阅读说明：本技术 一种含松脂素和环橄榄树脂素9-糖苷的组合物及其制备方法和应用 (Composition containing pinoresinol and cycloolivil 9-glucoside and preparation method and application thereof ) 是由 肖军平 李小锋 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,具体涉及一种含松脂素和环橄榄树脂素9-糖苷的组合物及其制备方法和应用。所述组合物,包括以下组分：环橄榄树脂素9-糖苷、绿原酸、松脂素、杜仲醇、槲皮素。该组合物能够有效提高组合物治疗膝关节炎的效果,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 pinoresinol and cycloolivil 9-glucoside, and a preparation method and application thereof. The composition comprises the following components: cycloolivil 9-glucoside, chlorogenic acid, pinoresinol, guttiferol, and quercetin. The composition can effectively improve the effect of the composition on treating the gonitis, has higher utilization rate of the medicine, effectively reduces the cost and reduces the side effect of the medicine.)

1. A composition containing cycloolivil 9-glucoside comprises the following components: cycloolivil 9-glucoside, chlorogenic acid, pinoresinol, guttiferol, and quercetin.

2. The cycloolivil 9-glycoside-containing composition according to claim 1, comprising the following components in parts by weight: 5-20 parts of cycloolivil 9-glucoside, 1-10 parts of chlorogenic acid, 5-15 parts of pinoresinol, 1-5 parts of gutta alcohol and 1-5 parts of quercetin.

3. The cycloolivil 9-glycoside-containing composition according to claim 1, comprising the following components in parts by weight: 8-12 parts of cycloolivil 9-glucoside, 5-7 parts of chlorogenic acid, 7-11 parts of pinoresinol, 2-4 parts of gutta alcohol and 2-3 parts of quercetin.

4. A process for the preparation of a cycloolivil 9-glycoside-containing composition according to any one of claims 1 to 3, comprising the following steps:

(1) adding surfactant A into distilled water, heating, dissolving cycloolivil 9-glucoside, chlorogenic acid and pinoresinol in sequence, and stirring to obtain solution 1;

(2) adding surfactant B into distilled water, and adding gutta alcohol and quercetin to form solution 2;

(3) and mixing the solution 1 and the solution 2, and stirring.

5. The method for preparing a cyclic olivil 9-glycoside-containing composition according to claim 4, wherein in step (1), the surfactant A is any one or more of soybean lecithin, octyl- β -D-glucopyranoside, dodecyl- β -D-maltopyranoside and N-dodecyl-N-methylglucamide; the addition amount of the surfactant A is 0.1-0.5% of the mass of the distilled water; the addition amount of the distilled water is 2-3 times of the total mass of the cycloolivil 9-glucoside, the chlorogenic acid and the pinoresinol.

6. The method for preparing the composition containing cycloolivil 9-glycoside according to claim 4, wherein in step (1), the surfactant A is a mixture of N-dodecyl-N-methylglucamide and soya lecithin in a mass ratio of 4-6: 1.

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 the gutta alcohol and quercetin.

8. The method of claim 7, wherein the tea saponin is prepared by the steps of: 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, 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;

preferably, the specific process of the ethanol aqueous solution elution is that the ethanol aqueous solution with the volume fraction of 80-95% is used for eluting to remove impurities, then 30-45% ethanol is used for eluting and collecting the eluent.

9. Use of a composition containing pinoresinol and cycloolivil 9-glycoside according to any one of claims 1 to 3 or a composition containing pinoresinol and cycloolivil 9-glycoside prepared by the preparation method according to any one of claims 4 to 8 for the preparation of a medicament for treating knee osteoarthritis.

10. The preparation of the composition containing pinoresinol and cycloolivil 9-glycoside according to any one of claims 1 to 3 or the composition containing pinoresinol and cycloolivil 9-glycoside prepared by the preparation method according to any one of claims 4 to 8, wherein the preparation is granules, tablets, capsules or oral liquid.

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to a composition containing pinoresinol and cycloolivil 9-glucoside, and a preparation method and application thereof.

Background

Gonarthritis is a disorder that is based on a degenerative change of pathology. The symptoms of the middle-aged and the elderly are manifested as red and sore knee, pain in ascending and descending stairs, and soreness and discomfort of knee when sitting up and standing. There are also symptoms of swelling, ringing, fluid accumulation, etc., which can cause joint deformity and disability if the patient is not treated in time. Joint diseases such as gonarthromeningitis, ligament injury, meniscus injury, loose knee joint body, popliteal cyst, patellar malacia, goose foot bursitis, and genu varum are also commonly suffered at the knee joint part. For example, Chinese patent application CN201910540161.2 provides a traditional Chinese medicine preparation for treating knee osteoarthritis, which comprises the following traditional Chinese medicine raw materials in parts by weight: 15-20 parts of eucommia ulmoides, 15-20 parts of epimedium herb, 5-10 parts of teasel root, 20-30 parts of medicinal cyathula root, 5-10 parts of fructus psoraleae, 15-20 parts of notopterygium root, 5-10 parts of bighead atractylodes rhizome, 15-20 parts of acanthopanax bark, 10-20 parts of angelica, 2-5 parts of platycladi seed, 5-10 parts of cassia bark, 5-10 parts of lycopodium clavatum, 15-20 parts of curcuma zedoary, 5-10 parts of medlar, 5-10 parts of notopterygium root, 2-5 parts of liquorice, 5-10 parts of rhizoma atractylodis, 1-2 parts of wrinkled gianthyssop, 5-10 parts of salvia miltiorrhiza, 10-20 parts of oyster, 10-20 parts of radix angelicae pubescentis, 2-5 parts of coix seed, 5-10 parts of pawpaw and 2-5 parts of glabrous greenbrier rhizome. The preparation can effectively relieve pain of patients and can simultaneously increase bone density.

Chinese patent application CN201610170627.0 discloses a combined traditional Chinese medicine for treating gonarthritis, which has the following formula: 12-18 parts of raw common monkshood mother root, 12-18 parts of raw kusnezoff monkshood root, 12-18 parts of manchurian wildginger herb, 12-18 parts of peach kernel, 12-18 parts of twotooth achyranthes root, 16-24 parts of incised notopterygium rhizome and root, 22-28 parts of common clubmoss herb, 22-28 parts of medicinal indianmulberry root, 22-28 parts of clematis root, 22-28 parts of garden balsam stem, 22-28 parts of rhubarb, 22-28 parts of Szechuan lovage rhizome, 20-30 parts of Chinese angelica, 20-30 parts of suberect spatholobus stem, 20-30 parts of erythrina bark and 20-30 parts of mulberry twig. Has effects in expelling pathogenic wind, removing dampness, promoting blood circulation, activating collateral flow, relieving pain, promoting blood circulation, and relieving pain.

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 the gonarthritis, 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 gonitis, which can effectively improve the effect of the composition for treating gonitis, has higher utilization rate of medicines, effectively reduces the cost and reduces the side effects of the medicines.

Disclosure of Invention

In order to overcome the technical problems, the invention provides a composition containing pinoresinol and cycloolivil 9-glucoside, a preparation method and application thereof. The composition has good effect of treating the gonitis, and simultaneously, the utilization rate of the medicine is high, so that the cost is effectively reduced, and the side effect of the medicine is reduced.

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

a composition containing cycloolivil 9-glucoside comprises the following components: cycloolivil 9-glucoside, chlorogenic acid, pinoresinol, guttiferol, and quercetin.

Preferably, the composition containing the cycloolivil 9-glucoside comprises the following components in parts by weight: 5-20 parts of cycloolivil 9-glucoside, 1-10 parts of chlorogenic acid, 5-15 parts of pinoresinol, 1-5 parts of gutta alcohol and 1-5 parts of quercetin.

Preferably, the composition containing the cycloolivil 9-glucoside comprises the following components in parts by weight: 8-12 parts of cycloolivil 9-glucoside, 5-7 parts of chlorogenic acid, 7-11 parts of pinoresinol, 2-4 parts of gutta alcohol and 2-3 parts of quercetin.

Another object of the present invention is to provide a method for preparing the composition containing cycloolivil 9-glycoside, comprising the steps of:

(1) adding surfactant A into distilled water, heating, dissolving cycloolivil 9-glucoside, chlorogenic acid and pinoresinol in sequence, and stirring to obtain solution 1;

(2) adding surfactant B into distilled water, and adding gutta alcohol and quercetin to form solution 2;

(3) mixing the solution 1 and the solution 2, and stirring to obtain the composition containing the pinoresinol and the cycloolivil 9-glucoside.

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

Preferably, in the step (1), the surfactant A is a mixture of N-dodecyl-N-methyl glucamide and soybean lecithin in a mass ratio of 4-6: 1.

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 9-glucoside, the chlorogenic acid and the pinoresinol.

Preferably, in step (1), the temperature of the heating is 30-45 ℃.

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 gutta alcohol 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 the pinoresinol and the cycloolivil 9-glucoside in preparing the knee osteoarthritis medicine.

The invention also aims to provide a preparation of the composition containing the pinoresinol and the cycloolivil 9-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 pinoresinol and cycloolivil 9-glucoside provided by the invention has a good effect of treating knee osteoarthritis, and simultaneously has a good synergistic effect with chlorogenic acid, pinoresinol, guttapercha 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 the eucommia alcohol and the 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 9-glucoside comprises the following components in parts by weight: 12 parts of cycloolivil 9-glucoside, 7 parts of chlorogenic acid, 7 parts of pinoresinol, 4 parts of gutta alcohol and 2 parts of quercetin.

The preparation method of the composition containing the cycloolivil 9-glucoside comprises the following steps:

(1) distilled water which is 2 times of the total mass of the cycloolivil 9-glucoside, the chlorogenic acid and the pinoresinol is taken, and 0.3 wt% of distilled water with the mass ratio of 5: 1, heating the mixture of N-dodecyl-N-methyl glucamide and soybean lecithin to 40 ℃, dissolving cycloolivil 9-glucoside, chlorogenic acid and pinoresinol in the mixture in turn, and stirring the mixture to form a solution 1;

(2) adding tea saponin into distilled water, wherein the adding amount of the tea saponin is 0.3 wt% of the total mass of the gutta alcohol and the quercetin, and then adding the gutta alcohol and the quercetin to form a solution 2;

(3) mixing the solution 1 and the solution 2, and stirring to obtain the composition containing the pinoresinol and the cycloolivil 9-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 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 macroporous adsorption resin, eluting by using 2BV of ethanol aqueous solution with the volume fraction of 80%, and removing impurities, wherein the elution rate is 2 BV/h; eluting with 4BV of 35% ethanol at an elution rate of 2BV/h, and collecting the eluate; drying the eluent to obtain the tea saponin.

Example 2

The composition containing cycloolivil 9-glucoside comprises the following components in parts by weight: 5 parts of cycloolivil 9-glucoside, 5 parts of chlorogenic acid, 11 parts of pinoresinol, 2 parts of gutta alcohol and 1 part of quercetin.

The preparation method of the composition containing the cycloolivil 9-glucoside comprises the following steps:

(1) taking distilled water with the weight 3 times of the total weight of the cycloolivine 9-glucoside, the chlorogenic acid and the pinoresinol, adding 0.5 wt% of a mixture of N-dodecyl-N-methyl glucamide and soybean lecithin with the mass ratio of 4:1 into the distilled water, heating to 30 ℃, then sequentially dissolving the cycloolivine 9-glucoside, the chlorogenic acid and the pinoresinol in the mixture, and stirring to form a solution 1;

(2) adding tea saponin into distilled water, wherein the adding amount of the tea saponin is 0.1 percent of the total mass of the gutta alcohol and the quercetin, and then adding the gutta alcohol and the quercetin to form a solution 2;

(3) mixing the solution 1 and the solution 2, and stirring to obtain the composition containing the pinoresinol and the cycloolivil 9-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 60 ℃, adding sodium carbonate with the mass of 5 wt% of the tea seed cakes, soaking and extracting for 3 hours, filtering, treating an extracting solution by using D4006 type macroporous adsorption resin, eluting and removing impurities by using an ethanol aqueous solution with the volume fraction of 1BV being 95%, wherein the elution rate is 1 BV/h; eluting with 5BV of 45% ethanol at an elution rate of 1BV/h, and collecting the eluate; drying the eluent to obtain the tea saponin.

Example 3

The composition containing cycloolivil 9-glucoside comprises the following components in parts by weight: 20 parts of cycloolivil 9-glucoside, 10 parts of chlorogenic acid, 11 parts of pinoresinol, 5 parts of gutta alcohol and 5 parts of quercetin.

The preparation method of the composition containing the cycloolivil 9-glucoside comprises the following steps:

(1) taking distilled water 2-3 times of the total mass of cycloolivil 9-glucoside, chlorogenic acid and pinoresinol, adding 0.1 wt% of a mixture of N-dodecyl-N-methyl glucamide and soybean lecithin with the mass ratio of 6:1 into the distilled water, heating to 45 ℃, sequentially dissolving the cycloolivil 9-glucoside, the chlorogenic acid and the pinoresinol in the mixture, and stirring to form a solution 1;

(2) adding tea saponin into distilled water, wherein the adding amount of the tea saponin is 1 percent of the total mass of the gutta alcohol and the quercetin, and then adding the gutta alcohol and the quercetin to form a solution 2;

(3) mixing the solution 1 and the solution 2, and stirring to obtain the composition containing the pinoresinol and the cycloolivil 9-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 with the mass of 10 wt% of the tea seed cakes, soaking and extracting for 4 hours, filtering, treating an extracting solution by using D4006 type macroporous adsorption resin, eluting by using 2BV of ethanol aqueous solution with the volume fraction of 80%, and removing impurities, wherein the elution rate is 3 BV/h; eluting with 30% ethanol of 3BV at an elution rate of 3BV/h, and collecting the eluate; drying the eluent to obtain the tea saponin.

Example 4

The composition containing cycloolivil 9-glucoside comprises the following components in parts by weight: 12 parts of cycloolivil 9-glucoside, 7 parts of chlorogenic acid, 7 parts of pinoresinol, 4 parts of gutta alcohol and 2 parts of quercetin.

The preparation method of the composition containing the cycloolivil 9-glucoside comprises the following steps:

(1) adding 0.3 wt% of soybean lecithin into distilled water which is 2 times of the total mass of cycloolivil 9-glucoside, chlorogenic acid and pinoresinol, heating to 40 ℃, dissolving the cycloolivil 9-glucoside, the chlorogenic acid and the pinoresinol in turn, and stirring to form a solution 1;

(2) adding tea saponin into distilled water, wherein the adding amount of the tea saponin is 0.3 wt% of the total mass of the gutta alcohol and the quercetin, and then adding the gutta alcohol and the quercetin to form a solution 2;

(3) mixing the solution 1 and the solution 2, and stirring to obtain the composition containing the pinoresinol and the cycloolivil 9-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 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 macroporous adsorption resin, eluting by using 2BV of ethanol aqueous solution with the volume fraction of 80%, and removing impurities, wherein the elution rate is 2 BV/h; eluting with 4BV of 35% ethanol at an elution rate of 2BV/h, and collecting the eluate; drying the eluent to obtain the tea saponin.

Example 5

The composition containing cycloolivil 9-glucoside comprises the following components in parts by weight: 12 parts of cycloolivil 9-glucoside, 7 parts of chlorogenic acid, 7 parts of pinoresinol, 4 parts of gutta alcohol and 2 parts of quercetin.

The preparation method of the composition containing the cycloolivil 9-glucoside comprises the following steps:

(1) taking distilled water 2 times of the total mass of cycloolivil 9-glucoside, chlorogenic acid and pinoresinol, adding 0.3 wt% of dodecyl-beta-D-maltopyranoside into the distilled water, heating to 40 ℃, dissolving the cycloolivil 9-glucoside, the chlorogenic acid and the pinoresinol in turn, and stirring to form a solution 1;

(2) adding tea saponin into distilled water, wherein the adding amount of the tea saponin is 0.3 wt% of the total mass of the gutta alcohol and the quercetin, and then adding the gutta alcohol and the quercetin to form a solution 2;

(3) mixing the solution 1 and the solution 2, and stirring to obtain the composition containing the pinoresinol and the cycloolivil 9-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 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 macroporous adsorption resin, eluting by using 2BV of ethanol aqueous solution with the volume fraction of 80%, and removing impurities, wherein the elution rate is 2 BV/h; eluting with 4BV of 35% ethanol at an elution rate of 2BV/h, and collecting the eluate; drying the eluent to obtain the tea saponin.

Comparative example 1

The difference compared to example 1 is the replacement of quercetin with gutta percha alcohol.

The composition containing cycloolivil 9-glucoside comprises the following components in parts by weight: 12 parts of cycloolivil 9-glucoside, 7 parts of chlorogenic acid, 7 parts of pinoresinol and 6 parts of gutta alcohol.

The preparation method of the composition containing the cycloolivil 9-glucoside comprises the following steps:

(1) distilled water which is 2 times of the total mass of the cycloolivil 9-glucoside, the chlorogenic acid and the pinoresinol is taken, and 0.3 wt% of distilled water with the mass ratio of 5: 1, heating the mixture of N-dodecyl-N-methyl glucamide and soybean lecithin to 40 ℃, dissolving cycloolivil 9-glucoside, chlorogenic acid and pinoresinol in the mixture in turn, and stirring the mixture to form a solution 1;

(2) adding tea saponin into distilled water, wherein the addition amount of the tea saponin is 0.3 wt% of the total mass of the gutta alcohol, and then adding the gutta alcohol to form a solution 2;

(3) mixing the solution 1 and the solution 2, and stirring to obtain the composition containing the pinoresinol and the cycloolivil 9-glucoside.

The preparation method of the tea saponin is the same as that of the example 1.

Comparative example 2

The difference compared to example 1 is the replacement of chlorogenic acid by citric acid.

The composition containing cycloolivil 9-glucoside comprises the following components in parts by weight: 12 parts of cycloolivil 9-glucoside, 7 parts of citric acid, 7 parts of pinoresinol, 4 parts of gutta alcohol and 2 parts of quercetin.

The preparation method of the composition containing the cycloolivil 9-glucoside comprises the following steps:

(1) distilled water which is 2 times of the total mass of the cycloolivil 9-glucoside, the citric acid and the pinoresinol is taken, and 0.3 wt% of distilled water with the mass ratio of 5: 1, heating the mixture of N-dodecyl-N-methyl glucamide and soybean lecithin to 40 ℃, dissolving cycloolivil 9-glucoside, citric acid and pinoresinol in the mixture in turn, and stirring the mixture to form a solution 1;

(2) adding tea saponin into distilled water, wherein the adding amount of the tea saponin is 0.3 wt% of the total mass of the gutta alcohol and the quercetin, and then adding the gutta alcohol and the quercetin to form a solution 2;

(3) mixing the solution 1 and the solution 2, and stirring to obtain the composition containing the pinoresinol and the cycloolivil 9-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 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 macroporous adsorption resin, eluting by using 2BV of ethanol aqueous solution with the volume fraction of 80%, and removing impurities, wherein the elution rate is 2 BV/h; eluting with 4BV of 35% ethanol at an elution rate of 2BV/h, and collecting the eluate; drying the eluent to obtain the tea saponin.

Comparative example 3

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

A composition containing cycloolivil 9-glycoside is prepared in the same manner as in example 1.

The preparation method of the composition containing the cycloolivil 9-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.

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

60 SPF grade healthy male Wistar rats with the body weight of 200 +/-20 g are divided into 10 groups, and 6 rats in each group are respectively a blank group, a positive drug group, examples 1-5 groups and comparative examples 1-3 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 the groups 1 to 5 and the groups 1 to 3 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 1.

TABLE 1 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 3 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.

2. Rat knee osteoarthritis model experiment

60 SD rats with a body weight of 200 + -20 g were randomly divided into 10 groups of 6 rats, namely a normal group, a model group, examples 1-5 groups and a comparative example-3 group.

Except for the normal group, rats replicated the KOA model by injecting papain through the joint cavity; the rats were prepared and sterilized, and after shaving the right anterior knee joint and the lower hair, the skin was sterilized with 75% ethanol, and then 0.2mL of 4% papain solution was injected into the right anterior knee joint cavity, once every 2 days, for 2 weeks. After 14d, successful replication was demonstrated if the following symptoms were observed in the rats: red swelling and pain of the right anterior knee joint accompanied by swelling of soft tissue or the appearance of fluid accumulation; stiffness and limited movement of the right anterior knee joint and surrounding areas; the pathological examination result of the right front knee joint shows that a large amount of exudates appear in the joint cavity; the cartilage surface is not smooth, and the arthritic pathology such as synovial hyperplasia or adhesion exists. On day 2 after successful molding, examples 1-5 and comparison were performed except that the normal and model groups were gazed with physiological salineExamples 1-3 groups of rats were administered the composition prepared in the corresponding group, and the composition was administered to the rats by gavage at a dose of 10mg/kg/d for intervention, and femoral artery blood was collected at 1500 rpm-min from each group of rats 4 weeks after the intervention^-1Centrifuging for 10min, collecting serum, and detecting IL-6 (interleukin-6), TNF-alpha (tumor necrosis factor-alpha), and IL-1 beta (interleukin-1 beta) levels by ELISA (enzyme-linked immunosorbent assay kit manufacturer: Wuhan Elirei Biotech). The results are shown in Table 2.

TABLE 2 levels of inflammatory factors

Test group	IL-6(ng·L-1)	TNF-α(ng·L-1)	IL-1β(ng·L-1)
				Blank group	18.32±2.37a	22.15±6.14f	6.01±1.09*
Model set	44.26±4.51b	41.34±4.11g	12.65±2.34#
				Example 1	23.57±5.34c	25.43±3.57h	7.46±0.87△
Example 2	24.61±3.08c	25.69±8.21h	7.29±1.54△
				Example 3	23.68±1.55c	26.07±6.47h	7.54±2.05△
Example 4	30.25±2.67d	28.34±5.22h	8.02±3.11▼
				Example 5	31.08±3.26d	29.10±4.28h	8.71±2.62▼
Comparative example 1	34.53±6.64e	33.76±9.14i	10.89±1.03◇
				Comparative example 2	37.28±4.09e	36.21±5.67i	11.32±5.43◇
Comparative example 3	33.56±5.12e	31.43±4.39i	9.86±2.19◇

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 2, the compositions provided in examples 1 to 5 and comparative examples 1 to 3 were effective in intervening serum inflammatory factor levels, compared to the blank group, indicating that the composition provided by the present invention has a superior effect of treating knee osteoarthritis. 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.