CN104257557B - The skin care compositions of a kind of anti-skin aging and the purposes in preparing cosmetics thereof - Google Patents
The skin care compositions of a kind of anti-skin aging and the purposes in preparing cosmetics thereof Download PDFInfo
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Abstract
The invention discloses the skin care compositions of a kind of anti-skin aging and the purposes in preparing cosmetics thereof.This ageing-resistant skin care composition contains palmitoleic acid (Palmitoleic Acid is called for short PA), and Flos Rosae Rugosae extract and Ligustrum japonicum Thunb.flower extract are as active component.Present invention discover that palmitoleic acid has protection epidermis and repairs the function of ultraviolet radiation damage.Cell experiment and zoopery show, Flos Rosae Rugosae extract, and Ligustrum japonicum Thunb.flower extract and PA compatibility have significant defying age and activate effect of Skin Cell, can effectively prevent application on human skin old and feeble, and protection application on human skin is not by ultraviolet damage.
Description
Technical field
The present invention relates to cosmetic field, be specifically related to a kind of anti-skin aging skin care compositions and
Prepare the purposes in cosmetics.
Background technology
The skin of people is as first protective barrier, and protection health vitals are by such as temperature or wet
Degree change, ultraviolet and the outside stimulus of pollution.But, skin is externally exposed environment and is easily subject to
The injury of outside stimulus.In these outside stimuluss, ultraviolet is to cause skin aging and epidermis cell to wither
Die death main stimulation.If skin is exposed to ultraviolet, such as DNA or protein may be damaged
Cell component, cause apoptosis and death.
The epidermis cell that basal layer in epidermis deep produces moves to outside direction, and it is new for always substituting
Cell, be referred to as the renewal of epidermis.In this process, epidermis cell is thin via basal cell, spine
Born of the same parents, granulocyte and the differentiation of horn cell four-stage, eventually form scurf, come off from skin surface.
Palmitoleic acid (Palmitoleic Acid is called for short PA) is a kind of containing Omega-7 position double bond
Unsaturated fatty acid, in micro-plan ball algae that PA natural origin is mainly in sea buckthorn fruit and marine microalgae,
Also it is a kind of material being naturally occurring in health skin.PA is to maintain healthy skin and health mucosa
Important element, can effectively eliminate the infringement of radical pair Skin Cell, and help hypertrophy ossein in order to avoid
Moisture loss, and elasticity of skin can be maintained.
Flos Rosae Rugosae is rosaceous plant Flos Rosae Rugosae (Rosa rugosa thunb) and polyphyll Flos Rosae Rugosae (Rosa rugosa
Thumb.var.plena Regel) dry flower.Flos Rosae Rugosae sweet in the mouth, micro-hardship, warm in nature, return liver, spleen
Warp.Flos Rosae Rugosae has resolving depression of regulating the flow of vital energy, effect of regulating menstruation by adjusting the flow of blood." property of medicine is examined " carries: " Flos Rosae Rugosae circulation of qi promoting
Removing mass, damage stasis of blood pain "；" detailed outline is picked up any lost article from the road " carries: " Flos Rosae Rugosae regulating and activating blood circulation, regulate the flow of vital energy, control migratory arthralgia "；
" book on Chinese herbal medicine is the newest " carries: " the strongly fragrant gas of soothing liver-QI gallbladder, spleen invigorating pathogenic fire reducing.Controlling cold type of pain in abdomen, gastral cavilty is long-pending cold, holds concurrently
Energy removing blood stasis ".Rose ethereal oil is applied relatively broad in cosmetics, and rose wate extract is rich in no food
Son acid, the plant polyphenol composition such as ellagic acid, general content dry weight for 8%-12%.
Ligustrum japonicum Thunb.flower is the dry flower of Oleaceae plants Fructus Ligustri Lucidi (Ligustrum lucidum Ait.), typically gathers
In 5-7 month.Ligustrum japonicum Thunb.flower aboundresources, but less to its research and development application.
Summary of the invention
The technical problem to be solved in the present invention is for lacking protection epidermis cell and preventing its ultraviolet from shining
Penetrate the defect of the material of damage, it is provided that the skin care compositions of a kind of anti-skin aging and preparing cosmetics
In purposes.
Present invention discover that palmitoleic acid (PA) has protection epidermis and repairs the function of ultraviolet radiation damage,
Can defying age, can be used in anti-apolexis composition or skin ultraviolet radiation damage remediation composition,
PA then has higher function after mixing with Flos Rosae Rugosae extract and Ligustrum japonicum Thunb.flower extract, but, rose
Rare flower extract, Ligustrum japonicum Thunb.flower extract itself but do not have this function, after being only used in combination with PA,
Just occur this function and make this function intensified, be i.e. Flos Rosae Rugosae extract and the mixture of Ligustrum japonicum Thunb.flower extract
It is the PA auxiliary agent that produces this function, thus completes the present invention.
Therefore, the invention provides the skin care compositions of a kind of anti-skin aging, containing palmitoleic acid, rose
Rare flower extract and Ligustrum japonicum Thunb.flower extract are as active component.
In the present invention, described palmitoleic acid (Palmitoleic Acid is called for short PA) is chemical formula (1)
Shown compound, molecular formula: CH3(CH2)5CH=CH (CH2)7COOH, molecular weight 254.408.
PA is the present invention active component for skin protection, has the apoptosis slowing down epidermis cell, prevents skin old
Change and keep the function of epidermis cell vigor.
In the present invention, described Flos Rosae Rugosae extract is the rose wate extract that this area is conventional, preferably
Be prepared from by the method comprised the following steps: dry Flos Rosae Rugosae alabastrum is pulverized, adds 10-15
Times of weight water, 85-100 DEG C is heated 60-90 minute, filters, takes filtrate；Filter cake adds 8-10 weight
Times water, 80-100 DEG C is heated 30-45 minute, filters, takes filtrate；Merge the filtrate of twice, concentrate,
It is dried, obtains Flos Rosae Rugosae extract (MGH).In the present invention, described Flos Rosae Rugosae is rosaceous plant
Flos Rosae Rugosae (Rosa rugosa thunb) and polyphyll Flos Rosae Rugosae (Rosa rugosa thumb.var.plena Regel)
Dry flower.
In the present invention, described Ligustrum japonicum Thunb.flower extract is the Ligustrum japonicum Thunb.flower alcohol water extract that this area is conventional, relatively
Good is prepared from by the method comprised the following steps: pulverized by dry Ligustrum japonicum Thunb.flower alabastrum, adds 10-15
Times of weight 40%-60% ethanol water, 20-60 DEG C is heated 60-90 minute, filters, takes filtrate；Filter
Cake adds 8-10 times of weight 40%-60% ethanol water, and 20-60 DEG C is heated 30-45 minute, filters,
Take filtrate；Merge the filtrate of twice, concentrate, be dried, obtain Ligustrum japonicum Thunb.flower extract (NZH).This
In bright, described Ligustrum japonicum Thunb.flower is the dry flower of Oleaceae plants Fructus Ligustri Lucidi (Ligustrum lucidum Ait.).
In the present invention, described pulverizing, filter, concentrate, the method that is dried is all this area conventional method.
In the present invention, described palmitoleic acid, Flos Rosae Rugosae extract and Ligustrum japonicum Thunb.flower extract, with skin protection group
On the basis of the gross weight of compound, total amount can be 0.0001%-10%, preferably 0.01%-1%.Described
In skin care compositions, surplus is conventional media, it is preferred that water.Described palmitoleic acid, Flos Rosae Rugosae carries
Take thing and Ligustrum japonicum Thunb.flower extract weight ratio can preferably 1 (15-25) (30-50), more preferably 1 20
40。
The apoptotic suppression that this skin care compositions of the present invention is preferably induced by ultraviolet radiation
Agent, by the keratin-14 of ultraviolet radiation down-regulation, keratin-17, ERK 1/2, cytokeratin-19
The regulator expressed.
The preparation method of skin care compositions of the present invention is that this area is conventional, each component is mixed in media as well
Close uniformly.
Present invention discover that PA is prevented from ultraviolet and irradiates the skin injury that causes thus prevent aging, and
Flos Rosae Rugosae extract and Ligustrum japonicum Thunb.flower extract and mixture strengthen this function of PA.Therefore, PA and containing
The compositions having PA, Flos Rosae Rugosae extract and Ligustrum japonicum Thunb.flower extract can be configured to external skin-care composition,
Such as cosmetic composition.But, purpose and the preparation of described skin care compositions are not limited thereto.Cause
This, the present invention also provides for described skin care compositions purposes in preparing cosmetics.Described cosmetics
It is made up of described skin care compositions and cosmetics routine carrier.The change of the preferred defying age of described cosmetics
Cosmetic or prevent or repair the cosmetics being exposed to UV-induced skin injury.
In the present invention, unless otherwise noted, described percentage ratio is percentage by weight.
On the basis of meeting common sense in the field, above-mentioned each optimum condition, can combination in any, i.e. get Ben Fa
Bright each preferred embodiments.
Agents useful for same of the present invention and raw material are the most commercially.
The most progressive effect of the present invention is: present invention discover that palmitoleic acid has protection epidermis and reparation
The function of ultraviolet radiation damage.Cell experiment and zoopery show, Flos Rosae Rugosae extract, Ligustrum japonicum Thunb.flower
Extract and PA compatibility have significant defying age and activate effect of Skin Cell, can effectively prevent
Application on human skin is old and feeble, repairs skin damage, and protection application on human skin is not by ultraviolet damage.
Accompanying drawing explanation
Fig. 1 is rose wate extractive HPLC spectrogram (main containing gallic acid and ellagic acid).
Fig. 2 is Ligustrum japonicum Thunb.flower extractive HPLC spectrogram.Chlorogenic acid (RT=9.59min)；Epicatechin
(RT=13.2min)；Echinacoside (RT=16.1min)；Rutin (RT=18.4min).
Fig. 3 is the natural growth curve of HaCaT cell.
The impact that HaCaT cell is grown by the PA of Fig. 4 difference extension rate.Sample initial concentration 2526
μm ol/L, the cell cultivation results after compareing as use same amount ethanol doubling dilution.
Fig. 5 shows the PA effect of vigor to ultraviolet irradiation injury HaCaT cell.Wherein, figure (A)
UV density: 1.185mJ/cm2；Figure (B) UV density: 2.37mJ/cm2。
Fig. 6 shows that HaCaT cell express keratin-14, keratin-17 and ERK 1/2 are affected by PA
Quantization.
Fig. 7 show PA to the HaCaT cell express keratin-14 of ultraviolet irradiation injury, keratin-17,
Cytokeratin-19 and the quantization of ERK 1/2 impact.
Fig. 8 shows that ultraviolet irradiation injury HaCaT cell viability is affected by PA and different extract combination.
Wherein, 1, blank group；2, PA groups；3, MGH groups；4, NZH groups；5, PA+MGH+NZH
1 group of mixture；6, PA+MGH+NZH 2 groups of mixture；7, PA+MGH+NZH mixture 3
Group；8, PA+MGH+NZH 4 groups of mixture.
Fig. 9 shows that PA and different extract combination are to the shadow of skin mean roughness Rz after ultraviolet irradiation
Figure 10 shows the change of mice skin tissue elastic fibers integral optical density (IOD).
Figure 11 shows the relative expression levels of MMP-1mRNA.
In Fig. 9-11,1, UV model control group, is to irradiate but at unused any external preparation with ultraviolet
The matched group of reason；2, PA groups；3, MGH groups；4, NZH groups；5, PA+MGH+NZH groups；
6, Normal group, is the matched group of unused ultraviolet irradiation the most unused external preparation process.
Detailed description of the invention
Further illustrate the present invention below by the mode of embodiment, but the most therefore limit the present invention to
Among described scope of embodiments.The experimental technique of unreceipted actual conditions in the following example, according to often
Rule method and condition, or select according to catalogue.
Raw material used in following example includes:
PA (Palmitoleic acid, palmitoleic acid), purchased from Sigma (article No. P9417, purity >=98.5%
(GC), molecular formula: CH3(CH2)5CH=CH (CH2)7COOH, molecular weight 254.408)；
HaCaT cell (people's immortalization epidermis cell), (military purchased from China typical culture collection center
Chinese university)；
DMEM high glucose medium (article No. C11995500BT) and hyclone (article No. 10099-141),
It is purchased fromLife Technologies；
Cell is cultivated disposable sterilized consumptive material and is purchased from。
UV-B type ultraviolet radiation meter, ring ground board, purchased from photoelectric instrument factory of Beijing Normal University.
The preparation of embodiment 1 Flos Rosae Rugosae extract
Dry Flos Rosae Rugosae alabastrum is pulverized, adds the solvent that 10 times of weight are certain, add at a certain temperature
Heat 70 minutes, filters, takes filtrate.Filter cake adds the solvent that 10 times of weight are identical, in identical heating temperature
The lower heating of degree 40 minutes, filters, takes filtrate.Merge twice filtrate, concentrate, be dried, obtain Flos Rosae Rugosae
Flower extract (MGH).
Table 1. Flos Rosae Rugosae different solvents extracts result of the test
| Solvent | Extract yield | Determination of Polyphenols in extract | |
| Embodiment 1 | Pure water (80-100 DEG C) | 45%-55% | 60%-70% |
| Comparative example 1 | 95% ethanol water (20-50 DEG C) | 20%-30% | 30%-40% |
Considering extraction ratio and Determination of Polyphenols, optimum extraction condition is pure water 80-100 DEG C extraction (being shown in Table 1).
This Flos Rosae Rugosae extract analyzes (see Fig. 1) through HPLC, and it is mainly composed of gallic acid and tan
Flower acid.
The preparation of embodiment 2-3 Flos Rosae Rugosae extract
Being pulverized by dry Flos Rosae Rugosae alabastrum, add constant weight times pure water, 90 DEG C are heated 70 minutes, mistake
Filter, takes filtrate.Filter cake adds constant weight times pure water, and 90 DEG C are heated 40 minutes, filter, take filtrate.
Merge twice filtrate, concentrate, be dried, obtain Flos Rosae Rugosae extract (MGH) (being shown in Table 2).
Table 2. Flos Rosae Rugosae difference ratio of water to material extracts result of the test
The preparation of embodiment 4-6 Flos Rosae Rugosae extract
Dry Flos Rosae Rugosae alabastrum is pulverized, adds 10 times of weight pure water, 100 DEG C of heating certain times, mistake
Filter, takes filtrate.Filter cake adds 10 times of weight pure water, 100 DEG C of heating certain times, filters, takes filtrate.
Merge twice filtrate, concentrate, be dried, obtain Flos Rosae Rugosae extract (MGH) (being shown in Table 3).
The table 3. Flos Rosae Rugosae different heating time extracts result of the test
The preparation of embodiment 7-8 Flos Rosae Rugosae extract
Dry Flos Rosae Rugosae alabastrum is pulverized, adds 10 times of weight pure water, at a certain temperature heating 70 points
Clock, filters, takes filtrate.Filter cake adds 10 times of weight pure water, at the same temperature heating 70 minutes,
Filter, take filtrate.Merge twice filtrate, concentrate, be dried, obtain Flos Rosae Rugosae extract (MGH) (see
Table 4).
Table 4. Flos Rosae Rugosae different heating temperature extracts result of the test
| Heating-up temperature/DEG C | Extract yield | Determination of Polyphenols in extract | |
| Embodiment 7 | 80 | 45% | 60% |
| Embodiment 8 | 100 | 55% | 70% |
| Comparative example 2 | 40 | 25% | 30% |
The preparation of embodiment 9-11 Ligustrum japonicum Thunb.flower extract
Dry Ligustrum japonicum Thunb.flower alabastrum is pulverized, adds the solvent that 10 times of weight are certain, heat 70 minutes, mistake
Filter, takes filtrate.Filter cake adds the solvent that 10 times of weight are identical, heats 40 minutes, filters, takes filtrate.
Merge twice filtrate, concentrate, be dried, obtain Ligustrum japonicum Thunb.flower extract (NZH) (being shown in Table 5).
Table 5. Ligustrum japonicum Thunb.flower different solvents condition extracts result of the test
| Solvent | Extract yield | Determination of Polyphenols in extract | |
| Comparative example 3 | Pure water (60-100 DEG C) | 35%-45% | 20%-30% |
| Embodiment 9 | 40% ethanol water (20-60 DEG C) | 30%-35% | 35%-45% |
| Embodiment 10 | 50% ethanol water (20-60 DEG C) | 30%-35% | 40%-50% |
| Embodiment 11 | 60% ethanol water (20-60 DEG C) | 30%-35% | 35%-45% |
Considering extraction ratio and Determination of Polyphenols, preferable extraction conditions is 40%-60% ethanol-water extraction, most preferably
Extraction conditions is 50% ethanol-water extraction.
This Ligustrum japonicum Thunb.flower 50% ethanol-water extraction thing analyzes (see Fig. 2) through HPLC, and it is mainly composed of green
Ortho acid, epicatechin, echinacoside, rutin etc..
The preparation of embodiment 12-13 Ligustrum japonicum Thunb.flower extract
Being pulverized by dry Ligustrum japonicum Thunb.flower alabastrum, add constant weight times 50% ethanol water, 40 DEG C are heated 70 points
Clock, filters, takes filtrate.Filter cake adds constant weight times 50% ethanol water, and 40 DEG C are heated 40 minutes,
Filter, take filtrate.Merge twice filtrate, concentrate, be dried, obtain Ligustrum japonicum Thunb.flower extract (NZH) (see
Table 6).
Table 6. Ligustrum japonicum Thunb.flower difference ratio of water to material extracts result of the test
The preparation of embodiment 14-16 Ligustrum japonicum Thunb.flower extract
Dry Ligustrum japonicum Thunb.flower alabastrum is pulverized, adds 10 times of weight 50% ethanol water, 40 DEG C of heating one timings
Between, filter, take filtrate.Filter cake adds 10 times of weight 50% ethanol water, and 40 DEG C are heated certain times,
Filter, take filtrate.Merge twice filtrate, concentrate, be dried, obtain Ligustrum japonicum Thunb.flower extract (NZH) (see
Table 7).
The table 7. Ligustrum japonicum Thunb.flower different heating time extracts result of the test
The preparation of embodiment 17-19 Ligustrum japonicum Thunb.flower extract
Dry Ligustrum japonicum Thunb.flower alabastrum is pulverized, adds 10 times of weight 50% ethanol water, add at a certain temperature
Heat 70 minutes, filters, takes filtrate.Filter cake adds 10 times of weight 50% ethanol water, at the same temperature
Heat 40 minutes, filter, take filtrate.Merge twice filtrate, concentrate, be dried, obtain Ligustrum japonicum Thunb.flower and carry
Take thing (NZH) (being shown in Table 8).
Table 8. Ligustrum japonicum Thunb.flower different heating temperature extracts result of the test
The preparation of embodiment 20 skin care compositions
The preparation DMEM high glucose medium containing 10% hyclone containing following additional thing: PA, rose
Rare flower extract and Ligustrum japonicum Thunb.flower extract.Wherein, each additional thing concentration in the medium is: 0.0025g/L
PA, 0.05g/L Flos Rosae Rugosae extract, 0.10g/L Ligustrum japonicum Thunb.flower extract (1:20:40).Wherein, described
Flos Rosae Rugosae extract, Ligustrum japonicum Thunb.flower extract is that embodiment 8,18 is prepared respectively.
Comparative example 4
The preparation 4 kinds DMEM high glucose medium containing 10% hyclone containing following additional thing respectively:
1、PA；
2, PA and Flos Rosae Rugosae extract；
3, PA and Ligustrum japonicum Thunb.flower extract；
4, blank.
Wherein, each additional thing concentration in the medium is: 0.0025g/L PA, 0.05g/L Flos Rosae Rugosae carries
Take thing, 0.10g/L Ligustrum japonicum Thunb.flower extract.
The preparation of embodiment 21 skin care compositions
The preparation DMEM high glucose medium containing 10% hyclone containing following additional thing:
Take raw material PA, Flos Rosae Rugosae extract and Ligustrum japonicum Thunb.flower extract in the DMEM containing 10% hyclone
In high glucose medium.Wherein, each raw material concentration in the medium is: 0.0025g/L PA, 0.05g/L
Flos Rosae Rugosae extract, 0.0075g/L Ligustrum japonicum Thunb.flower extract (1:20:30).Wherein, described Flos Rosae Rugosae carries
Taking thing, Ligustrum japonicum Thunb.flower extract is that embodiment 4,17 is prepared respectively.
The preparation of embodiment 22 skin care compositions
Take raw material PA, Flos Rosae Rugosae extract and Ligustrum japonicum Thunb.flower extract in the DMEM containing 10% hyclone
In high glucose medium.Wherein, each raw material concentration in the medium is: 0.0025g/L PA, 0.0625g/L
Flos Rosae Rugosae extract, 0.125g/L Ligustrum japonicum Thunb.flower extract (1:25:50).Wherein, described Flos Rosae Rugosae carries
Taking thing, Ligustrum japonicum Thunb.flower extract is that embodiment 2,12 is prepared respectively.
The preparation of embodiment 23 skin care compositions
Take raw material PA, Flos Rosae Rugosae extract and Ligustrum japonicum Thunb.flower extract in the DMEM containing 10% hyclone
In high glucose medium.Wherein, each raw material concentration in the medium is: 0.0025g/L PA, 0.0375g/L
Flos Rosae Rugosae extract, 0.075g/L Ligustrum japonicum Thunb.flower extract (1:15:30).Wherein, described Flos Rosae Rugosae carries
Taking thing, Ligustrum japonicum Thunb.flower extract is that embodiment 6,13 is prepared respectively.
The preparation of embodiment 24 skin care compositions
Take raw material PA, Flos Rosae Rugosae extract and Ligustrum japonicum Thunb.flower extract in water, be configured to aqueous solution.Its
In, each raw material concentration in aqueous is: 0.002g/L PA, 5g/L Flos Rosae Rugosae extract, 10g/L
Ligustrum japonicum Thunb.flower extract.Wherein, described Flos Rosae Rugosae extract, Ligustrum japonicum Thunb.flower extract be respectively embodiment 7,
15 are prepared.
HaCaT cell growth effect is tested by embodiment 25PA
1 step
The light using BioTek company of the U.S. absorbs microplate reader and measures the light absorption value of tetrazolium bromide-MTT experiment.
Use MTT cell proliferation and detection method for cytotoxicity of gas-liquid, measure the first a ceremonial jade-ladle, used in libation generated at 490nm
Light absorption value, reflects number of viable cells.Lucifuge operation during experiment.
1.1PA test sample dilutes
100mg PA is dissolved in 0.75ml dehydrated alcohol, standby after 0.45 μM of micro-pore-film filtration is degerming.Take
10 μ l PA solution add in 2mlDMEM high glucose medium (containing 10% hyclone), and concentration is 2625
μm ol/L, then is used for testing with culture medium 2 doubling dilution, and experiment extension rate used and concentration are shown in Table
9。
Table 9.PA solution extension rate and final test concentration
1.2 inoculating cell
It is made into individual cells suspension, with every hole about with the DMEM high glucose medium containing 10% hyclone
5,000 HaCaT cells are inoculated into 96 orifice plates, every pore volume 100 μ l.Add after 37 DEG C of overnight incubation
Entering test sample, each concentration samples sets 5 holes, and uses after same content ethanol doubling dilution as right
According to.
1.3 in normal complexion colorimetric
After cultivating 24 hours, every hole adds MTT solution 10 μ l.Continue to hatch 4 hours, terminate cultivating.
Every hole adds 100 μ l DMSO, hatches and shakes 10min and fully melt to the purple crystal of first a ceremonial jade-ladle, used in libation.Select
490nm wavelength, measures each hole light absorption value in microplate reader.Respectively group institute value, with different disposal
Group is abscissa, and light absorption value is that vertical coordinate is drawn, and calculates IC50Value.
2 results
The natural growth curve of 2.1HaCaT cell
For checking the proliferative conditions of HaCaT cell, first do this cell propagation under experimental conditions bent
Line.This cell can be with normal proliferative under experiment condition of culture as seen from Figure 3.
The impact that HaCaT cell is grown by 2.2PA
Referring to Fig. 4, result of the test shows, under the low concentration of 5-41 μm ol/L, occurs promoting cell
The effect of growth.Calculate the IC to HaCaT cell50Value is 354 μm ol/L.Visible, PA is to HaCaT
Cell growth has a significant impact, and illustrates that toxicity is the least.
The repair to HaCaT cell ultraviolet irradiation injury of embodiment 26PA
The test sample of the culture medium containing PA is prepared with embodiment 25.
Take well-grown HaCaT cell, when cell paving is to culture dish surface about 80%, remove
Culture medium, after PBS washing, in the presence of PBS, respectively to record intensity for 1.185 at 254nm
And 2.37mJ/cm2Ultraviolet be irradiated, then remove PBS, add the PA containing respective concentration
Culture medium (the DMEM high glucose medium containing 10% hyclone), continue 37 DEG C and hatch 24 hours
After, the vigor of cell is investigated with the mtt assay that embodiment 25 is identical.
Result is as it is shown in figure 5, record the PA of 10,20 and 40 μMs to the work by UV injury cell
Power has and necessarily promotes restitution.Although the effect promoted is not reaching to significant level, but points out cell
It may happen that the change of inherence, the test of cell expressing protein change can be carried out.
The impact of HaCaT cell growth associated protein is tested by embodiment 27PA
Keratin-17 can promote tissue repair and rapid regeneration (Kim, Wong et al.2006).Angle egg
-17 it is also possible to relevant to the growth cycle of hair (Tong and Coulombe 2006) in vain, lacks keratin
-17 can cause alopecia (McGowan, Tong et al.2002).Keratin-14 participates in the propagation of epidermis cell
Rear early differentiation, maintain basal confluent monolayer cells differentiation and proliferation in have important function (Alam,
Sehgal et al.2011).ERK then can reduce UVA and irradiate the cytoactive oxygen content caused, for skin
Skin reparation provides help (Henri, Beaumel et al.2012).
1 step
Add PA test sample when HaCaT cell is cultivated, after cultivating a period of time, collect cell, broken
Protein content to cell pyrolysis liquid carries out quantitatively afterwards, uses Western after adjustment protein concentration is consistent
Blotting method compares Keratin 14, the content of Keratin 17 and ERK.
1.1PA test sample dilutes
100mg PA is dissolved in 0.75ml dehydrated alcohol, standby after 0.45 μM of micro-pore-film filtration is degerming.Take
6.7 μ l PA solution add in 10ml culture medium (containing 10% hyclone), final concentration of 350 μm ol/L.
The culture medium of comparison adds the ethanol of corresponding content.
1.2 cell incubation
Cultivate HaCaT cell with the DMEM high glucose medium containing 10% hyclone, treat that cell spreads extremely
Change liquid by the culture medium containing test sample when of culture dish surface 50%, continue 37 DEG C and cultivate cell 24
After hour, collecting cell, after crushing, the protein content to cell pyrolysis liquid carries out quantitatively, adjusts albumen
Concentration unanimously rear use, the expression of Western Blot detection cellular associated proteins.
2 results
Western Blotting result shows, applied sample amount during each group electrophoresis is basically identical.Utilize ImageJ
Image is analyzed by software, and using α-Tubulin content as benchmark, it is right to carry out each group of image quantifying
Ratio, result is shown in Fig. 6.In HaCaT cell, after PA processes, 3 kinds of expressing quantities all have increasing
Adding, wherein keratin-14 increases by 19.4%, and keratin-17 increases by 12.1%, and ERK 1/2 increases by 13.1%.
Visible, PA can promote keratin-14 in HaCaT cell, keratin-17, ERK 1/2 expression
Increase.
Embodiment 28PA is real on the impact of the reparation associated protein of HaCaT cell ultraviolet irradiation injury
Test
The test sample of the culture medium containing PA dilutes with embodiment 27.
Cultivate HaCaT cell with the DMEM high glucose medium containing 10% hyclone, take well-grown
HaCaT cell, until cell paving to culture dish surface about 80% time, remove culture medium, PBS washes
After washing, in the presence of PBS, it is 1.185 and 2.37mJ/cm to record intensity at 254nm respectively2's
Ultraviolet is irradiated, and then removes PBS, adds the culture medium of the PA containing respective concentration, continues
After 37 DEG C hatch 24 hours, collecting cell, after crushing, the protein content to cell pyrolysis liquid carries out determining
Amount, adjusts protein concentration unanimously rear use, the expression of Western Blot detection cellular associated proteins.
After ultraviolet irradiates, there is apoptosis in part cell；Non-apoptotic cell shows as growth to be pressed down
System, reduces (see Fig. 7) with the expression of proliferation-associated protein such as ERK 1/2, keratin-17.But it is purple
After external exposure, immediately to contain the PA culture medium incubated cell 24 hours of 10 and 40 μMs respectively, send out
Existing cell expresses ERK 1/2 and keratin-17 improves, and additionally characterizes the angle egg of epidermis cell mitotic activity
In vain-14 and cytokeratin-19 be also improved, wherein 40 μMs PA process the most obvious.Visible, at purple
In outside line irradiation injury HaCaT cell, PA can promote keratin-14 in HaCaT cell, angle egg
In vain-17, cytokeratin-19, ERK 1/2 expression bring up to Normocellular level, promote the life of cell
Long recovery.Thus PA has the function promoting epidermal growth with reparation.
The test cell line of embodiment 29 skin care compositions defying age
Cultivate HaCaT cell with the DMEM high glucose medium containing 10% hyclone, take well-grown
HaCaT cell, until cell paving to culture dish surface about 80% time, remove culture medium, PBS washes
After washing, to record intensity as 2.37mJ/cm at 254nm2Ultraviolet irradiate HaCaT cell, then
It is separately added into containing PA, Flos Rosae Rugosae extract, Ligustrum japonicum Thunb.flower extract, PA-Flos Rosae Rugosae extract-Ligustrum japonicum Thunb.flower
Extract-mixture 1, PA-Flos Rosae Rugosae extract-Ligustrum japonicum Thunb.flower extract-mixture 2, PA-Flos Rosae Rugosae carry
Take thing-Ligustrum japonicum Thunb.flower extract-mixture 3, PA-Flos Rosae Rugosae extract-Ligustrum japonicum Thunb.flower extract-mixture 4,
And blank totally 8 groups of culture medium, continue 37 DEG C and hatch 24 hours, MTT as described in Example 25
Method surveys the vigor by UV injury cell.Wherein, in each described culture medium, the content of each component is as follows:
Front several culture medium contain respectively: 0025g/L PA, 0.05g/L Flos Rosae Rugosae extract, 0.10g/L Fructus Ligustri Lucidi
Flower extract (sees comparative example 4)；Culture medium containing mixture 1 contains: 0.0025g/L PA, 0.05g/L
Flos Rosae Rugosae extract, 0.10g/L Ligustrum japonicum Thunb.flower extract (sees embodiment 20)；Cultivation containing mixture 2
Base contains: 0.0025g/L PA, 0.05g/L Flos Rosae Rugosae extract, 0.0075g/L Ligustrum japonicum Thunb.flower extract (ginseng
See embodiment 21)；Culture medium containing mixture 3 contains: 0.0025g/L PA, 0.0625g/L Flos Rosae Rugosae
Extract, 0.125g/L Ligustrum japonicum Thunb.flower extract (sees embodiment 22)；In culture medium containing mixture 4
Containing: 0.0025g/L PA, 0.0375g/L Flos Rosae Rugosae extract, 0.075g/L Ligustrum japonicum Thunb.flower extract (sees
Embodiment 23).
The impact of ultraviolet irradiation injury HaCaT cell viability is shown in Fig. 8 by PA and different extract combination.
Wherein, 1, blank group；2, PA groups；3, MGH groups；4, NZH groups；5, PA+MGH+NZH
Mixture 1；6, PA+MGH+NZH mixture 2；7, PA+MGH+NZH mixture 3；8,
PA+MGH+NZH mixture 4.Test shows, PA group, and PA mixing Flos Rosae Rugosae extract and
Ligustrum japonicum Thunb.flower extract group has significantly to be recovered because of the effect of irradiation damage HaCaT cell viability.Flos Rosae Rugosae
Extract, Ligustrum japonicum Thunb.flower extract group have no positive effect.And PA mixing Flos Rosae Rugosae extract and Ligustrum japonicum Thunb.flower
It is significant that extract group compares the improvement in effect of the PA group.Visible, PA and PA and Flos Rosae Rugosae
Extract, the compositions of Ligustrum japonicum Thunb.flower extract can be repaired and be exposed to UV-induced cell injury, tool
There is obvious anti-senescence function.
Embodiment 30 skin care compositions skin anti-aging zoopery
Ultraviolet irradiates hairless mouse, irradiates height 40cm.Irradiate once the next day of first 4 weeks, shine every time
Penetrate 2 hours.Latter 4 weeks, irradiation 5 days weekly, each 2 hours, accumulated dose 5.9J/cm2.Ultraviolet is shone
Penetrate front local application PA, Flos Rosae Rugosae extract, Ligustrum japonicum Thunb.flower extract and PA mixing Flos Rosae Rugosae to extract
Thing and the aqueous solution of Ligustrum japonicum Thunb.flower extract.Normal group is left intact.Each component is in aqueous
Concentration be: 0.0025g/L PA, 0.05g/L Flos Rosae Rugosae extract, 0.10g/L Ligustrum japonicum Thunb.flower extract,
Aqueous solution preparation process sees embodiment 24.
The impact of skin mean roughness Rz after ultraviolet irradiation is shown in figure by PA, each extract and combinations thereof
9.Figure 10 is shown in the change of mice skin tissue elastic fibers integral optical density (IOD).MMP-1mRNA
Relative expression levels see Figure 11.Experiment shows, hairless mouse is after ultraviolet irradiates 8 weeks, and skin goes out
The most coarse, dermatoglyph is deepened, is widened, and Elastic fibers reduces, Fibroblast collagenase (MMP-1)
Expression rise.In extract pretreated group, PA group, PA mixing Flos Rosae Rugosae extract and Ligustrum japonicum Thunb.flower
Extract group can significantly improve the degree of roughness of skin, increases spandex content, suppresses MMP-1
Expression.Flos Rosae Rugosae extract, Ligustrum japonicum Thunb.flower extract group have no positive effect.And PA mixing rose
It is significant that rare flower extract compares the improvement in effect of the PA group with Ligustrum japonicum Thunb.flower extract group.It is visible,
The compositions of PA and PA and Flos Rosae Rugosae extract, Ligustrum japonicum Thunb.flower extract can be repaired and is exposed to ultraviolet
The skin injury caused, has obvious anti-senescence function.
Should be understood that, after the foregoing having read the present invention, those skilled in the art can be to this
Bright making various changes or modifications, these equivalent form of values fall within the application appended claims equally and are limited
Scope.
Claims (10)
1. the skin care compositions of an anti-skin aging, it is characterised in that it contains chemical formula such as (1)
Shown palmitoleic acid, Flos Rosae Rugosae extract and Ligustrum japonicum Thunb.flower extract as active component,
Described palmitoleic acid, the weight ratio of Flos Rosae Rugosae extract and Ligustrum japonicum Thunb.flower extract is 1 (15-25)
(30-50).
2. skin care compositions as claimed in claim 1, it is characterised in that described Flos Rosae Rugosae is extracted
Thing is rose wate extract, and described Ligustrum japonicum Thunb.flower extract is Ligustrum japonicum Thunb.flower alcohol water extract.
3. skin care compositions as claimed in claim 1, it is characterised in that described Flos Rosae Rugosae is extracted
Thing is prepared from by the method comprised the following steps: pulverized by dry Flos Rosae Rugosae alabastrum, adds 10-15 weight
Amount times water, 85-100 DEG C is heated 60-90 minute, filters, takes filtrate；Filter cake adds 8-10 times of weight
Water, 80-100 DEG C is heated 30-45 minute, filters, takes filtrate；Merge the filtrate of twice, concentrate, dry
Dry, obtain Flos Rosae Rugosae extract；Or, described Ligustrum japonicum Thunb.flower extract is by the method comprised the following steps
It is prepared from: dry Ligustrum japonicum Thunb.flower alabastrum is pulverized, adds 10-15 times of weight 40%-60% ethanol water,
20-60 DEG C is heated 60-90 minute, filters, takes filtrate；Filter cake adds 8-10 times of weight 40%-60% second
Alcohol-water solution, 20-60 DEG C is heated 30-45 minute, filters, takes filtrate；Merge the filtrate of twice, concentrate,
It is dried, obtains Ligustrum japonicum Thunb.flower extract.
4. skin care compositions as claimed in claim 1, it is characterised in that described palmitoleic acid,
Flos Rosae Rugosae extract and Ligustrum japonicum Thunb.flower extract weight ratio are 1 20 40.
5. skin care compositions as claimed in claim 1, it is characterised in that described palmitoleic acid,
Flos Rosae Rugosae extract and Ligustrum japonicum Thunb.flower extract, on the basis of the gross weight of skin care compositions, total amount is
0.0001%-10%, described percentage ratio is percentage by weight.
6. skin care compositions as claimed in claim 5, it is characterised in that described palmitoleic acid,
Flos Rosae Rugosae extract and Ligustrum japonicum Thunb.flower extract, on the basis of the gross weight of skin care compositions, total amount is
0.01%-1%, described percentage ratio is percentage by weight.
7. the skin care compositions as described in claim 5 or 6, it is characterised in that described skin protection group
The surplus of compound is water.
8. skin care compositions as claimed in claim 1, it is characterised in that described Flos Rosae Rugosae is rose
Common vetch section plant Flos Rosae Rugosae (Rosa rugosa thunb) or polyphyll Flos Rosae Rugosae (Rosa rugosa thumb.var.
Plena Regel), described Ligustrum japonicum Thunb.flower is Oleaceae plants Fructus Ligustri Lucidi (Ligustrum lucidum Ait.).
The skin care compositions of anti-skin aging the most as claimed in claim 1 use in preparing cosmetics
On the way.
10. purposes as claimed in claim 9, it is characterised in that described cosmetics are defying age
Cosmetics or prevent or repair the cosmetics being exposed to UV-induced skin injury.
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|---|---|---|---|---|
| JPH08259429A (en) * | 1995-03-20 | 1996-10-08 | Shiseido Co Ltd | Oil-based powder cosmetic |
| JP2000273014A (en) * | 1999-03-23 | 2000-10-03 | Kansai Koso Kk | Cosmetic composition |
| CN102846587A (en) * | 2011-06-29 | 2013-01-02 | 邢军武 | Fatty acid composition, plant extract, pharmaceutical preparation, and application thereof |
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2014
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH08259429A (en) * | 1995-03-20 | 1996-10-08 | Shiseido Co Ltd | Oil-based powder cosmetic |
| JP2000273014A (en) * | 1999-03-23 | 2000-10-03 | Kansai Koso Kk | Cosmetic composition |
| CN102846587A (en) * | 2011-06-29 | 2013-01-02 | 邢军武 | Fatty acid composition, plant extract, pharmaceutical preparation, and application thereof |
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| 女贞花化学成分研究;王军宪，等;《中国中药杂志》;19900331;第15卷(第3期);第40-42、63页 * |
| 女贞花的化学成分研究;龙飞等;《华西药学杂志》;20111231;第26卷(第2期);第97-100页 * |
| 护肤品配方中的天然活性成分;宋永波;《日用化学品科学》;20130331;第36卷(第3期);第44-46页 * |
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