CN114983923A - A topical composition for resisting endogenous and exogenous aging - Google Patents
A topical composition for resisting endogenous and exogenous aging Download PDFInfo
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- CN114983923A CN114983923A CN202210819003.2A CN202210819003A CN114983923A CN 114983923 A CN114983923 A CN 114983923A CN 202210819003 A CN202210819003 A CN 202210819003A CN 114983923 A CN114983923 A CN 114983923A
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- aging
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- fermentation product
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- 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
- A61K8/99—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from microorganisms other than algae or fungi, e.g. protozoa or bacteria
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- 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
- A61K8/9728—Fungi, e.g. yeasts
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- A—HUMAN NECESSITIES
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- A61Q19/08—Anti-ageing preparations
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
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- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
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- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/85—Products or compounds obtained by fermentation, e.g. yoghurt, beer, wine
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Abstract
The invention discloses a skin external composition capable of simultaneously resisting endogenous and exogenous aging and a preparation method thereof. The skin external composition comprises the following raw material components: 0.1-30% of anti-aging active substance, 0.1-15% of humectant, 0.1-15% of emollient, 0.1-12% of emulsifier, 0.1-12% of filler, 0.1-12% of thickener, 0.01-5% of preservative, 0.01-1% of pH regulator, 0.01-1% of daily essence and the balance of deionized water. The main components of the anti-aging active comprise: mirabilis jalapa active plant cell, yeast/rice fermentation product filtrate and split yeast fermentation product lysate. The components of the anti-aging active substance in the formula are used simultaneously, so that the anti-aging active substance has a synergistic anti-aging effect, can resist endogenous aging, and can repair skin damage caused by exogenous aging.
Description
Technical Field
The invention belongs to the technical field of daily chemicals, and relates to a skin external composition capable of resisting endogenous and exogenous aging simultaneously and a preparation method thereof.
Background
For skin aging, there are various theories regarding skin aging, such as genetics, free radical theories, nonenzymatic glycosylation theories, and the like. In general, there are two main causes of skin aging, namely, intrinsic aging and extrinsic aging.
The intrinsic aging is generally determined by genetic factors and is an irreversible process of natural aging of our skin. With age, DNA in the body is gradually damaged to cause aging and damage of cells, when mitochondrial dysfunction occurs in cells, ATP supply is reduced, and when energy is lost in cell activities, problems occur in all physiological activities, such as proliferation and differentiation of skin cells, and reduction of levels of collagen, elastin and other proteins, and finally, the structure of the skin is degraded to form aged wrinkles. Exogenous aging is mainly caused by environmental factors and life style, such as ultraviolet irradiation, PM2.5, staying up all night, eating habits, psychological stress and other problems, wherein the longest is ultraviolet irradiation, the phenomenon of skin aging occurs due to the reduction of skin collagen fibers and elastic fibers caused by long-term ultraviolet irradiation, and when the skin loses elasticity and retractive force, the skin becomes more and more lax.
Because skin aging is caused by the combined action of endogenous factors and exogenous factors, it is generally necessary to pay attention to both aspects of skin care in order to delay the aging process.
For example, chinese patent CN113244156B discloses an anti-aging composition, which comprises the following components in percentage by weight: 0.01-0.07% of humus extract, 0.001-0.05% of polyphenol antioxidant, 0.01-0.8% of mussel extract, 0.05-2% of sodium hyaluronate, 0.05-2% of sodium polyglutamate and the balance of deionized water. The patent achieves the anti-aging effect through the following four aspects: (1) the generation of skin Reactive Oxygen Species (ROS) is reduced through effective shielding of ultraviolet rays and high-energy blue light; (2) the generated excessive free radicals are effectively eliminated; (3) performing anti-inflammation repair on the damaged skin; (4) the water and nutrition are supplemented in time, and the composition is reasonably designed, so that the components can fully play the self-action, the synergistic interaction effect can be generated, and the effective anti-aging effect is realized. Chinese patent CN112932988B discloses an anti-aging composition containing polydeoxyribonucleic acid, which comprises honey and polydeoxyribonucleic acid; the disclosed anti-aging composition and skin care products including the same enhance skin cell regeneration and anti-aging effects through the synergistic effect of honey and polydeoxyribonucleic acid. Chinese patent CN108392436B discloses a skin care composition for whitening skin and resisting aging, which comprises the following components in percentage by weight: 10-35% of butanediol, 0.1-5% of glucan, 0.1-50% of lotus leaf flavone, 0.1-5% of nicotinamide and the balance of distilled water. The contents of heavy metals and microorganisms are low, and the whitening cream has remarkable whitening effect, obvious anti-aging performance and capability of removing DPPH free radicals. Chinese patent CN113893203A discloses an external skin composition containing anti-aging liposome and a preparation method thereof. The skin external composition comprises the following raw material components: 0.1-15% of anti-aging efficacy liposome, 1-30% of humectant, 1-22.5% of emollient, 1-21% of emulsifier, 0.1-10% of filler, 1-12.5% of thickener, 0.1-3.5% of preservative, 0.01-1% of pH regulator, 0.01-1% of daily essence and the balance of deionized water. The anti-aging efficacy liposome comprises the following main components: hexapeptide-11, Dunaliella salina extract, Epilobium freudenrich extract, and Gynostemma pentaphyllum extract. The patent can increase the synthesis of skin collagen, enhance the activity of superoxide dismutase, clear excessive free radicals in the skin, relieve the peroxidation damage of tissues or cells, enhance the skin barrier and strengthen the skin structure to resist skin aging and relaxation by compounding the components at the same time.
Therefore, the related technologies of the current anti-aging skin care products are various, and various active ingredients are randomly matched, so that how to compound an effective composition capable of resisting endogenous and exogenous aging simultaneously is a key and focus content of research in the cosmetics industry all the time.
Disclosure of Invention
The invention aims to provide a novel skin external composition capable of resisting endogenous and exogenous aging simultaneously and a preparation method thereof.
The purpose of the invention can be realized by the following technical scheme:
in order to achieve the purpose, the invention adopts the following technical scheme:
the invention firstly provides an anti-aging active composition, which comprises the following components: the culture medium comprises Mirabilis jalapa active plant cells, yeast/rice fermentation product filtrate and split yeast fermentation product lysate, wherein the mass ratio of the Mirabilis jalapa active plant cells to the yeast/rice fermentation product filtrate to the split yeast fermentation product lysate is (0.5-5): (1-10): (0.5 to 5), preferably 1: (1-4): 1.
in one embodiment of the invention, the mirabilis jalapa active plant cell is prepared by the following preparation method:
1) carrying out surface disinfection on the Mirabilis jalapa leaves, crushing, and culturing in a solid culture medium to obtain leaf callus;
2) separating the leaf callus from the Mirabilis jalapa when the leaf callus grows to the diameter of 1-3 cm, carrying out suspension culture on the leaf callus, placing the leaf callus in continuous fluorescent lamp irradiation, and placing the leaf callus in a shaking table at 24-26 ℃ for subculture to obtain a subculture suspension culture;
3) and separating the subculture suspension, removing tissue masses, dispersing the filtered cell sap in a fresh culture medium, and continuously culturing to obtain the mirabilis jalapa active plant cell.
In one embodiment of the present invention, the composition of the solid medium in step 1) is: adding 2mg/mL NAA, 1mg/mL 6-BA and 1mg/mL 2,4-D into the common MS culture medium.
In one embodiment of the present invention, the culture medium used for suspension culture in step 2) is: sterilized MS medium, and adding 2mg/mL NAA, 1mg/mL 6-BA, 1mg/mL 2, 4-D.
In one embodiment of the invention, the intensity of the continuous fluorescent lamp irradiation in step 2) is 4000-.
In one embodiment of the present invention, the specific method for performing suspension culture in step 2) is: taking out callus (the callus is soft and loose in structure), placing into a bottle, slightly crushing, and shake culturing in a shaking table at 24-26 deg.C for sterilized MS culture at 120 rpm.
In one embodiment of the present invention, the subculture in step 2) means passaging every 7 d.
In one embodiment of the invention, the fresh medium in step 3) is: sterilized MS culture medium, and adding 2mg/mL NAA, 1mg/mL 6-BA, 1mg/mL 2, 4-D.
In one embodiment of the invention, the operation of step 3) is mainly for the filtration and expansion of the cells: the suspension culture obtained at the time of passage was filtered from the large tissue mass with a 50-mesh stainless steel sieve, and the filtered cell suspension was dispersed in a fresh medium to continue the culture.
In one embodiment of the invention, the yeast/rice fermentation product filtrate is a relatively conventional additive substance in the field of cosmetics, and can be obtained by a conventional preparation method or directly purchased.
In one embodiment of the invention, the split yeast fermentation product lysate is a bifidobacterium extract containing metabolites, cytoplasmic components, cell wall components and polysaccharide complexes.
The invention also provides a skin external composition capable of resisting endogenous and exogenous aging at the same time, which comprises the anti-aging active composition and also comprises an auxiliary agent acceptable in the skin external product.
In one embodiment of the present invention, the present invention specifically provides a skin external composition capable of simultaneously resisting endogenous and exogenous aging, which comprises the following components by weight:
0.1-30% of anti-aging active substance, 0.1-15% of humectant, 0.1-15% of emollient, 0.1-12% of emulsifier, 0.1-12% of filler, 0.1-12% of thickener, 0.01-5% of preservative, 0.01-1% of pH regulator, 0.01-1% of daily essence and the balance of deionized water, wherein the anti-aging active substance is the anti-aging active composition.
Preferably, the external composition for skin capable of simultaneously resisting endogenous and exogenous aging consists of the following components in percentage by weight of the total composition: 0.1-8% of Mirabilis jalapa active plant cells, 0.1-14% of yeast/rice fermentation product filtrate, 0.1-8% of schizosaccharomyces cerevisiae fermentation product lysate, 0.1-15% of humectant, 0.1-15% of emollient, 0.1-12% of emulsifier, 0.1-12% of filler, 0.1-12% of thickener, 0.01-5% of preservative, 0.01-1% of pH regulator, 0.01-1% of daily essence, and the balance of deionized water.
Preferably, the external composition for skin capable of simultaneously resisting endogenous and exogenous aging consists of the following components in percentage by weight of the total composition:
0.1-20% of anti-aging active substance, 0.1-10% of humectant, 1-15% of emollient, 1-15% of emulsifier, 0.1-5% of filler, 0.1-8% of thickener, 0.01-3% of preservative, 0.01-0.8% of pH regulator, 0.01-0.8% of daily essence and the balance of deionized water, wherein the anti-aging active substance is the anti-aging active composition.
Preferably, the external composition for skin capable of simultaneously resisting endogenous and exogenous aging consists of the following components in percentage by weight of the total composition:
0.1-5% of Mirabilis jalapa active plant cells, 0.1-10% of yeast/rice fermentation product filtrate, 0.1-5% of schizosaccharomyces cerevisiae fermentation product lysate, 0.1-10% of humectant, 1-15% of emollient, 1-15% of emulsifier, 0.1-5% of filler, 0.1-8% of thickener, 0.01-3% of preservative, 0.01-0.8% of pH regulator, 0.01-0.8% of daily essence, and the balance of deionized water. Preferably, the skin external composition capable of simultaneously resisting endogenous and exogenous aging consists of the following components in percentage by weight of the total composition:
0.1-15% of an anti-aging active comprising: 0.1-3% Mirabilis jalapa active plant cells, 0.1-8% yeast/rice fermentation product filtrate, and 0.1-3% yeast fermentation product lysate;
0.1-8% of humectant, 2-15% of emollient, 2-15% of emulsifier, 0.1-2% of filler, 0.1-5% of thickener, 0.01-2% of preservative, 0.01-0.5% of pH regulator, 0.01-0.4% of daily essence and the balance of deionized water.
Preferably, the skin external composition capable of simultaneously resisting endogenous and exogenous aging comprises the following moisturizers: one or more of glycerol, butanediol, propylene glycol, pentanediol, phytosterol, dipropylene glycol, hexanediol, polyethylene glycol, panthenol, xylitol, sodium hyaluronate, glyceryl polyether, methyl glucitol polyether, and the like.
The emollient is selected from: one or more of methyl glucose sesquistearate, squalane, beeswax, myristyl alcohol, cetostearyl alcohol, caprylic/capric triglyceride, shea butter, hydrogenated castor oil, jojoba oil, canola oil, olive oil, sunflower seed oil, isohexadecane, etc.;
the emulsifier is selected from: one or more of glyceryl stearate, PEG-100 stearate, PEG400 dilaurate, PEG-400 monolaurate, PEG-12 lauryl alcohol ether, PEG20 sorbitan monostearate, cetearyl glucoside, etc.;
the filler is selected from: one or more of polymethylsilsesquioxane, kaolin, silica powder, aluminum octyl starch succinate, talc powder and the like;
the thickening agent is selected from: one or more of carbomer, polyethylene, sodium polyacrylate, sodium acrylate/sodium acryloyldimethyl taurate copolymer, acrylic acid (ester)/C10-30 alkanol acrylate cross-linked polymer and the like;
the preservative is selected from: one or more of phenoxyethanol, ethylhexyl glycerol, p-hydroxyacetophenone, sodium benzoate and potassium sorbate;
the pH regulator is one or more of lactic acid, arginine and sodium hydroxide.
Preferably, the preparation method of the skin external composition containing the anti-aging liposome comprises the following steps:
a method for preparing a composition for external application on the skin capable of simultaneously resisting endogenous and exogenous aging comprises the following steps:
1) stirring the humectant and deionized water, heating to 70-80 deg.C, keeping the temperature, and stirring for 10-30 min to obtain water phase material.
2) Dissolving emollient and emulsifier, stirring, dispersing, heating to 70-80 deg.C, and dissolving completely to obtain oil phase material.
3) Slowly adding oil phase raw materials into the water phase material under the homogenization state of 3000-.
4) Stirring the mixed solution in the step 3), cooling to 40-50 ℃, adding the anti-aging active substance and essence, and homogenizing at 1500-;
5) stirring and stirring at the speed of 200-300r/min and cooling to 30-40 ℃ after homogenizing is finished, thus obtaining the skin external composition for simultaneously resisting endogenous and exogenous aging.
The invention also provides the application of the anti-aging active composition capable of resisting endogenous and exogenous aging simultaneously, and the anti-aging active composition capable of resisting endogenous and exogenous aging simultaneously is used for preparing cosmetics.
In one embodiment of the present invention, the cosmetic comprises the form of a cream, eye cream, lotion, essence, or the like.
The invention discloses a skin external composition capable of simultaneously resisting endogenous and exogenous aging. The main anti-aging active ingredients of the skin external composition capable of simultaneously resisting endogenous and exogenous aging comprise: mirabilis jalapa active plant cells, yeast/rice fermentation filtrate, split yeast fermentation lysate, by preferred combination and efficacy testing, the most preferred composition ratios were found to achieve synergistic anti-senescence repair efficacy.
(1) Mirabilis jalapa active plant cell: mirabilis jalapa is a traditional drug widely used in many parts of the world for treating various diseases. Due to the development of plant tissue culture technology, the market begins to develop new mirabilis jalapa active ingredients, which are derived from the culture of dedifferentiated plant cells, and utilize leaves, stems or roots thereof to culture a large amount of bioactive compounds with sustainable sources. Mirabilis jalapa L.active plant cells cultured from plant cells are proved to enhance the malic acid-aspartic acid shuttle activity, so that the energy supply function of mitochondria is enhanced, and therefore, the Mirabilis jalapa L.active plant cells can display strong anti-aging capability by enhancing the cell metabolic process required by healthy cells for physiological activities, so that the skin becomes firmer and younger.
(2) Yeast/rice fermentation product filtrate: the yeast/rice fermentation product filtrate contains more than 90 nutrient components, the types comprise fruit acid, sugar, amino acid, polyamine, nucleic acid and the like, and the most important component is rich in alpha-ketoglutaric acid, can directly participate in tricarboxylic acid cycle to enhance cell energy metabolism and provide energy for the physiological activity of cells; at the same time, mRNA expression of protein is accurately regulated and controlled, and the cell structure is strengthened from the gene level, thereby solving the problem of aging.
(3) Split yeast fermentation lysate: is a Bifidobacterium extract containing metabolites, cytoplasmic components, cell wall components and polysaccharide complexes. When the skin receives ultraviolet radiation, the balance between IL-10 and IL-12 is broken, so that the cellular DNA is damaged, the lysate of the yeast fermentation product of the schizosaccharomyces can prevent the release of the immunosuppressant IL-10 and simultaneously prevent the IL-12 from being inhibited, so that the lysate can play a role in promoting the repair mechanism of the cellular DNA, help epidermal cells and dermal cells to resist the harmful effect of the ultraviolet radiation, and keep the skin in a young and active appearance.
By means of the technical scheme, the invention has the following beneficial effects:
1) the inventive anti-aging actives of the present invention mainly comprise: mirabilis jalapa active plant cell, yeast/rice fermentation product filtrate, and split yeast fermentation product lysate. The Mirabilis jalapa active plant cell and yeast/rice fermentation product filtrate can promote the tricarboxylic acid cycle of mitochondria, maintain the energy supply of cells so as to resist endogenous aging, and the schizosaccharomyces fermentation product lysate can help epidermal cells and dermal cells to resist the damage of external ultraviolet rays, and specifically support the self protection and repair mechanism of skin so as to resist skin premature senility.
2) The innovative anti-aging active substance can play a role in synergistic anti-aging by effectively compounding the components, can resist endogenous aging, and can repair skin damage caused by exogenous aging.
Drawings
FIG. 1 is a graph of the change in levels of malate/aspartate shuttle activity (young cells) in the presence of different concentrations of example 1;
FIG. 2 is a graph of the change in levels of malate/aspartate shuttle activity (senescent cells) in the presence of different concentrations of example 1;
FIG. 3 is a graph of the level of Malondialdehyde (MDA) in the presence of different concentrations of example 1.
Detailed Description
The invention discloses a skin external composition capable of simultaneously resisting endogenous and exogenous aging. The main anti-aging active ingredients of the skin external composition comprise: mirabilis jalapa active plant cells, yeast/rice fermentation filtrate, split yeast fermentation lysate, by preferred combination and efficacy testing, the most preferred composition ratios were found to achieve synergistic anti-senescence repair efficacy.
The present invention is illustrated by the following examples, which are provided for the purpose of further illustration only and are not intended to limit the scope of the present invention. Other insubstantial modifications and adaptations of the present invention can be made without departing from the scope of the present invention.
Unless otherwise specified, the following percentages (%) are mass percentages. All the raw material components of the invention are commercial products.
Examples 1 to 3
An external composition for skin capable of simultaneously resisting endogenous and exogenous aging is prepared into an anti-aging cream, and the proportion and the content of the raw material components are shown in the following table 1.
TABLE 1 raw material component ratios (% by mass) of examples 1 to 3
The process for preparing the cream composition of examples 1-3 above, which is capable of simultaneously resisting endogenous and exogenous aging, comprises the following steps:
1) and stirring the humectant and the deionized water together, heating to 80 ℃, and keeping the temperature and stirring for 30 minutes to obtain a water-phase material.
2) And dissolving the emollient and the emulsifier, stirring and dispersing uniformly, and heating to 80 ℃ until the emollient and the emulsifier are completely dissolved to obtain the oil phase material.
3) Slowly adding oil phase raw materials into water phase material under homogenizing state of 4500r/min, continuing homogenizing for 6min, and adding pH regulator, antiseptic, thickener, and filler.
4) Stirring the mixed solution in the step 3), cooling to 45 ℃, adding the anti-aging active substance and the essence, and homogenizing for 5min at 3000 r/min.
5) Stirring and cooling to 37 ℃ at the speed of 200r/min after homogenizing, thus obtaining the cream composition containing the anti-aging active substances.
Test example for Effect verification
1. Safety test
Test samples: example 1;
the test method comprises the following steps: and (4) testing the safety by adopting a third-party detection mechanism. The selection area is not more than 50mm 2 In a suitable patch tester with a depth of about 1mm, the test substance is added in an amount of about 0.02-0.025ml (g) in a closed patch test. The patch tester is attached to the curved side of the forearm of the subject, the subject is removed 24h later, the skin reaction is observed 0.5h, 24h and 48h after removal, and the result is recorded according to the skin reaction grading standard in the technical regulation for cosmetic safety 2015.
TABLE 2 Patch test results
Wherein, the control group is tested by purified water, and the control group has no stimulation or anaphylactic reaction.
Remarking: grading standard of skin reaction
Level 0: negative reaction;
level 1: carrying out suspicious reaction; only faint erythema;
and 2, stage: weak positive reaction (erythema reaction); erythema, infiltration, edema, and possibly papules;
and 3, level: strong positive reaction (herpes reaction); erythema, infiltration, edema, pimples, herpes; the reaction may be beyond the test area;
and 4, stage 4: very strong response (fusional herpes response); erythema, severe infiltration, edema, fusional herpes, and response beyond the test area.
As can be seen from table 2 above, the cream prepared showed no irritation and passed safety. Examples 2 and 3 differ from example 1 only in the amount used, and are generally safe and non-irritating as no other irritating ingredients are added.
2. Stability test
Test samples: example 1;
the test method comprises the following steps: the internal control standard, stability examination of the prepared cosmetic compositions was conducted by placing the compositions in the following conditions, respectively, and CYCLE means that the temperature was cycled once a day from 45 ℃ to-15 ℃ and the appearance color, viscosity, and pH were observed for three months, and the results are shown in table 3.
Table 3 stability test results
As can be seen from Table 3, the prepared cream has no significant changes in color, viscosity and pH value within three months under the conditions of high and low temperatures or illumination, which indicates that the cream product of example 1 has better stability.
3. Efficacy evaluation of anti-aging actives
Test samples: anti-aging actives of example 1
The test method comprises the following steps:
(1) malic acid/aspartic acid shuttle assay
Using an enzymatic method, human keratinocyte primary cultures were obtained from 35-and 62-year-old donors, the human keratinocytes were seeded on a polycarbonate semipermeable membrane and incorporated into a cell culture system, and placed in a standard well plate, which placed the cultures at the gas-liquid interface (medium in well plate is MCDB 153 serum-free medium), and the cells were cultured at the gas-liquid interface for 14 days, with medium changes every two days. The epidermal cells thus formed were used for the study starting from day 14 of culture. The anti-aging active substance in example 1 was co-cultured with human keratinocytes at various concentrations (0.5%, 1%, 2.5% of the anti-aging active substance by MASs of the culture system, respectively) for 24h, and then mitochondria were isolated to determine the activity of malate-aspartate shuttle (MAS). Specifically, 50. mu.l of the mitochondrial suspension was mixed with 2ml of a test solution (300mM mannitol, 10mM potassium phosphate, 10mM Tris buffer, 10mM potassium chloride, 5mM magnesium chloride, 2mM aspartic acid, 2mM ADP, 0.14mM NADH, 3U/ml Malate Dehydrogenase (MDH), 2U/ml aspartate aminotransferase mitochondria) at pH 7.4. The oxidation of NADH was monitored at 37 ℃ for 4 minutes at a constant temperature of 340 nm. 4mM malic acid and 4mM glutamic acid (final concentration) were added to initiate the malate-aspartate shuttle activity. The dose of malate/aspartate shuttle activity was measured at 340nm for 4 minutes.
(2) Assessment of lipid peroxidation by Malondialdehyde (MDA)
After the cell suspension was treated after co-culturing the anti-aging actives of example 1 with human keratinocytes at different concentrations (0.5%, 1%, 2.5%) for 24h, MDA levels were measured using the malondialdehyde content assay kit.
And (3) testing results:
(1) malic acid/aspartic acid shuttle assay: results in young groups of cells (cells from 35 year old donors) the anti-senescence active at concentrations of 0.5%, 1%, 2.5% can increase malate/aspartate shuttle activity by 20%, 23%, 25%, respectively, as shown in figure 1; results in senescent groups of cells (cells from 62 year old donors) the anti-senescence actives increased the malate/aspartate shuttle activity by 18%, 24%, 28% at concentrations of 0.5%, 1%, 2.5%, respectively, as shown in figure 2. In conclusion, the anti-aging active substance can effectively promote the functions of mitochondria, thereby realizing the anti-aging function.
(2) Malondialdehyde (MDA) evaluation of lipid peroxidation degree: the end product of lipid peroxidation in organisms is malondialdehyde, which can cause cross-linking polymerization of life macromolecules such as protein, nucleic acid and the like, and has cytotoxicity, the content of the malondialdehyde is an important parameter reflecting the potential oxidation resistance of the organisms, can reflect the lipid peroxidation rate and strength of the organisms, and can also indirectly reflect the tissue peroxidation damage degree, and the results shown in the result in fig. 3 show that the anti-aging active substance in the example 1 has a remarkable protective effect, and the anti-physiological lipid peroxidation is 24%, 30% and 33% respectively at the concentration of 0.5%, 1.0% and 2.5%.
4. Verification of synergistic effect of skin external composition
Test samples: the experimental group is example 1, and the specific composition of the control group is shown in table 4.
Table 4 raw material composition ratio (% by mass) of example 1 and comparative examples 1 to 7
The preparation process of the cream compositions of the above comparative examples 1 to 7 includes the following steps:
1) and stirring the humectant and the deionized water together, heating to 80 ℃, and keeping the temperature and stirring for 30 minutes to obtain a water-phase material.
2) And dissolving the emollient and the emulsifier, stirring and dispersing uniformly, and heating to 80 ℃ until the emollient and the emulsifier are completely dissolved to obtain the oil phase material.
3) Slowly adding oil phase raw materials into water phase material under homogenizing state of 4500r/min, continuing homogenizing for 6min, and adding pH regulator, antiseptic, thickener, and filler.
4) Stirring the mixed solution in the step 3), cooling to 45 ℃, adding the anti-aging active substance and the essence, and homogenizing at 3000r/min for 5 min.
5) Stirring and cooling to 37 ℃ at the speed of 200r/min after homogenizing, thus obtaining the cream composition containing the anti-aging active substances.
The test method comprises the following steps: after cleaning every day, a proper amount of the experimental group sample is smeared on one random side, and the control group sample is smeared on the other random side. The sunscreen product is applied to the whole face every morning. The results were tested at the following 4 time periods, respectively: before use of the sample (D0), after use of the sample for 14 days (D14), after use of the sample for 28 days (D28) and after use of the sample for 56 days (D56). The test was carried out in a room at 20-22 ℃ and 40-60% relative humidity, and the subjects had to be acclimated for 15 minutes in the same environment.
The detection items are as follows: primos is used for facial image acquisition and skin wrinkle analysis;
the test effects are shown in table 5 below.
TABLE 5 synergistic test group Effect
As seen from the results in table 5, the anti-wrinkle ability comparison results are: example 1> comparative examples 5-7> comparative examples 1-3, which illustrate that Mirabilis jalapa active plant cells, yeast/rice fermentation filtrate, and split yeast fermentation lysate, which are selected as the active ingredients of the present invention, can effectively reduce facial wrinkles of skin by 15.76% when they are used in a simultaneous combination, and have better anti-aging effect compared with the use of a single component or two components, which illustrates that the components of the anti-aging active in the formulation of the present invention can be used simultaneously, and have synergistic anti-aging effect. In addition, the experimental results of the functional components show that the anti-aging active substance can effectively enhance the activity of mitochondria and provide an energy source for the physiological activity of skin cells. In conclusion, the composition for external application to skin of the present invention has a significant anti-aging effect, can effectively improve the aging condition of skin, can delay the aging of skin when a facial cream prepared from the composition is used for a long time, and can also make people of the same age look more young and energetic.
The embodiments described above are described to facilitate an understanding and use of the invention by those skilled in the art. It will be readily apparent to those skilled in the art that various modifications to these embodiments may be made, and the generic principles described herein may be applied to other embodiments without the use of the inventive faculty. Therefore, the present invention is not limited to the above embodiments, and those skilled in the art should make modifications and alterations without departing from the scope of the present invention.
Claims (10)
1. An anti-aging active composition, characterized by consisting of: the culture medium comprises Mirabilis jalapa active plant cells, yeast/rice fermentation product filtrate and split yeast fermentation product lysate, wherein the mass ratio of the Mirabilis jalapa active plant cells to the yeast/rice fermentation product filtrate to the split yeast fermentation product lysate is (0.5-5): (1-10): (0.5-5).
2. The anti-aging active composition according to claim 1, wherein the weight ratio of the mirabilis jalapa active plant cells, the yeast/rice fermentation product filtrate and the schizosaccharomyces cerevisiae fermentation product lysate is 1: (1-4): 1.
3. the anti-aging active composition according to claim 1, wherein the Mirabilis jalapa active plant cells are prepared by the following preparation method:
1) carrying out surface disinfection on the Mirabilis jalapa leaves, crushing, and culturing in a solid culture medium to obtain leaf callus;
2) separating the leaf callus from the Mirabilis jalapa when the leaf callus grows to the diameter of 1-3 cm, carrying out suspension culture on the leaf callus, placing the leaf callus in continuous fluorescent lamp irradiation, and placing the leaf callus in a shaking table at 24-26 ℃ for subculture to obtain a subculture suspension culture;
3) and separating the subculture suspension, removing tissue masses, dispersing the filtered cell sap in a fresh culture medium, and continuously culturing to obtain the mirabilis jalapa active plant cell.
4. A composition for external application to the skin which is simultaneously resistant to endogenous and exogenous aging, comprising the anti-aging active composition according to any one of claims 1 to 3, and further comprising an auxiliary agent acceptable for use in a skin external product.
5. The composition for external application on skin, which is capable of simultaneously resisting endogenous and exogenous aging, according to claim 4, is characterized by comprising the following components in percentage by weight:
0.1-30% of anti-aging active substance, 0.1-15% of humectant, 0.1-15% of emollient, 0.1-12% of emulsifier, 0.1-12% of filler, 0.1-12% of thickening agent, 0.01-5% of preservative, 0.01-1% of pH regulator, 0.01-1% of daily essence and the balance of deionized water, wherein the anti-aging active substance is the anti-aging active composition of any one of claims 1-3.
6. The composition for external application on skin for simultaneously resisting endogenous and exogenous aging as claimed in claim 5, wherein the composition for external application on skin for simultaneously resisting endogenous and exogenous aging comprises the following components in percentage by weight:
0.1-8% of Mirabilis jalapa active plant cells, 0.1-14% of yeast/rice fermentation product filtrate, 0.1-8% of schizosaccharomyces cerevisiae fermentation product lysate, 0.1-15% of humectant, 0.1-15% of emollient, 0.1-12% of emulsifier, 0.1-12% of filler, 0.1-12% of thickener, 0.01-5% of preservative, 0.01-1% of pH regulator, 0.01-1% of daily essence, and the balance of deionized water.
7. The composition for external application on skin capable of simultaneously resisting endogenous and exogenous aging according to claim 4, which is characterized by comprising the following components in percentage by weight:
0.1-20% of anti-aging active substance, 0.1-10% of humectant, 1-15% of emollient, 1-15% of emulsifier, 0.1-5% of filler, 0.1-8% of thickener, 0.01-3% of preservative, 0.01-0.8% of pH regulator, 0.01-0.8% of daily essence and the balance of deionized water, wherein the anti-aging active substance is the anti-aging active composition of any one of claims 1-3.
8. The composition for external application on skin for simultaneously resisting endogenous and exogenous aging as claimed in claim 7, which comprises the following components in percentage by weight:
0.1-5% of Mirabilis jalapa active plant cells, 0.1-10% of yeast/rice fermentation product filtrate, 0.1-5% of schizosaccharomyces cerevisiae fermentation product lysate, 0.1-10% of humectant, 1-15% of emollient, 1-15% of emulsifier, 0.1-5% of filler, 0.1-8% of thickener, 0.01-3% of preservative, 0.01-0.8% of pH regulator, 0.01-0.8% of daily essence and the balance of deionized water.
9. The composition for external application on skin for simultaneously resisting endogenous and exogenous aging as claimed in claim 4, wherein the composition for external application on skin for simultaneously resisting endogenous and exogenous aging comprises the following components in percentage by weight:
0.1-3% of Mirabilis jalapa active plant cells, 0.1-8% of yeast/rice fermentation product filtrate, 0.1-3% of schizosaccharomyces cerevisiae fermentation product lysate, 0.1-8% of humectant, 2-15% of emollient, 2-15% of emulsifier, 0.1-2% of filler, 0.1-5% of thickener, 0.01-2% of preservative, 0.01-0.5% of pH regulator, 0.01-0.4% of daily essence, and the balance of deionized water.
10. The composition for external application to the skin, which is capable of simultaneously combating extrinsic aging, according to any one of claims 4 to 9, wherein the composition for external application to the skin, which is capable of simultaneously combating extrinsic aging, is a cosmetic.
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