CN115364003A - Application of herba Terminaliae extract in preventing and/or repairing ultraviolet induced skin injury - Google Patents

Application of herba Terminaliae extract in preventing and/or repairing ultraviolet induced skin injury Download PDF

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CN115364003A
CN115364003A CN202110558860.7A CN202110558860A CN115364003A CN 115364003 A CN115364003 A CN 115364003A CN 202110558860 A CN202110558860 A CN 202110558860A CN 115364003 A CN115364003 A CN 115364003A
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不公告发明人
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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
    • A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
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    • A61K8/9789—Magnoliopsida [dicotyledons]
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    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/53—Lamiaceae or Labiatae (Mint family), e.g. thyme, rosemary or lavender
    • A—HUMAN NECESSITIES
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    • A—HUMAN NECESSITIES
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    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q17/00—Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
    • A61Q17/04—Topical preparations for affording protection against sunlight or other radiation; Topical sun tanning preparations
    • 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
    • A61Q19/004—Aftersun preparations
    • 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
    • A61Q19/005—Preparations for sensitive skin
    • 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
    • A61Q19/08—Anti-ageing preparations

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Abstract

The invention provides an application of a turner grass extract in preventing and/or repairing ultraviolet-induced skin injury. Specifically, the turner grass extract has good protection effect on cells before and after ultraviolet irradiation, is very suitable to be used as an active ingredient of cosmetics, and is particularly suitable to be used for preparing cosmetics for preventing sun.

Description

Application of herba Terminaliae extract in preventing and/or repairing ultraviolet induced skin injury
Technical Field
The invention relates to the field of cosmetics, in particular to a cell protection effect of a turnabout extract.
Background
The skin problems induced by ultraviolet rays are the main causes of exogenous damage and skin aging. The damage of the skin by ultraviolet rays includes suntan, sunburn, decomposition and denaturation of collagen and elastin, inflammation aggravation, free radical formation induction, skin immunity inhibition, actinic dermatosis induction, pigmented spots aggravation, and skin cancer induction.
Ultraviolet damage can be classified into acute damage and chronic damage, wherein acute damage includes sunburn and the like, and skin aging or accelerated aging (i.e., photoaging) is caused by cumulative damage after the skin is irradiated with ultraviolet rays in the sun for a long time.
Therefore, the prevention and repair of ultraviolet-induced skin damage is an effective means for protecting skin and delaying aging. There is a need in the art to provide further products having the effect of preventing and/or repairing uv-induced skin damage.
Disclosure of Invention
The invention aims to provide a tonaliella extract which has the effect of preventing and/or repairing ultraviolet-induced skin damage.
In a first aspect of the invention, there is provided the use of an extract of tnatopia maternita for the preparation of a pharmaceutical or cosmetic composition for the prevention and/or repair of uv-induced skin damage.
In another preferred embodiment, the uv-induced skin damage is selected from the group consisting of: sunburn (e.g., ultraviolet-induced erythema), sunburn (e.g., pigmentation), photoaging, phototoxic and photosensitizing reactions, skin tumors (e.g., squamous carcinoma of the skin, basal cell carcinoma), or combinations thereof.
In another preferred embodiment, the ultraviolet light is selected from the group consisting of: UVA (320-400 nm), UVB (290-320 nm), or a combination thereof.
In another preferred example, the sunburn includes: erythema, DNA damage, uv-induced epidermal cell death, skin surface roughness, increased keratinization, impaired skin barrier, or a combination thereof.
In another preferred example, the photo-aging includes: wrinkles, skin laxity, telangiectasia, uneven skin pigmentation, disruption of collagen and/or elastic fibers.
In another preferred example, the uv-induced skin damage comprises: epidermal layer damage (e.g., stratum corneum) and dermal layer damage.
In another preferred embodiment, the pharmaceutical or cosmetic composition is used for sun protection and/or anti-photoaging, anti-oxidation.
In another preferred embodiment, the turner grass extract is an extract of the branches, leaves, roots, flowers and/or seeds of a turner grass plant.
In another preferred embodiment, the tnaline grass extract comprises an alcohol extract, an alcohol-water solvent extract, an organic solvent extract, or a combination thereof, preferably a methanol extract.
In another preferred embodiment, the turner grass extract is an extract of leaves.
In another preferred embodiment, the extract contains one or more components selected from the group consisting of: luteolin, apigenin, vitexin, and their glycosides, arbutin, echinocandin, pinocembrin, or their combination.
In another preferred embodiment, the extract is obtained by solvent extraction and/or chromatography.
In another preferred embodiment, the extractant of the extract is selected from the group consisting of: alcohols (preferably C1-C4 alcohols such as methanol, ethanol, propanol), aqueous alcohol solvents (such as alcohol content of 40-80wt% of the total solvent weight), ethyl acetate, n-hexane, or mixtures thereof.
In another preferred example, the extract is obtained by the following extraction method, and the method comprises the following steps:
(a) Leaching the Terna herb with 80-100wt% alcohol water solvent for 1-5 times, mixing extractive solutions, and concentrating to obtain crude extract; and
(b) Separating chloroform-soluble part of the crude extract on silica gel column chromatography, sequentially washing with dichloromethane and eluting with ethyl acetate, and collecting ethyl acetate part to obtain herba Trifolii Pratentis extract.
In another preferred embodiment, in step (a), the alcohol-water solvent contains alcohol in an amount of 90 to 100wt%, such as 95wt%, based on the total solvent.
In another preferred example, in step (a), in the alcohol-water solvent, the alcohol is a C1-C4 alcohol, such as methanol, ethanol, propanol, isopropanol, or a combination thereof, and more preferably, methanol.
In another preferred embodiment, in step (a), the leaching has one or more of the following characteristics:
(1) The leaching times are 1, 2, 3 or 4;
(2) The leaching temperature is 10-40 deg.C, preferably 20-30 deg.C;
(3) The time of each leaching is 1-3h; and/or
(4) The dosage ratio of the Terna herb and the alcohol-water solvent is 50-300g/L, preferably 100-250g/L, such as 200g/L.
In another preferred example, in step (b), the separation has one or more of the following characteristics:
(1) The average particle size of the silica gel column is 100-400 meshes, preferably 200-300 meshes;
(2) The ratio of the column diameter to the height of the resin is 1:10;
(3) The washing volume of dichloromethane is 1 to 3 times of column volume, preferably 1.5 to 2.5 times of column volume; and/or
(4) The elution volume of ethyl acetate is 1 to 3 column volumes, preferably 1.5 to 2.5 column volumes.
In another preferred example, the ethyl acetate fraction may further include a step of passing through a steaming agent.
In another preferred embodiment, the pharmaceutical or cosmetic composition further comprises an additional component selected from the group consisting of: a whitening or spot-removing component, an anti-inflammatory component, an antioxidant component, an anti-ultraviolet component, or a combination thereof.
In another preferred embodiment, the cosmetically acceptable carrier or excipient is selected from the group consisting of: humectant, antioxidant, anti-ultraviolet agent, antiseptic, film forming agent, oil soluble gelling agent, organic modified clay mineral, resin, antibacterial agent, essence, salt, pH regulator, chelating agent, algefacient, antiinflammatory agent, skin beautifying component, vitamin, amino acid, nucleic acid, hormone, inclusion compound or their combination.
In another preferred embodiment, the pharmaceutical composition comprises: an ointment, cream, gel or paste.
In another preferred embodiment, the cosmetic composition is in the form of a solid dosage form, a semi-solid dosage form, or a liquid dosage form, such as a solution, gel, cream, lotion, spray, ointment, cream, paste, cake, powder, patch, and the like.
In a second aspect of the present invention, there is provided a cosmetic composition comprising: an extract of the plant of the species Terna, and a cosmetically acceptable carrier.
In another preferred embodiment, the content of the tner grass extract (dried) in the cosmetic composition is 0.0001 to 15wt%, preferably 0.0001 to 10wt% or 0.001 to 5wt%, such as 0.01wt%, 0.1wt%, 0.2wt%, 0.5wt%, 1wt%, 2wt% or 4wt%, based on the total weight of the cosmetic composition.
In another preferred embodiment, the cosmetic composition is in the form of a solid, semi-solid, or liquid dosage form, such as a solution, gel, emulsion, spray, ointment, cream (e.g., sunscreen), paste, cake, powder, patch, and the like.
In a third aspect of the present invention, there is provided a method for preventing and/or repairing uv-induced skin damage comprising the steps of: administering to the skin of a subject in need thereof an effective amount of a tnalina grass extract for topical application, thereby preventing and/or repairing uv-induced skin damage.
In another preferred embodiment, the subject is a human, especially a human population of sensitive muscles, such as a human whose skin is sensitive to light.
It is to be understood that within the scope of the present invention, the above-described features of the present invention and those specifically described below (e.g., in the examples) may be combined with each other to form new or preferred embodiments. Not to be reiterated herein, but to the extent of space.
Drawings
FIG. 1 shows the UV protection of cells by different treatments. Each value is the mean ± standard error from three independent experiments,. + -. P <0.0001,. + -. P <0.01.
FIG. 2. Effect of different treatments on the level of oxidative stress of cells. Each value is from the mean ± sem of three independent experiments, # p <0.001, # p <0.01.
Detailed Description
The inventor of the invention has conducted extensive and intensive studies, and through a large number of screening and tests, the application of the trinexandrium chinense extract in preventing and/or repairing ultraviolet-induced skin damage is discovered for the first time. Experiments prove that the skin keratinocytes treated by the trinexapac-ethyl extract can effectively resist ultraviolet injury, so that the skin keratinocytes can be used for preventing and/or repairing ultraviolet-induced skin injury. The present invention has been completed based on this finding.
Term(s) for
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.
As used herein, the term "about" when used in reference to a specifically recited value means that the value may vary by no more than 1% from the recited value. For example, as used herein, the expression "about 100" includes 99 and 101 and all values in between (e.g., 99.1, 99.2, 99.3, 99.4, etc.).
As used herein, the term "comprising" or "includes" can be open, semi-closed, and closed. In other words, the term also includes "consisting essentially of or" consisting of 823030A ".
As used herein, the term "room temperature" or "ambient temperature" means a temperature of 4-40 ℃, preferably, 25 ± 5 ℃.
All-grass of Terna
Turnera diffusa (Turnera diffusa) belongs to the family passionfaceae and is a shrub native to southern texas, central america, mexico, south america and the caribbean sea, usa. The leaf of Terna comprises volatile oil, and mainly contains caryophyllene, caryophyllene oxide, juniperberry terpene, elemene and eucalyptol. The non-volatile components mainly comprise flavone compounds, including luteolin, apigenin, vitexin, and their glycosides, and arbutin, echinacolin, etc. Terna is considered to be useful as a yang-strengthening agent and tonic, and its irritant and tonic properties also contribute to the treatment of mild depression; it also has a strong aroma and a slightly bitter taste, and its leaves are used in mexico as a substitute for tea leaves and as a flavor for liqueurs.
Ultraviolet induced skin damage
According to the wavelength, the sunlight can be divided into three parts of visible light, ultraviolet light and infrared light, and the ultraviolet light can be divided into three parts of long wave (UVA), medium wave (UVB) and short wave (UVC). The long wave ultraviolet wavelength is 320-400nm, the medium wave ultraviolet wavelength is 280-320nm, and the short wave ultraviolet wavelength is 100-280nm. The main harm to skin is medium-wave ultraviolet rays and long-wave ultraviolet rays, which can cause a great amount of death of epidermal cells, lead to rough skin surface and increase of cutin, and are easy to accelerate skin aging, generate wrinkles, sunburn and the like, and bring a series of skin problems. Wherein, the medium-wave ultraviolet ray can cause sunburn or suntan of the skin, but can be repaired after nursing; the long-wave ultraviolet rays can directly reach the dermis layer of the skin, collagen and elastic fibers in the skin are damaged, the aging of the skin is accelerated, the damage is irreversible, the skin becomes fragile and fragile after the skin is accumulated for a long time, and skin problems can also appear on the epidermis layer of the skin.
In another preferred embodiment, the uv-induced skin damage is selected from the group consisting of: sunburn (e.g., ultraviolet-induced erythema), sunburn (e.g., pigmentation), photoaging, phototoxic and photosensitizing reactions, skin tumors (e.g., squamous carcinoma of the skin, basal cell carcinoma), or combinations thereof. (see Typhaea minor, et al, damage to skin and protection from ultraviolet light, proceedings of Ningxia medical college, vol 25, no. 4, 8 months 2003)
Extract and preparation method thereof
The invention provides a tenala extract. The extract can be used as an active ingredient for preventing and/or repairing ultraviolet-induced skin damage.
In a preferred embodiment of the invention, the preparation of the active ingredient is carried out by solvent extraction, supercritical extraction and/or chromatography.
In the present invention, the number of extraction times may be one or more (e.g., 2 times, 3 times, 4 times, 5 times). When the extraction is carried out for multiple times, the extracts obtained from multiple times can be combined for subsequent treatment.
In the present invention, as for the solvent extraction method, the solvent used is not particularly limited, and representative examples include (but are not limited to): one or a mixture of water, ethanol, methanol and alcohol-water (mixture) solvent.
In the present invention, the alcohol-water solvent refers to a mixed solvent of water and other solvents such as alcohols (especially C1-C4 alcohols such as methanol, ethanol, propanol). Generally, the alcohol is present in an amount of 40 to 100wt%, preferably 60 to 95wt%, such as 80wt%, 90wt%, 98wt%, etc., based on the total weight of the alcohol-water solvent.
In the present invention, as for the solvent extraction method, the solvent used therefor is not particularly limited, and representative examples include (but are not limited to): one or more of n-butyl alcohol, dichloromethane, chloroform, C5-C7 alkane, cyclohexane and petroleum ether. The number of extractions may be one or more.
In the present invention, for the supercritical extraction, the solvent used is not particularly limited, and representative examples include (but are not limited to): carbon dioxide or propane.
In the present invention, as for column chromatography, column chromatography thereof is not particularly limited, and representative examples include (but are not limited to): one or more of activated carbon, silica gel, reversed phase silica gel, macroporous resin and sephadex.
In a preferred embodiment of the present invention, the preparation method of the tner grass extract comprises: extracting herba Trifolii Pratentis (dried leaf) with sufficient amount of alcohol (such as 1-20 times of water) or alcohol-water mixture solvent (50-100% alcohol, preferably 80-100% methanol) for 1-5 times to obtain alcoholic extract. In the present invention, the fat-soluble component can be further obtained by column separation (silica gel column) from the above alcohol extract to obtain a tnaline grass extract having a higher content (or relative content) of the fat-soluble component, or a tnaline grass extract mainly containing the fat-soluble component.
Composition and application
As used herein, the term "pharmaceutical or cosmetic composition" includes (a) an extract of tnalinia according to the invention; and (b) a pharmaceutically or cosmetically acceptable carrier.
In addition, the cosmetic composition includes a skin care product.
It is possible to prepare the tnatopsis tetragonoloba extract of the present invention into pharmaceutical compositions such as ointments, creams, gels, pastes, patches, etc. The drug can be prepared by a generally known preparation technique, and a suitable pharmaceutical additive can be added to the drug.
Examples of the pharmaceutical additives include excipients, binders, disintegrating agents, lubricants, flow aids, suspending agents, emulsifiers, stabilizers, warming (wetting) agents, preservatives, solvents, solubilizers, preservatives, flavoring agents, sweeteners, dyes, flavors, propellants and the like, and these pharmaceutical additives may be selected and added in an appropriate amount within a range not affecting the effect of the present invention.
It is possible to prepare the tnatopia mater extract of the present invention into a cosmetic composition, a solid dosage form, a semi-solid dosage form, or a liquid dosage form such as a solution, a gel, a cream, an emulsion, a spray, an ointment, a cream, a paste, a cake, a powder, a patch, etc.
Other ingredients used in general cosmetics, for example, film-forming agents, oil-soluble gelling agents, organically modified clay minerals, resins, moisturizing agents, preservatives, antibacterial agents, fragrances, salts, antioxidants, pH adjusting agents, chelating agents, cooling agents, anti-inflammatory agents, skin beautifying ingredients (whitening agents, cell activating agents, skin roughness improving agents, blood circulation promoting agents, skin astringents, anti-seborrheic agents, etc.), vitamins, amino acids, nucleic acids, hormones, inclusion compounds, and the like may be added to the cosmetic of the present invention within a range that does not impair the effects of the present invention.
The oil-soluble gelling agent is selected from metal soaps such as aluminum stearate, magnesium stearate, and zinc myristate; amino acid derivatives such as N-lauroyl-L-glutamic acid, alpha, gamma-di-N-butylamine, and the like; cyclodextrin fatty acid esters such as cyclodextrin palmitate, cyclodextrin stearate, and cyclodextrin 2-ethylhexanoate palmitate; sucrose fatty acid esters such as sucrose palmitate and sucrose stearate; benzylidene derivatives of sorbitol such as monobenzylidene sorbitol and dibenzylidene sorbitol; one or two or more kinds of gelling agents such as an organically modified clay mineral such as dimethylbenzyl dodecylammonium montmorillonite clay or dimethyl octacosyl ammonium montmorillonite clay may be used as required.
The humectant comprises: glycerin, sorbitol, propylene glycol, dipropylene glycol, 1, 3-butylene glycol, glucose, xylitol, maltitol, polyethylene glycol, hyaluronic acid, chondroitin sulfate, pyrrolidone carboxylate, polyoxyethylene methyl glucoside, polyoxypropylene methyl glucoside, and the like.
The antibacterial preservative comprises: alkyl parabens, benzoic acid, sodium benzoate, sorbic acid, potassium sorbate, phenoxyethanol, and the like, and the antibacterial agents are: benzoic acid, salicylic acid, carbolic acid, sorbic acid, alkyl parabens, parachloro-metacresol, hexachlorophene, benzalkonium chloride, chlorhexidine chloride, trichloro-carbanilide, triclosan, a photosensitizer, phenoxyethanol, and the like.
The antioxidant is as follows: tocopherol, butyl hydroxy anisole, dibutyl hydroxy toluene, phytic acid and the like, and the pH regulator comprises: lactic acid, citric acid, glycolic acid, succinic acid, tartaric acid, dl-malic acid, potassium carbonate, sodium bicarbonate, ammonium bicarbonate, and the like, as chelating agents, alanine, sodium ethylenediaminetetraacetate, sodium polyphosphate, sodium metaphosphate, phosphoric acid, and the like, as cooling agents: l-menthol, camphor, etc., and the anti-inflammatory agents include: allantoin, glycyrrhetinic acid, glycyrrhizic acid, tranexamic acid, and Azulene (Azulene).
The skin beautifying components are as follows: whitening agent such as placenta extract, arbutin, glutathione, and herba Saxifragae extract; cell activator such as Lac Regis Apis, photosensitizer, cholesterol derivative, calf blood extractive solution, etc.; an agent for improving rough skin; blood circulation promoters such as valerian nonanoate, benzyl nicotinate, beta-butoxyethyl nicotinate, capsaicin, zingerone, cantharides tincture, ichthammol, caffeine, tannic acid, alpha-borneol, tocopherol nicotinate, inositol hexanicotinate, cyclamate, cinnarizine, tolazoline, acetylcholine, verapamil, cepharanthin, and gamma-oryzanol; skin astringents such as zinc oxide and tannic acid; sulfur, and antilipidemic agents, and the like, and vitamins include: vitamin A oil, rosin oil, acetic acid rosin oil, palmitic acid rosin oil, and the like; vitamin B2 compounds such as riboflavin, riboflavin butyrate, and flavin adenine nucleotide; vitamin B6 such as pyridoxine hydrochloride, pyridoxine dicaprylate, and pyridoxine tripalmitate, vitamin B12 and its derivatives, and vitamin B15 and its derivatives; vitamin C compounds such as L-ascorbic acid, L-ascorbic acid dipalmitate, L-ascorbic acid-2-sodium sulfate, and L-ascorbic acid phosphoric acid diester dipotassium; vitamin D compounds such as ergocalciferol and cholecalciferol; vitamin E compounds such as alpha-tocopherol, beta-tocopherol, gamma-tocopherol, dl-alpha-tocopherol acetate, dl-alpha-tocopherol nicotinate, and dl-alpha-tocopherol succinate; vitamin H; a vitamin P; nicotinic acids such as nicotinic acid, benzyl nicotinate and nicotinamide; pantothenic acids such as calcium pantothenate, D-panthenol, panthenyl ethyl ether, and acetyl panthenyl ethyl ether; biotin, and the like.
The amino acids are: glycine, valine, leucine, isoleucine, serine, threonine, phenylalanine, arginine, lysine, aspartic acid, glutamic acid, cystine, cysteine, methionine, tryptophan, etc., nucleic acids include deoxyribonucleic acid, etc., and hormones include estradiol, vinylestradiol, etc.
Preferred examples of the cosmetic of the present invention include: skin care cosmetics, makeup cosmetics, and ultraviolet-shielding cosmetics. Such as basic cosmetics such as milky lotion, cream, lotion, sunscreen agent, mask material, face toilet, essence, etc.; makeup cosmetics such as foundation, powdery skin, blush, etc.
The form of the product is not particularly limited, and may be liquid, emulsion, cream, solid, paste, gel, powder, multi-layer, mousse (mousse), spray, or the like.
The present invention also provides a skin care method, comprising the steps of: administering said tnnisetum sinese extract of the invention or the composition of the invention to a subject in need thereof. Generally, the method is a method for preventing and/or repairing uv-induced skin damage.
In another preferred embodiment, the effective concentration of the Terna extract ranges from 1. Mu.g/ml to 500mg/ml, such as 5. Mu.g/ml, 10. Mu.g/ml, 20. Mu.g/ml, 50. Mu.g/ml, 100. Mu.g/ml or 200. Mu.g/ml.
As used herein, the term "effective dose" refers to any amount that, when used alone or in combination with another therapeutic agent or cosmetic, promotes disease regression as manifested by a decrease in the severity of disease symptoms, an increase in the frequency and duration of disease symptom-free periods, or prevention of a disorder or disability resulting from the disease. A "therapeutically effective dose" of a drug of the invention also includes a "prophylactically effective dose," which is any amount of a drug that, when administered alone or in combination with another therapeutic agent or cosmetic, inhibits the occurrence or recurrence of a disease or symptom in a subject at risk of developing the disease or symptom or suffering from the recurrence of the disease or symptom.
In the present invention, the use of sunburn, sunburn (e.g., ultraviolet-induced erythema), sunburn (e.g., pigmentation), photoaging, phototoxic and photosensitizing reactions, skin tumors, and the like, includes both prophylactic and post-hoc applications. For example, for sunburn, the application of the tnnisetum extract or composition of the invention before, during, and/or after sunburn is included to prevent and/or repair sunburn.
The main advantages of the invention include:
(1) The invention discovers for the first time that the extract of the tonaliella is capable of preventing and/or repairing skin damage induced by ultraviolet, and is very suitable to be used as an active ingredient of cosmetics, especially sunscreen products.
(2) The herba te nandinae extract can obviously prevent and/or repair ultraviolet-induced skin injury at low concentration, has high biological activity, and can exert the activity by adding a small amount.
The invention is further illustrated with reference to specific embodiments. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. Experimental procedures without specific conditions noted in the following examples, generally following conventional conditions, such as Sambrook et al, molecular cloning: conditions described in a Laboratory Manual (New York: cold Spring Harbor Laboratory Press, 1989), or according to the manufacturer's recommendations. Unless otherwise indicated, percentages and parts are by weight.
Example 1
1. Preparation of plant extract
Dried and chopped leaves of tnatopia mater (90 g) were soaked in methanol (400 mL) at room temperature and extracted repeatedly at room temperature for 1h three times. Mixing the three methanol lixiviums, and evaporating under reduced pressure to obtain a crude methanol extract of the turnabout grass. Subjecting chloroform-soluble fraction of the crude methanol extract to column chromatography on silica gel column (60 mmx600mm column chromatography, 200-300 mesh silica gel), sequentially separating with 2 times column volume of dichloromethane, ethyl acetate: methanol (1), and methanol to give four different fractions, the experimental extract being the second fraction, namely the ethyl acetate fraction.
2. Cell culture, UVA irradiation and extract treatment
Human primary keratinocytes were purchased from ATCG, inc. (Rockville, md., USA). Only the keratinocytes of the 1 st to 5 th passages were selected, and cultured in a 37 ℃ incubator containing 5% carbon dioxide by adding a keratinocyte growth kit to a dermal cell culture medium. UV irradiation was carried out using a UVA fluorescent lamp (Vilber Lourmat, france) and the total UVA dose used in the experiment was 4.5J/cm 2 . Before UV irradiation, cells were washed with PBS solution and a trace amount of PBS solution was retained to keep the cells moist during the experiment. All cells were treated identically throughout the experiment. After the UVA irradiation was completed, the remaining PBS solution was removed, the previously prepared culture solution containing the extract of Terna grass (5. Mu.g/mL, based on the dry weight of the extract) was added to the experimental group, the same amount of the culture solution containing no plant extract component was added to the UV control group, and the cells were returned to the incubator to continue the culture.
3. Cell activity detection method
And after the cells are continuously cultured for 24 hours, the cells added with the corresponding culture solution after the ultraviolet irradiation are subjected to cell activity detection by using a CCK-8 kit. The cells were washed three times with a PBS solution, followed by addition of a culture solution to which a 10-percent CCK-8 solution was added, and after incubating the cells in a thermostat for 2 hours, they were detected with a microplate reader and absorbance was measured at 450 nm.
The experimental results are shown in fig. 1, and the treatment of human primary keratinocytes with a turner grass extract after ultraviolet irradiation can effectively improve the cell activity. This suggests that the extract of Terna has an ultraviolet protective effect on skin cells and a repairing effect on ultraviolet damage.
4. Cellular oxidative stress (ROS) measurement
The method measures intracellular ROS levels by DCFA and corrects by cellular activity to accurately assess the damage caused by Ultraviolet (UVA) radiation to cells. Human primary keratinocytes were cultured at a rate of 1.5X 10 4 The density of individual/well was seeded in 96-well plates. After 24 hours of inoculation, the cell culture medium of the experimental group was changed to the previously prepared culture medium containing the extract of Terna grass (10. Mu.g/mL, dry weight of the extract), and the UV control group and blank control group were changed to the same amount of fresh culture medium without any plant extract. After 24 hours of pretreatment, cells were washed three times with PBS solution and a trace amount of PBS solution was retained so that the cells could be kept wet during the experiment. Using 4.5J/cm 2 After UVA irradiation of the cells, a PBS solution containing 25uM DCFA was added, and the cells were placed back in the incubator for 30 minutes and subjected to fluorescence measurement using an excitation wavelength of 485nm and an emission wavelength of 535nm.
5. Cellular oxidative stress (ROS) level measurement
Experimental results show that ultraviolet radiation is carried out after the Terna extract is used for pretreating human primary keratinocytes, the increase of intracellular oxidative stress level caused by ultraviolet radiation can be effectively inhibited, and the Terna extract has obvious antioxidation and anti-aging effects.
All documents referred to herein are incorporated by reference into this application as if each were individually incorporated by reference. Furthermore, it should be understood that various changes and modifications of the present invention can be made by those skilled in the art after reading the above teachings of the present invention, and these equivalents also fall within the scope of the present invention as defined by the appended claims.

Claims (10)

1. Use of a trinexapac-ethyl extract for the preparation of a pharmaceutical or cosmetic composition for the prevention and/or repair of uv-induced skin damage.
2. The use of claim 1, wherein the uv-induced skin damage is selected from the group consisting of: sunburn (e.g., ultraviolet-induced erythema), sunburn (e.g., pigmentation), photoaging, phototoxic and photosensitizing reactions, skin tumors (e.g., squamous carcinoma of the skin, basal cell carcinoma), or combinations thereof.
3. The use of claim 1, wherein the sunburn comprises: erythema, DNA damage, uv-induced epidermal cell death, skin surface roughness, increased keratinization, impaired skin barrier, or a combination thereof.
4. The use of claim 1, wherein the turnabout extract is a leaf extract.
5. The use as claimed in claim 1, wherein said extract is obtained by an extraction process comprising the steps of:
(a) Leaching the Terna herb with 80-100wt% alcohol water solvent for 1-5 times, mixing extractive solutions, and concentrating to obtain crude extract; and
(b) Separating chloroform-soluble part of the crude extract on silica gel column chromatography, sequentially washing with dichloromethane and eluting with ethyl acetate, and collecting ethyl acetate part to obtain herba Trifolii Pratentis extract.
6. The use according to claim 1, wherein in step (a) the leaching has one or more of the following characteristics:
(1) The leaching times are 1, 2, 3 or 4;
(2) The leaching temperature is 10-40 ℃, preferably 20-30 ℃;
(3) The time of each leaching is 1-3h; and/or
(4) The dosage ratio of the Terna herb and the alcohol-water solvent is 50-300g/L, preferably 100-250g/L, such as 200g/L.
7. The use of claim 1, wherein in step (b), the separation has one or more of the following characteristics:
(1) The average particle size of the silica gel column is 100-400 meshes, preferably 200-300 meshes;
(2) The ratio of the column diameter to the height of the resin is 1:10;
(3) The washing volume of dichloromethane is 1 to 3 times of the column volume, preferably 1.5 to 2.5 times of the column volume; and/or
(4) The elution volume of ethyl acetate is 1 to 3 column volumes, preferably 1.5 to 2.5 column volumes.
8. Use according to claim 1, wherein the cosmetic composition is in the form of a solid, semi-solid, or liquid dosage form, such as a solution, gel, emulsion, spray, ointment, cream, paste, cake, powder, patch, and the like.
9. A cosmetic composition, comprising: an extract of the plant Terna (T. Nala), and a cosmetically acceptable carrier.
10. Use according to claim 1, wherein the cosmetic composition is in the form of a solid, semi-solid, or liquid dosage form, such as a solution, gel, emulsion, spray, ointment, cream (e.g. sun cream), paste, cake, powder, patch, etc.
CN202110558860.7A 2021-05-21 2021-05-21 Application of herba Terminaliae extract in preventing and/or repairing ultraviolet induced skin injury Pending CN115364003A (en)

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