CN113440538A - Polysaccharide composition with effect of relieving atopic dermatitis and skin external medical auxiliary preparation - Google Patents
Polysaccharide composition with effect of relieving atopic dermatitis and skin external medical auxiliary preparation Download PDFInfo
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/72—Cosmetics or similar toiletry preparations characterised by the composition containing organic macromolecular compounds
- A61K8/73—Polysaccharides
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0014—Skin, i.e. galenical aspects of topical compositions
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/04—Antipruritics
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/06—Fungi, e.g. yeasts
- A61K36/07—Basidiomycota, e.g. Cryptococcus
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/28—Asteraceae or Compositae (Aster or Sunflower family), e.g. chamomile, feverfew, yarrow or echinacea
- A61K36/285—Aucklandia
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/88—Liliopsida (monocotyledons)
- A61K36/898—Orchidaceae (Orchid family)
- A61K36/8984—Dendrobium
Abstract
The invention discloses a polysaccharide composition with an effect of relieving atopic dermatitis and a skin external medical auxiliary preparation. The polysaccharide composition obtained by matching the tremella polysaccharide, the dendrobium officinale polysaccharide, the bamboo fungus polysaccharide and the saussurea involucrate polysaccharide has good immunoregulation effect on skin, has good effect of relieving atopic dermatitis, and has the drug effect obviously superior to that of the single use of each component or the matching use of the two components; the four polysaccharides are extracted from pure natural Chinese medicines, have low toxic and side effects, reduce the harm to human bodies, and have relatively low price. The polysaccharide composition can be used as an active ingredient for preparing a skin external medical auxiliary preparation, such as a skin external medical auxiliary medicament or a cosmetic.
Description
Technical Field
The invention relates to the technical field of dermatological drugs, and in particular relates to a polysaccharide composition with an effect of relieving atopic dermatitis and an external skin medical auxiliary preparation.
Background
Atopic Dermatitis (AD) is a chronic, recurrent, inflammatory skin disease with intractable itching, dry skin, eczematous lesions as the main clinical manifestations, and its exact pathogenesis is not clear. Atopic dermatitis is characterized by repeated onset and difficulty in healing, and can damage the physical health of patients, seriously affect the mood of patients and reduce the quality of life of patients. The symptoms of skin damage after the disease occurs, and the beauty of the skin of a patient is affected.
The main means of the prior art for dealing with atopic dermatitis is topical treatment, and topical glucocorticoid preparations are the main drugs for the treatment of atopic dermatitis. Besides, oral antibiotics and immunomodulators also have a certain relieving effect on atopic dermatitis. However, topical glucocorticoids, oral antibiotics and immunomodulators have toxic and side effects, and new drugs are needed to be developed so as to improve the drug effect, reduce the dosage and reduce the toxic and side effects. In addition, the existing drugs for treating atopic dermatitis are expensive.
Therefore, there is an urgent need to develop a drug which has a good effect of relieving atopic dermatitis, a low dosage, less toxic and side effects, and a relatively low price.
Disclosure of Invention
The polysaccharide composition has the advantages of immunoregulation effect, good effect of relieving atopic dermatitis, low toxic and side effects and relatively low price.
Another object of the present invention is to provide a process for producing the above polysaccharide composition.
Still another object of the present invention is to provide the use of the above polysaccharide composition for the preparation of a medicament for alleviating atopic dermatitis.
In order to solve the technical problems, the invention adopts the technical scheme that:
a polysaccharide composition with effect of relieving atopic dermatitis comprises Tremella polysaccharide, herba Dendrobii officinalis polysaccharide, caulis Bambusae in Taeniam polysaccharide and herba Saussureae Involueratae polysaccharide.
The research of the inventor finds that the tremella polysaccharide, the dendrobium officinale polysaccharide, the dictyophora indusiata polysaccharide and the saussurea involucrate polysaccharide have a synergistic effect on skin immunoregulation, the polysaccharide composition obtained by matching the four polysaccharides has a good immunoregulation effect on skin, the effect of relieving atopic dermatitis is good, and the drug effect is obviously superior to that of the single use of each component or the matching use of the two components. The four polysaccharides are all extracted from pure natural traditional Chinese medicines, have low toxic and side effects, reduce the harm to human bodies, and have relatively low price.
Preferably, the mass ratio of the tremella polysaccharide to the dendrobium officinale polysaccharide to the bamboo fungus polysaccharide to the saussurea involucrate polysaccharide is (1-5) to (1-5).
More preferably, the mass ratio of the tremella polysaccharide to the dendrobium officinale polysaccharide to the bamboo fungus polysaccharide to the saussurea involucrate polysaccharide is (1-2) to (1-1.5) to 1.
Optionally, the mass ratio of the tremella polysaccharide, the dendrobium officinale polysaccharide, the bamboo fungus polysaccharide and the saussurea involucrate polysaccharide is 1: 1; or the mass ratio of the tremella polysaccharide to the dendrobium officinale polysaccharide to the bamboo fungus polysaccharide to the saussurea involucrate polysaccharide is 2: 1; or the mass ratio of the tremella polysaccharide to the dendrobium officinale polysaccharide to the bamboo fungus polysaccharide to the saussurea involucrate polysaccharide is 1.5: 1: 1.5: 1; or the mass ratio of the tremella polysaccharide to the dendrobium officinale polysaccharide to the bamboo fungus polysaccharide to the saussurea involucrate polysaccharide is 1.5: 1.
Further preferably, the mass ratio of the tremella polysaccharide, the dendrobium officinale polysaccharide, the bamboo fungus polysaccharide and the saussurea involucrate polysaccharide is 1: 1.
Researches show that the tremella polysaccharide, the dendrobium officinale polysaccharide, the bamboo fungus polysaccharide and the saussurea involucrate polysaccharide are combined according to the mass ratio of 1:1, so that the medicine has a better immunoregulation effect, has a good inhibition effect on a mouse atopic dermatitis model induced by dinitrofluorobenzene, and can be safely applied to skin external medical auxiliary medicines or cosmetics with an effect of relieving the atopic dermatitis.
The invention also provides a preparation method of the polysaccharide composition, wherein the preparation method comprises the steps of respectively taking tremella, dendrobium officinale, dictyophora phalloidea and saussurea involucrate as raw materials, preparing tremella polysaccharide, dendrobium officinale polysaccharide, dictyophora phalloidea polysaccharide and saussurea involucrate polysaccharide by a water extraction and alcohol precipitation method, and mixing to obtain the polysaccharide composition.
In the field, generally, the polysaccharide content in the tremella polysaccharide, the dendrobium officinale polysaccharide, the bamboo fungus polysaccharide and the saussurea involucrate polysaccharide is more than or equal to 80%.
In one embodiment, the tremella polysaccharide is prepared by taking dried tremella, removing insect or deterioration parts, pulverizing with a pulverizer, and sieving with a 60-mesh sieve. Taking medicinal powder, adding a 75% ethanol solution according to a material-to-liquid ratio of 1: 10-1: 40, ultrasonically leaching for 30min at 50 ℃, heating for 2 hours, centrifuging the obtained solution for 15min at 4000r/min, taking supernatant, filtering, performing reduced pressure rotary evaporation to remove ethanol, and freeze-drying for 48 hours to obtain the tremella polysaccharide.
In one embodiment, the dendrobium officinale polysaccharide is prepared by taking dried dendrobium officinale, removing insect-containing or metamorphic parts, crushing by using a powder grinding machine, and sieving by using a 60-mesh sieve. Taking medicinal material powder, adding a 75% ethanol solution according to a material-liquid ratio of 1: 10-1: 40, performing ultrasonic extraction at 50 ℃ for 30min, heating for 2 h, centrifuging the obtained solution at 4000r/min for 15min, taking supernate, filtering, performing reduced pressure rotary evaporation to remove ethanol, and freeze-drying for 48h to obtain the dendrobium officinale polysaccharide.
In one embodiment, the dictyophora indusiata polysaccharide is prepared by taking dried dictyophora indusiata, removing insect-containing or deteriorated parts, crushing by using a powder grinding machine, and sieving by using a 60-mesh sieve. Adding a 75% ethanol solution into medicinal material powder according to a material-liquid ratio of 1: 10-1: 40, performing ultrasonic extraction at 50 ℃ for 30min, heating for 2 h, centrifuging the obtained solution at 4000r/min for 15min, filtering supernate, performing rotary evaporation under reduced pressure to remove ethanol, and freeze-drying for 48h to obtain the dictyophora indusiata polysaccharide.
In one embodiment, the saussurea involucrate polysaccharide is prepared by drying saussurea involucrate, removing insect or deterioration parts, pulverizing with a pulverizer, and sieving with a 60-mesh sieve. Adding a 75% ethanol solution into medicinal material powder according to a material-liquid ratio of 1: 10-1: 40, performing ultrasonic extraction at 50 ℃ for 30min, heating for 2 h, centrifuging the obtained solution at 4000r/min for 15min, filtering supernatant, performing reduced pressure rotary evaporation to remove ethanol, and freeze-drying for 48h to obtain the saussurea involucrata polysaccharide. The invention also protects the application of the polysaccharide composition in preparing a medicament for relieving atopic dermatitis.
The application of the polysaccharide composition in preparing the medicine for relieving atopic dermatitis induced by dinitrofluorobenzene is also within the protection scope of the invention.
The invention also provides an external skin medical auxiliary preparation, and the active ingredient of the external skin medical auxiliary preparation is the polysaccharide composition.
As one embodiment, the medical auxiliary preparation for external application to the skin is a medical auxiliary drug or cosmetic for external application to the skin.
The skin external medical auxiliary drug or the cosmetic is a preparation prepared according to certain formulation requirements, and mainly has the effect of relieving atopic dermatitis by being applied to the skin surface or acting on the skin surface in other modes.
Preferably, the external medical auxiliary preparation for skin comprises auxiliary materials and the polysaccharide composition.
Preferably, the formulation of the medical auxiliary preparation for external application to the skin is liquid, cream, gel, spray, freeze-dried preparation, granule, tablet or capsule.
As one embodiment, the external medical auxiliary drug for skin is in the form of cream, and the auxiliary materials comprise squash, Montanov S, deionized water, glycerol and phenoxyethanol.
The cosmetic can be in the form of cream, lotion, essence, gel or spray. The auxiliary materials of the cosmetics comprise a base material and additives which are commonly used in the cosmetics.
Compared with the prior art, the invention has the beneficial effects that:
the polysaccharide composition obtained by matching the tremella polysaccharide, the dendrobium officinale polysaccharide, the bamboo fungus polysaccharide and the saussurea involucrate polysaccharide has good immunoregulation effect on skin, has good effect of relieving atopic dermatitis, and has the drug effect obviously superior to that of the single use of each component or the matching use of the two components. The four polysaccharides are all extracted from pure natural traditional Chinese medicines, have low toxic and side effects, reduce the harm to human bodies, and have relatively low price.
The polysaccharide composition can be used as active ingredient for preparing skin external medical auxiliary preparation, such as skin external medical auxiliary medicine or cosmetic.
Drawings
Fig. 1 is a graph showing the results of a test of the transdermal water loss rate of the medical auxiliary preparation for external use for skin prepared in example 1 of the present invention and in comparative examples 1 to 10.
FIG. 2 is a graph showing the results of the test of the relative thickness of the epidermis of the medical auxiliary preparation for external use on skin prepared in example 1 of the present invention and in comparative examples 1 to 10.
FIG. 3 is a graph showing the results of a test for TNF-. alpha.inhibition of the medical auxiliary for external application to the skin prepared in example 1 of the present invention and in comparative examples 1 to 10.
FIG. 4 is a graph showing the results of cytotoxicity test of polysaccharide extracts used in examples and comparative examples.
Detailed Description
The present invention will be further described with reference to the following embodiments.
The raw materials in the examples are all commercially available;
reagents, methods and apparatus used in the present invention are conventional in the art unless otherwise indicated.
In the following examples and comparative examples, tremella polysaccharide, dendrobium officinale polysaccharide, bamboo fungus polysaccharide and saussurea involucrate polysaccharide are respectively prepared by the following preparation methods:
the preparation method of the tremella polysaccharide comprises the following steps: (1) pretreatment of tremella: taking dried tremella, removing insect or deterioration parts, pulverizing with a pulverizer, and sieving with a 60-mesh sieve; (2) taking medicinal powder, adding a 75% ethanol solution according to a material-to-liquid ratio of 1: 10-1: 40, performing ultrasonic extraction at 50 ℃ for 30min, and heating for 2 hours; (3) centrifuging the tremella extract obtained in the step (2) at 4000r/min for 15min, and filtering supernate; (4) and (4) carrying out reduced pressure rotary evaporation on the tremella extract obtained in the step (3) to remove ethanol, and freeze-drying for 48 hours to finally obtain the extract, namely tremella polysaccharide.
The preparation method of the dendrobium officinale polysaccharide comprises the following steps: (1) pretreatment of the dendrobium officinale: taking the dried dendrobium officinale, removing insect-containing or metamorphic parts, crushing by using a powder grinding machine, and sieving by using a 60-mesh sieve; (2) taking medicinal powder, adding a 75% ethanol solution according to a material-to-liquid ratio of 1: 10-1: 40, performing ultrasonic extraction at 50 ℃ for 30min, and heating for 2 hours; (3) centrifuging the dendrobium officinale extract obtained in the step (2) at 4000r/min for 15min, and filtering supernate; (4) and (4) carrying out reduced pressure rotary evaporation on the dendrobium officinale extract obtained in the step (3) to remove ethanol, and freeze-drying for 48 hours to finally obtain the extract, namely the dendrobium officinale polysaccharide.
The preparation method of the bamboo fungus polysaccharide comprises the following steps: (1) pretreatment of bamboo fungus: taking dried bamboo fungus, removing insect-containing or deteriorated parts, pulverizing with a pulverizer, and sieving with a 60-mesh sieve; (2) taking medicinal powder, adding a 75% ethanol solution according to a material-to-liquid ratio of 1: 10-1: 40, performing ultrasonic extraction at 50 ℃ for 30min, and heating for 2 hours; (3) centrifuging the Dictyophora Indusiata extract obtained in step (2) at 4000r/min for 15min, and filtering the supernatant; (4) and (4) carrying out reduced pressure rotary steaming on the dictyophora indusiata extract obtained in the step (3) to remove ethanol, and freeze-drying for 48 hours to finally obtain an extract, namely the dictyophora indusiata polysaccharide.
The preparation method of the saussurea involucrate polysaccharide comprises the following steps: (1) pretreatment of the saussurea involucrate: taking sun-dried saussurea involucrate, removing insect-containing or deteriorated parts, pulverizing with a pulverizer, and sieving with a 60-mesh sieve; (2) taking medicinal powder, adding a 75% ethanol solution according to a material-to-liquid ratio of 1: 10-1: 40, performing ultrasonic extraction at 50 ℃ for 30min, and heating for 2 hours; (3) centrifuging the saussurea involucrate extract obtained in the step (2) at 4000r/min for 15min, and filtering supernatant; (4) and (4) carrying out reduced pressure rotary evaporation on the saussurea involucrate extract obtained in the step (3) to remove ethanol, and freeze-drying for 48 hours to finally obtain the extract, namely the saussurea involucrate polysaccharide.
Example 1
A polysaccharide composition with effect of relieving atopic dermatitis comprises Tremella polysaccharide, herba Dendrobii polysaccharide, caulis Bambusae in Taeniam polysaccharide and herba Saussureae Involueratae polysaccharide. In the embodiment, the mass ratio of the tremella polysaccharide, the dendrobium officinale polysaccharide, the bamboo fungus polysaccharide and the saussurea involucrate polysaccharide is 1: 1.
The polysaccharide composition of the present example was used as an active ingredient to prepare an auxiliary preparation for external use for skin treatment. The skin external medical auxiliary preparation comprises auxiliary materials and a polysaccharide composition.
The preparation method of the external skin medical aid preparation of the embodiment comprises the following steps:
(1) mixing and pre-dissolving the raw materials of the component A, and uniformly stirring at the temperature of 130 ℃ at 5000 r/min;
(2) stirring and cooling, adding the raw material of the component B at 90 ℃, and stirring uniformly;
(3) taking 0.05 percent of polysaccharide composition according to mass percentage; adding 19.85% water, heating to 90 deg.C, slowly adding A, B components mixture, and stirring;
(4) stirring and cooling, adding the component C at the temperature of below 40 ℃, and stirring uniformly;
(5) stirring and cooling, and discharging at 38 ℃.
The ratio of each raw material in component A, B, C to the total mass is shown in table 1.
TABLE 1 ratio of the raw materials in component A, B, C to the total mass
Example 2
The embodiment provides a polysaccharide composition with an effect of relieving atopic dermatitis, and a skin external medical auxiliary preparation prepared from the same. In the polysaccharide composition of the embodiment, the mass ratio of the tremella polysaccharide, the dendrobium officinale polysaccharide, the bamboo fungus polysaccharide and the saussurea involucrate polysaccharide is 2: 1.
The method for preparing the medical preparation for external application to the skin of this example is the same as in example 1.
Example 3
The embodiment provides a polysaccharide composition with an effect of relieving atopic dermatitis, and a skin external medical auxiliary preparation prepared from the same. In the polysaccharide composition of the embodiment, the mass ratio of the tremella polysaccharide, the dendrobium officinale polysaccharide, the bamboo fungus polysaccharide and the saussurea involucrate polysaccharide is 3: 2: 3: 2.
The method for preparing the medical preparation for external application to the skin of this example is the same as in example 1.
Example 4
The embodiment provides a polysaccharide composition with an effect of relieving atopic dermatitis, and a skin external medical auxiliary preparation prepared from the same. In the polysaccharide composition of the embodiment, the mass ratio of the tremella polysaccharide, the dendrobium officinale polysaccharide, the bamboo fungus polysaccharide and the saussurea involucrate polysaccharide is 3: 2.
The method for preparing the medical preparation for external application to the skin of this example is the same as in example 1.
Comparative example 1
This comparative example provides a medical auxiliary preparation for external use on skin, which is different from example 1 in that it contains only one polysaccharide, i.e., tremella polysaccharide;
other raw materials and preparation method are the same as example 1.
Comparative example 2
This comparative example provides a medical auxiliary preparation for external application to skin, which is different from example 1 in that it contains only one polysaccharide, i.e., dendrobium officinale polysaccharide;
other raw materials and preparation method are the same as example 1.
Comparative example 3
This comparative example provides a medical auxiliary preparation for external use on skin, which is different from example 1 in that it contains only one polysaccharide, i.e., dictyophora indusiata polysaccharide;
other raw materials and preparation method are the same as example 1.
Comparative example 4
This comparative example provides a medical auxiliary preparation for external application to skin, which is different from example 1 in that it contains only one polysaccharide, namely saussurea involucrate polysaccharide;
other raw materials and preparation method are the same as example 1.
Comparative example 5
The present comparative example provides a polysaccharide composition having an effect of alleviating atopic dermatitis, and a skin external medical aid preparation prepared therefrom. The polysaccharide composition of the comparative example only contains two polysaccharides, namely tremella polysaccharide and dendrobium officinale polysaccharide, and the mass ratio of the tremella polysaccharide to the dendrobium officinale polysaccharide is 1: 1.
The procedure for preparing the medical auxiliary preparation for external application to the skin of this comparative example was the same as in example 1.
Comparative example 6
The present comparative example provides a polysaccharide composition having an effect of alleviating atopic dermatitis, and a skin external medical aid preparation prepared therefrom. The polysaccharide composition of the comparative example only contains two polysaccharides, namely tremella polysaccharide and saussurea involucrate polysaccharide, and the mass ratio of the tremella polysaccharide to the saussurea involucrate polysaccharide is 1: 1.
The procedure for preparing the medical auxiliary preparation for external application to the skin of this comparative example was the same as in example 1.
Comparative example 7
The present comparative example provides a polysaccharide composition having an effect of alleviating atopic dermatitis, and a skin external medical aid preparation prepared therefrom. The polysaccharide composition of the comparative example only contains two polysaccharides, namely the dendrobium officinale polysaccharide and the saussurea involucrate polysaccharide, and the mass ratio of the dendrobium officinale polysaccharide to the saussurea involucrate polysaccharide is 1: 1.
The procedure for preparing the medical auxiliary preparation for external application to the skin of this comparative example was the same as in example 1.
Comparative example 8
The present comparative example provides a polysaccharide composition having an effect of alleviating atopic dermatitis, and a skin external medical aid preparation prepared therefrom. The polysaccharide composition of the comparative example only contains two polysaccharides, namely dendrobium officinale polysaccharide and bamboo fungus polysaccharide, and the mass ratio of the dendrobium officinale polysaccharide to the bamboo fungus polysaccharide is 1: 1.
The procedure for preparing the medical auxiliary preparation for external application to the skin of this comparative example was the same as in example 1.
Comparative example 9
The present comparative example provides a polysaccharide composition having an effect of alleviating atopic dermatitis, and a skin external medical aid preparation prepared therefrom. The polysaccharide composition of the comparative example only contains two polysaccharides, namely bamboo fungus polysaccharide and tremella polysaccharide, and the mass ratio of the bamboo fungus polysaccharide to the tremella polysaccharide is 1: 1.
The procedure for preparing the medical auxiliary preparation for external application to the skin of this comparative example was the same as in example 1.
Comparative example 10
The present comparative example provides a polysaccharide composition having an effect of alleviating atopic dermatitis, and a skin external medical aid preparation prepared therefrom. The polysaccharide composition of the comparative example only contains two polysaccharides, namely bamboo fungus polysaccharide and saussurea involucrate polysaccharide, and the mass ratio of the bamboo fungus polysaccharide to the saussurea involucrate polysaccharide is 1: 1.
The procedure for preparing the medical auxiliary preparation for external application to the skin of this comparative example was the same as in example 1.
Test for efficacy in alleviating atopic dermatitis
1. Test method
100 healthy Babl/c mice, 6 weeks old, 15-23 g. After 3 days of acclimatization, the animals were randomly divided into 20 groups. 24 hours before the first application, mice were shaved 2cm by 4cm on their backs. Sensitizing in the first week, smearing 0.25% DNFB (prepared from acetone and olive oil at a volume ratio of 3: 1) on the shaved part of the back for 100ul, and smearing once every three days for two times; in the second week, the back was applied with 0.2% DNFB 100ul once every three days, twice, to prepare an atopic dermatitis model. In this application, DNFB refers to 2, 4-dinitrofluorobenzene.
Dosing was started 24h after the second sensitization, 1 time per day for 8 days. The blank mice were not treated after depilating, the model mice were not treated after molding successfully, the positive group was smeared with 35ul prednisolone cream, and the experimental group was smeared with 35ul sample cream. On day 15 of the experiment, the transdermal water loss rate (TWEL) was tested using a skin moisture loss tester (MCP 2, CK, germany).
The skin of the back was removed after the mice were sacrificed and the tissue was fixed with 4% paraformaldehyde. HE stained and the morphological changes of the skin tissue were observed and the relative thickness of the epidermis was measured using the software Image J.
After the mice were sacrificed, the skin of the back was removed, 100mg of tissue was taken, and blood stain was washed with 1 XPBS. Cut into small pieces and placed in a tissue grinder, and 1mL of tissue lysate (Thermo Fisher Scientific Inc., USA) is added to make a homogenate. Centrifuging the tissue homogenate at 2-8 deg.C and 15000r/min for 5min to obtain supernatant, and collecting appropriate amount of supernatant for immediate experiment, or packaging and storing the supernatant at-80 deg.C. And centrifuging the thawed sample again, and detecting the TNF-alpha by using an ELISA kit to obtain the expression quantity of the TNF-alpha.
The cytotoxicity of TPS was tested using the 3- (4, 5-dimethylthiazole-2) -2, 5-diphenyltetrazolium bromide (MTT) method. RAW264.7 macrophages were cultured in DMEM medium containing 10% FBS and 1% diabody and placed in an incubator at 37 ℃ and 5% carbon dioxide. Seeding RAW cells (5X 10) in 96-well plates4Cells/well) were incubated in an incubator for 24 hours. Then the supernatant is discarded and 100 mul of tremella polysaccharide, dendrobium officinale polysaccharide, bamboo fungus polysaccharide and saussurea involucrate polysaccharide with different concentrations (6.25 mug/mL, 25 mug/mL and 100 mug/mL) are added. After 24 hours, 0.5mg/mL MTT was added to each well, incubated in the incubator for 4 hours, the supernatant discarded, and 150. mu.L DMSO was added. After 15 minutes, the OD was measured at 570nm using a microplate reader. Each set was provided with 4 replicate wells and replicated 3 times.
2. Test results
The experimental results are shown in the following table, wherein table 1 shows the results of the transdermal water loss rate, the relative thickness of the epidermis and the expression level of TNF-alpha, and table 2 shows the results of the polysaccharide cytotoxicity test.
TABLE 1 test results of transdermal water loss rate, epidermal relative thickness and TNF-alpha expression
Note:*compared with the model group, P is less than 0.1,**compared with the model group, P is less than 0.01,***compared with the model group, P is less than 0.001,****compared with the model group, P is less than 0.0001,
△compared with the embodiment 1, P is less than 0.1,△△compared with the embodiment 1, P is less than 0.01,△△△compared with the embodiment 1, P is less than 0.001,△△△△p < 0.0001 compared with example 1.
TABLE 2 polysaccharide cytotoxicity test results
Note:***p is less than 0.001 compared with the model group.
The results of the test on the percutaneous water loss rate of the medical auxiliary preparations for external use for skin prepared in examples 1 to 4 and comparative examples 1 to 10 are shown in fig. 1, and it can be seen that the medical auxiliary preparations for external use for skin prepared in examples 1 to 4 and comparative examples 1 to 10 can improve the water loss on the skin surface of the atopic dermatitis mouse. The examples 1 to 4 are obviously superior to the comparative examples 1 to 10, and it can be seen that the skin external medical auxiliary preparation prepared by mixing the tremella polysaccharide, the dendrobium officinale polysaccharide, the dictyophora phalloidea polysaccharide and the saussurea involucrate polysaccharide according to the mass ratio of the invention can more effectively reduce the skin percutaneous water loss rate.
The results of the test on the relative thickness of the epidermis of the medical preparation for external application to skin prepared in examples 1 to 4 and comparative examples 1 to 10 are shown in fig. 2, and it can be seen from fig. 2 that the medical preparation for external application to skin prepared in examples 1 to 4 and comparative examples 1 to 10 can reduce the relative thickness of the epidermis of the skin. Moreover, the embodiments 1-4 are significantly superior to the comparative examples 1-10, so that the skin external medical auxiliary preparation prepared by mixing the tremella polysaccharide, the dendrobium officinale polysaccharide, the dictyophora phalloidea polysaccharide and the saussurea involucrate polysaccharide according to the mass ratio of the invention can effectively reduce the relative thickness of the skin epidermis.
FIG. 3 is a graph showing the results of the test for the inhibition of TNF- α by the medical auxiliary preparations for external application to the skin prepared in examples 1 to 4 and comparative examples 1 to 10, and it can be seen from the results in FIG. 3 that the medical auxiliary preparations for external application to the skin prepared in examples 1 to 4 and comparative examples 1 to 10 can inhibit the production of TNF- α. In addition, the skin external medical auxiliary preparations of examples 1 to 4 have the best inhibition effect, more effectively inhibit the generation of TNF-alpha, and are obviously superior to the skin external medical auxiliary preparations prepared in comparative examples 1 to 10.
Fig. 4 is a graph showing the cytotoxicity test results of the polysaccharide extracts used in the examples and comparative examples, and it can be found from the results in fig. 4 that tremella polysaccharide, dendrobium officinale polysaccharide, dictyophora indusiata polysaccharide and saussurea involucrate polysaccharide have no cytotoxicity under the action of high dose (100 μ g/ml), so that a medium dose (50 μ g/ml) is adopted in the medical adjuvant for external use on skin, the dosage is small, and no toxic or side effect is caused.
It should be understood that the above-described embodiments of the present invention are merely examples for clearly illustrating the present invention, and are not intended to limit the embodiments of the present invention. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. And are neither required nor exhaustive of all embodiments. Any modification, equivalent replacement, and improvement made within the spirit and principle of the present invention should be included in the protection scope of the claims of the present invention.
Claims (10)
1. A polysaccharide composition with effect of relieving atopic dermatitis is characterized by comprising tremella polysaccharide, dendrobium officinale polysaccharide, bamboo fungus polysaccharide and saussurea involucrate polysaccharide.
2. The polysaccharide composition of claim 1, wherein the mass ratio of the tremella polysaccharide to the dendrobium officinale polysaccharide to the bamboo fungus polysaccharide to the saussurea involucrate polysaccharide is (1-5) to (1-5).
3. The polysaccharide composition of claim 2, wherein the mass ratio of the tremella polysaccharide to the dendrobium officinale polysaccharide to the bamboo fungus polysaccharide to the saussurea involucrate polysaccharide is (1-2) to (1-1.5) to 1.
4. The polysaccharide composition of claim 3, wherein the mass ratio of the tremella polysaccharide to the dendrobium officinale polysaccharide to the bamboo fungus polysaccharide to the saussurea involucrate polysaccharide is 1: 1.
5. The preparation method of the polysaccharide composition according to any one of claims 1 to 4, wherein tremella, dendrobium officinale, dictyophora phalloidea and saussurea involucrata are respectively used as raw materials, tremella polysaccharide, dendrobium officinale polysaccharide, dictyophora phalloidea polysaccharide and saussurea involucrata polysaccharide are prepared by a water extraction and alcohol precipitation method, and are mixed to obtain the polysaccharide composition.
6. Use of the polysaccharide composition of any one of claims 1 to 4 in the manufacture of a medicament for the alleviation of atopic dermatitis.
7. Use of the polysaccharide composition according to any one of claims 1 to 4 in the preparation of a medicament for alleviating atopic dermatitis induced by dinitrofluorobenzene.
8. A medical preparation for external application to the skin, characterized in that the active ingredient of the medical preparation for external application to the skin is the polysaccharide composition according to any one of claims 1 to 5.
9. The medical preparation for external application to skin as claimed in claim 8, wherein the medical preparation for external application to skin comprises an adjuvant and the polysaccharide composition as claimed in any one of claims 1 to 5.
10. The medical preparation for external application on skin as claimed in claim 8 or 9, wherein the dosage form of the medical preparation for external application on skin is liquid, cream, gel, spray, freeze-dried preparation, granule, tablet or capsule.
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Application publication date: 20210928 |