relates to a medicine for treating early and middle stage knee osteoarthritis, in particular to a traditional Chinese medicine preparation prepared with Miao medicine and traditional Chinese medicine as raw materials. Background Art

[0002] Knee osteoarthritis (KOA) is a chronic degenerative disease caused by multiple factors. It primarily manifests as degeneration of the articular cartilage structure and long-term chronic inflammation, involving pathological changes in multiple structures, including the subchondral bone, synovium, and joint capsule. This process ultimately leads to cartilage erosion, osteophyte formation, and joint space narrowing. Its primary clinical symptoms are pain and limited joint motion. In severe cases, OA can lead to severe damage. Effective treatments for KOA, aside from surgery, are limited. Currently, oral analgesics are the primary treatment for patients with intractable pain in the early and middle stages of knee OA, which can provide effective pain relief. However, the disease is long-lasting, and long-term medications can have a significant impact on the patient's health. Furthermore, the disease is prone to relapse. Therefore, effective methods to alleviate severe pain, actively prevent relapse, and improve treatment outcomes have become key clinical research priorities, and the search for more effective and safe treatment options remains crucial.

[0003] In recent years, ethnic medicine has achieved remarkable results in the prevention and treatment of KOA. It has the advantages of multiple pathways, multiple channels, multiple targets and few side effects, and has received widespread attention. For the treatment of early and middle-stage knee osteoarthritis, compared with Western medicine, ethnic medicine is low in price, has few toxic side effects and significant efficacy.

[0004] For example, Chinese patent CN103230484B discloses a drug for treating knee osteoarthritis in patients under 40 years of age. The drug contains 20 grams of Belamcanda chinensis, 20 grams of Polygonum multiflorum root, 30 grams of Longan pulp, 12 grams of Ziziphus jujuba seeds, 12 grams of Cinnamomum cassia twig, 12 grams of Atractylodes macrocephala, 6 grams of Polygala tenuifolia, 18 grams of Acorus calamus, 15 grams of Ephedra sinica, 10 grams of Morus alba bark, and 45 grams of Sargassum. The drug is mixed and evenly ground into a 180-mesh fine powder. The drug is taken orally twice daily, 10 grams each time, with warm water. A course of treatment is three months.

[0005] Chinese patent CN103705673B discloses a therapeutic drug for treating knee osteoarthritis. The components of the drug, calculated by weight, are as follows: 90-100 parts of black snake, 60-70 parts of star anise, 60-70 parts of perilla seeds, 60-70 parts of white mustard seeds, 20-30 parts of safflower, 50-60 parts of Chinese rhubarb, 50-60 parts of radix polygoni avicularis, 45-50 parts of frankincense, 45-50 parts of myrrh, 40-50 parts of arisaema, 20-25 parts of processed strychnine seeds, 40-50 parts of asarum, and 40-50 parts of ephedra. The present invention adopts traditional Chinese medicine and heat therapy. The warming effect of the Chinese medicine heat therapy and the Chinese medicine contained in the present invention can dilate local capillaries, improve and increase the blood supply around the knee joint, and promote the absorption and dissipation of pathological products such as chronic inflammation, adhesion, infiltration, exudate, etc. in the joint and its surroundings, thereby achieving the effects of promoting blood circulation and removing blood stasis, removing dampness and relieving inflammation, unblocking meridians and relieving pain, and restoring function.

[0006] The above patents each have their own merits, but the inventor has prepared a Chinese medicine preparation (commonly known as "Miao medicine joint and network drink") based on the theory of traditional Chinese medicine Miao medicine using Miao medicine and Chinese medicine as raw materials to treat early and middle stage knee osteoarthritis, and the components are fewer than those in the above two patents. Summary of the Invention

[0007] The purpose of the present invention is to provide a traditional Chinese medicine preparation for treating early and middle stage knee osteoarthritis, so as to address the shortcomings of existing treatment options, alleviate the pain caused by knee osteoarthritis in patients, reduce drug side effects, reduce the recurrence rate, and alleviate the financial burden on patients.

[0008] The technical solution of the present invention: A traditional Chinese medicine preparation for treating early-to-mid-stage knee osteoarthritis, comprising the following components, measured in dosages of: 20g of Herba Cibotii, 20g of Herba Cibotii, 15g of Herba Epimedii, 15g of Rhizoma Chuanxiong, 15g of Eucommia ulmoides, 15g of Corydalis Vinegar, 20g of Atractylodes macrocephala, 12g of Codonopsis pilosula, 20g of Radix Angelicae Sinensis, and 15g of Achyranthes bidentata; Preparation method: put the above components into a container, add an appropriate amount of cold water, the amount of water should be enough to immerse the medicinal materials and be 2 to 3 cm higher, and the soaking time is 30 minutes to 1 hour; put the soaked medicinal materials together with the soaking water into the decoction container, and then add a small amount of water, first boil it over high heat, then turn to low heat, keep it in a slightly boiling state and simmer for 20 to 30 minutes, pour out the medicinal liquid after the first decoction, and add an appropriate amount of cold water again, the amount of water should be enough to immerse the medicinal residue and be 1 to 2 cm higher, and also boil it over high heat first, then turn to low heat and simmer for 20 to 30 minutes; pour out the medicinal liquids of the first and second decoctions separately, and combine them together.

[0009] Usage: One dose three times a day after meals.

[0010] In the formula, Herba Epimedii and Herba Jiujiecha are the main medicines for removing blood stasis, relieving pain, clearing away heat and reducing swelling; Herba Epimedii, Herba Achyranthis Bidentatae, and Herba Eucommiae are the kidney-strengthening and bone-strengthening, and nourishing the kidney and replenishing essence; Herba Angelicae Sinensis, Herba Chuanxiong, and Herba Corydalis Vinegar are the assistant medicines for promoting blood circulation, dispersing blood stasis, replenishing blood and activating blood circulation; most patients with knee osteoarthritis are obese patients with internal accumulation of phlegm and dampness, and Atractylodes macrocephala is added to protect the spleen and stomach, dry dampness and resolve phlegm, and is an adjuvant; in this formula, Codonopsis pilosula replenishes the Qi of the spleen and lungs, is more conducive to protecting the Qi of the spleen and stomach, and is easy for drug absorption, and is the guiding medicine. The above formulas work together to have the effects of promoting blood circulation, removing blood stasis, unblocking meridians and relieving pain, replenishing the liver and kidneys, and strengthening tendons and bones, and can treat early and middle stage knee osteoarthritis.

[0011] The beneficial effects of the present invention are: the present invention selects natural plant medicines, and each component complies with the provisions of the pharmaceutical law. In vitro experiments show that the prescription can have a repairing effect on cartilage cell damage. Therefore, the combined effects of various medicines can be further promoted to treat early and middle-stage knee osteoarthritis. The prescription contains no chemical preparations, is non-toxic and harmless to the human body, has no side effects, is safe and reliable, has high efficacy, is low in price, and does not cause any discomfort. BRIEF DESCRIPTION OF THE DRAWINGS

[0012] Figure 1 These are four groups of chondrocyte morphology images (100×).

[0013] The groups included: A. Blank serum group; B. IL-1β group; C. Miao Yao Joint and Luoyin medicated serum group; D. Miao Yao Joint and Luoyin medicated serum + Compound C group. (Compound C is an inhibitor of the AMPK / mTOR / ULK1 signaling pathway. Group D was established primarily to test whether Miao Yao Joint and Luoyin promote chondrocyte repair by regulating the AMPK / mTOR / ULK1 signaling pathway.) DETAILED DESCRIPTION

[0014] The present invention will be further described below through examples, experiments and drawings.

[0015] Example: A traditional Chinese medicine preparation for treating early and middle stage knee osteoarthritis, comprising the following components, in terms of dosage per dose: 20g of Herba Cibotii, 20g of Herba Cibotii, 15g of Herba Epimedii, 15g of Rhizoma Chuanxiong, 15g of Eucommia ulmoides, 15g of Corydalis Vinegar, 20g of Atractylodes macrocephala, 12g of Codonopsis pilosula, 20g of Radix Angelicae Sinensis, and 15g of Achyranthes bidentata; Preparation method: put the above components into a container, add an appropriate amount of cold water, the amount of water should be enough to immerse the medicinal materials and be 2 to 3 cm higher, and the soaking time is 30 minutes to 1 hour; put the soaked medicinal materials together with the soaking water into the decoction container, and then add a small amount of water, first boil it over high heat, then turn to low heat, keep it in a slightly boiling state and simmer for 20 to 30 minutes, pour out the medicinal liquid after the first decoction, and add an appropriate amount of cold water again, the amount of water should be enough to immerse the medicinal residue and be 1 to 2 cm higher, and also boil it over high heat first, then turn to low heat and simmer for 20 to 30 minutes; pour out the medicinal liquids of the first and second decoctions separately, and combine them together.

[0016] Usage: One dose three times a day after meals.

[0017] In this formula, Herba Cibotii and Herba Corydalis are the main herbs, dissolving blood stasis, relieving pain, clearing heat, and reducing swelling. Herba Epimedii, Herba Achyranthis Bidentatae, and Herba Eucommiae are known to strengthen the kidneys and bones, nourishing the kidneys and replenishing essence. Radix Angelicae Sinensis, Rhizoma Chuanxiong, and Rhizoma Corydalis in vinegar promote blood circulation, dissipate blood stasis, and invigorate blood circulation, assisting Herba Cibotii and Herba Corydalis as the assistant herbs. Knee osteoarthritis patients are often obese with internal phlegm and dampness, so Atractylodes macrocephala is added as an adjuvant to strengthen the spleen and stomach, dry dampness, and resolve phlegm. Codonopsis pilosula is the guiding herb, nourishing the spleen and lungs, further protecting the spleen and stomach, and facilitating drug absorption. These ingredients work together to promote blood circulation, dissipate blood stasis, dredge meridians, relieve pain, nourish the liver and kidneys, and strengthen tendons and bones, effectively treating early and late-stage knee osteoarthritis.

[0018] To demonstrate the therapeutic effect of the drug of the present invention on knee osteoarthritis, in vitro experiments have confirmed that the drug can repair cartilage cell damage. The specific steps are: 1 Materials and Instruments 1.1 Experimental Animals Ten SPF male Sprague-Dawley rats, 6-8 weeks old, weighing (200 ± 20) g, were purchased from Beijing Sibeifu Biotechnology Co., Ltd. (Laboratory Animal Production License No. SCXK (Beijing) 2024-0001) and housed under conventional procedures. The animal husbandry and ethical review procedures were approved by the Ethics Committee of the Bayesian Center for Model Organisms (BSMS Animal (Fu) No. 2024-12-25C).

[0019] 1.2 Chondrocytes Primary rat articular chondrocytes were purchased from Wuhan Huayan Biotechnology Co., Ltd.

[0020] 1.3 Drugs and Reagents The Miao medicine Joint and Meridian Drink is composed of three Miao herbs: 20g of Xiantaocao, 20g of Jiujiecha, and 15g of Epimedium, supplemented by 15g of Chuanxiong, 15g of Eucommia, 15g of Vinegar Corydalis, 20g of Atractylodes macrocephala, 12g of Codonopsis pilosula, 20g of Angelica sinensis, and 15g of Achyranthes bidentata. All experimental drugs were provided by the pharmacy of the Second Affiliated Hospital of Guizhou University of Traditional Chinese Medicine. The ingredients in the formula were authenticated by Associate Professor Wu Zhikun of the Department of Traditional Chinese Medicine Authentication at Guizhou University of Traditional Chinese Medicine. The above herbs were added to a container containing five times the amount of boiling water and soaked for 2 hours. The soaking solution and the herbs were then boiled over high heat, then heated to 100°C for 30 minutes over low heat. The solution was filtered to obtain the medicinal solution. The medicinal residue was then added to five times the amount of water and heated at 100°C for 30 minutes. The residue was filtered to obtain the medicinal solution. The two solutions were mixed and concentrated to 167mL, resulting in a crude drug content of 1g / mL. The solution was then placed in a bottle and stored in a refrigerator at 4°C. CCK8 cell proliferation and cytotoxicity detection kits were provided by Wuhan Huiyucheng Biotechnology Co., Ltd.; rat chondrocyte complete culture medium and Trypsin-EDTA (0.25%) were provided by Wuhan Punosai Biotechnology Co., Ltd.

[0021] 1.5 Main instruments Multiskan FC microplate reader, Thermo Fisher Scientific, USA; H1-16KR desktop high-speed refrigerated centrifuge, Hunan Kecheng Instrument Equipment Co., Ltd.; SPX70BIII biochemical incubator, Feisifu Instrument (Hebei) Co., Ltd.

[0022] 2 Experimental methods 2.1 Cell recovery and passaging The rat chondrocytes were removed from liquid nitrogen and placed in a 37°C water bath. After dissolving, the cells were transferred to a centrifuge tube and collected by centrifugation. The cells were centrifuged at 1000 rpm for 5 min at room temperature and the supernatant was discarded. The cells were resuspended in complete culture medium containing fetal bovine serum. When the cell density reached 80%, the culture medium was discarded and the cells were washed with PBS. 0.25% trypsin was added to digest the cells. After digestion, the trypsin was discarded and complete culture medium was added. The cells were pipetted to make a single-cell suspension. The suspension was passaged at a ratio of 1:3 and expanded and cultured at 37°C and 5% CO2 saturated humidity.

[0023] 2.2 Preparation of drug-containing serum After acclimating for one week, male SD rats were randomly divided into a blank serum group (n=5) and a drug-containing serum group (n=5). The equivalent dose for rats was calculated to be approximately 6.3 times that for humans. Therefore, the drug-containing serum group received 15.02 g / kg of the Miao medicine Jiejiaoheluoyin (Shangguan Heluo Yin) via gavage twice daily for seven days. The blank serum group received the corresponding volume of normal saline via gavage twice daily for seven days. Blood was collected 2-3 hours after the last drug gavage. Rats were anesthetized with continuous isoflurane inhalation. The abdominal cavity was opened, and blood was collected from the abdominal aorta. The blood was placed in a centrifuge tube, allowed to rest for 30 minutes, and then centrifuged at 3000 rpm for 10 minutes. The supernatant serum was aspirated and frozen. The separated serum was inactivated at 56°C for 30 minutes, filtered, and stored at -80°C.

[0024] 2.3 CCK8 assay to detect the effects of different concentrations of drug-containing serum on the proliferation activity of degenerative chondrocytes Normal rat chondrocytes were cultured for three generations in a 37°C incubator and then divided into six groups: control group, model group, 5% Miaoyao Joint and Luoyin medicated serum group, 10% Miaoyao Joint and Luoyin medicated serum group, 15% Miaoyao Joint and Luoyin medicated serum group, and 20% Miaoyao Joint and Luoyin medicated serum group. The control group was cultured with DEME medium only, the model group was cultured with culture medium containing 10 ng / mL IL-1β for 24 hours, and the 5%, 10%, 15%, and 20% Miaoyao Joint and Luoyin medicated serum groups were pretreated with culture medium containing 10 ng / mL IL-1β for 24 hours and then cultured with 5%, 10%, 15%, and 20% Miaoyao Joint and Luoyin medicated serum for 24 hours. After the required cell culture time, 10 μl of cck8 was added to each well and the cells were cultured at 37°C for 1-4 hours. The absorbance OD450 of each well was measured using a microplate reader. The optimal concentration of 10% drug-containing serum was selected for subsequent experiments, and the results are shown below.

[0025] 2.4 Grouping and treatment of degenerated chondrocytes The blank serum group (only 10% blank serum was added to the culture medium for 24 h), IL-1β group (10 ng / mL IL-1β + 10% blank serum), Miao medicine joint and Luoyin medicated serum group (10 ng / mL IL-1β + 10% Miao medicine joint and Luoyin medicated serum), Miao medicine joint and Luoyin medicated serum + AMPK inhibitor Compound C group (10 ng / mL IL-1β + 10% Miao medicine joint and Luoyin medicated serum + 50μmol / L Compound C), among which 10 ng / mL IL-1β was added to the IL-1β group, Miao medicine joint and Luoyin medicated serum group, and Miao medicine joint and Luoyin medicated serum + AMPK inhibitor Compound C group. After 24 hours of IL-1β pretreatment, 10% of the Miao medicine joint and Luoyin medicated serum group and the Miao medicine joint and Luoyin medicated serum + AMPK inhibitor Compound C group were added respectively. The Miao medicine joint and Luoyin medicated serum + AMPK inhibitor Compound C group was added with 10% of the medicated serum and 50 μmol / L Compound C for 24 hours. 2.5 Observation of chondrocyte morphology in different groups The morphology of chondrocytes in each group was observed using an inverted microscope, and the chondrocytes and culture supernatant were collected for subsequent experiments.

[0026] in conclusion: like Figure 1 As shown, the chondrocytes in the blank serum group (A) have regular morphology and are oval. The cells are closely arranged and are basically the same in size, resembling paving stones. In the IL-1β group (B), the Miao medicine joint and Luoyin medicated serum group (C), and the Miao medicine joint and Luoyin medicated serum + Compound C group (D), the chondrocytes had irregular morphology, such as spindle-shaped or irregular triangles, with significantly enlarged intercellular spaces, a decreased number of chondrocytes, and an increased number of dead cells; Compared with the IL-1β group (B), the number of chondrocytes in the Miao medicine joint and Luoyin medicated serum group (C) increased, and the cells were arranged more evenly. The number of cells in the Miao medicine joint and Luoyin medicated serum group + Compound C group (D) decreased compared with the medicated serum group, and the proportion of irregular cells increased significantly.

[0027] This indicates that the "Miao medicine joint and meridian drink" of the present invention can promote chondrocyte proliferation, reduce chondrocyte damage and cell apoptosis, and this process may be related to the regulation of AMPK / mTOR / ULK1 signaling pathway.

[0028] In summary, the present invention selects natural herbal medicines, and each component complies with the provisions of pharmaceutical laws and regulations. In vitro experiments show that this prescription can have a repairing effect on cartilage cell damage. Therefore, the combined effects of various medicines can be further promoted to treat early and middle-stage knee osteoarthritis. This prescription does not contain chemical preparations, is non-toxic and harmless to the human body, has no side effects, is safe and reliable, has high efficacy, is low in price, and does not cause any discomfort.

[0029] The above description is merely a preferred embodiment of the present invention and does not constitute any form of limitation to the present invention. Any simple modification, equivalent change and modification made to the above embodiment based on the technical essence of the present invention without departing from the content of the technical solution of the present invention shall still fall within the scope of the technical solution of the present invention.

Claims

1. A Chinese medicine preparation for treating early and middle stage knee osteoarthritis, characterized by: The invention comprises the following components, calculated in terms of the amount for one dose: 20g of Herba Cibotii, 20g of Herba Cibotii, 15g of Herba Epimedii, 15g of Rhizoma Chuanxiong, 15g of Eucommia ulmoides, 15g of Corydalis yanhusuo, 20g of Atractylodes macrocephala, 12g of Codonopsis pilosula, 20g of Angelica sinensis, and 15g of Achyranthes bidentata.

2. A Chinese medicine preparation for treating early and middle stage knee osteoarthritis according to claim 1, characterized in that: The preparation method comprises the following steps: placing the components into a container, adding an appropriate amount of cold water, wherein the amount of water is sufficient to immerse the medicinal materials and protrude by 2 to 3 centimeters, and the soaking time is 30 minutes to 1 hour; placing the soaked medicinal materials together with the soaking water into a decoction container, adding an appropriate amount of water, first boiling over high heat, then reducing heat, and decocting for 20 to 30 minutes while maintaining a slight boiling state; pouring out the medicinal liquid after the first decoction, and re-adding an appropriate amount of cold water, wherein the amount of water is sufficient to immerse the medicinal residue and protrude by 1 to 2 centimeters, and similarly first boiling over high heat, then reducing heat and decocting for 20 to 30 minutes; and pouring out the medicinal liquids from the first and second decoctions respectively, and combining them together to prepare the preparation.

3. A Chinese medicine preparation for treating early and middle stage knee osteoarthritis according to claim 2, characterized in that: Usage: One dose three times a day after meals.

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