CN104257557A - Skincare composition for resisting skin aging and usage of skincare composition in preparation of cosmetics - Google Patents
Skincare composition for resisting skin aging and usage of skincare composition in preparation of cosmetics Download PDFInfo
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- CN104257557A CN104257557A CN201410529115.XA CN201410529115A CN104257557A CN 104257557 A CN104257557 A CN 104257557A CN 201410529115 A CN201410529115 A CN 201410529115A CN 104257557 A CN104257557 A CN 104257557A
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Abstract
The invention discloses a skincare composition for resisting skin aging and usages of the skincare composition in preparation of cosmetics. The skincare composition for resisting the skin aging contains PA (Palmitoleic Acid), rose extracts and privet extracts serve as active ingredients. The PA has the functions of protecting the epidermis and repairing the ultraviolet irradiation damage. Cell experiments and animal experiments show that the compatibility of the rose extracts, the privet extracts and the PA has the remarkable effects of resisting aging and activating skin cells, the skin aging of a user can be effectively prevented, and the skin of the user is protected from ultraviolet irradiation.
Description
Technical field
The present invention relates to cosmetic field, be specifically related to a kind of skin care compositions of anti-skin aging and preparing the purposes in cosmetics.
Background technology
The skin of people is as first protective barrier, and protection health vitals are by the outside stimulus of the change of such as temperature or humidity, ultraviolet and pollution.But Dermal exposure is easily subject to the injury of outside stimulus in external environment condition.In these outside stimuluss, ultraviolet is the main stimulation causing skin aging and apoptosis in epidermal cell death.If Dermal exposure is in ultraviolet, the cell component of such as DNA or protein may be damaged, cause apoptosis and death.
The epidermis cell produced at the basal layer in the most deep of epidermis externally moves in direction, always substitutes for new cell, is referred to as the renewal of epidermis.In this process, epidermis cell, via the differentiation of basal cell, spine cell, granulocyte and horn cell four-stage, finally forms scurf, comes off from skin surface.
Palmitoleic acid (Palmitoleic Acid, be called for short PA) be a kind of unsaturated fatty acid containing Omega-7 position double bond, in micro-plan ball algae of PA natural origin mainly in sea buckthorn fruit and marine microalgae, it is also a kind of natural material being present in health skin.PA is the important element maintaining healthy skin and health mucosa, effectively can eliminate the infringement of radical pair Skin Cell, and helps hypertrophy ossein in order to avoid moisture loss, and can maintain elasticity of skin.
Flos Rosae Rugosae is the dry flower of rosaceous plant Flos Rosae Rugosae (Rosa rugosa thunb) and polyphyll Flos Rosae Rugosae (Rosa rugosa thumb.var.plena Regel).Flos Rosae Rugosae sweet in the mouth, micro-hardship, warm in nature, return liver, spleen channel.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 new again " carries: " the strongly fragrant gas of soothing liver-QI gallbladder, spleen invigorating pathogenic fire reducing.Control cold type of pain in abdomen, gastral cavilty is long-pending cold, energy removing blood stasis of holding concurrently ".Rose ethereal oil is applied comparatively extensive in cosmetics, and rose wate extract is rich in gallic acid, the plant polyphenol compositions such as ellagic acid, general content dry weight be 8%-12%.
Ligustrum japonicum Thunb.flower is the dry flower of Oleaceae plants Fructus Ligustri Lucidi (Ligustrum lucidum Ait.), is generally gathered 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 protects epidermis cell and the defect preventing the material of its ultraviolet radiation damage for lacking, and provides a kind of skin care compositions of anti-skin aging and preparing the purposes in cosmetics.
The present invention finds 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 capable, PA then has stronger function after mixing with Flos Rosae Rugosae extract and Ligustrum japonicum Thunb.flower extract, but, Flos Rosae Rugosae extract, Ligustrum japonicum Thunb.flower extract itself does not but have this function, only have with PA conbined usage after, just occur this function and make this function intensified, namely the mixture of Flos Rosae Rugosae extract and Ligustrum japonicum Thunb.flower extract is the auxiliary agent that PA produces this function, thus complete the present invention.
Therefore, the invention provides a kind of skin care compositions of anti-skin aging, containing palmitoleic acid, Flos Rosae Rugosae 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 the compound shown in chemical formula (1), molecular formula: CH
3(CH
2)
5cH=CH (CH
2)
7cOOH, molecular weight 254.408.PA is the active component of the present invention for skin protection, has the apoptosis slowing down epidermis cell, prevents skin aging and keeps the function of epidermis cell vigor.
In the present invention, described Flos Rosae Rugosae extract is the rose wate extract of this area routine, is preferably prepared from by the method comprised the following steps: pulverized by dry Flos Rosae Rugosae alabastrum, add 10-15 times of weight water, 85-100 DEG C of heating 60-90 minute, filters, gets filtrate; Add 8-10 times of weight water in filter cake, 80-100 DEG C of heating 30-45 minute, filters, gets filtrate; Merge the filtrate of twice, concentrated, dry, obtain Flos Rosae Rugosae extract (MGH).In the present invention, described Flos Rosae Rugosae is the dry flower of rosaceous plant Flos Rosae Rugosae (Rosa rugosa thunb) and polyphyll Flos Rosae Rugosae (Rosa rugosa thumb.var.plena Regel).
In the present invention, described Ligustrum japonicum Thunb.flower extract is the Ligustrum japonicum Thunb.flower alcohol water extract of this area routine, preferably be prepared from by the method comprised the following steps: dry Ligustrum japonicum Thunb.flower alabastrum is pulverized, add 10-15 times of weight 40%-60% ethanol water, 20-60 DEG C of heating 60-90 minute, filter, get filtrate; Filter cake adds 8-10 times of weight 40%-60% ethanol water, and 20-60 DEG C of heating 30-45 minute, filters, get filtrate; Merge the filtrate of twice, concentrated, dry, obtain Ligustrum japonicum Thunb.flower extract (NZH).In the present invention, described Ligustrum japonicum Thunb.flower is the dry flower of Oleaceae plants Fructus Ligustri Lucidi (Ligustrum lucidum Ait.).
In the present invention, described pulverizing, filtration, concentrated, dry method are all this area conventional methods.
In the present invention, described palmitoleic acid, Flos Rosae Rugosae extract and Ligustrum japonicum Thunb.flower extract, with the gross weight of skin care compositions for benchmark, total amount can be 0.0001%-10%, preferred 0.01%-1%.In described skin care compositions, surplus is conventional media, preferably water.Described palmitoleic acid, Flos Rosae Rugosae extract and Ligustrum japonicum Thunb.flower extract weight ratio can preferred 1 ﹕ (15-25) ﹕ (30-50), more preferably 1 ﹕ 20 ﹕ 40.
The apoptotic inhibitor that this skin care compositions of the present invention is preferably induced by ultraviolet radiation, the regulator of being expressed by the keratin-14 of ultraviolet radiation down-regulation, keratin-17, ERK 1/2, cytokeratin-19.
The preparation method of skin care compositions of the present invention is that this area is conventional, by each component mix homogeneously in media as well.
The present invention finds the skin injury that PA can prevent ultraviolet radiation from causing thus prevents aging, and Flos Rosae Rugosae extract and Ligustrum japonicum Thunb.flower extract and mixture strengthen this function of PA.Therefore, PA and the compositions containing PA, Flos Rosae Rugosae extract and Ligustrum japonicum Thunb.flower extract can be mixed with external skin-care composition, such as cosmetic composition.But object and the preparation of described skin care compositions are not limited thereto.Therefore, the present invention also provides described skin care compositions preparing the purposes in cosmetics.Described cosmetics are made up of described skin care compositions and the conventional carrier of cosmetics.Preferred antidotal cosmetics of described cosmetics 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 meeting this area general knowledge, above-mentioned each optimum condition, can combination in any, obtains the preferred embodiments of the invention.
Agents useful for same of the present invention and raw material are all commercially.
Positive progressive effect of the present invention is: the present invention finds 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, repair 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 self-sow curve of HaCaT cell.
The PA of the different extension rate of Fig. 4 is on the impact of HaCaT Growth of Cells.Sample initial concentration 2526 μm of ol/L, contrast the cell culture result into using after same amount ethanol doubling dilution.
Fig. 5 shows the effect of vigor of PA to ultraviolet radiation injury HaCaT cell.Wherein, (A) UV density is schemed: 1.185mJ/cm
2; Figure (B) UV density: 2.37mJ/cm
2.
Fig. 6 shows the quantification that PA affects HaCaT cellular expression keratin-14, keratin-17 and ERK 1/2.
Fig. 7 shows the quantification of PA on HaCaT cellular expression keratin-14, keratin-17, cytokeratin-19 and ERK 1/2 impact that ultraviolet radiation injures.
Fig. 8 shows PA and different extract combination affects ultraviolet radiation injury HaCaT cell viability.Wherein, 1, blank group; 2, PA group; 3, MGH group; 4, NZH group; 1 group, 5, PA+MGH+NZH mixture; 2 groups, 6, PA+MGH+NZH mixture; 3 groups, 7, PA+MGH+NZH mixture; 4 groups, 8, PA+MGH+NZH mixture.
Fig. 9 shows PA and different extract combination to the shadow of skin mean roughness Rz after ultraviolet radiation
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 with ultraviolet radiation but not with the matched group of any external preparation process; 2, PA group; 3, MGH group; 4, NZH group; 5, PA+MGH+NZH group; 6, Normal group, be not with ultraviolet radiation also not with the matched group of external preparation process.
Detailed description of the invention
Mode below by embodiment further illustrates the present invention, but does not therefore limit the present invention among described scope of embodiments.The experimental technique of unreceipted actual conditions in the following example, conventionally and condition, or selects according to catalogue.
Raw material used in following examples comprises:
PA (Palmitoleic acid, palmitoleic acid), purchased from Sigma (article No. P9417, purity>=98.5% (GC), molecular formula: CH
3(CH
2)
5cH=CH (CH
2)
7cOOH, molecular weight 254.408);
HaCaT cell (people's immortalization epidermis cell), purchased from China typical culture collection center (Wuhan University);
DMEM high glucose medium (article No. C11995500BT) and hyclone (article No. 10099-141), purchased from
life Technologies;
The disposable sterilized consumptive material of cell culture 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, heat 70 minutes at a certain temperature, filter, get filtrate.Filter cake adds the identical solvent of 10 times of weight, heats 40 minutes under identical heating-up temperature, filters, gets filtrate.Merge twice filtrate, concentrated, dry, obtain Flos Rosae Rugosae 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％ |
Consider extraction ratio and Determination of Polyphenols, optimum extraction condition is pure water 80-100 DEG C and extracts (see table 1).
This Flos Rosae Rugosae extract analyzes (see Fig. 1) through HPLC, and its main component is gallic acid and ellagic acid.
The preparation of embodiment 2-3 Flos Rosae Rugosae extract
Pulverized by dry Flos Rosae Rugosae alabastrum, add constant weight times pure water, 90 DEG C are heated 70 minutes, filter, get filtrate.Filter cake adds constant weight times pure water, and 90 DEG C are heated 40 minutes, filter, get filtrate.
Merge twice filtrate, concentrated, dry, obtain Flos Rosae Rugosae extract (MGH) (see table 2).
The different ratio of water to material of table 2. Flos Rosae Rugosae extracts result of the test
The preparation of embodiment 4-6 Flos Rosae Rugosae extract
Pulverized by dry Flos Rosae Rugosae alabastrum, add 10 times of weight pure water, 100 DEG C of heating certain hours, filter, get filtrate.Filter cake adds 10 times of weight pure water, and 100 DEG C of heating certain hours, filter, get filtrate.Merge twice filtrate, concentrated, dry, obtain Flos Rosae Rugosae extract (MGH) (see 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, heat 70 minutes at a certain temperature, filter, get filtrate.Filter cake adds 10 times of weight pure water, heats 70 minutes at the same temperature, filters, gets filtrate.Merge twice filtrate, concentrated, dry, 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, filter, get filtrate.Filter cake adds the identical solvent of 10 times of weight, heats 40 minutes, filters, gets filtrate.Merge twice filtrate, concentrated, dry, obtain Ligustrum japonicum Thunb.flower extract (NZH) (see 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％ |
Consider extraction ratio and Determination of Polyphenols, better extraction conditions is 40%-60% ethanol-water extraction, and optimum extraction condition is 50% ethanol-water extraction.
This Ligustrum japonicum Thunb.flower 50% ethanol-water extraction thing analyzes (see Fig. 2) through HPLC, and its main component is chlorogenic acid, epicatechin, echinacoside, rutin etc.
The preparation of embodiment 12-13 Ligustrum japonicum Thunb.flower extract
Pulverized by dry Ligustrum japonicum Thunb.flower alabastrum, add constant weight doubly 50% ethanol water, 40 DEG C are heated 70 minutes, filter, get filtrate.Filter cake adds constant weight doubly 50% ethanol water, and 40 DEG C are heated 40 minutes, filter, get filtrate.Merge twice filtrate, concentrated, dry, obtain Ligustrum japonicum Thunb.flower extract (NZH) (see table 6).
The different ratio of water to material of table 6. Ligustrum japonicum Thunb.flower extracts result of the test
The preparation of embodiment 14-16 Ligustrum japonicum Thunb.flower extract
Pulverized by dry Ligustrum japonicum Thunb.flower alabastrum, add 10 times of weight 50% ethanol water, 40 DEG C of heating certain hours, filter, get filtrate.Filter cake adds 10 times of weight 50% ethanol water, and 40 DEG C of heating certain hours, filter, get filtrate.Merge twice filtrate, concentrated, dry, 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, heat 70 minutes at a certain temperature, filter, get filtrate.Filter cake adds 10 times of weight 50% ethanol water, heats 40 minutes at the same temperature, filters, gets filtrate.Merge twice filtrate, concentrated, dry, obtain Ligustrum japonicum Thunb.flower extract (NZH) (see 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 DMEM high glucose medium containing 10% hyclone of preparation containing following additional thing: PA, Flos Rosae Rugosae extract and Ligustrum japonicum Thunb.flower extract.Wherein, each additional thing concentration is in the medium: 0.0025g/LPA, 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 and obtains respectively.
Comparative example 4
Prepare 4 kinds of DMEM high glucose mediums 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 is in the medium: 0.0025g/L PA, 0.05g/L Flos Rosae Rugosae extract, 0.10g/L Ligustrum japonicum Thunb.flower extract.
The preparation of embodiment 21 skin care compositions
The DMEM high glucose medium containing 10% hyclone of preparation containing following additional thing:
Get raw material PA, Flos Rosae Rugosae extract and Ligustrum japonicum Thunb.flower extract in the DMEM high glucose medium containing 10% hyclone.Wherein, each raw material concentration is in the medium: 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 extract, Ligustrum japonicum Thunb.flower extract is that embodiment 4,17 is prepared and obtains respectively.
The preparation of embodiment 22 skin care compositions
Get raw material PA, Flos Rosae Rugosae extract and Ligustrum japonicum Thunb.flower extract in the DMEM high glucose medium containing 10% hyclone.Wherein, each raw material concentration is in the medium: 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 extract, Ligustrum japonicum Thunb.flower extract is that embodiment 2,12 is prepared and obtains respectively.
The preparation of embodiment 23 skin care compositions
Get raw material PA, Flos Rosae Rugosae extract and Ligustrum japonicum Thunb.flower extract in the DMEM high glucose medium containing 10% hyclone.Wherein, each raw material concentration is in the medium: 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 extract, Ligustrum japonicum Thunb.flower extract is that embodiment 6,13 is prepared and obtains respectively.
The preparation of embodiment 24 skin care compositions
Get raw material PA, Flos Rosae Rugosae extract and Ligustrum japonicum Thunb.flower extract in water, be mixed with aqueous solution.Wherein, each raw material concentration is in aqueous: 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 is that embodiment 7,15 is prepared and obtains respectively.
Embodiment 25PA is on HaCaT Growth of Cells impact test
1 step
The light absorption microplate reader of BioTek company of the U.S. is adopted to measure the light absorption value of tetrazolium bromide-MTT experiment.Use MTT cell proliferation and detection method for cytotoxicity of gas-liquid, the first a ceremonial jade-ladle, used in libation measuring generation, in the light absorption value at 490nm place, reflects number of viable cells.Lucifuge operation during experiment.
1.1PA tests diluted sample
100mg PA is dissolved in 0.75ml dehydrated alcohol, and 0.45 μM of micro-pore-film filtration is degerming rear for subsequent use.Get 10 μ l PA solution and add (containing 10% hyclone) in 2mlDMEM high glucose medium, concentration is 2625 μm of ol/L, is then used for experiment with culture medium 2 doubling dilution, tests extension rate used and concentration in table 9.
Table 9.PA solution dilution multiple and final test concentration
1.2 inoculating cell
Be made into individual cells suspension with the DMEM high glucose medium containing 10% hyclone, with every hole about 5,000 HaCaT cell is inoculated into 96 orifice plates, every pore volume 100 μ l.Add test sample after 37 DEG C of overnight incubation, each concentration samples establishes 5 holes, and after using same content ethanol doubling dilution in contrast.
1.3 in normal complexion colorimetric
Cultivate after 24 hours, every hole adds MTT solution 10 μ l.Continue to hatch 4 hours, stop cultivating.Every hole adds 100 μ l DMSO, hatches to jolt 10min and fully melt to the purple crystal of first a ceremonial jade-ladle, used in libation.Select 490nm wavelength, microplate reader measures each hole light absorption value.Respectively organize institute's value, with different disposal group for abscissa, light absorption value is that vertical coordinate is drawn, and calculates IC
50value.
2 results
The self-sow curve of 2.1HaCaT cell
For the proliferative conditions of inspection HaCaT cell, first do this cell growth curve under experimental conditions.This cell can normal proliferative under experiment condition of culture as seen from Figure 3.
2.2PA is on the impact of HaCaT Growth of Cells
Refer to Fig. 4, result of the test shows, under the low concentration of 5-41 μm of ol/L, occurs the effect of Promote cell's growth.Calculate the IC to HaCaT cell
50value is 354 μm of ol/L.Visible, PA has appreciable impact to HaCaT Growth of Cells, illustrates that toxicity is very little.
The repair that embodiment 26PA injures HaCaT cell ultraviolet radiation
The test sample preparation of the culture medium containing PA is with embodiment 25.
Getting well-grown HaCaT cell, when cell paving is to culture dish surface about 80%, remove culture medium, after PBS washing, under PBS exists, is 1.185 and 2.37mJ/cm to record intensity at 254nm respectively
2ultraviolet irradiate, then remove PBS, add the culture medium (the DMEM high glucose medium containing 10% hyclone) of the PA containing respective concentration, continue 37 DEG C hatch 24 hours after, investigate the vigor of cell with the identical mtt assay of embodiment 25.
As shown in Figure 5, the PA recording 10,20 and 40 μMs has the vigor injuring cell by UV and necessarily promotes restitution result.Although the effect promoted does not reach significant level, may there is inherent change in prompting cell, can carry out the test of cellular expression albumen change.
Embodiment 27PA tests the impact of HaCaT Growth of Cells associated protein
Keratin-17 can promote tissue repair and rapid regeneration (Kim, Wong et al.2006).Keratin-17 also may relevant to the growth cycle of hair (Tong and Coulombe 2006), lacks keratin-17 and can cause alopecia (McGowan, Tong et al.2002).Early differentiation after the propagation of keratin-14 participation epidermis cell, has important function (Alam, Sehgal et al.2011) in the differentiation and proliferation maintaining basal confluent monolayer cells.ERK then can reduce UVA and irradiate the cytoactive oxygen content caused, for skin repair offers help (Henri, Beaumel et al.2012).
1 step
Add PA during HaCaT cell culture and test sample, collecting cell after cultivation a period of time, carry out quantitatively to the protein content of cell pyrolysis liquid after fragmentation, adjustment protein concentration unanimously compares the content of Keratin 14, Keratin 17 and ERK afterwards by Western Blotting method.
1.1PA tests diluted sample
100mg PA is dissolved in 0.75ml dehydrated alcohol, and 0.45 μM of micro-pore-film filtration is degerming rear for subsequent use.Get 6.7 μ l PA solution and add (containing 10% hyclone) in 10ml culture medium, final concentration is 350 μm of ol/L.The ethanol of corresponding content is added in the culture medium of contrast.
1.2 cell incubation
HaCaT cell is cultivated with the DMEM high glucose medium containing 10% hyclone, until cell paving to culture dish surface 50% time standby containing test sample culture medium change liquid, continue 37 DEG C of cultured cells after 24 hours, collecting cell, carry out quantitatively to the protein content of cell pyrolysis liquid after fragmentation, adjustment protein concentration is rear use unanimously, and Western Blot detects the expression of cellular associated proteins.
2 results
Western Blotting result shows, and applied sample amount during each group electrophoresis is basically identical.Utilize ImageJ software to analyze image, using α-Tubulin content as benchmark, each group of image is carried out quantification contrast, the results are shown in Figure 6.In HaCaT cell, after PA process, 3 kinds of expressing quantities all have increase, and wherein keratin-14 increases by 19.4%, and keratin-17 increases by 12.1%, ERK 1/2 and increases by 13.1%.Visible, PA can promote the increase of keratin-14 in HaCaT cell, keratin-17, ERK 1/2 expression.
Embodiment 28PA tests the impact of the reparation associated protein that HaCaT cell ultraviolet radiation injures
The test diluted sample of the culture medium containing PA is with embodiment 27.
Cultivate HaCaT cell with the DMEM high glucose medium containing 10% hyclone, get well-grown HaCaT cell, when cell paving is to culture dish surface about 80%, remove culture medium, after PBS washing, under PBS exists, be 1.185 and 2.37mJ/cm to record intensity at 254nm respectively
2ultraviolet irradiate, then PBS is removed, add the culture medium of the PA containing respective concentration, continue 37 DEG C hatch 24 hours after, collecting cell, carry out quantitatively to the protein content of cell pyrolysis liquid after fragmentation, adjustment protein concentration is rear use unanimously, and Western Blot detects the expression of cellular associated proteins.
After ultraviolet radiation, there is apoptosis in part cell; Non-apoptotic cell shows as growth and is suppressed, with proliferation-associated protein as the expression of ERK 1/2, keratin-17 reduces (see Fig. 7).But after ultra-vioket radiation, immediately with the PA culture medium incubated cell 24 hours respectively containing 10 and 40 μMs, find that cellular expression ERK 1/2 and keratin-17 improve, the keratin-14 and the cytokeratin-19 that characterize epidermis cell mitotic activity in addition are also improved, and wherein the PA process of 40 μMs is comparatively obvious.Visible, in ultraviolet radiation injury HaCaT cell, PA can promote that keratin-14 in HaCaT cell, keratin-17, cytokeratin-19, ERK 1/2 expression bring up to Normocellular level, promotes the growth recovery of cell.Thus PA has the function promoting epidermal growth and reparation.
The antidotal test cell line of embodiment 29 skin care compositions
Cultivate HaCaT cell with the DMEM high glucose medium containing 10% hyclone, get well-grown HaCaT cell, when cell paving is to culture dish surface about 80%, remove culture medium, after PBS washing, to record intensity at 254nm for 2.37mJ/cm
2ultraviolet radiation HaCaT cell, then add 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 extract-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 respectively, continue 37 DEG C and hatch 24 hours, the mtt assay as described in embodiment 25 surveys the vigor injuring cell by UV.Wherein, in each described culture medium, the content of each component is as follows: contain respectively in front several culture medium: 0025g/L PA, 0.05g/L Flos Rosae Rugosae extract, 0.10g/L Ligustrum japonicum Thunb.flower extract (see comparative example 4); Contain in the culture medium of contain mixtures 1: 0.0025g/L PA, 0.05g/L Flos Rosae Rugosae extract, 0.10g/L Ligustrum japonicum Thunb.flower extract (see embodiment 20); Contain in the culture medium of contain mixtures 2: 0.0025g/L PA, 0.05g/L Flos Rosae Rugosae extract, 0.0075g/L Ligustrum japonicum Thunb.flower extract (see embodiment 21); Contain in the culture medium of contain mixtures 3: 0.0025g/L PA, 0.0625g/L Flos Rosae Rugosae extract, 0.125g/L Ligustrum japonicum Thunb.flower extract (see embodiment 22); Contain in the culture medium of contain mixtures 4: 0.0025g/L PA, 0.0375g/L Flos Rosae Rugosae extract, 0.075g/L Ligustrum japonicum Thunb.flower extract (see embodiment 23).
PA and different extract combination are shown in Fig. 8 to the impact of ultraviolet radiation injury HaCaT cell viability.Wherein, 1, blank group; 2, PA group; 3, MGH group; 4, NZH group; 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 have the effect significantly recovered because 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 to compare the improvement of PA group in effect with Ligustrum japonicum Thunb.flower extract group be significant.Visible, the compositions of PA and PA and Flos Rosae Rugosae extract, Ligustrum japonicum Thunb.flower extract can be repaired and is exposed to UV-induced cell injury, has obvious anti-senescence function.
The zoopery of embodiment 30 skin care compositions skin anti-aging
Ultraviolet radiation hairless mouse, irradiates height 40cm.Irradiate once the next day of first 4 weeks, each irradiation 2 hours.Latter 4 weeks, irradiate 5 days weekly, each 2 hours, accumulated dose 5.9J/cm
2.The aqueous solution of local application PA before ultra-vioket radiation, Flos Rosae Rugosae extract, Ligustrum japonicum Thunb.flower extract and PA mixing Flos Rosae Rugosae extract and Ligustrum japonicum Thunb.flower extract.Normal group is left intact.Each component concentration is in aqueous: 0.0025g/L PA, 0.05g/L Flos Rosae Rugosae extract, and 0.10g/L Ligustrum japonicum Thunb.flower extract, aqueous solution preparation process is see embodiment 24.
PA, each extract and combination thereof are shown in Fig. 9 to the impact of skin mean roughness Rz after ultraviolet radiation.Figure 10 is shown in the change of mice skin tissue elastic fibers integral optical density (IOD).The relative expression levels of MMP-1mRNA sees Figure 11.Experiment shows, hairless mouse is through ultraviolet radiation after 8 weeks, and skin occurs that coarse, dermatoglyph is deepened, widened, and Elastic fibers reduces, and the expression of Fibroblast collagenase (MMP-1) rises.In extract pretreated group, PA group, PA mixing Flos Rosae Rugosae extract and Ligustrum japonicum Thunb.flower extract group significantly can improve the degree of roughness of skin, increase spandex content, suppress the expression of MMP-1.Flos Rosae Rugosae extract, Ligustrum japonicum Thunb.flower extract group have no positive effect.And PA mixing Flos Rosae Rugosae extract to compare the improvement of PA group in effect with Ligustrum japonicum Thunb.flower extract group be significant.Visible, the compositions of PA and PA and Flos Rosae Rugosae extract, Ligustrum japonicum Thunb.flower extract can be repaired and is exposed to UV-induced skin injury, has obvious anti-senescence function.
Should be understood that those skilled in the art can make various changes or modifications the present invention after having read foregoing of the present invention, these equivalent form of values fall within the application's appended claims limited range equally.
Claims (10)
1. a skin care compositions for anti-skin aging, is characterized in that, it contains the palmitoleic acid of chemical formula as shown in (1), Flos Rosae Rugosae extract and Ligustrum japonicum Thunb.flower extract as active component,
2. skin care compositions as claimed in claim 1, it is characterized in that, described Flos Rosae Rugosae extract 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 characterized in that, described Flos Rosae Rugosae extract is prepared from by the method comprised the following steps: pulverized by dry Flos Rosae Rugosae alabastrum, add 10-15 times of weight water, 85-100 DEG C of heating 60-90 minute, filters, gets filtrate; Add 8-10 times of weight water in filter cake, 80-100 DEG C of heating 30-45 minute, filters, gets filtrate; Merge the filtrate of twice, concentrated, dry, obtain Flos Rosae Rugosae extract; Or described Ligustrum japonicum Thunb.flower extract is prepared from by the method comprised the following steps: pulverized by dry Ligustrum japonicum Thunb.flower alabastrum, add 10-15 times of weight 40%-60% ethanol water, 20-60 DEG C of heating 60-90 minute, filters, gets filtrate; Filter cake adds 8-10 times of weight 40%-60% ethanol water, and 20-60 DEG C of heating 30-45 minute, filters, get filtrate; Merge the filtrate of twice, concentrated, dry, obtain Ligustrum japonicum Thunb.flower extract.
4. skin care compositions as claimed in claim 1, it is characterized in that, described palmitoleic acid, the weight ratio of Flos Rosae Rugosae extract and Ligustrum japonicum Thunb.flower extract is 1 ﹕ (15-25) ﹕ (30-50).
5. skin care compositions as claimed in claim 1, it is characterized in that, described palmitoleic acid, Flos Rosae Rugosae extract and Ligustrum japonicum Thunb.flower extract weight ratio are 1 ﹕ 20 ﹕ 40.
6. skin care compositions as claimed in claim 1, is characterized in that, described palmitoleic acid, Flos Rosae Rugosae extract and Ligustrum japonicum Thunb.flower extract, with the gross weight of skin care compositions for benchmark, total amount is 0.0001%-10%, preferred 0.01%-1%, described percentage ratio is percentage by weight.
7. skin care compositions as claimed in claim 6, it is characterized in that, the surplus of described skin care compositions is water.
8. skin care compositions as claimed in claim 1, it is characterized in that, described Flos Rosae Rugosae is rosaceous plant Flos Rosae Rugosae (Rosa rugosa thunb) or polyphyll Flos Rosae Rugosae (Rosa rugosa thumb.var.plena Regel), and described Ligustrum japonicum Thunb.flower is Oleaceae plants Fructus Ligustri Lucidi (Ligustrum lucidum Ait.).
9. the skin care compositions of anti-skin aging as claimed in claim 1 is preparing the purposes in cosmetics.
10. purposes as claimed in claim 9, it is characterized in that, described cosmetics are antidotal cosmetics or prevent or repair the cosmetics being exposed to UV-induced skin injury.
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| CN107540756A (en) * | 2017-09-15 | 2018-01-05 | 黄河科技学院 | One kind promotees blood coagulation Ligustrum japonicum Thunb.flower polysaccharide and its purification methods and uses |
| CN115105448A (en) * | 2022-07-14 | 2022-09-27 | 广东瀚润生物科技有限公司 | Preparation method and application of raw materials of anti-radiation, blood-activating, face-beautifying and anti-aging skin care product |
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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 |
| CN102846587B (en) * | 2011-06-29 | 2022-05-06 | 邢军武 | Fatty acid composition and plant extract and pharmaceutical preparation and application thereof |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107540756A (en) * | 2017-09-15 | 2018-01-05 | 黄河科技学院 | One kind promotees blood coagulation Ligustrum japonicum Thunb.flower polysaccharide and its purification methods and uses |
| CN107540756B (en) * | 2017-09-15 | 2020-06-02 | 黄河科技学院 | Blood coagulation promoting privet flower polysaccharide and extraction and separation method and application thereof |
| CN115105448A (en) * | 2022-07-14 | 2022-09-27 | 广东瀚润生物科技有限公司 | Preparation method and application of raw materials of anti-radiation, blood-activating, face-beautifying and anti-aging skin care product |
| CN116270345A (en) * | 2022-07-14 | 2023-06-23 | 广东瀚润生物科技有限公司 | Natural herbal cosmetic with effects of radiation protection, blood circulation activating, skin nourishing and anti-aging and application thereof |
| CN116270345B (en) * | 2022-07-14 | 2024-04-26 | 北京娅熙妍国际贸易有限公司 | Natural herbal cosmetic with effects of radiation protection, blood circulation activating, skin nourishing and anti-aging and application thereof |
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