CN105662905B - Purposes of the dihydromyricetin in preparing prevention dermal photodamage skin care item or drug - Google Patents
Purposes of the dihydromyricetin in preparing prevention dermal photodamage skin care item or drug Download PDFInfo
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- CN105662905B CN105662905B CN201610036173.8A CN201610036173A CN105662905B CN 105662905 B CN105662905 B CN 105662905B CN 201610036173 A CN201610036173 A CN 201610036173A CN 105662905 B CN105662905 B CN 105662905B
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- dihydromyricetin
- sun screen
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- skin care
- uva
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- KJXSIXMJHKAJOD-LSDHHAIUSA-N (+)-dihydromyricetin Chemical compound C1([C@@H]2[C@H](C(C3=C(O)C=C(O)C=C3O2)=O)O)=CC(O)=C(O)C(O)=C1 KJXSIXMJHKAJOD-LSDHHAIUSA-N 0.000 title claims abstract description 58
- 230000008832 photodamage Effects 0.000 title claims abstract description 32
- KQILIWXGGKGKNX-UHFFFAOYSA-N dihydromyricetin Natural products OC1C(=C(Oc2cc(O)cc(O)c12)c3cc(O)c(O)c(O)c3)O KQILIWXGGKGKNX-UHFFFAOYSA-N 0.000 title claims abstract description 30
- 230000002500 effect on skin Effects 0.000 title claims abstract description 21
- 230000002265 prevention Effects 0.000 title claims abstract description 11
- 239000003814 drug Substances 0.000 title abstract description 14
- 229940079593 drug Drugs 0.000 title abstract description 13
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 claims abstract description 33
- 239000000516 sunscreening agent Substances 0.000 claims abstract description 31
- 230000000475 sunscreen effect Effects 0.000 claims abstract description 28
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims abstract description 22
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 12
- 229910019142 PO4 Inorganic materials 0.000 claims abstract description 11
- 125000000118 dimethyl group Chemical group [H]C([H])([H])* 0.000 claims abstract description 11
- 235000011187 glycerol Nutrition 0.000 claims abstract description 11
- LXCFILQKKLGQFO-UHFFFAOYSA-N methylparaben Chemical compound COC(=O)C1=CC=C(O)C=C1 LXCFILQKKLGQFO-UHFFFAOYSA-N 0.000 claims abstract description 11
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 claims abstract description 11
- 239000010452 phosphate Substances 0.000 claims abstract description 11
- 229920005573 silicon-containing polymer Polymers 0.000 claims abstract description 11
- UIVPNOBLHXUKDX-UHFFFAOYSA-N 3,5,5-trimethylhexyl 3,5,5-trimethylhexanoate Chemical compound CC(C)(C)CC(C)CCOC(=O)CC(C)CC(C)(C)C UIVPNOBLHXUKDX-UHFFFAOYSA-N 0.000 claims abstract description 8
- 229940100554 isononyl isononanoate Drugs 0.000 claims abstract description 8
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 claims abstract description 7
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 claims abstract description 7
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 claims abstract description 7
- 239000000463 material Substances 0.000 claims description 14
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- TWNIBLMWSKIRAT-VFUOTHLCSA-N levoglucosan Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@H]2CO[C@@H]1O2 TWNIBLMWSKIRAT-VFUOTHLCSA-N 0.000 claims description 10
- 238000002360 preparation method Methods 0.000 claims description 9
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- FYGDTMLNYKFZSV-URKRLVJHSA-N (2s,3r,4s,5s,6r)-2-[(2r,4r,5r,6s)-4,5-dihydroxy-2-(hydroxymethyl)-6-[(2r,4r,5r,6s)-4,5,6-trihydroxy-2-(hydroxymethyl)oxan-3-yl]oxyoxan-3-yl]oxy-6-(hydroxymethyl)oxane-3,4,5-triol Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1[C@@H](CO)O[C@@H](OC2[C@H](O[C@H](O)[C@H](O)[C@H]2O)CO)[C@H](O)[C@H]1O FYGDTMLNYKFZSV-URKRLVJHSA-N 0.000 abstract 1
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- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 13
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- IKMDFBPHZNJCSN-UHFFFAOYSA-N Myricetin Chemical compound C=1C(O)=CC(O)=C(C(C=2O)=O)C=1OC=2C1=CC(O)=C(O)C(O)=C1 IKMDFBPHZNJCSN-UHFFFAOYSA-N 0.000 description 1
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- 229940116852 myricetin Drugs 0.000 description 1
- PCOBUQBNVYZTBU-UHFFFAOYSA-N myricetin Natural products OC1=C(O)C(O)=CC(C=2OC3=CC(O)=C(O)C(O)=C3C(=O)C=2)=C1 PCOBUQBNVYZTBU-UHFFFAOYSA-N 0.000 description 1
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Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/49—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds
- A61K8/4973—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with oxygen as the only hetero atom
- A61K8/498—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with oxygen as the only hetero atom having 6-membered rings or their condensed derivatives, e.g. coumarin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
- A61K31/352—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/87—Vitaceae or Ampelidaceae (Vine or Grape family), e.g. wine grapes, muscadine or peppervine
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/33—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
- A61K8/34—Alcohols
- A61K8/345—Alcohols containing more than one hydroxy group
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/33—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
- A61K8/37—Esters of carboxylic acids
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/60—Sugars; Derivatives thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/72—Cosmetics or similar toiletry preparations characterised by the composition containing organic macromolecular compounds
- A61K8/73—Polysaccharides
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/72—Cosmetics or similar toiletry preparations characterised by the composition containing organic macromolecular compounds
- A61K8/84—Cosmetics or similar toiletry preparations characterised by the composition containing organic macromolecular compounds obtained by reactions otherwise than those involving only carbon-carbon unsaturated bonds
- A61K8/89—Polysiloxanes
- A61K8/891—Polysiloxanes saturated, e.g. dimethicone, phenyl trimethicone, C24-C28 methicone or stearyl dimethicone
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q17/00—Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
- A61Q17/04—Topical preparations for affording protection against sunlight or other radiation; Topical sun tanning preparations
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Epidemiology (AREA)
- Birds (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Pharmacology & Pharmacy (AREA)
- Medicinal Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Emergency Medicine (AREA)
- Dermatology (AREA)
- Biotechnology (AREA)
- Botany (AREA)
- Medical Informatics (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Alternative & Traditional Medicine (AREA)
- Engineering & Computer Science (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention belongs to medical cosmetic fields, disclose purposes of the dihydromyricetin in preparing prevention dermal photodamage skin care item or drug.The dihydromyricetin is sun screen preparing the skin care item of prevention dermal photodamage, and the group of wherein sun screen is divided into：Water, propylene glycol, glycerine, trehalose, oat beta glucan, dihydromyricetin, C20‑22Alcohol phosphate, dimethyl silicone polymer, isononyl isononanoate and methyl hydroxybenzoate.The present invention induces the experiment of HaCaT cell models by UVA, prove that dihydromyricetin can effectively inhibit cell ROS to generate and lower the expression of antiapoptotic factors, inflammatory factor, mitigate cell oxidative damage, reduce cellular inflammation reaction and Apoptosis, achievees the effect that prevent dermal photodamage.Dihydromyricetin has extensive development prospect in the skin care item or drug for preparing prevention dermal photodamage.
Description
Technical field
The invention belongs to medical cosmetic fields, and in particular to dihydromyricetin is preparing prevention dermal photodamage skin care item
Or the purposes in drug.
Background technology
As atmospheric ozone layer is destroyed by getting worse, ultraviolet radiation intensity enhancing, and living standard gradually carries
The chance of height, outdoor sports, sunbath increases, and people is caused to receive excessive ultraviolet light irradiation, and light damage dermatoses are gradual
Increase.Ultraviolet light can be divided into long wave ultraviolet (UVA, 320-400nm), ultraviolet B radiation (UVB, 280- by its wavelength difference
320nm) and three kinds of short wave ultraviolet (UVC, 200-280nm).Wherein UVC almost all is inhaled by the ozone in atmospheric advection layer
Receive, thus to human body work mainly UVA and UVB.Previous studies think that UVB has energy more stronger than UVA, therefore
UVB is considered as the main reason for causing skin injury.But in recent years recent studies have shown that UVA is leading to skin injury and light
The influence that should not be underestimated also is generated in terms of aging.Because for UVB, contents of the UVA in ultraviolet radiation accounts for
95-98%；And UVA has penetration capacity more stronger than UVB, and can go directly dermal layer of the skin；Last UVA can be in 1 year
The four seasons can generate radiation effect to the mankind, and the radiation effect of UVB is concentrated mainly on summer.
The mechanism for causing light sensitive dermatoses at present is fully aware of not yet, but more it is clear that its morbidity is anti-with inflammation
It answers, the correlations such as immune response and Apoptosis, wherein again with the onset relation of apoptosis of keratinocytes and light sensitive dermatoses
The most closely.
Keratinocyte（HaCaT）It is the main cell for constituting epidermis, body can be protected from extraneous physics, chemistry
And the infringement of microorganism, maintain the stabilization of organismic internal environment.In addition to this modern research shows that keratinocyte also takes part in
Various biological processes are such as immune, inflammation, hyperplasia and tumor transformation etc..It can cause oxygen radical after UVA irradiation skins
Increase, lead to the ROS excess generations in skin.The a large amount of ROS generated in vivo can result in oxidative system and antioxidant system
It is unbalance, oxide have little time remove accumulation response to oxidative stress occurs in vivo, cause neutrophil leucocyte inflammatory infiltration and inflammation
A large amount of releases of sex factor, this is considered as a key factor for causing disease and aging to occur.On the other hand, excessive
ROS can also be by destroying cell DNA, and the activity of inducing cell apoptosis destructive enzyme destroys mitochondria, its function is caused to damage.
In conclusion the cell oxidative damage caused by ROS is the important mechanisms that UVA causes skin photoage.Therefore in order to protect
Keratinocyte safeguards its normal function, and it is an important aspect to mitigate UV-induced oxidative damage.
Dihydromyricetin (Dihydromyricelin, DHM or DMY) is the extract of vitis spp vine tea, is vine tea
Middle main active flavone compound, molecular formula C15H12O8.Modern pharmacology research shows that DHM has and removes freely
Base, anti-oxidant, anti-inflammatory, antithrombotic, the various biologicals activity such as antitumor, decompression, lipid-loweringing, analgesia.So far, there is not DHM
Relevant report with uva-resistant skin injury caused effect.
Invention content
For overcome the deficiencies in the prior art, the purpose of the present invention is to provide dihydromyricetins to prepare prevention skin light
Damage the purposes in skin care item or drug.
The technical solution that the present invention solves above-mentioned technical problem is as follows：
The dihydromyricetin of the present invention is skin care item preparing the purposes in preventing dermal photodamage product, the product
Or drug.
Further, the skin care item are mildy wash, lotion, essence, toner, face cream, facial mask, sun screen or original
Liquid.
Further, the drug is powder, granule, tablet, capsule, pill, oral solution, injection, powder-injection
Or ointment.
Further, skin care item of the present invention are sun screen.
Further, the sun screen is grouped as by the group of following percentages：Propylene glycol 2-6%, glycerine 2-6%, sea
Algae sugar 2-4%, avenabeta glucosan 2-4%, dihydromyricetin 2-6%, C20-22 Alcohol phosphate 1-4%, dimethyl silicone polymer 1-
4%, isononyl isononanoate 1-4%, methyl hydroxybenzoate 0.1-0.2%, surplus are water.
Preferably, the sun screen is grouped as by the group of following percentages：Propylene glycol 4%, glycerine 4%, trehalose 4%,
Avenabeta glucosan 4%, dihydromyricetin 6%, C20-22 Alcohol phosphate 4%, dimethyl silicone polymer 3%, isononyl isononanoate 3%,
Methyl hydroxybenzoate 0.1%, surplus are water.
Correspondingly, the preparation method of the sun screen, includes the following steps：
S1：It takes dihydromyricetin, propylene glycol, glycerine, trehalose and water to be placed in emulsification pot, is heated to 75-80 DEG C, stirring
Uniformly to get material A；
S2：Take C20-22 Alcohol phosphate, dimethyl silicone polymer and isononyl isononanoate are placed in oil cauldron, are heated to 75-80
DEG C, it stirs evenly to get material B；
S3：The material B is added in the emulsification pot of S1 and is stirred with material A, homogeneous 10-15min is cold
But；
S4：40 DEG C are cooled to, avenabeta glucosan and methyl hydroxybenzoate is added, stirs, it is cooling to get isolation
Frost.
In addition, the composition containing dihydromyricetin, which is claimed, in the present invention is preparing prevention dermal photodamage skin care item or medicine
Purposes in product.
The present invention induces the experiment of HaCaT cell models by UVA, it was demonstrated that the HaCaT that DHM can be improved after UVA irradiations is thin
The survival rate of born of the same parents makes the apoptosis of HaCaT cells reduce, and reduces the expression of inflammatory factor.The DHM is caused by reducing UVA
The ROS of HaCaT cells generate, protect the proliferation activity of cell, mitigate oxidative damage, and to damage HaCaT thin by intervening UVA
The ROS-MAPKs-NF- κ B accesses of born of the same parents, to reduce the expression of inflammatory factor；In addition, excessive lifes of the DHM by reduction ROS
At, regulate and control Bcl-2 family proteins on mitochondrial membrane and expresses, recovery mitochondrial membrane potential, the cracking of reduction caspases and PARP
Activation inhibits the Apoptosis caused by UVA, has the function that prevent dermal photodamage.
Purposes of the dihydromyricetin of the present invention in preparing prevention dermal photodamage skin care item or drug, with existing skill
Art is compared, and is had the advantage that：
（1）Dihydromyricetin is the extract of vitis spp vine tea, and the content in vine tea may be up to 30%, is pure
Natural component, abundance, be easy obtain and it is safe and non-stimulating；
（2）Dihydromyricetin can effectively inhibit UVA induction HaCaT cells ROS and generate, and mitigate oxidative damage, reduce scorching
The expression of sex factor, antiapoptotic factors inhibits Apoptosis, has the function that prevent dermal photodamage, can be widely applied to prevent
In the skin care item or drug of dermal photodamage；
（3）Clinical test show the sun screen containing dihydromyricetin can significantly decrease light injury patient skin itch,
The incidence of the malaise symptoms such as redness, pain, and it can be effective against the generation of erythema after illumination, have for dermal photodamage
There is preferable preventive and therapeutic effect, is suitble to prolonged application in confrontation dermal photodamage.
Description of the drawings
Fig. 1 is the cell survival rate of the HaCaT cells of different UVA dose irradiations；
Fig. 2 is that DHM acts on influences of the 6h later to the cell survival rate of the HaCaT cells of UVA irradiations；
Fig. 3 is that DHM acts on influences of the 6h later to the HaCaT intracellular ROS levels of UVA irradiations；
Fig. 4 is that DHM acts on HaCaT intracellular cytokines Cox-2, IL-1 β, the IL-6 gene irradiated to UVA after 6h
The influence of expression；
Fig. 5 is that DHM acts on HaCaT intracellular cytokines Bax, the Bcl-2 gene expression dose irradiated to UVA after 6h
Variation.
Specific implementation mode
It will be understood by those skilled in the art that technology disclosed in following embodiment represent the inventors discovered that in this hair
The technology of good action is played in bright practice.However, many changes can be made in disclosed specific embodiment, and
Still it obtains same or analogous as a result, without departing from the spirit and scope of the present invention.
Influences of 1 DHM of embodiment to the cell survival rate of the UVA HaCaT cells irradiated
1. solution is prepared
（1）The preparation of DHM：The dihydromyricetin powder of 2.1mg is taken to be dissolved in 65.6 μ LDMSO（The final volume of DMSO does not surpass
0.1%）, it is configured to the storing solution of 100mmol/L, is stored in the PCR pipe of 200 μ L, is kept in dark place at -20 DEG C with 10 μ L/ pipes
Refrigerator.It is taken out in advance when use, according to the needs of experiment various concentration, respective concentration is diluted to using DMEM.
（2）HaCaT cell culture mediums（Complete medium）：DMEM culture mediums（High sugar serum free medium）+ 10%FBS
（South America fetal calf serum）, its ratio be 9:1；It is added in culture medium dual anti-（Penicillin and streptomysin）, keep its final concentration of
100mg/L；Sealed membrane seals, and 4 DEG C save backup.
2. experimental method
The Survival Effects of 2.1 various dose UVA irradiation HaCaT cells
（1）The cell of logarithmic phase growth is collected according to 1x105The cell density of cells/ml is inoculated in 96 orifice plates, per hole
100 μ L, 10 experimental ports of every group of setting are inoculated with, while cell controls group, 5%CO are set2, 37 DEG C of incubations.
（2）For 24 hours, cell is grown to single layer for culture, and suction abandons culture medium and adds PBS to be distinguished to cell, experimental group is covered
Give 7.5min, 15min, 30min, 60min, 90min, the UVA irradiations of 120min, 150min；
（3）Continue to cultivate after 100 μ l complete mediums are added per hole after irradiation, after illumination for 24 hours in every hole
10 μ LMTT solution are added, continue to cultivate 4h；
（4）Culture is terminated, culture solution in hole is carefully sucked, sets 96 orifice plates after 100 μ L dimethyl sulfoxide (DMSO)s are added per hole
In low-speed oscillation 15min on shaking table, crystal is made fully to dissolve.Each hole is measured under the wavelength condition of 490nm using microplate reader
Light absorption value.
（5）Calculate the survival rate of each group UVA radiation：Cell survival rate (%)=(ODExperimental group-ODBlank well)/(ODCell controls group-
ODBlank well)×100%
Influences of 2.2 DHM to the UVA HaCaT cell survivals irradiated
（1）The cell of logarithmic phase growth is collected according to 1x105The cell density of cells/ml is inoculated in 96 orifice plates, per hole
100 μ L are inoculated with, while cell controls group is set.Every group is respectively provided with 4 experimental ports.5%CO2,37 DEG C of incubations.
（2）Cell is grown to single layer after culture for 24 hours, and the DHM of various concentration is added in experimental group（Concentration is set as 30 μ
Mol/L, 20 μm of ol/L, 10 μm of ol/L, 5 μm of ol/L, 1 μm of ol/L, 0.5 μm of ol/L, 0.1 μm of ol/L, 0 μm of ol/L）And continue to train
Support 6h.It is inhaled after 6h and abandons culture medium and add PBS to cell is covered, give UVA irradiations 60min；
（3）Irradiation inhales the drug-treated before abandoning PBS and restoring irradiation and continues to be incubated for 24 hours after completing；
（4）10 μ LMTT solution are added in every hole for 24 hours after UVA irradiations, continue to cultivate 4h；
（5）Culture is terminated, the culture medium in hole is carefully sucked.96 orifice plates are set after 100 μ L dimethyl sulfoxide (DMSO)s are added per hole
In low-speed oscillation 15min on shaking table, crystal is made fully to dissolve.Each hole is measured under the wavelength condition of 490nm using microplate reader
Light absorption value.Cell survival rate (%)=(ODExperimental group-ODBlank well)/(ODCell controls group-ODBlank well)×100%
3. data processing
Data are indicated with mean+SD, are analyzed using Student ' s t-test, and P ＜ 0.05 indicate data tool
Significant difference, P ＜ 0.01 indicate that data have pole significant difference.
4. experimental result
The survival condition of the HaCaT cells of 4.1 various dose UVA irradiations
The results are shown in Figure 1.Mtt assay determination data is shown, with the survival of the increase HaCaT cells of UVA exposure intensities
Rate is gradually reduced, and shows certain dose relationship.And in 10J/cm2When cell survival rate drop to 50% or so, therefore
We choose 10J/cm2Exposure intensity of the UVA intensity as subsequent experimental.
Influences of 4.2 DHM to the UVA HaCaT cell survivals irradiated
The results are shown in Figure 2.When the concentration of DHM is in 1 μm of ol/L or less, DHM does not increase significantly UVA（10J/
cm2）The survival rate of HaCaT cells for 24 hours after irradiation, but can significantly inhibit when the concentration of DHM is in 1~30 μm of ol/L
The decline of the survival rate of the caused HaCaT cells of UVA irradiations,And reach ceiling effect in 10 μm of ol/L.
Influences of 2 DHM of embodiment to the UVA HaCaT intracellular ROS levels irradiated
1. experimental method
（1）DCFH-DA is prepared：DCFH-DA is diluted with serum-free medium, makes its final concentration of 10 μm of ol/L；
（2）By the HaCaT cells of logarithmic phase according to 1x105The cell density of cells/ml is inoculated in 35mm Tissue Culture Dish
It is interior, per ware 2ml；
（3）Old culture solution is discarded after culture for 24 hours, and the culture medium containing various concentration DHM is added and continues to be incubated
6h；
（4）PBS is abandoned and is changed into culture medium suction after 6h, then ultraviolet irradiation 60min；
（5）PBS is changed into culture mediums containing serum after ultraviolet irradiation and continues to be incubated 30min；
（6）It inhales after 30min and abandons supernatant, wash cell with PBS is added the DCFH-DA diluted 3 times later, often ware 2ml
And continue to be incubated 30min at 37 DEG C；
（7）It inhales after 30min and abandons supernatant, after wash cell 3 times with PBS, using Laser Scanning Confocal Microscope detection ROS fluorescence
Intensity, 488nm is as excitation wavelength, and 568nm is as launch wavelength.
2. data processing
Data are indicated with mean+SD, are analyzed using Student ' s t-test, and P ＜ 0.05 indicate data tool
Significant difference, P ＜ 0.01 indicate that data have pole significant difference.
3. experimental result
The results are shown in Figure 3.HaCaT is generated largely into the cell after can be seen that UVA irradiations by total focusing results
ROS, and DHM can significantly reduce the generation of ROS after ultraviolet radioactive, but do not show certain dose dependent.
Influences of 3 DHM of embodiment to the UVA intracellular inflammatory factor gene expression levels of HaCaT irradiated.
1. experimental method
（1）The cell of logarithmic phase growth is collected according to 1x105The cell density of cells/ml is inoculated in the cell training of 35mm
It supports in ware, per ware 2ml, is placed in 37 DEG C, 5%CO2Culture in incubator；
（2）It waits for discarding old culture solution after cell is grown to single layer after culture for 24 hours, various concentration is added in experimental group
Dihydromyricetin（A concentration of 10 μm of ol/L, 5 μm of ol/L, 2.5 μm of ol/L, 1.25 μm of ol/L, 0 μm of ol/L）, cell controls group adds
Add and continue to be incubated 6h after the culture medium containing serum at 37 DEG C, is inhaled after 6h and abandon culture medium and add PBS to cell is covered, it is then purple
External exposure 60min, ultraviolet irradiation is inhaled after terminating to be abandoned PBS and adds the culture medium containing serum again, and it is different to then proceed to culture
Time；
（3）20min after ultraviolet irradiation, 40min, 60min, 80min, 100min receive cell and are carried according to Trizol methods
Take total serum IgE.
（4）The variation of the intracellular inflammatory factor gene expression dose of different time is detected using Q-PCR.
2. data processing
Data are indicated with mean+SD, are analyzed using Student ' s t-test, and P ＜ 0.05 indicate data tool
Significant difference, P ＜ 0.01 indicate that data have pole significant difference.
3. experimental result
The results are shown in Figure 4.It can be seen that DHM is capable of the reduction of concentration dependent due to ultraviolet irradiation by Q-PCR results
The expression of caused inflammatory factor COX-2, IL-1, IL-6.
Influences of 4 DHM of embodiment to the UVA intracellular antiapoptotic factors expressions of HaCaT irradiated.
1. experimental method
（1）The cell of logarithmic phase growth is collected according to 1x105The cell density of cells/ml is inoculated in the cell training of 35mm
It supports in ware, per ware 2ml, is placed in 37 DEG C, 5%CO2Culture in incubator；
（2）It waits for discarding old culture solution after cell is grown to single layer after culture for 24 hours, various concentration is added in experimental group
DHM（A concentration of 10 μm of ol/L, 5 μm of ol/L, 2.5 μm of ol/L, 1.25 μm of ol/L, 0 μm of ol/L）, the addition of cell controls group is containing serum
Culture medium after continue to be incubated 6h at 37 degree, inhale after 6h and abandon culture medium and add PBS to cell is covered, then ultraviolet irradiation
60min, ultraviolet irradiation is inhaled after terminating to be abandoned PBS and adds the culture medium containing serum again, and culture different time is then proceeded to；
（3）2h after ultraviolet irradiation, 4h, 6h, 8h, 10h, 12h, 14h collect cell and are extracted according to Trizol methods total
RNA。
（4）The variation of the intracellular antiapoptotic factors gene expression dose of different time is detected using Q-PCR.
2. data processing
Data are indicated with mean+SD, are analyzed using Student ' s t-test, and P ＜ 0.05 indicate data tool
Significant difference, P ＜ 0.01 indicate that data have pole significant difference.
3. experimental result
The results are shown in Figure 5.DHM is capable of rush apoptogene Bax of the downward of concentration dependent caused by UVA irradiations
Rising, while the expression of suppression apoptogene Bcl-2 can also be raised.
The preparation of the 5 anti-dermal photodamage sun screen of the present invention of embodiment
The sun screen of the anti-dermal photodamage of the present invention is grouped as by the group of following percentages：Propylene glycol 4%, glycerine 4%, sea
Algae sugar 4%, avenabeta glucosan 4%, dihydromyricetin 6%, C20-22 Alcohol phosphate 4%, dimethyl silicone polymer 3%, isononanoic acid are different
Nonyl ester 3%, methyl hydroxybenzoate 0.1%, surplus are water.
Preparation method：
S1：It takes dihydromyricetin, propylene glycol, glycerine, trehalose and water to be placed in emulsification pot, is heated to 80 DEG C, stirring is equal
It is even to get material A；
S2：Take C20-22 Alcohol phosphate, dimethyl silicone polymer and isononyl isononanoate are placed in oil cauldron, are heated to 80 DEG C,
It stirs evenly to get material B；
S3：The material B is added in the emulsification pot of S1 and is stirred with material A, homogeneous 15min is cooling；
S4：40 DEG C are cooled to, avenabeta glucosan and methyl hydroxybenzoate is added, stirs, it is cooling to get isolation
Frost.
The preparation of the 6 anti-dermal photodamage sun screen of the present invention of embodiment
The sun screen of the anti-dermal photodamage of the present invention is grouped as by the group of following percentages：Propylene glycol 2%, glycerine 2%, sea
Algae sugar 2%, avenabeta glucosan 2%, dihydromyricetin 2%, C20-22 Alcohol phosphate 1%, dimethyl silicone polymer 1%, isononanoic acid are different
Nonyl ester 1%, methyl hydroxybenzoate 0.1%, surplus are water.
Preparation method is the same as embodiment 5.
The preparation of the 7 anti-dermal photodamage sun screen of the present invention of embodiment
The sun screen of the anti-dermal photodamage of the present invention is grouped as by the group of following percentages：Propylene glycol 6%, glycerine 6%, sea
Algae sugar 4%, avenabeta glucosan 4%, dihydromyricetin 6%, C20-22 Alcohol phosphate 4%, dimethyl silicone polymer 4%, isononanoic acid are different
Nonyl ester 4%, methyl hydroxybenzoate 0.2%, surplus are water.
Preparation method is the same as embodiment 5.
Through examining 5 ~ 7 product of embodiment to meet the pertinent regulations requirement of QB/T 1857-2013, sense index（Color and luster,
Fragrance, structure）Meet industry regulation, physical and chemical index（PH, heat-resisting, cold-resistant, active matter）Meet industry regulation.
The anti-dermal photodamage sun screen Product Safety evaluation of test example one, the present invention（Human skin patch is tested）
1. test drug：The anti-dermal photodamage sun screen that embodiment 5-7 is prepared.
2. subjects and grouping：240 skin healths, the tested volunteer of women of no skin disease allergies, 25-55
Year, average age 34 years old are randomly divided into 3 groups, every group of 80 people.
3. test method：Qualified spot examination device is selected to take the tested material of about soya bean size with closed patch experiment method
Be placed in spot examination device in, external application special adhesive tape is covered on subject back, and tested material is removed after 24 hours, respectively remove after 2,4,
6, observation dermoreaction in 12,24,48 hours, according to《Cosmetics health specification》Middle dermoreaction grade scale records its result.
4. test result：Human skin patch result is shown：Each group subject is refreshing by the antiallergy of embodiment 5 ~ 7
The patch test of skin water observed dermoreaction at 2,4,6,12,24,48 hours and illustrates this hair without there is skin adverse reaction
Bright sun screen obtained is safe to use.
Test example two, the anti-dermal photodamage sun screen clinic drug efficacy study of the present invention
1. subjects：
The patient of 320 skin healths is screened as research object, all research object skin types are similar, patient's signature
Informed consent form voluntarily participates in experiment, excludes the heart, liver, kidney serious disease patient, excludes photosensitization patient, and research object is complied with
Property is good.Wherein man 120, female 200 are age 18-55 Sui, 35 years old average.Research object is put down using random digits table
It is divided into experiment A, B, C group and comparative example group, every group of 80 people.For these patients in traditionally equal no significant difference, partner treatment is poor
It is different not statistically significant, it is comparable.
2. test method：
Comparative example group uses commercially available Estee Lauder red pomegranate suncream；Testing A, B, component C does not use embodiment 5-7 obtained
Anti- dermal photodamage sun screen.After experimenter's cleaning skin, takes sun screen to be uniformly applied to skin exposure portion surface, applying
Eyes are avoided enter into during smearing, four groups of patients are carrying out ultraviolet light irradiation, wavelength 320-400 nm simultaneously, and irradiated site is to grind
Study carefully subjects face, irradiation time is half an hour, compares four groups of patients and occurs that erythema, a situation arises for uncomfortable reaction.
3. observation index：
Compare four groups of patient skins and the case where uncomfortable reaction occur, includes mainly itch, pain and redness, compare four groups of trouble
There is the case where erythema in person's skin, respectively without erythema, light red spot and apparent erythema.Apparent erythema is 2 cm of erythema area ＞,
Light red spot is erythema area 1-2 cm, and no erythema is 1 cm of erythema area ＜, and erythema is assessed by same evaluator, according to trouble
Person's different situations are assessed, and light red spot is visible a small amount of erythema and paler colour, hence it is evident that erythema refers to that erythema area is big and face
Color depth.
4. test result
1 each group patient skin of table is inadaptable, and there is a situation where compare
There is the comparison of erythema situation in 2 each group patient skin of table
Dermal photodamage mainly since action of ultraviolet radiation is in skin, destroys skin surface cuticula, cell is met an urgent need
Reaction ultimately causes the itch, redness, heat pain of skin surface, forms erythema, or even destroy skin integrity.It is commercially available commercially available
Estee Lauder red pomegranate suncream contains the ingredients such as red pomegranate essence, green tea extractive liquor, vitamin, titanium dioxide, have it is sun-proof,
The effect of isolation, moisturizing, by the present invention in that as a comparison case with Estee Lauder red pomegranate suncream, investigating the present invention and containing dihydro
The effect of the anti-dermal photodamage of sun screen of myricetin.
By upper table 1 it is found that four groups of patients are after light irradiates, the skin of experiment A, B, C group is not suitable with incidence and is respectively
2.5%, 3.75%, 3.75%, hence it is evident that be not suitable with incidence 8.75% less than control group, wherein with test A groups i.e. embodiment 5 be made every
It is minimum that skin from frost is not suitable with incidence；By upper table 2 it is found that the erythema of experiment A, B, C group is not suitable with incidence difference
Be 2.5%, 3.75%, 3.75%, hence it is evident that be not suitable with incidence 10% less than control group, wherein with test A groups i.e. embodiment 5 be made every
Erythema incidence from frost is minimum.The above result shows that dihydromyricetin can obviously reduce patient skin itch, redness,
The incidence of the malaise symptoms such as pain can be effective against the generation of erythema after illumination, have for dermal photodamage preferable
Preventive and therapeutic effect, and it is best with the effect of sun screen made from embodiment 5.
The above-described embodiments merely illustrate the principles and effects of the present invention, and is not intended to limit the present invention.It is any ripe
The personage for knowing this technology can all carry out modifications and changes to above-described embodiment without violating the spirit and scope of the present invention.Cause
This, institute is complete without departing from the spirit and technical ideas disclosed in the present invention by those of ordinary skill in the art such as
At all equivalent modifications or change, should by the present invention claim be covered.
Claims (2)
1. a kind of sun screen of the prevention dermal photodamage containing dihydromyricetin, it is characterised in that：The sun screen is by following
The group of percentages is grouped as：Propylene glycol 2-6%, glycerine 2-6%, trehalose 2-4%, avenabeta glucosan 2-4%, dihydromyricetin
2-6%, C20-22 alcohol phosphate 1-4%, dimethyl silicone polymer 1-4%, isononyl isononanoate 1-4%, 0 .1-0 of methyl hydroxybenzoate
.2%, surplus is water；The preparation method of the sun screen includes the following steps：
S1：It takes dihydromyricetin, propylene glycol, glycerine, trehalose and water to be placed in emulsification pot, is heated to 75-80 DEG C, stirring is equal
It is even to get material A；
S2：It takes C20-22 alcohol phosphate, dimethyl silicone polymer and isononyl isononanoate to be placed in oil cauldron, is heated to 75-80
DEG C, it stirs evenly to get material B；
S3：The material B is added in the emulsification pot of S1 and is stirred with material A, homogeneous 10-15min is cooling；
S4：40 DEG C are cooled to, avenabeta glucosan and methyl hydroxybenzoate is added, stirs, it is cooling to get sun screen.
2. the sun screen of prevention dermal photodamage as described in claim 1, which is characterized in that the sun screen is by following percentage
Group than meter is grouped as：Propylene glycol 4%, glycerine 4%, trehalose 4%, avenabeta glucosan 4%, dihydromyricetin 6%, C20-22 alcohol
Phosphate 4%, dimethyl silicone polymer 3%, isononyl isononanoate 3%, 0 .1% of methyl hydroxybenzoate, surplus is water.
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| CN112076103A (en) * | 2020-09-28 | 2020-12-15 | 嘉文丽（福建）化妆品有限公司 | A sensitive skin care product containing dihydromyricetin |
| DE102020213959A1 (en) * | 2020-11-06 | 2022-05-12 | Beiersdorf Aktiengesellschaft | Use of dihydromyricetin as an epigenetic active substance for cosmetic or dermatological skin care |
| CN114432296A (en) * | 2022-03-16 | 2022-05-06 | 中国农业大学 | Application of eupatilin in preparation of product for resisting skin photodamage |
| CN114848536A (en) * | 2022-05-13 | 2022-08-05 | 上海好肌茵生物科技有限公司 | Skin photodamage repairing composition, preparation method and application thereof |
| CN115252496B (en) * | 2022-08-18 | 2024-08-23 | 云南贝泰妮生物科技集团股份有限公司 | Preparation method of phyllanthus emblica extract and application of phyllanthus emblica extract in preventing and treating ultraviolet injury |
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| JPS63208506A (en) * | 1987-02-24 | 1988-08-30 | Nonogawa Shoji:Kk | Cosmetic |
| JPH01175932A (en) * | 1987-12-28 | 1989-07-12 | Nonogawa Shoji:Kk | Antimutagenic agent |
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2016
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103156786A (en) * | 2013-03-06 | 2013-06-19 | 张朝群 | Non-additivated antisenescence cosmetic |
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