阅读说明：本技术 一种含环橄榄树脂素4’-糖苷的组合物及其制备方法和应用 (Composition containing cycloolivil 4' -glucoside and preparation method and application thereof ) 是由 肖军平 陈梁 毛金娣 刘胜 张小梅 旷春兰 张洪军 于 2021-08-31 设计创作，主要内容包括：本发明属于医药技术领域,具体涉及一种含环橄榄树脂素4’-糖苷的组合物及其制备方法和应用。所述含环橄榄树脂素4’-糖苷的组合物,包括以下组分：环橄榄树脂素4’-糖苷、橄榄素二糖苷、丁香素、β-萘黄酮、槲皮素。该组合物制备方法简单,能够有效提高组合物治疗颈椎病的效果,同时,药物的利用率较高,在有效降低成本的同时减少药物副作用。(The invention belongs to the technical field of medicines, and particularly relates to a composition containing cycloolivil 4' -glucoside, and a preparation method and application thereof. The composition containing cycloolivil 4' -glucoside comprises the following components: cycloolivil 4' -glucoside, olivil diglycoside, syringin, beta-naphthalene flavone, and quercetin. The preparation method of the composition is simple, the effect of the composition in treating cervical spondylosis can be effectively improved, meanwhile, the utilization rate of the medicine is high, the cost is effectively reduced, and meanwhile, the side effects of the medicine are reduced.)

1. A composition containing cycloolivil 4' -glucoside comprises the following components: cycloolivil 4' -glucoside, olivil diglycoside, syringin, beta-naphthalene flavone, and quercetin.

2. The cycloolivil 4' -glycoside-containing composition according to claim 1, characterized by comprising the following components in parts by weight: 5-10 parts of cycloolivil 4' -glucoside, 1-10 parts of olivine diglycoside, 5-15 parts of syringin, 1-10 parts of beta-naphthoflavone and 1-5 parts of quercetin.

3. The cycloolivil 4' -glycoside-containing composition according to claim 1, characterized by comprising the following components in parts by weight: 5-8 parts of cycloolivil 4' -glucoside, 1-5 parts of olivine diglycoside, 5-7 parts of syringin, 2-4 parts of beta-naphthoflavone and 2-3 parts of quercetin.

4. A process for the preparation of a cycloolivil 4' -glycoside-containing composition according to any one of claims 1 to 3, comprising the following steps:

(1) adding a surfactant A into distilled water to prepare a water-phase solvent;

(2) under the condition of heating, dissolving cycloolivil 4' -glucoside, olivine diglycoside and syringin in an aqueous phase solvent in sequence, and stirring to form a solution 1;

(3) adding a surfactant B into the solution 1, and then adding beta-naphthoflavone and quercetin to form a solution 2; stirring to obtain the composition containing cycloolivil 4' -glucoside.

5. The method for preparing a composition containing cycloolivil 4' -glycoside according to claim 4, wherein in step (1), the amount of surfactant A added is 0.1-0.5% by mass of distilled water; the surfactant A is any one or more of octyl-beta-D-glucopyranoside, dodecyl-beta-D-maltopyranoside and N-dodecyl-N-methyl glucamide.

6. The method for preparing the composition containing cycloolivil 4' -glycoside according to claim 4, wherein in the step (1), the surfactant A is a mixture of surfactants in a mass ratio of 1-3: 1 of octyl- β -D-glucopyranoside and N-dodecyl-N-methylglucamide.

7. The method of claim 4, wherein the surfactant B is tea saponin, and the amount of tea saponin added is 0.1-1% of the total weight of β -naphthoflavone and quercetin.

8. The method of claim 7, wherein the tea seed cake is pulverized, soaked in hot water for extraction, filtered, the extractive solution is treated with macroporous adsorbent resin, eluted with aqueous ethanol solution, and dried to obtain tea saponin;

preferably, the temperature of the hot water is 60-80 ℃, and the extraction time is 3-4 h;

preferably, sodium carbonate is added in the extraction process, and the adding amount of the sodium carbonate is 5-10% of the mass of the tea seed cake;

preferably, the macroporous adsorption resin is D3520 or D4006 type macroporous adsorption resin;

the specific process of the ethanol water solution elution is that 80-95% of ethanol water solution is used for eluting to remove impurities, 30-45% of ethanol is used for eluting and collecting eluent.

9. Use of the cycloolivil 4 '-glycoside-containing composition according to any one of claims 1 to 3 or the cycloolivil 4' -glycoside-containing composition prepared by the process according to any one of claims 4 to 8 for the preparation of a medicament for the treatment of cervical spondylosis.

10. A formulation of the cycloolivil 4' -glycoside-containing composition according to any one of claims 1 to 3 or the preparation method of any one of claims 4 to 8, wherein the formulation is a granule, tablet, capsule or oral liquid.

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to a composition containing cycloolivil 4' -glucoside, and a preparation method and application thereof.

Background

Cervical spondylosis is also known as cervical syndrome, is a general name of cervical osteoarthritis, proliferative cervical spondylitis, cervical radicular syndrome and cervical disc herniation, and is a disease based on degenerative pathological changes. Mainly, due to long-term strain of cervical vertebrae, hyperosteogeny, or prolapse of intervertebral disc and ligament thickening, the spinal cord, nerve root or vertebral artery of cervical vertebrae are pressed, and a series of clinical syndromes of dysfunction appear. Instability and looseness of the vertebral segment; herniation or herniation of the nucleus pulposus; forming bone spurs; ligament hypertrophy, secondary spinal stenosis and the like irritate or press tissues such as adjacent nerve roots, spinal cords, vertebral arteries, cervical sympathetic nerves and the like, and cause a series of symptoms and signs.

In the prior art, the medicines for treating cervical spondylosis mainly comprise anti-inflammatory and analgesic medicines; related medicines are various, for example, Chinese patent application CN01109396.X discloses a new medicine for treating cervical spondylosis, which takes red sage root, largehead atractylodes rhizome, white paeony root, pawpaw, eucommia bark, saffron, notopterygium root and kudzuvine root as raw materials, and the raw materials are respectively ground, decocted in water, extracted, dried, sterilized and the like according to different drug properties of each traditional Chinese medicine, and are prepared into the medicine by screening and proportioningThe pill of (1); clinical examination shows that the medicine has obvious curative effect.

Chinese patent application CN201710535200.0 discloses a traditional Chinese medicine formula for treating lumbar cervical spondylosis, which comprises: 20-25 parts of wolfberry fruit, 10-15 parts of pit viper, 8-15 parts of rhizoma polygonati, 20-30 parts of Chinese yam, 10-15 parts of mulberry, 6-10 parts of cinnamon and 10-20 parts of poria cocos. The optimal weight part ratio of the medicines in the formula is as follows: 24 parts of wolfberry fruit, 12 parts of pit viper, 12 parts of rhizoma polygonati, 24 parts of Chinese yam, 12 parts of mulberry, 8 parts of cinnamon and 15 parts of poria cocos. The traditional Chinese medicine formula is suitable for patients with deficiency of the lower-jiao and obstruction of governor vessel, the patients are basically asymptomatic after lying in bed and have comfortable feeling in the morning, and the patients only have ache, numbness and distending pain after noon or physical exertion.

Most of the raw materials of the medicament in the prior art are more in variety, and the medicament has a certain effect of relieving cervical spondylosis, but the utilization efficiency of the medicament is not effectively improved, the curative effect still needs to be improved, the interaction among the raw materials is unclear, and the side effect is greater in the using process of a patient.

The invention aims to provide a composition for treating cervical spondylosis, which can effectively improve the effect of the composition for treating the cervical spondylosis, has higher utilization rate of medicaments, effectively reduces the cost and simultaneously reduces the side effects of the medicaments.

Disclosure of Invention

In order to overcome the technical problems, the invention provides the composition containing cycloolivil 4' -glucoside, which has the advantages of good effect of treating cervical spondylosis, simple preparation method, low cost and contribution to popularization and application.

In order to achieve the above purpose, the technical scheme provided by the invention is as follows:

a composition containing cycloolivil 4' -glucoside comprises the following components: cycloolivil 4' -glucoside, olivil diglycoside, syringin, beta-naphthalene flavone, and quercetin.

Preferably, the composition containing cycloolivil 4' -glucoside comprises the following components in parts by weight: 5-10 parts of cycloolivil 4' -glucoside, 1-10 parts of olivine diglycoside, 5-15 parts of syringin, 1-10 parts of beta-naphthoflavone and 1-5 parts of quercetin.

Preferably, the composition containing cycloolivil 4' -glucoside comprises the following components in parts by weight: 5-8 parts of cycloolivil 4' -glucoside, 1-5 parts of olivine diglycoside, 5-7 parts of syringin, 2-4 parts of beta-naphthoflavone and 2-3 parts of quercetin.

Another object of the present invention is to provide a method for preparing the composition containing cycloolivil 4' -glycoside, comprising the following steps:

(1) adding a surfactant A into distilled water to prepare a water-phase solvent;

(2) under the condition of heating, dissolving cycloolivil 4' -glucoside, olivine diglycoside and syringin in an aqueous phase solvent in sequence, and stirring to form a solution 1;

(3) adding a surfactant B into the solution 1, and then adding beta-naphthoflavone and quercetin to form a solution 2; stirring to obtain the composition containing cycloolivil 4' -glucoside.

Preferably, in the step (1), the surfactant a is any one or more of octyl- β -D-glucopyranoside, dodecyl- β -D-maltopyranoside and N-dodecyl-N-methylglucamide.

Preferably, in the step (1), the surfactant A is prepared from the following components in a mass ratio of 1-3: 1 of octyl- β -D-glucopyranoside and N-dodecyl-N-methylglucamide.

Preferably, the addition amount of the surfactant A is 0.1-0.5% of the mass of the distilled water.

Preferably, the addition amount of the distilled water is 2-3 times of the total mass of the cycloolivil 4' -glucoside, the olivine diglycoside and the syringin.

Preferably, the surfactant B is tea saponin, and the addition amount of the tea saponin is 0.1-1% of the total mass of the beta-naphthoflavone and the quercetin.

Preferably, the preparation method of the tea saponin comprises the following steps:

pulverizing tea seed cake, soaking in hot water for extraction, filtering, treating the extractive solution with macroporous adsorbent resin, eluting with ethanol water solution, and drying to obtain tea saponin.

Preferably, the temperature of the hot water is 60-80 ℃, and the extraction time is 3-4 h.

Preferably, sodium carbonate is added in the extraction process, and the adding amount of the sodium carbonate is 5-10% of the mass of the tea seed cake.

Preferably, the macroporous adsorption resin is D3520 or D4006 type macroporous adsorption resin.

The specific process of the ethanol water solution elution is that 80-95% of ethanol water solution is used for eluting to remove impurities, 30-45% of ethanol is used for eluting and collecting eluent.

Preferably, the volume fraction of 80-95% has an elution rate of 1-3BV/h and is used in an amount of 1-2 BV.

Preferably, the elution rate of 30-45% by volume is 1-3BV/h, and the amount is 3-5 BV.

The invention also aims to provide application of the composition containing cycloolivil 4' -glucoside in preparation of a cervical spondylosis drug.

The invention also aims to provide a preparation of the composition containing cycloolivil 4' -glucoside, wherein the preparation is granules, tablets, capsules or oral liquid.

Compared with the prior art, the invention has the technical advantages that:

(1) the composition containing cycloolivil 4' -glucoside provided by the invention has a good effect of treating cervical spondylosis, and simultaneously has a good synergistic effect with olivine diglycoside, syringin, beta-naphthalene flavone and quercetin.

(2) In the invention, the surfactant is added in the process of preparing the composition, so that the storage stability of the composition can be improved on one hand, and the stability of the drug effect can be improved on the other hand.

(3) The tea saponin is added in the preparation process, so that the stability of the composition can be effectively promoted, and the tea saponin can also be synergistically acted with beta-naphthoflavone and quercetin to promote the drug effect of the composition.

Detailed Description

The present invention will be described below with reference to specific examples to make the technical aspects of the present invention easier to understand and grasp, but the present invention is not limited thereto. The experimental methods described in the following examples are all conventional methods unless otherwise specified; the reagents and materials are commercially available, unless otherwise specified.

Example 1

The composition containing cycloolivil 4' -glucoside comprises the following components in parts by weight: 8 parts of cycloolivil 4' -glucoside, 5 parts of olivine diglycoside, 7 parts of syringin, 4 parts of beta-naphthoflavone and 3 parts of quercetin.

The preparation method of the composition containing cycloolivil 4' -glucoside comprises the following steps:

(1) distilled water with the total mass of cycloolivil 4' -glucoside, olivine diglycoside and syringin being 3 times is taken, and distilled water with the mass of 0.3 wt% is added into the distilled water, and the mass ratio is 2: 1 octyl-beta-D-glucopyranoside and N-dodecyl-N-methyl glucamide to prepare an aqueous phase solvent;

(2) under the condition of heating, dissolving cycloolivil 4' -glucoside, olivine diglycoside and syringin in an aqueous phase solvent in sequence, and stirring to form a solution 1;

(3) adding tea saponin with the total mass of beta-naphthoflavone and quercetin being 0.5 wt% into the solution 1, and then adding the beta-naphthoflavone and quercetin to form a solution 2; stirring to obtain the composition containing cycloolivil 4' -glucoside.

The preparation method of the tea saponin comprises the following steps:

crushing tea seed cakes, adding the crushed tea seed cakes into hot water of 70 ℃, adding sodium carbonate with the mass of 6 wt% of the tea seed cakes, soaking and extracting for 3 hours, filtering, treating an extracting solution by using D3520 type macroporous adsorption resin, eluting and removing impurities by using an ethanol water solution with the volume fraction of 2BV being 90% according to the elution rate of 2BV/h, eluting and collecting eluent by using ethanol with the volume fraction of 4BV being 40% according to the elution rate of 2BV/h, and drying to obtain the tea saponin.

Example 2

The composition containing cycloolivil 4' -glucoside comprises the following components in parts by weight: 5 parts of cycloolivil 4' -glucoside, 10 parts of olivine diglycoside, 5 parts of syringin, 1 part of beta-naphthoflavone and 5 parts of quercetin.

The preparation method of the composition containing cycloolivil 4' -glucoside comprises the following steps:

(1) distilled water which is 2 times of the total mass of the cycloolivil 4' -glucoside, the olivine diglycoside and the syringin is taken, and 0.1 wt% of the mass of the distilled water is added into the distilled water, and the mass ratio of the distilled water is 1: 1 octyl-beta-D-glucopyranoside and N-dodecyl-N-methyl glucamide to prepare an aqueous phase solvent;

(2) under the condition of heating, dissolving cycloolivil 4' -glucoside, olivine diglycoside and syringin in an aqueous phase solvent in sequence, and stirring to form a solution 1;

(3) adding tea saponin with the total mass of beta-naphthoflavone and quercetin being 0.1 wt% into the solution 1, and then adding the beta-naphthoflavone and quercetin to form a solution 2; stirring to obtain the composition containing cycloolivil 4' -glucoside.

The preparation method of the tea saponin comprises the following steps:

crushing tea seed cakes, adding the crushed tea seed cakes into hot water of 60 ℃, adding sodium carbonate accounting for 5 wt% of the tea seed cakes, soaking and extracting for 3 hours, filtering, treating an extracting solution by D4006 type macroporous adsorption resin, eluting and removing impurities by using 1BV of ethanol aqueous solution with the volume fraction of 80% according to the elution rate of 1BV/h, eluting and collecting eluent by using 3BV of ethanol with the volume fraction of 30% according to the elution rate of 1BV/h, and drying to obtain the tea saponin.

Example 3

The composition containing cycloolivil 4' -glucoside comprises the following components in parts by weight: 10 parts of cycloolivil 4' -glucoside, 1 part of olivine diglycoside, 15 parts of syringin, 10 parts of beta-naphthalene flavone and 1 part of quercetin.

The preparation method of the composition containing cycloolivil 4' -glucoside comprises the following steps:

(1) distilled water which is 2 times of the total mass of the cycloolivil 4' -glucoside, the olivine diglycoside and the syringin is taken, and the distilled water is added with the distilled water with the mass ratio of 0.5 wt% of the distilled water being 3: 1 octyl-beta-D-glucopyranoside and N-dodecyl-N-methyl glucamide to prepare an aqueous phase solvent;

(2) under the condition of heating, dissolving cycloolivil 4' -glucoside, olivine diglycoside and syringin in an aqueous phase solvent in sequence, and stirring to form a solution 1;

(3) adding tea saponin with the total mass of 1 wt% of beta-naphthoflavone and quercetin into the solution 1, and adding the beta-naphthoflavone and quercetin to form a solution 2; stirring to obtain the composition containing cycloolivil 4' -glucoside.

The preparation method of the tea saponin comprises the following steps:

crushing tea seed cakes, adding the crushed tea seed cakes into hot water at 80 ℃, adding sodium carbonate accounting for 10 wt% of the weight of the tea seed cakes, soaking and extracting for 4 hours, filtering, treating an extracting solution by using D3520 type macroporous adsorption resin, eluting and removing impurities by using 2BV ethanol water solution with the volume fraction of 95% according to the elution rate of 3BV/h, eluting and collecting eluent by using 5BV ethanol with the volume fraction of 45% according to the elution rate of 3BV/h, and drying to obtain the tea saponin.

Example 4

The composition containing cycloolivil 4' -glucoside comprises the following components in parts by weight: 8 parts of cycloolivil 4' -glucoside, 5 parts of olivine diglycoside, 7 parts of syringin, 4 parts of beta-naphthoflavone and 3 parts of quercetin.

The preparation method of the composition containing cycloolivil 4' -glucoside comprises the following steps:

(1) taking distilled water with 3 times of total mass of cycloolivil 4' -glucoside, olivine diglycoside and syringin, and adding octyl-beta-D-glucopyranoside with the mass of 0.3 wt% of the distilled water into the distilled water to prepare an aqueous phase solvent;

(2) under the condition of heating, dissolving cycloolivil 4' -glucoside, olivine diglycoside and syringin in an aqueous phase solvent in sequence, and stirring to form a solution 1;

(3) adding tea saponin with the total mass of beta-naphthoflavone and quercetin being 0.5 wt% into the solution 1, and then adding the beta-naphthoflavone and quercetin to form a solution 2; stirring to obtain the composition containing cycloolivil 4' -glucoside.

The preparation method of the tea saponin comprises the following steps:

crushing tea seed cakes, adding the crushed tea seed cakes into hot water of 70 ℃, adding sodium carbonate with the mass of 6 wt% of the tea seed cakes, soaking and extracting for 3 hours, filtering, treating an extracting solution by using D3520 type macroporous adsorption resin, eluting and removing impurities by using an ethanol water solution with the volume fraction of 2BV being 90% according to the elution rate of 2BV/h, eluting and collecting eluent by using ethanol with the volume fraction of 4BV being 40% according to the elution rate of 2BV/h, and drying to obtain the tea saponin.

Example 5

The composition containing cycloolivil 4' -glucoside comprises the following components in parts by weight: 8 parts of cycloolivil 4' -glucoside, 5 parts of olivine diglycoside, 7 parts of syringin, 4 parts of beta-naphthoflavone and 3 parts of quercetin.

The preparation method of the composition containing cycloolivil 4' -glucoside comprises the following steps:

(1) taking distilled water with 3 times of total mass of cycloolivil 4' -glucoside, olivine diglycoside and syringin, adding N-dodecyl-N-methyl glucamide with the mass of 0.3 wt% of the distilled water into the distilled water to prepare a water phase solvent;

(2) under the condition of heating, dissolving cycloolivil 4' -glucoside, olivine diglycoside and syringin in an aqueous phase solvent in sequence, and stirring to form a solution 1;

(3) adding tea saponin with the total mass of beta-naphthoflavone and quercetin being 0.5 wt% into the solution 1, and then adding the beta-naphthoflavone and quercetin to form a solution 2; stirring to obtain the composition containing cycloolivil 4' -glucoside.

The preparation method of the tea saponin comprises the following steps:

crushing tea seed cakes, adding the crushed tea seed cakes into hot water of 70 ℃, adding sodium carbonate with the mass of 6 wt% of the tea seed cakes, soaking and extracting for 3 hours, filtering, treating an extracting solution by using D3520 type macroporous adsorption resin, eluting and removing impurities by using an ethanol water solution with the volume fraction of 2BV being 90% according to the elution rate of 2BV/h, eluting and collecting eluent by using ethanol with the volume fraction of 4BV being 40% according to the elution rate of 2BV/h, and drying to obtain the tea saponin.

Comparative example 1

The difference compared to example 1 is the replacement of the olivinediglycoside by the cycloolivoresin 4' -glycoside.

The composition containing cycloolivil 4' -glucoside comprises the following components in parts by weight: 13 parts of cycloolivil 4' -glucoside, 7 parts of syringin, 4 parts of beta-naphthoflavone and 3 parts of quercetin.

The preparation method of the composition containing cycloolivil 4' -glucoside comprises the following steps:

(1) distilled water with the total mass of the cycloolivil 4' -glucoside and the syringin being 3 times is taken, and the distilled water with the mass of 0.3 wt% is added into the distilled water, and the mass ratio is 2: 1 octyl-beta-D-glucopyranoside and N-dodecyl-N-methyl glucamide to prepare an aqueous phase solvent;

(2) under the condition of heating, dissolving cycloolivil 4' -glucoside and syringin in turn in an aqueous phase solvent, and stirring to form a solution 1;

(3) adding tea saponin with the total mass of beta-naphthoflavone and quercetin being 0.5 wt% into the solution 1, and then adding the beta-naphthoflavone and quercetin to form a solution 2; stirring to obtain the composition containing cycloolivil 4' -glucoside.

Wherein, the preparation method of the tea saponin is the same as the example 1.

Comparative example 2

The difference compared to example 1 is the replacement of cycloolivil 4' -glycoside with olivil diglycoside.

The composition comprises the following components in parts by weight: 13 parts of olivine diglycoside, 7 parts of syringin, 4 parts of beta-naphthalene flavone and 3 parts of quercetin.

The preparation method of the composition comprises the following steps:

(1) distilled water which is 3 times of the total mass of the olivine diglycoside and the syringin is taken, and the distilled water is added with the distilled water with the mass ratio of 0.3 wt% to 2: 1 octyl-beta-D-glucopyranoside and N-dodecyl-N-methyl glucamide to prepare an aqueous phase solvent;

(2) under the condition of heating, dissolving the olivine diglycoside and the syringin in the water phase solvent in sequence, and stirring to form a solution 1;

(3) adding tea saponin with the total mass of beta-naphthoflavone and quercetin being 0.5 wt% into the solution 1, and then adding the beta-naphthoflavone and quercetin to form a solution 2; stirring to obtain the composition.

Wherein, the preparation method of the tea saponin is the same as the example 1.

Comparative example 3

Compared with example 1, the difference lies in the preparation method of the tea saponin.

A composition containing cycloolivil 4' -glycoside is prepared in the same manner as in example 1.

The preparation method of the composition containing cycloolivil 4' -glucoside comprises the following steps:

(1) - (3) same as in example 1.

The preparation method of the tea saponin comprises the following steps:

crushing tea seed cakes, soaking and extracting the tea seed cakes in hot water at 70 ℃ for 3 hours, filtering, treating an extracting solution by using D3520 type macroporous adsorption resin, eluting and removing impurities by using an ethanol water solution with the volume fraction of 2BV being 90% according to the elution rate of 2BV/h, eluting and collecting eluent by using ethanol with the volume fraction of 4BV being 40% according to the elution rate of 2BV/h, and drying to obtain the tea saponin.

Comparative example 4

Compared with example 1, the difference lies in the preparation method of the tea saponin.

A composition containing cycloolivil 4' -glycoside is prepared in the same manner as in example 1.

The preparation method of the composition containing cycloolivil 4' -glucoside comprises the following steps:

(1) - (3) same as in example 1.

The preparation method of the tea saponin comprises the following steps: pulverizing tea seed cake, soaking in 70 deg.C hot water for 3 hr, filtering, and concentrating to obtain concentrated solution; precipitating by using 90 wt% ethanol, wherein the volume ratio of the ethanol to the concentrated solution is 4: 1, precipitating the tea saponin at the alcohol precipitation temperature of 60 ℃ for 3 hours, filtering, and drying the precipitate to obtain the tea saponin.

Evaluation of efficacy

1. Vertebral artery type cervical spondylosis model

72 SPF grade healthy male Wistar rats with the weight of 200 +/-20 g are divided into 12 groups, and 6 rats in each group are respectively a model group, a sham operation group, a positive medicine group, examples 1-5 groups and comparative examples 1-4 groups. The anesthetized rat is fixed on a fixing plate, the abdomen of the rat is required to be tightly attached to the fixing plate, four limbs of chalk are fixed on two sides, and the back of the rat faces upwards. The hair at the back of the neck was cut, sterilized with iodophor, and then the skin was incised longitudinally at the midline of the back of the neck of the rat with a scalpel, the incision length being 2.5 cm. After the surgical incision of the sham group rats, the rats are subjected to conventional hemostatic suture. The model group, the groups of examples 1 to 5, the groups of comparative examples 1 to 4 and the positive drug control group were subjected to blunt muscle separation layer by means of a vascular clamp or the like, and the superficial and deep muscle tissues of the neck were cut. After the operation, the rat is treated by conventional hemostasis and suture treatment, and the rat is observed and continuously raised.

After 1 month of molding, the sham-operated rats were fixed on a daily basis without intervention by other means. The model group rats were not subjected to any intervention after successful model building.

The intervention method comprises the following steps: groups 1-5 and comparative groups 1-4 are performed by intragastric administration at a dose of 50mg/kg/d effective content using the compositions prepared by the methods of the respective corresponding examples or comparative examples; after the positive drug group rats are successfully molded, the rats are subjected to intragastric administration intervention by using radix angelicae sinensis and astragalus blood activating capsules (Lunanqianpu pharmaceutical Co., Ltd., specification: 0.53g multiplied by 36 capsules/box), the dosage is 1 capsule/d, and the continuous intervention is carried out for 4 weeks;

the rats are weighed respectively, are subjected to lethal abdominal anesthesia, and after the rats are determined to be in place, relevant experimental operations are carried out under the low-temperature condition. The rat dentate vertebrae were retained and the broken ends after occipital bone were treated.

The skin is longitudinally cut along the median line of the back of the rat neck by using a scalpel, each layer of muscle and soft tissue are separated in a blunt manner, C2-7 cervical vertebrae are taken down, and the integrity of intervertebral discs and cartilages is guaranteed in the operation process. And cleaning the taken tissue by adopting PBS liquid, and fixing part of intervertebral disc tissue for 1-2 days by using 4% paraformaldehyde for immunohistochemical detection. The remaining tissue was frozen using a liquid nitrogen vacuum flask and stored in a freezer at-80 ℃ as soon as possible.

IL-6 and IL-8 contents were detected by immunohistochemical method using IL-6 and IL-8 immunohistochemical SP kit (BIO-RAD, USA). The results are shown in Table 1:

table 1 intervention effect data

In the same list, the different letters or symbols have significant difference, and P is less than 0.05.

As can be seen from Table 1, compared with the model group, the compositions provided by the groups of examples 1-5 and the groups of comparative examples 1-4 can effectively control the IL-6 and IL-8 levels of the chondrocytes of the cervical intervertebral disc of the rat, which shows that the composition provided by the invention can effectively prevent and treat and intervene cervical spondylosis. Meanwhile, the raw material composition and the preparation method of the composition have great influence on the effect of the composition.

2. Experiment of anti-inflammatory effect of xylene on swelling of auricle of rat

66 SPF grade healthy male Wistar rats with the body weight of 200 +/-20 g are divided into 11 groups, 6 rats are respectively a blank group, a positive drug group, examples 1-5 groups and comparative examples 1-4 groups. Xylene is smeared on the two sides of the right ear of each rat, 50 ul/mouse, and the drug is administrated according to the following groups after 30 min: blank group was not given any drug; the compositions prepared in the corresponding groups are respectively administered to groups 1 to 5 and groups 1 to 4 according to the effective content of 0.5mg/kg, and the compositions are properly massaged after the right ears of rats are smeared on the two sides to promote the absorption of the medicaments; the positive drug group was prepared by applying commercially available loxoprofen sodium gel ointment (prepared in Jiedian of Hunan) on both sides of the rat's right ear and then massaging the rat's right ear appropriately to promote drug absorption. After 1h, the animals were sacrificed by cervical dislocation, round ears were respectively punched at the same positions by using a 6mm diameter punch at both sides of the ears, the mass was weighed by using a precision torsion balance, and the swelling degree and swelling rate were calculated, and the results are shown in table 2.

TABLE 2 swelling degree and swelling Rate data

In the same list, the different symbols have significant difference, and P is less than 0.05.

As can be seen from Table 1, the compositions provided in examples 1 to 5 and comparative examples 1 to 4 reduced swelling of auricles of rats caused by xylene compared with the blank group, which indicates that the composition provided by the present invention has a superior anti-inflammatory effect. Meanwhile, the raw material composition and the preparation method of the composition have great influence on the effect of the composition.

The above detailed description is specific to one possible embodiment of the present invention, and the embodiment is not intended to limit the scope of the present invention, and all equivalent implementations or modifications without departing from the scope of the present invention should be included in the technical scope of the present invention.