阅读说明：本技术 灵芝酸a在治疗银屑病药物中的应用 (Application of ganoderic acid A in medicine for treating psoriasis ) 是由 肖成根 陈翔 刘洪� 周星辰 陈军臣 李旺 欧家芮 于 2021-10-28 设计创作，主要内容包括：本发明公开了灵芝酸A在治疗银屑病中的应用以及灵芝酸A的提取方法,灵芝酸A直接从灵芝中提取获得,成分天然,无毒,无副作用,且灵芝酸对于银屑病有显著的治疗作用,可长期使用。(The invention discloses application of ganoderic acid A in treating psoriasis and an extraction method of ganoderic acid A, wherein ganoderic acid A is directly extracted from ganoderma, is natural in component, non-toxic and free of side effect, has a remarkable treatment effect on psoriasis, and can be used for a long time.)

1. Application of ganoderic acid A in medicine for treating psoriasis is provided.

2. The use of ganoderic acid a in a medicament for treating psoriasis according to claim 1 wherein the medicament is an oral medicament.

3. The use of ganoderic acid a in a medicament for treating psoriasis according to claim 2, wherein the oral medicament comprises granules, capsules, tablets.

4. The use of ganoderic acid A in a medicament for treating psoriasis according to claim 1, wherein the medicament is a topical medicament.

5. The use of ganoderic acid A in medicine for treating psoriasis according to claim 4, wherein the external medicine comprises ointment, cream or milky lotion.

6. The use of ganoderic acid A in a medicament for treating psoriasis according to claim 1, wherein the extraction method of ganoderic acid A comprises the following steps:

s1: taking the lucid ganoderma and fully drying the lucid ganoderma in a dryer;

s2: crushing the dried lucid ganoderma to 200-300 meshes by using a crusher;

s3: leaching the pulverized Ganoderma powder with ethanol water solution for 3 times, collecting leaching solution each time, and mixing the leaching solutions for 3 times;

s4: concentrating the leaching solution obtained in S3 under reduced pressure at 60-70 deg.C and-0.1 MPa to obtain ganoderic acid extract;

s5: mixing the solution obtained in S4Mixing ganoderic acid extract with water at a ratio of 1:5-1:7 to obtain mixed solution, and adding NaHCO 5 wt% of the mixed solution₃Uniformly stirring and filtering, and adjusting the pH value of the filtrate to 2 by using a hydrochloric acid aqueous solution with the mass percentage concentration of 10%;

s6: extracting the filtrate with pH of 2 in S5 with 5 times volume of chloroform for 3 times, collecting chloroform layer each time, and mixing the chloroform layers for 3 times; concentrating at 40-50 deg.C under-0.1 MPa to obtain concentrated solution;

s7: the concentrated solution obtained in S6 was purified by silica gel column chromatography using chloroform as a column-packing solvent, chloroform: gradient washing with acetone at a volume ratio of 10:1-3:1, and collecting to obtain ganoderic acid A crude product;

s8: completely eluting the crude product of ganoderic acid A with 51% methanol water solution on LH-20 normal pressure gel column, and concentrating at 40 deg.C and-0.1 MPa to obtain ganoderic acid A with purity of more than 99%;

s9: dissolving 99% ganoderic acid A obtained in S8 with chloroform, adding methanol, standing at 4 deg.C until white crystal is separated out, and filtering to obtain ganoderic acid A.

Technical Field

The invention belongs to the technical field of psoriasis treatment, and particularly relates to application of ganoderic acid A in a medicament for treating psoriasis.

Background

Psoriasis, commonly known as "psoriasis," is a common highly recurrent skin disease. Psoriasis has the characteristics of long course of disease, difficult recovery, and the like, can cause disability and even endanger the life of a patient when being serious, and causes double physiological-psychological burdens to the patient. However, the pathogenesis of psoriasis is complex, and no radical treatment medicine is available in clinical practice until now.

At present, oral and external medicines for psoriasis mainly block an overactivated immune system, but a small part of patients with psoriasis still have slight response, ineffectiveness or serious adverse reaction accompanied with the existing biological preparation, most of the patients with effective treatment have the problems of drug withdrawal recurrence and secondary drug resistance, and because the medicines can only temporarily relieve the psoriasis symptoms, and secondary infection, liver and kidney function damage and other toxic and side effects are large, the price is high, the long-term maintenance is difficult, the search for a medicine which can be maintained for a long time is urgent.

Disclosure of Invention

In order to solve the technical problems, the invention provides the ganoderic acid A for treating psoriasis, and the medicinal components are natural and non-toxic, and have remarkable effect.

In order to achieve the technical effects, the invention is realized by the following technical scheme:

application of ganoderic acid A in medicine for treating psoriasis;

preferably, the medicament is an oral medicament;

preferably, the oral medicament comprises granules, capsules and tablets;

preferably, the medicament is an external medicament;

preferably, the external quasi-drug comprises ointment, cream and cream;

preferably, the method for extracting ganoderic acid A comprises the following steps:

s1: taking the lucid ganoderma and fully drying the lucid ganoderma in a dryer;

s2: crushing the dried lucid ganoderma to 200-300 meshes by using a crusher;

s3: leaching the pulverized Ganoderma powder with ethanol water solution for 3 times, collecting leaching solution each time, and mixing the leaching solutions for 3 times;

s4: concentrating the leaching solution obtained in S3 under reduced pressure at 60-70 deg.C and-0.1 MPa to obtain ganoderic acid extract;

s5: mixing the ganoderic acid extract in S4 with water according to the proportion of 1:5-1:7 to obtain a mixed solution, and then adding NaHCO with the mass of 5% of the mixed solution₃Uniformly stirring and filtering, and adjusting the pH value of the filtrate to 2 by using a hydrochloric acid aqueous solution with the mass percentage concentration of 10%;

s6: extracting the filtrate with pH of 2 in S5 with 5 times volume of chloroform for 3 times, collecting chloroform layer each time, and mixing the chloroform layers for 3 times; concentrating at 40-50 deg.C under-0.1 MPa to obtain concentrated solution;

s7: the concentrated solution obtained in S6 was purified by silica gel column chromatography using chloroform as a column-packing solvent, chloroform: gradient washing with acetone at a volume ratio of 10:1-3:1, and collecting to obtain ganoderic acid A crude product;

s8: completely eluting the crude product of ganoderic acid A with 51% methanol water solution on LH-20 normal pressure gel column, and concentrating at 40 deg.C and-0.1 MPa to obtain ganoderic acid A with purity of more than 99%;

s9: dissolving 99% ganoderic acid A obtained in S8 with chloroform, adding methanol, standing at 4 deg.C until white crystal is separated out, and filtering to obtain ganoderic acid A.

The invention has the beneficial effects that:

according to the invention, the psoriasis is treated by using the ganoderic acid A, the ganoderic acid A is directly extracted from the ganoderma lucidum, the components are natural and non-toxic, no toxic or side effect is caused to a human body, and the ganoderic acid has a remarkable treatment effect on the psoriasis; has no toxicity, can be taken for a long time, and is not easy to relapse.

Drawings

FIG. 1 is a picture of a real object of an animal experiment result;

FIG. 2 is a graph showing the results of observation of HE stained sections;

FIG. 3 is a data statistics diagram of the results of cell experiments.

Detailed Description

In order to further embody the aspects and effects of the present invention, the following detailed description will be given by way of specific examples.

Example 1

Application of ganoderic acid A in medicine for treating psoriasis;

preferably, the medicament is a cream.

Example 2

The extraction method of the ganoderic acid A comprises the following steps:

s1: taking the lucid ganoderma and fully drying the lucid ganoderma in a dryer;

s2: pulverizing the dried Ganoderma lucidum to 200 meshes by a pulverizer;

s3: leaching the pulverized Ganoderma powder with ethanol water solution for 3 times, collecting leaching solution each time, and mixing the leaching solutions for 3 times;

s4: concentrating the leaching solution obtained in S3 under reduced pressure at 60 deg.C and-0.1 MPa to obtain ganoderic acid extract;

s5: mixing the ganoderic acid extract in S4 with water at a ratio of 1:5 to obtain a mixed solution, and adding NaHCO at an amount of 5% of the mixed solution₃Uniformly stirring and filtering, and adjusting the pH value of the filtrate to 2 by using a hydrochloric acid aqueous solution with the mass percentage concentration of 10%;

s6: extracting the filtrate with pH of 2 in S5 with 5 times volume of chloroform for 3 times, collecting chloroform layer each time, and mixing the chloroform layers for 3 times; concentrating at 50 deg.C and-0.1 MPa to obtain concentrated solution;

s7: the concentrated solution obtained in S6 was purified by silica gel column chromatography using chloroform as a column-packing solvent, chloroform: gradient washing with acetone at a volume ratio of 10:1-3:1, and collecting to obtain ganoderic acid A crude product;

s8: completely eluting the crude product of ganoderic acid A with 51% methanol water solution on LH-20 normal pressure gel column, and concentrating at 40 deg.C and-0.1 MPa to obtain ganoderic acid A with purity of more than 99%;

s9: dissolving 99% ganoderic acid A obtained in S8 with chloroform, adding methanol, standing at 4 deg.C until white crystal is separated out, and filtering to obtain ganoderic acid A.

Example 3

To further verify the therapeutic effect of the cream containing ganoderic acid a in example 1 on psoriasis, the following animal experiments were designed;

the preparation of an animal model of psoriasis: selecting 40 female mice with the body weight of 18-20g and the age of 6-8 weeks, and feeding the female mice in a standard clean-grade animal feeding room; depilating the mouse with depilatory cream, wherein the depilatory area is 2cm x 4 cm;

uniformly applying 5% imiquimod cream on the unhaired part according to the dose of 62.5 mg/mouse, and forming a model of the psoriasis mouse after 5-7 days;

grouping experiments: a normal control group, a psoriasis + solvent control group, a psoriasis + ganoderic acid A treatment group; 10 mice per group;

the experimental method comprises the following steps: gavage with ganoderic acid A at 2mg/kg 1 time per day, and not treating the normal control group, and adding solvent and no drug to the solvent control group.

The experimental results are as follows: the results are shown in figure 1, where a is the normal control group, b is the psoriasis non-treated group, c is the psoriasis + solvent control group, d is the psoriasis + ganoderic acid a group;

psoriasis no treatment group: the affected part skin has obvious erythema flakiness, much scale shedding, large-area obvious hyperkeratosis, thick spinous layer and abnormal folds on the skin surface;

psoriasis + solvent control group: compared with psoriasis group, the medicine has no obvious difference;

psoriasis + ganoderic acid group a: erythema on the affected part of the skin basically subsides, the thickness of skin folds becomes thinner, and scales are reduced.

Example 4

Cell section staining experiment;

experimental reagent: fixing liquid: 95% ethanol and glacial acetone; hematoxylin staining solution: weighing 0.5g of hematoxylin and 24g of ammonium alum, dissolving in 70ml of distilled water, weighing 31g of NaCl and 5ml of water, adding 30ml of glycerol and 2ml of glacial acetic acid, uniformly mixing, and filtering to obtain filtrate to obtain hematoxylin staining solution; eosin dye liquor: weighing 0.5g of water-soluble eosin dye solution, and dissolving in 100ml of distilled water; dilute hydrochloric acid ethanol solution: preparing 1% hydrochloric acid by using 75% ethanol;

preparation of the experiment: taking back skin of each group of mice, and preparing into slices;

the experimental steps are as follows: s1: paraffin section dewaxing: sequentially placing the slices in xylene I10 min-xylene II 10 min-absolute ethyl alcohol I5 min-absolute ethyl alcohol II 5 min-95% ethyl alcohol 5 min-90% ethyl alcohol 5 min-80% ethyl alcohol 5 min-70% ethyl alcohol 5 min-distilled water washing;

s2: hematoxylin staining of cell nucleus: slicing into Harris hematoxylin, dyeing for 3-8min, washing with tap water, differentiating with 1% hydrochloric acid alcohol for several seconds, washing with tap water, returning blue with 0.6% ammonia water, and washing with running water;

s3: eosin staining of cytoplasm: placing the slices in eosin dye solution for dyeing for 1-3 min;

s4: dewatering and sealing: placing the slices in 95% alcohol I5 min-95% alcohol II 5 min-absolute ethanol I5 min-absolute ethanol II 5 min-xylene I5 min-xylene II 5min to dehydrate and transparent, taking out the slices from xylene, air drying, and sealing with neutral gum;

s5: microscopic examination and image acquisition and analysis;

the experimental results are as follows: as shown in fig. 2, wherein a is a normal control group, b is a psoriasis non-treatment group, c is a psoriasis + ointment group, and d is a psoriasis + ganoderic acid a group;

and (3) analyzing a slicing result: the epidermis of the psoriasis group is obviously thickened, inflammatory cell infiltration of the dermis layer is obvious, and after the treatment of the ganoderic acid A, the proliferation of the epidermis is obviously improved, and the inflammatory cell infiltration is reduced (as the direction indicated by an arrow in figure 2).

Example 5

Cell experiments;

preparation of the experiment: immortalized human Hacat keratinocytes were treated with different ganoderic acids A (5. mu.M/10. mu.M/20. mu.M/40. mu.M)

The experimental method comprises the following steps: recovering human Hacat keratinocyte, when the growth vigor is good, placing culture dishes of 5000 cells in a CO2 incubator, standing and culturing at 37 ℃ for 24 hours by 5% CO2, adding culture solution of ganoderic acid A with different concentrations, continuously culturing for 24 hours, removing the culture solution, incubating for 1-3 hours after CCK8, and detecting the inhibition effect of the ganoderic acid A on keratinocyte proliferation by 450nm degrees.

The experimental results are as follows: as shown in FIG. 3, ganoderic acid A significantly inhibited keratinocyte proliferation and was concentration dependent.

And (4) analyzing results: ganoderic acid A can obviously inhibit keratinocyte proliferation.