Tibetan medicine spray for diminishing inflammation and relieving pain and preparation method thereof

Abstract

The invention discloses a Tibetan medicine spray for diminishing inflammation and relieving pain and a preparation method thereof. The preparation method comprises the following steps: 1, washing and airing; 2, pretreatment: soaking the Artemisia villosa in the highland barley wine, and supplementing with the fructus chebulae; 3, taking ethanol with the concentration of 55%-80%, and soaking the Artemisia villosa obtained in the step 2 and the oxytropis into an ethanol solution; 4, ultrasonic extraction and ethanol volatilization; and 5, blending to a proper concentration to obtain the spray. The Tibetan two-ingredient black medicine spray has the advantages that 1, the Tibetan two-ingredient black medicine spray has the specific effects of relieving arthralgia and swelling, improving joint functions, reducing inflammation indexes and the like on rheumatoid arthritis; 2, immune inflammatory response can be regulated, abnormal proliferation of joint synovial cells is inhibited, and cartilage repair is promoted;

Description

Technical Field

[0001] The invention belongs to the technical field of medicine, and particularly relates to an anti-inflammatory and analgesic Tibetan medicine spray and a preparation method thereof. Background Art

[0002] Rheumatoid arthritis (RA) is a common autoimmune disease characterized by erosive arthritis. In Tibetan medicine, RA is called "Zhenbu," a category encompassing "yellow water disease" and "Bacon disease." Li Qiuyue et al. believe RA is an inflammatory disease that causes redness, swelling, pain, and other symptoms in the joints, muscles, bones, and tendons. Shen Peng believes that Tibetan areas, situated on a plateau with drastic climate changes, are prone to RA, making it a common and recurring disease in the high-altitude, cold regions of the Qinghai-Tibet Plateau. Most research on Tibetan medicine for the treatment of rheumatoid arthritis focuses on compound formulas, such as those containing Oxytropis vulgaris, Bangga, Tibetan Acorus calamus, and Broad-tendoned Vine for treating joint inflammation. Among them, Oxytropis vulgaris and Bangga have anti-inflammatory and analgesic effects, while Acorus calamus can invigorate the mind and resolve phlegm, regulate qi and activate blood circulation, and Rhizoma Cynanchifolia can relax tendons and activate meridians, dispel wind and relieve pain. Combined, these herbs effectively dispel wind and dampness, unblock meridians, and relieve pain. They are commonly used to treat inflammatory joint diseases such as rheumatoid arthritis and rheumatoid arthritis, relieving symptoms such as joint pain, swelling, and difficulty flexing and extending. Studies exploring the monomeric components of certain Tibetan medicines have found that some have anti-inflammatory and immunomodulatory effects, potentially showing potential therapeutic value for rheumatoid arthritis. Researchers both domestically and internationally have conducted extensive research on the pathogenesis of rheumatoid arthritis using transcriptomics techniques, identifying many key genes and signaling pathways involved in the development and progression of the disease. However, in-depth research on specific Tibetan medicine sprays and transcriptomics remains lacking.

[0003] RA is characterized by a long course and progressive worsening of symptoms, severely compromising patients' health and reducing their physical function and quality of life. Commonly used treatments include disease-modifying antirheumatic drugs, glucocorticoids, nonsteroidal anti-inflammatory drugs (NSAIDs), and herbal remedies, but these drugs pose challenges such as high cost, severe adverse reactions, and drug resistance. The present invention addresses this issue by providing a spray for rheumatoid arthritis. Summary of the Invention

[0004] The present invention aims to provide a Tibetan medicine spray for reducing inflammation and relieving pain and a preparation method thereof, which specifically comprises the following steps:

[0005] 1) Prepare the herbs: First, prepare Banggai (Banggai), Oxytropis chinensis, and Terminalia chebula. Soak Banggai in highland barley wine, supplemented with Terminalia chebula, for 24 hours. Banggai is derived from the whole herb, including the roots, of the buttercup plants Aconitum carmichaelii or Aconitum gansuensis. It is bitter, cooling, and slightly toxic. It enters the liver, stomach, and lung meridians, clearing heat, detoxifying, and promoting diuresis. Terminalia chebula can detoxify Banggai.

[0006] 2) Preliminary dissolution: Take out the bangga obtained in step 1, prepare ethanol with a concentration of 55%-80%, put the bangga and oxytropis into the ethanol solution, and allow the active ingredients to dissolve in the ethanol.

[0007] 3) Extraction of active ingredients: Use ultrasonic technology to extract the active ingredients in medicinal materials. This process can extract the active ingredients more efficiently.

[0008] 4) Adjust the concentration: Add water to the desired concentration of the spray to ensure the ideal concentration for the desired effect.

[0009] 5) Finished product production: bottling to obtain spray.

[0010] Furthermore, the ethanol concentration in step 2 is 70%.

[0011] Furthermore, the ratio of the scutellaria baicalensis to the oxtera vine is 1:1.

[0012] Furthermore, the ultrasonic extraction in step 3 is specifically as follows: 1) placing the pretreated sample and the extraction solvent into ultrasonic cleaning;

[0013] 2) Adjust the parameters of the ultrasonic generator to a frequency within the range of 20-50 kHz and an extraction time of 0.5-1 hour;

[0014] 3) Start the ultrasonic generator. The ultrasonic wave will generate strong eddy currents and oscillation effects, which will fully mix and contact the sample and solvent, thereby promoting the dissolution and diffusion of the target components.

[0015] 4) After ultrasonic extraction, the mixture is separated into an upper solution layer and a lower residue layer. This separation can be achieved by centrifugation, filtration, or evaporation.

[0016] 5) The extract is further purified, such as by using macroporous resin adsorption technology or ceramic membrane impurity removal operation, to obtain high-purity active ingredients.

[0017] The anti-inflammatory and analgesic Tibetan medicine spray is prepared by the method.

[0018] In summary, the present invention has the following beneficial effects: 1. The Tibetan Erwei Heiyao spray has specific effects on rheumatoid arthritis in terms of relieving joint pain and swelling, improving joint function, and reducing inflammatory indicators; 2. In terms of mechanism of action: it can regulate immune inflammatory response, inhibit abnormal proliferation of synovial cells, promote cartilage repair, etc., and explain its therapeutic effect at the molecular and cellular levels. DETAILED DESCRIPTION

[0019] The present invention is further described in detail below in conjunction with the embodiments:

[0020] Example 1

[0021] 1) Prepare the herbs: First, prepare the bangga, oyster bean, and terminalia chebula. Soak the bangga in highland barley wine, along with the terminalia chebula, for 24 hours.

[0022] 2) Preliminary dissolution: Take out the bangga obtained in step 1, prepare 70% ethanol, put the bangga and oxytropis into the ethanol solution, and allow the active ingredients to dissolve in the ethanol.

[0023] 3) Extraction of active ingredients: Use ultrasonic technology to extract the active ingredients in the medicinal materials, adjust the parameters of the ultrasonic generator, the frequency is within the range of 50 kHz, and the extraction time is 0.5 hours.

[0024] 4) Adjust the concentration: Add water to the required concentration of the spray (1:2.6~1:3.0) to ensure the ideal concentration.

[0025] 5) Finished product production: bottling to obtain spray.

[0026] Example 2

[0027] (1) Study on the efficacy of Tibetan Erwei Heiyao spray on rats with rheumatoid arthritis

[0028] ① Establishing a Collagen-Induced Arthritis (CIA) Rat Model: Equal volumes of bovine type II collagen (CII) and complete Freund's adjuvant (CFA) were mixed to create an emulsion containing 1 mg / mL of CII. Wistar rats were intradermally injected with 200 μL of this emulsion at the base of their tails. Seven days later, rats were re-immunized with 100 μL of the same emulsion. A normal control group was injected with saline. Arthritis index (AI) scores were assessed 14 days after the initial injection. A successful model was considered established if the AI ​​score in a single paw was greater than 2.

[0029] ② Grouping and drug administration

[0030] ③Behavioral and representational assessment

[0031] ④Ankle joint histopathology

[0032] Thymus and spleen indexes, serum inflammatory index were measured.

[0033] (2) Study on the mechanism of action of Tibetan Erwei Heiyao spray in the treatment of rheumatoid arthritis based on transcriptomics technology

[0034] ① Transcriptomic analysis

[0035] ②Verification of the mechanism of action

[0036] According to the results of transcriptomic analysis, the RT-qPCR method was used to detect the expression levels of differentially expressed gene mRNAs, and the Western blot method was used to detect the expression levels of related pathway proteins, which preliminarily clarified the mechanism of action of Tibetan Erwei Heiyao spray in the treatment of rheumatoid arthritis.

[0037] The therapeutic efficacy of the spray of the present invention was tested, and the test results are as follows:

[0038] Comparison of toe swelling in rats across experimental groups: Compared with the normal control group, the toe joints of rats in the model group were significantly red and swollen, with joint deformation and limited mobility. Compared with the model group, the joint swelling, redness, and deformation symptoms were alleviated to varying degrees in the Erwei Heiyao and diclofenac sodium groups. The results are shown in Table 1.

[0039] Table 1 Comparison of toe swelling in rats of each group on day 0 / 14 / 28

[0040] Group / Days n 0 14 28 P Normal control group 8 5.72±0.35 5.66±0.38 5.78±0.32 Model Group 8 8.73±0.78 9.82±0.87 10.25±0.79 Two-flavor black medicine group 8 8.62±0.35 7.63±0.41 7.03±0.28 Diclofenac sodium group 8 8.35±0.47 7.35±0.75 7.01±0.18

[0041] 2. Comparison of SOD, TNF-α, and IL-6 Levels in Rats Among Groups: Compared with the normal control group, the serum SOD level of rats in the model group was significantly decreased; compared with the model group, the SOD level in the Erwei Heiyao group and the diclofenac sodium group was significantly increased. Compared with the normal control group, the serum TNF-α and IL-6 levels of rats in the model group were significantly increased; compared with the model group, the TNF-α and IL-6 levels in the Erwei Heiyao group and the diclofenac sodium group were significantly decreased. The results are shown in Table 2.

[0042] Table 2 Effects of Erwei Heiyao spray on SOD, TNF-α and IL-6 levels in rats

[0043]

[0044] As can be seen from Tables 1 and 2 above, animal experiments have shown that the two-flavor black medicine spray of the present invention can alleviate the symptoms of redness, swelling and deformation of the toe joints of rats to varying degrees, increase the level of superoxide dismutase in the serum to varying degrees, and exert an antioxidant function; and exert an anti-inflammatory effect by downregulating the levels of tumor necrosis factor and interleukin-6.

[0045] This specific embodiment is merely an explanation of the present invention and is not intended to limit the present invention. After reading this specification, those skilled in the art may make non-creative modifications to this embodiment as needed. However, as long as such modifications are within the scope of the claims of the present invention, they are protected by patent law.

Claims

1. A method for preparing an anti-inflammatory and analgesic Tibetan medicine spray, characterized in that: The specific steps include: 1) Prepare the herbs: First, prepare the Chinese angelica root, Oxytropis chinensis, and Terminalia chebula. Soak the Chinese angelica root in highland barley wine, supplemented with Terminalia chebula, for 24 hours. 2) Preliminary Dissolution: Take the scutellaria baicalensis obtained in step 1, prepare 55%-80% ethanol, place the scutellaria baicalensis and oxytropis oxytropis into the ethanol solution, and allow the active ingredients to dissolve in the ethanol; 3) Extraction of active ingredients: Use ultrasonic technology to extract active ingredients from medicinal materials; 4) Adjust the concentration: add water to the desired concentration of the spray; 5) Finished product production: bottling to obtain spray.

2. The method for preparing an anti-inflammatory and analgesic Tibetan medicine spray according to claim 1, wherein: The ethanol concentration in step 2 is 70%.

3. The method for preparing a Tibetan medicine spray for anti-inflammatory and analgesic purposes according to claim 1, wherein: The ratio of the scutellaria baicalensis to the oxtera prawn is 1:

1.

4. The method for preparing an anti-inflammatory and analgesic Tibetan medicine spray according to claim 1, wherein: The ultrasonic extraction in step 3 specifically comprises: 1) placing the pretreated sample and the extraction solvent in an ultrasonic cleaning chamber; 2) Adjust the parameters of the ultrasonic generator to a frequency within the range of 20-50 kHz and an extraction time of 0.5-1 hour; 3) Start the ultrasonic generator. The ultrasonic wave will generate strong eddy currents and oscillation effects, which will fully mix and contact the sample and solvent, thereby promoting the dissolution and diffusion of the target components. 4) After the ultrasonic extraction is completed, the mixed solution is separated into an upper solution layer and a lower residue layer, which can be separated by centrifugation, filtration or evaporation; 5) The extract is further purified.

5. The anti-inflammatory and analgesic Tibetan medicine spray obtained by the preparation method as claimed in claims 1-4.

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