CN106520753A - LED technology-based method for regulating melanocyte pigment synthesis - Google Patents
LED technology-based method for regulating melanocyte pigment synthesis Download PDFInfo
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Abstract
The invention belongs to the application field of biotechnology and phototherapy and specifically relates to a novel light emitting diode (LED) technology-based method for regulating melanocyte pigment synthesis. Through determined high-purity single spectrum LED light, melanocyte pigment synthesis is regulated, and proliferation and apoptosis of melanocyte are not affected. According to the method, LED yellow light with wavelength of 585+/-5 nm, light intensity of 18 mw/cm<2> and energy range of 20J/cm<2> is adopted to continuously irradiate melanocyte. Then, tyrosinase activity of melanocyte can be inhibited, melanosome maturation is inhibited to reduce mature pigment granules in the melanocyte III-IV stage, expression of genes and protein related to melanogenesis is inhibited, melanin content is reduced, and proliferation and apoptosis of melanocyte are not affected. The method is safe and effective, can be used for preparation of an in vitro study model of melanocyte, is helpful for making intervening measures for treatment and adjuvant treatment of pigment disorders, and is used for making intervening measures for non-ablative medical cosmetology.
Description
Technical field
The invention belongs to biotechnology and phototherapy application, and in particular to a kind of new based on light emitting diode (LED) technology
The method for adjusting melanophore pigment synthesis, the LED light of the high-purity unicast spectrum in the present invention by determining, adjusts melanocyte
Pigment synthesis, while do not affect melanocytic Proliferation and apoptosis.This method can be provided newly for the in vitro study of hyperpigmentation
Model, and new thinking can be provided for the clinical treatment of hyperpigmentation.
Background technology
Prior art discloses melanocyte is a kind of dendritic cell for being settled in the true epidermal junction of skin, and which produces and conveys
The color of skin and hair is determined to the melanin in horn cell around.Research report, melanocyte can cause species extremely
Various pigmented dermatosis, hypopigmentation such as vitiligo, amelanotic nevuses, piebaldism etc., pigment deepen as chloasma, freckle,
Pigmentation etc. after inflammation.Practice shows that described pigmented dermatosis sickness rate is high, and the physical and mental health of people has been brought very
Big to affect, at the same time, as its cause of disease is complicated, definite pathogenesis still do not understand, and current therapeutic effect still people not to the utmost
Meaning, therefore melanocyte biological characteristicses are further elucidated, explore the influence factor for adjusting melanophore pigment synthesis, it has also become
The focus of the art research worker, at present, still lack effectively for hyperpigmentation in clinical research practice
The cell model of in vitro study.
LED is referred to as forth generation lighting source, green light source, the features such as with energy-saving and environmental protection, life-span length, small volume,
Jing is widely used in the fields such as various instructions, display, decoration, backlight, general lighting and landscape light in city.In the world
Some developed countries expand fierce technology contest around the development of LED.500,000,000 dollars are invested from 2000 in the U.S.
Implement " National Semiconductor illumination plan ", European Union also announces to start similar " rainbow plan " in July, 2000.The Department of Science and Technology of China
Under the support that " 863 " plan, in June, 2003 proposes to develop semiconductor lighting plan first.For many years, LED illumination with
The advantage of its energy-saving and environmental protection, is paid attention to by country and governments at all levels, and relevant policies are launched respectively in various places and LED is accelerated in behave
The development of lamp.
In dermatological treatment field, aid in for technology frequently with light source, the light source adopted in current clinical research practice with laser,
Based on intense pulsed light (IPL), LED also receives much concern as a kind of new light source in recent years.LED light tune is used as near several
The study hotspot in year, in its antiinflammatory action, promotion hair growth, promotion wound healing, treatment acne, treatment cicatrix lump
The many aspects such as carbuncle are generally satisfactory.Multiple studies have shown that, the effect of light tune can be by increasing collage synthesis, reducing metal
The generation of protease (MMP) is improving skin aging.In the clinical trial that Weiss etc. is carried out, 90 patient's applications
590nm pulseds LED are treated, and treatment cycle is 2 times a week, totally 8 weeks, as a result to show, 90% patient's photoaging disease
Shape improves, and shows as skin quality improvement, and all wrinkles of socket of the eye are reduced, and skin erythema, pigment substantially mitigate.Another research discovery, non-heat effect
LED and non-exfoliative combined laser therapy photoaging, can not only produce synergism, and by LED antiinflammatory actions, alleviate
The erythematous response occurred after laser operation.In the clinical trial that Alster etc. is carried out, 20 patients receive full face 1550nm and mix
Erbium optical-fiber laser changes skin treatment, and treatment rear portion point facial zone gives 590nm LED illuminations immediately, after as a result showing 24 hours, 20
Example patient's LED area for treatment erythema is reduced compared with check plot, wherein 6 patients for receiving the treatment of higher energy density dot matrix, and 48
After hour, LED area for treatment erythema is further reduced compared with check plot, points out the effect of LED light tune to produce after reducing dot matrix laser operation
The degree of raw erythema and persistent period, and mitigate the inflammation infectious-related complication after laser therapy.
At present LED light adjusts the application acted in pigment disorders to be rarely reported, with regard to the effect of light tune research mainly for into fibre
Dimension cell, leukocyte, macrophage, mastocyte, keratinocyte, therefore, the development and application based on LED technology,
And LED illumination is for the present situation of skin pigment influence research, present inventor intends providing a kind of regulation melanophore pigment
The new method of synthesis, further, studies impact of the LED light to melanocyte biological characteristicses, so as to hinder for setting up a kind of pigment
The new model of impenetrability disease in vitro study, and important support is provided in terms of the treatment of hyperpigmentation and medical and beauty treatment.
The content of the invention
The purpose of the present invention is for the present situation of skin pigment influence research for illumination in prior art, there is provided a kind of to adjust melanocyte
The method of cytochrome synthesis, especially a kind of method for adjusting melanophore pigment synthesis based on LED technology.Present invention side
The LED light of the high-purity unicast spectrum in method by determining, adjusts melanophore pigment synthesis, while not affecting melanocytic increasing
Grow apoptosis.
Using value of the LED light in scientific research and clinic can be further expanded using this method, be that melanocytic in vitro study is carried
For new study model, for treatment and the auxiliary treatment of hyperpigmentation, to provide theory and practice basic, is non-to strip off medical treatment
The development of beauty provides new reference data.
Specifically, the invention discloses a kind of method for adjusting melanophore pigment synthesis based on LED technology, wherein, adopts
The LED gold-tinteds of specific wavelength, and specific shot dosage, Continuous irradiation are separated, culture, and the human melanocytes of purification are adjusted black
Plain cytochrome synthesis, while do not affect melanocytic Proliferation and apoptosis.
Invention further provides this method is for preparing purposes in melanocytic in vitro study model, and it is being used for
Work out the purposes in treatment and the intervening measure of auxiliary treatment hyperpigmentation, and non-medical and beauty treatment intervention is stripped off for working out
Purposes in measure.
Tyrosinase activity in melanocyte can be suppressed using the inventive method, suppress melanosome maturation (melanocyte III-IV
Phase maturation pigment granule is reduced), melanin content is reduced, suppresses the expression of melanin genesis related gene and albumen, while not affecting
Melanocytic Proliferation and apoptosis.
In the present invention, by step, step screening, determine its wavelength of LED gold-tinteds for 585 ± 5nm；
In the present invention, it is preferred to use the light intensity of LED gold-tinteds is 18mw/cm2, exposure dose is 20J/cm2Within；
In the present invention, tested, using the LED gold-tinteds of specific wavelength, specific shot dosage, Continuous irradiation is separated, training
The foster, human melanocytes of purification, as a result show, adjust melanophore pigment synthesis by this method, can suppress in melanocyte
Tyrosinase activity, suppresses melanosome ripe (melanocyte III-IV phases maturation pigment granule is reduced), reduces melanin content,
Suppress the expression of melanin genesis related gene and albumen, while not affecting melanocytic Proliferation and apoptosis.
More specifically, a kind of method for adjusting melanophore pigment synthesis based on LED technology of the invention, it is characterised in that
Which includes that, using the LED gold-tinteds of specific wavelength, using specific shot dosage, Continuous irradiation is separated, and culture, the people of purification are black
Plain cell, adjusts melanophore pigment synthesis；By following step：
1) separation, culture, Purification of Human Normal primary melanocyte：
By improvement Zheng Shi legal systems for melanocyte single cell suspension, it is inoculated in culture bottle, by passing on twice, obtains pure melanocyte
Cell；
2) LED light safety screening：
(1) LED light intervention：Unicast spectrum, highly purified HONGGUANG, gold-tinted is used respectively, and blue light, different-energy gradient continuously shine
Penetrate the melanocyte of culture acquisition 5 days；
In embodiments of the invention, the unicast for using respectively is composed, highly purified HONGGUANG is, 635 ± 5nm, 115mw/cm2, 0-103J；
Gold-tinted is, 585 ± 5nm, 18mw/cm2, 0-40J；Blue light is, 420 ± 5nm, 75mw/cm2, 0-30J；
(2) LED light is screened on melanocyte proliferation and apoptosis without the wave spectrum and energy range for affecting：
Using observation by light microscope cellular morphology, CCK-8 methods, Flow cytometry determine melanocytic propagation and apoptosis,
Impact of the different wave spectrum LED lights irradiation to the melanocyte activity of In vitro culture is evaluated, is filtered out to melanocyte proliferation and apoptosis
Without the wave spectrum and the LED light of energy range for affecting it is：LED gold-tinteds, 585 ± 5nm, 0-20J/cm2；LED blue lights, 420 ± 5nm,
0-10J/cm2；
3) according to step 2) the selection result, further screen：
Carry out LED gold-tinteds and blue light different-energy value and adjust the experiment of pigment synthesis dependency：LED gold-tinteds, 585 ± 5nm,
18mw/cm2, 0J, 5J, 10J, 20J；LED blue lights, 420 ± 5nm, 75mw/cm2, 0J, 5J, 10J；It is continuous respectively
Irradiation melanocyte 5 days, carry out observing by change of the transmission electron microscope to melanocyte melanosome quantity and Maturity,
Data statisticss, then melanophore pigment synthesis associated protein MITF, TYR, TRP-1 are surveyed using PCR, western blot
Fixed, screening determines and effectively adjusts LED light that melanophore pigment reduces and energy value scope is LED 585 ± 5nm of gold-tinted, light intensity
For 18mw/cm2, exposure dose is 20J/cm2Within.
The invention provides a kind of new method for adjusting melanophore pigment synthesis based on LED technology, in the embodiment of this method,
It is preferred that adopting wavelength for 585 ± 5nm, light intensity is 18mw/cm2,Energy range is in 20J/cm2LED gold-tinted Continuous irradiation melanocytes
Cell, can suppress tyrosinase activity in melanocyte, suppress ripe (the melanocyte III-IV phases maturation pigment of melanosome
Grain is reduced), suppress the expression of melanin genesis related gene and albumen, reduce melanin content, while not affecting melanocytic increasing
Grow apoptosis；The inventive method safely and effectively, can be used to prepare melanocytic in vitro study model.
On the other hand, based in current clinical practice for pigment increase property disease, such as pigmentation etc. after chloasma, inflammation
Therapeutic Method is various, but offer limited effectiveness, and such as chloasma at present based on prevention (sun-proof) may also be combined with Drug therapy and carry out
Laser therapy, but conventional laser therapy is usual in the side effect, therefore clinical practice such as pigmentation and recurrence after inflammation because often producing
The line therapeutic intervention for chloasma is not recommended；The present invention is carried out using the described method for adjusting melanophore pigment synthesis
Clinical trial, as a result shows, using LED gold-tinteds (585 ± 5nm) 20J/cm2, once in a week, irradiate within continuous 10 weeks, it is right
There is good improvement and curative effect in part patient with chloasma, show that the inventive method can be used for disease phototherapy and medical cosmetology field,
It is particularly useful to work out the intervening measure for the treatment of and auxiliary treatment hyperpigmentation, and does for working out the non-medical and beauty treatment that strips off
Pre- measure.
Advantages of the present invention has：
1, the gentle nature of phototherapy technology of low energy narrowed light source LED of employing, area are big, Small side effects.
2, for the cell model for setting up hyperpigmentation in vitro study can provide technical support.
3, treatment hyperpigmentation can be widely used in, clinic can not only be applied individually to any, moreover it is possible to cooperate with other laser beautifying skills
Art, strengthens its curative effect, and improves the inflammatory reaction after laser operation, accelerates the reparation for damaging.
4th, the therapeutic intervention new for formulation hyperpigmentation and assistant interventional therapeutic scheme have vital meaning.
In order to make it easy to understand, will be described in detail to the present invention by specific drawings and Examples below.Needs are referred in particular to
Go out, instantiation and accompanying drawing are merely to explanation, it is clear that one of ordinary skill in the art can according to illustrating herein,
Various amendments and change is made to the present invention in the scope of the present invention, and the scope of the present invention is also included in these amendments and change
It is interior.
Description of the drawings
Fig. 1：The primary melanocyte morphological feature of observer under optical microscope, wherein,
A：Amplification × 100, B：Amplification × 200.
Fig. 2：Flow cytometer is examined. survey the impact that LED is red, yellow, blue light illumination is to people's epidermal melanophore apoptosis.
Fig. 3：Impact of the electric Microscopic observation different-energy LED gold-tinteds (585 ± 5nm) to melanosome, wherein, A1, A2 are
Without the melanocyte of illumination, B1, B2 are Jing gold-tinted 10J/cm2Melanocyte after irradiation, C1, C2 are Jing gold-tinteds
20J/cm2Melanocyte after irradiation, I-II phases are the not immaturity melanosome in black, dye in figure black for III-IV
The melanosome of phase maturation.
Fig. 4：Impact of the different-energy LED gold-tinteds to the important protein-tyrosine expression of enzymes amount of melanin genesis, wherein, A is RT-PCR
The expression of tryrosinase mRNA is surveyed, B measures the expression of tyrosinase protein for westen-blot.
Fig. 5：LED gold-tinteds (585 ± 5nm) clinical testing treatment chloasma efficacy analysis, wherein, before A is for treatment, B is to control
After treatment.
Specific embodiment
Embodiment 1, the test for adjusting melanophore pigment synthesis based on LED technology
1st, the primary melanocytic separation of normal person and culture
Prepare the primary melanocyte single cell suspension of normal person:The in vitro normal human skin tissues for obtaining are in 75% ethanol solution
Middle immersion 30sec, then with PBS 3-4 all over to depletion of blood, aseptic condition goes down except subcutaneous tissue, blood vessel and part are true
Skin, accomplishes the slice of 1mm × 5mm, moves to 4 DEG C of digestion 16h of 0.5%dispase, and serum terminates digestion, separated epidermis
And corium, corium is removed, the epidermis for obtaining is placed in into -0.02%EDTA37 DEG C of incubation 10min of 0.25% pancreatin, epidermis base is beaten
Bottom, blows and beats into single cell suspension with suction pipe；200 μm of screen filtrations, 1500r/min centrifugation 7min, abandon supernatant, add
Medium-254s of the 5ml containing 1%HMGS, by 5 × 105The density of/bottle is inoculated in 25cm2In culture bottle, 37 DEG C, 5%CO2
Culture, changes liquid 1 time for 3 days；
It is melanocytic to pass on and purification culture：When the nearly 70%-80% of melanocyte growth merges, the melanocyte in culture bottle is discarded
Cell culture fluid, Jing after PBS liquid is rinsed twice, adds the 0.25% trypsin-EDTA Digestive systems of 2ml, 37 DEG C of conditions
Lower incubation 3-5min；Under phase contrast microscope observe melanocyte metamorphosis, when most cells dendron shorten, cell space that
When this separates, cell rounding suspends, add 10%FBS serum to terminate digestion, by cell suspension move in centrifuge tube 2200g from
After heart 5min, abandoning supernatant, PBS solution are rinsed twice, add melanocyte culture fluid, the uniform simultaneously cell counting of piping and druming
After move to Tissue Culture Flask, 37 DEG C, 5%CO2Culture, changes liquid 1 time for 3 days；Pass on 2 times and obtain pure melanocyte；
2nd, different wave length LED light Continuous irradiation melanocyte
LED light is intervened：Using unicast spectrum, highly purified HONGGUANG (635 ± 5nm, 115mw/cm2) (0-103J), gold-tinted
(585 ± 5nm, 18mw/cm2) (0-40J), blue light (42 ± 50nm, 75mw/cm2) (0-30J), different-energy gradient,
Melanocyte, Continuous irradiation 5 days is irradiated respectively；
The experiment of CCK-8 cytoactives, Annexin V/PI Bicolor-codes method detection melanocyte proliferation, apoptosis situation：
Annexin V-APC/PI Bicolor-codes method detects melanocyte apoptosis situation：Collect the black of LED intervention groups and matched group
Plain cell, adjusts cell density to be 106Individual/ml；PBS washing centrifugation 2000rpm/5min, totally 2 times, abandon supernatant, cell weight
Be suspended from the labelling buffer of 500 μ l, often pipe adds AnnexinV-APC solution and each 5 μ l of PI solution respectively, mix after
Lucifuge incubation 15min under room temperature；Flow cytomery apoptosis situation；10,000 cells of each sample detection, use
Cellquest softwares carry out Apoptosis assay；
CCK-8 methods detect cell-proliferation activity：The melanocyte of LED intervention groups and matched group, is seeded to 96 orifice plates respectively,
Per 1 Χ 10 of hole3It is individual, 3 multiple holes are set per group, blank well is not added with cell, only adds culture medium.5%CO2, 37 DEG C of incubator trainings
Foster 48h abandoning supernatants, the PBS without Ca2+ and Mg2+ of PH7.4 wash 2 times；The addition CCK-850ul per hole, 5%
CO2, 37 DEG C of incubator culture 4h；Put microplate reader absorbing light shading value (A values) is surveyed in 450nm wavelength；Cell proliferation rate=(A450
Irradiation group-A450 blank groups)/(A450 matched group-A450 blank groups) × 100%, it is repeated 3 times；
As a result show, after continuous 5 days irradiation melanocytes of different wave length LED light, gold-tinted (585 ± 5nm) 20J/cm2Within,
Blue light (420 ± 5nm) low energy 5,10J/cm2Apoptosis are had no significant effect with (as shown in Figure 2)；
3, electron microscopic observation melanosome
The melanocyte after LED illumination is taken, the MC culture medium of purification is sucked, PBS is washed 3 times, pancreatin digestion, 100r/min,
Centrifugation 5min, abandons supernatant, takes cell mass, adds the 2.5% glutaraldehyde 0.5ml of pre-cooling of new configuration, 4 DEG C of fixed 1.5h or
For more time, 20min × 3 time are rinsed with 0.1M phosphate buffers, adds 2% osmic acid fixative and fix 1.5h, use
0.1M phosphate buffers rinse 15min × 3 time, sequentially add ethanol by serial dehydration, 30% ethanol 5min, 50% second
Alcohol 5min, 70% ethanol 10min, 80% ethanol 15min, 95% ethanol 15min, 100% ethanol (anhydrous sodium sulfate process)
20min × 2 time, 100% acetone (anhydrous sodium sulfate process) 15min × 3 time.Pure acetone+embedding liquid (1:2) room temperature
Sample is embedded in embedding groove with 812 embedding mediums by 1-1.5h, pure embedding liquid ambient temperature overnight, and 48h in 60 DEG C of baking ovens is ultra-thin to cut
Piece machine section 70nm, the double dyeing of 2% acetic acid uranium-lead citrate, transmission electron microscope observing, film making；
Fig. 3 shows electron microscopic observation result, and wherein A1, A2 is the melanocyte without illumination, and B1, B2 are Jing gold-tinted 10J/cm2
Melanocyte after irradiation, C1, C2 are Jing gold-tinted 20J/cm2Melanocyte after irradiation, the I-II phases be not in black not into
Ripe melanosome, for III-IV phase ripe melanosome of the dyeing in black；As a result show, without in the melanocyte of illumination
Ripe melanosome is more, accounts for 79 ± 8%；Jing gold-tinted 10J/cm2The melanosome of irradiation after ripening is reduced to 51 ± 8%；And
The gold-tinted energy that acts on for suppressing melanosome ripe reaches 20J/cm2When become apparent from, III-IV phase ripe melanosome ratio
Example is down to 24 ± 7%；
4, Western blot detect different-energy LED gold-tinteds to the important protein-tyrosine expression of enzymes amount of melanin genesis
Electrophoresis：SDS-PAGE running gels are prepared, adds sample-loading buffer in sample, after being cooled to room temperature, by protein sample
Sample carries out electrophoresis to gel；
Transferring film：Electrophoresis after terminating cuts adhesive tape to suitable size, is balanced with transferring film buffer, 5min × 3 time；
By obtained with an equal amount of filter paper of adhesive tape and NC films, 10min in immersion transferring film buffer, membrane-transferring device from down to
On press successively carbon anode plate, 24 metafiltration paper, NC films, gel, 24 metafiltration paper, negative electrode carbon plate order place, filter paper,
Gel, NC film Accurate aligns, each step go bubble removing, upper ballast to blot liquid unnecessary on carbon plate；Switch on power,
Constant current 300mA, after transfer 1.5h, takes film；
Antibody incubation：Film bar to be measured being cut, film being washed with 0.01M PBS, 5min × 3 time add confining liquid, steady to shake,
Room temperature 2h, abandons confining liquid, washes film, 5min × 3 time with 0.01M PBS；Add one anti-2ml (1 of rabbit anti-human igg:1000),
4 DEG C of placement more than 12h；Negative control, replaces one to resist with 1%BSA, and remaining step is identical with experimental group；Abandon one it is anti-and
1%BSA, washes film, 5min × 4 time respectively with 0.01M PBS；Add the two of horseradish peroxidase to resist, steadily shake
It is dynamic, room temperature 2h；Abandon two to resist, film, 5min × 4 time are washed with 0.01M PBS；
Colour developing：Nitrite ion is added, film in darkroom, is prepared；
Fig. 4 shows testing result of the different-energy LED gold-tinteds to the important protein-tyrosine expression of enzymes amount of melanin genesis, wherein,
A surveys the expression of tryrosinase mRNA for RT-PCR, and B measures the expression of tyrosinase protein for westen-blot,
According to band shade, LED gold-tinted 20J/cm2After irradiation, in melanocyte, expression quantity of tyrosinase is decreased obviously；
Test result indicate that：Wavelength is 585 ± 5nm, and energy range is in 20J/cm2Within LED gold-tinteds, Continuous irradiation is black
After plain cell, melanosome can be suppressed ripe, reduce III-IV phases maturation pigment granule in melanocyte, suppress melanocyte to close
Into the expression of related gene and albumen, suppress tyrosinase activity in melanocyte, reduce melanin content, while not affecting black
The Proliferation and apoptosis of plain cell, safely and effectively.
2 clinical experiment of embodiment
Chloasma 1 year female middle-aged patient of medical history is chosen, LED gold-tinteds (585 ± 5nm), 20J/cm is given2, it is weekly to shine
Penetrate, after 10 weeks, chloasma is clearly better continuous therapeutic intervention at patient's bilateral buccal, achieves good curative effect, point out
The inventive method contributes to the intervening measure for working out the pigmentation diseases such as chloasma.
Claims (9)
1. a kind of method that regulation melanophore pigment based on LED technology synthesizes, it is characterised in which includes, using spy
The LED gold-tinteds of standing wave length, and specific shot dosage, Continuous irradiation are separated, culture, and the human melanocytes of purification adjust melanocyte
Cytochrome synthesizes, while not affecting melanocytic Proliferation and apoptosis；
Described its wavelength of LED gold-tinteds is 585 ± 5nm；Its exposure dose is 20J/cm2Within.
2. the method as described in claim 1, it is characterised in which includes step：
1) separation, culture, Purification of Human Normal primary melanocyte：
By improvement Zheng Shi legal systems for melanocyte single cell suspension, it is inoculated in culture bottle, by passing on twice, obtains the black of purification
Plain cell；
2) LED light safety screening：
(1) LED light intervention：Unicast spectrum, highly purified HONGGUANG, gold-tinted is used respectively, and blue light, different-energy gradient continuously shine
Penetrate the melanocyte that culture is obtained；
(2) LED light is screened on melanocyte proliferation and apoptosis without the wave spectrum and energy range for affecting：
Using observation by light microscope cellular morphology, CCK-8 methods, Flow cytometry determine melanocytic propagation and apoptosis,
Impact of the different wave spectrum LED lights irradiation to the melanocyte activity of In vitro culture is evaluated, is filtered out to melanocyte proliferation and apoptosis
Without the wave spectrum and energy range that affect；
3) according to step 2) the selection result, further screen：
Carry out LED gold-tinteds and blue light different-energy value and adjust the experiment of pigment synthesis dependency：LED gold-tinteds, 585 ± 5nm,
18mw/cm2, 0J, 5J, 10J, 20J；LED blue lights, 420 ± 5nm, 75mw/cm2, 0J, 5J, 10J；It is continuous respectively
Irradiation melanocyte 5 days, carry out observing by change of the transmission electron microscope to melanocyte melanosome quantity and Maturity,
Data statisticss, then melanophore pigment synthesis associated protein MITF, TYR, TRP-1 are surveyed using PCR, western blot
Fixed, screening determines LED light and the energy value scope for effectively adjusting that melanophore pigment is reduced.
3. the method as described in claim 2, it is characterised in that the step 2) in (1)：What Continuous irradiation culture was obtained
Melanocyte 5 days.
4. the method as described in claim 2, it is characterised in that the step 2) in (1)：Use respectively unicast spectrum,
Highly purified HONGGUANG is, 635 ± 5nm, 115mw/cm2, 0-103J；Gold-tinted is, 585 ± 5nm, 18mw/cm2, 0-40J；
Blue light is, 420 ± 5nm, 75mw/cm2, 0-30J.
5. the method as described in claim 2, it is characterised in that the step 2) in (2)：That what is screened increases to melanocyte
Grow and with LED light of the apoptosis without the wave spectrum and energy range that affect be：LED gold-tinteds, 585 ± 5nm, 0-20J/cm2；LED blue lights,
420 ± 5nm, 0-10J/cm2。
6. the method as described in claim 2, it is characterised in that the step 3) in further screening determine and effectively adjust black
The LED light and energy value scope that plain cytochrome is reduced is LED 585 ± 5nm of gold-tinted, 20J/cm2Within.
7. the method for adjusting melanophore pigment synthesis based on LED technology described in claim 1 is thin for preparing melanocyte
Purposes in the in vitro study model of born of the same parents.
8. described in claim 1 based on LED technology adjust melanophore pigment synthesis method for work out treatment with
Purposes in the intervening measure of auxiliary treatment hyperpigmentation.
9. the method for adjusting melanophore pigment synthesis based on LED technology described in claim 1 is for working out non-stripping off
Purposes in medical and beauty treatment intervening measure.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109735486A (en) * | 2019-01-31 | 2019-05-10 | 中国疾病预防控制中心辐射防护与核安全医学所 | A kind of primary melanocyte cultural method causing early ageing for studying UVB irradiation |
| CN115521900A (en) * | 2022-07-23 | 2022-12-27 | 复旦大学附属华山医院 | Visible light control angiogenesis method based on diode light modulation technology and application |
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2015
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Non-Patent Citations (1)
| Title |
|---|
| JEONG MO KIM等: "Light-emitting Diodes at 830 and 850 nm Inhibit Melanin Synthesis In vitro", 《ACTA DERM VENEREOL》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109735486A (en) * | 2019-01-31 | 2019-05-10 | 中国疾病预防控制中心辐射防护与核安全医学所 | A kind of primary melanocyte cultural method causing early ageing for studying UVB irradiation |
| CN115521900A (en) * | 2022-07-23 | 2022-12-27 | 复旦大学附属华山医院 | Visible light control angiogenesis method based on diode light modulation technology and application |
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