阅读说明：本技术 一种缓解关节骨炎炎症反应的中药组合物及其制备方法 (Traditional Chinese medicine composition for relieving arthritis inflammatory reaction and preparation method thereof ) 是由 黎一江 郭弘妍 邢婉丽 程京 于 2019-12-10 设计创作，主要内容包括：本发明公开了一种缓解关节骨炎炎症反应的中药组合物及其制备方法。所述中药组合物由下述质量份的组分制成：伸筋草100～140份；透骨草100～140份；独活100～140份；威灵仙100～140份；当归100～140份；冰片100～140份；川牛膝100～140份。本发明中药组合物中不含有药典标明的毒性中药材成分,并且在肝肾细胞中对该药物组合进行了细胞毒性评价,发现对肝肾细胞的活性无明显的抑制作用,可长期服用。(The invention discloses a traditional Chinese medicine composition for relieving arthritis inflammatory reaction and a preparation method thereof. The traditional Chinese medicine composition is prepared from the following components in parts by mass: 100-140 parts of lycopodium clavatum; 100-140 parts of garden balsam stem; 100-140 parts of radix angelicae pubescentis; 100-140 parts of clematis root; 100-140 parts of angelica; 100-140 parts of borneol; 100-140 parts of medicinal cyathula root. The traditional Chinese medicine composition does not contain toxic traditional Chinese medicine components indicated by pharmacopoeia, and the cytotoxicity evaluation is carried out on the medicine composition in liver and kidney cells, so that the traditional Chinese medicine composition has no obvious inhibition effect on the activity of the liver and kidney cells, and can be taken for a long time.)

1. A traditional Chinese medicine composition for relieving osteoarthritis inflammatory reaction is prepared from the following components in parts by mass:

100-140 parts of lycopodium clavatum;

100-140 parts of garden balsam stem;

100-140 parts of radix angelicae pubescentis;

100-140 parts of clematis root;

100-140 parts of angelica;

100-140 parts of borneol;

100-140 parts of medicinal cyathula root.

2. The traditional Chinese medicine composition according to claim 1, characterized in that: the traditional Chinese medicine composition is prepared from the following components in parts by mass:

100 parts of lycopodium clavatum;

100 parts of garden balsam stem;

100 parts of radix angelicae pubescentis;

100 parts of clematis root;

100 parts of angelica;

100 parts of borneol;

100 parts of medicinal cyathula root.

3. A method for preparing the Chinese medicinal composition of claim 1 or 2, comprising the steps of: weighing lycopodium clavatum, garden balsam stem, doubleteeth pubescent angilica root, clematis root, Chinese angelica, borneol and medicinal cyathula root, mixing the raw materials, then carrying out reflux extraction by adopting ethanol water solution or water, filtering the obtained extract, and concentrating to obtain an extract, namely the traditional Chinese medicine composition.

4. The production method according to claim 3, characterized in that:

in the preparation method, the volume fraction of ethanol in the ethanol aqueous solution is 85-95%;

the volume consumption of the ethanol water solution is 10-15 times of the mass of the raw materials, and the unit is mL/g;

the volume consumption of the water is 10-15 times of the mass of the raw materials, and the unit is mL/g.

5. The method of claim 3 or 4, wherein: the reflux extraction time is 2-3 h;

the concentration temperature is 40-45 ℃; the concentration multiple is 0.2-1.

6. A medicament for relieving osteoarthritis inflammation reaction, wherein the active ingredients of the medicament are the traditional Chinese medicine composition as claimed in claim 1 or 2.

7. The medicament of claim 6, wherein: the dosage form of the medicine is tablets, pills, paste, capsules or granules.

8. Use of a Chinese medicinal composition according to claim 1 or 2 in the manufacture of a medicament for alleviating an inflammatory response to osteoarthritis.

9. Use according to claim 8, characterized in that: the application appears to reduce the relative expression of COX-2 in chondrocytes.

Technical Field

The invention relates to a traditional Chinese medicine composition for relieving osteoarthritis inflammatory reaction and a preparation method thereof, belonging to the technical field of traditional Chinese medicine preparations.

Background

Osteoarthritis (OA) is a degenerative joint disease with an increasing incidence of age, and is pathologically characterized by primary or secondary degeneration of articular cartilage. Relevant research shows that the incidence rate of osteoarthritis is about 5 percent in population over 50 years old, the incidence rate of female over 60 years old is 25 percent, and the incidence rate of male is 5 percent in China. Cyclooxygenase (COX) catalyzes the synthesis of prostaglandins, which are involved in a variety of pathological and physiological processes in the body. COX has two isoenzymes, COX-1 and COX-2, the former is related to normal physiological functions of the body, such as protecting the gastric mucosa and controlling platelet aggregation, and the latter is related to inflammation or tumor occurrence. COX-1 is secreted in most cells in normal tissues, and COX-2 is expressed only when stimulated by pathological conditions. Proinflammatory cytokines promote COX-2 transcription and expression by modulating transcription factor activity. COX-2 promotes the conversion of arachidonic acid to PGE2, increases PGE2 and collagenase secreted by chondrocytes and synoviocytes, promotes chondrocytes to synthesize various matrix metalloproteinases, accelerates the degradation of proteoglycan and type II collagen in cartilage matrix, and inhibits the synthesis thereof, thereby causing destruction of bone joints and aggravating inflammatory response of osteoarthritis.

Therapeutic agents for osteoarthritis are largely divided into two categories: western medicines and Chinese medicines. Non-steroidal anti-inflammatory drugs (NSAIDs) are the most commonly used western medicines in clinic for treating osteoarthritis symptoms, and have the effects of resisting inflammation, relieving pain and allaying fever. Some adverse reactions may be caused by using the non-steroidal anti-inflammatory drugs, and if the drugs are taken for a long time, the drugs may cause digestive tract ulcer and renal function damage. Studies have found that 1% to 4% of people taking NSAIDs annually develop severe gastrointestinal complications, including bleeding, perforation and infarction. The risk of severe gastrointestinal complications after intake of NSAIDs is 3-5 times higher than that of non-taken patients. The traditional Chinese medicine considers that the main pathogenesis of the knee osteoarthritis is deficiency and excess, flaccidity and arthralgia; the main treatment formulas are tonifying and wind-dispelling dampness-eliminating formulas, because the liver stores blood, governs tendons, kidneys store essence, governs bones, and is deficient in liver and kidney, and essence and blood are deficient, so that muscles and bones are not nourished, the striae are empty and deficient, and wind-cold-dampness is easily induced to cause arthralgia. Most of the traditional Chinese medicine formulas for treating osteoarthritis contain medicinal materials such as monkshood, kusnezoff monkshood root, monkshood and the like containing highly toxic aconitine and other medicinal materials marked with high toxicity in other traditional Chinese medicine pharmacopoeias, and are not suitable for oral administration. Meanwhile, the specific left and right mechanisms of most prescriptions are not exact. Therefore, aiming at the defects of the existing therapeutic drugs, a new traditional Chinese medicine composition is provided.

Disclosure of Invention

The invention aims to provide a traditional Chinese medicine composition for relieving osteoarthritis inflammatory reaction and a preparation method thereof, the traditional Chinese medicine composition is prepared by compounding lycopodium clavatum, garden balsam stem, pubescent angelica root, clematis root, Chinese angelica, borneol and medicinal cyathula root according to a specific proportion, and the anti-inflammatory effect is achieved by reducing the expression of related genes of COX-2 (Cyclo-oxidative gene 2, cyclooxygenase 2).

Aiming at the problem of adverse reaction of the non-steroidal anti-inflammatory drugs, the invention adopts liver and kidney cells to evaluate the cytotoxicity of the developed composition; aiming at the problems that the traditional Chinese medicine prescription contains toxic prescription and the curative effect mechanism of the traditional Chinese medicine is not clear, the traditional Chinese medicine composition selects medicinal materials without toxicity in pharmacopoeia, adopts a transcriptome sequencing method to evaluate the expression of key genes in inflammatory reaction, and finally screens out a nontoxic and effective composition.

Specifically, the traditional Chinese medicine composition for relieving osteoarthritis inflammatory reaction provided by the invention is prepared from the following components in parts by mass:

100-140 parts of lycopodium clavatum;

100-140 parts of garden balsam stem;

100-140 parts of radix angelicae pubescentis;

100-140 parts of clematis root;

100-140 parts of angelica;

100-140 parts of borneol;

100-140 parts of medicinal cyathula root.

Specifically, the traditional Chinese medicine composition is prepared from the following components in parts by mass:

100 parts of lycopodium clavatum;

100 parts of garden balsam stem;

100 parts of radix angelicae pubescentis;

100 parts of clematis root;

100 parts of angelica;

100 parts of borneol;

100 parts of medicinal cyathula root.

The traditional Chinese medicine composition is prepared by the following method:

weighing lycopodium clavatum, garden balsam stem, doubleteeth pubescent angilica root, clematis root, Chinese angelica, borneol and medicinal cyathula root, mixing the raw materials, then carrying out reflux extraction by adopting ethanol water solution or water, filtering the obtained extract, and concentrating to obtain an extract, namely the traditional Chinese medicine composition.

In the preparation method, the volume fraction of ethanol in the ethanol aqueous solution is 85-95%;

the volume consumption of the ethanol water solution is 10-15 times of the mass of the raw materials, and the unit is mL/g;

the volume consumption of the water is 10-15 times of the mass of the raw materials, and the unit is mL/g.

In the preparation method, the reflux extraction time is 2-3 h;

the concentration temperature is 40-45 ℃; the concentration multiple is 0.2-1.

The traditional Chinese medicine composition can be prepared into a medicine for relieving osteoarthritis inflammatory reaction by taking the traditional Chinese medicine composition as an active ingredient, and the dosage form of the medicine can be tablets, pills, paste, capsules or granules.

The invention adopts transcriptome sequencing technology to evaluate the cell pharmacology of the traditional Chinese medicine composition, and finally obtains the traditional Chinese medicine composition by evaluating the expression of a key gene COX-2 influencing inflammatory reaction, and the traditional Chinese medicine composition has the characteristic of remarkably reducing the related expression of COX-2 in chondrocytes.

The traditional Chinese medicine composition does not contain toxic traditional Chinese medicine components indicated by pharmacopoeia, and the cytotoxicity evaluation is carried out on the traditional Chinese medicine composition in liver and kidney cells, so that no obvious inhibition effect on the activity of the liver and kidney cells is found.

The traditional Chinese medicine composition has definite anti-inflammatory effect, has no hepatorenal cytotoxicity, and is suitable for long-term administration.

Compared with the overlapped parts in the prescription reported previously, the prescription of the invention has better anti-inflammatory effect. Meanwhile, the effect of the prescription is compared with that of partial prescription schemes (a first comparison prescription comprising lycopodium clavatum, garden balsam stem, radix angelicae pubescentis and radix cyathulae and a second comparison prescription comprising radix clematidis, angelica sinensis and radix angelicae pubescentis), and the reversion effect of the comparison prescription on inflammation is remarkably lower than that of the scheme through relevant cell experiments, so that the prescription has better osteoarthritis relieving effect.

Drawings

FIG. 1 shows the cytotoxicity results of the formulations of the present invention.

Figure 2 is the pathway enrichment results for the osteoarthritic cell model (normal cells vs osteoarthritic cells).

Figure 3 shows the results of pathway enrichment in the dosing model (dosing of osteoarthritic cells vs osteoarthritic cells).

FIG. 4 is an analysis of inflammatory genes (COX-2) in a dosing model.

FIG. 5 shows the results of channel enrichment for dosing models of the inventive and comparative formulations.

Detailed Description

The experimental procedures used in the following examples are all conventional procedures unless otherwise specified.

Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.