CN102028759A - Chinese medicine hair-darkening empirical formula and preparation method thereof - Google Patents
Chinese medicine hair-darkening empirical formula and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a Chinese medicine hair-darkening empirical formula which comprises the following Chinese medicine compositions in parts by weight: 1-40 parts of tuber fleeceflower root, 1-40 parts of black sesame, 1-40 parts of rhizoma drynariae, 1-40 parts of eclipta alba, 1-30 parts of fruit of glossy privet, 1-30 parts of dodder, 1-30 parts of selfheal, 1-30 parts of herba lycopi, 1-30 parts of mint, 1-30 parts of chuanxiong rhizome and 1-30 parts of rhizoma zingiberis. A lotion and a liniment/tincture as two preparation forms for external use can be prepared by adding a certain amount of water or ethanol into the Chinese medicine hair-darkening empirical formula for extraction. The Chinese medicine hair-darkening empirical formula has the main effects of nourishing liver and kidney, cooling the blood to stop bleeding, removing dampness to relieve itching, penetrating deeply into hair follicle to improve the activity of tyrosinase, promoting the proliferation of melanoma cells and generating more melanins to achieve the purpose of darkening the hair. A Chinese medicine compound is adopted by the invention, all the Chinese medicines are synergetic, and compared with a single Chinese medicine, the Chinese medicine hair-darkening empirical formula has the characteristics of multisystem, multi-target and multilevel exertion of all-round medicine actions and radically solves the problem that the black hair is whitened.
Description
Technical field
The invention belongs to cosmetic field, disclose a kind of Chinese medicine black hair through proved recipe and preparation method thereof.
Background technology
Along with Chinese economic development, the enhancing of people's consumption consciousness and the raising of level of consumption thereof, the hair caring product becomes people's current consumption necessary from luxury goods gradually.On the one hand, 21 century enters aging society, and the Chinese aged will reach 1.3 hundred million; Simultaneously current social youngster dog-eat-dog, psychentonia and the poliosis that causes increases.Hair dyeing, the black hair cosmetics market demand are increased day by day, have huge development space and potential market.On the other hand, because the rapid progress of beauty treatment science, people no longer satisfy the dyeing of hair dyeing cosmetic to hair, but the nutrient substance in the pursuit cosmetics is participated in metabolism, the nursing of pursuit science of hair directly.
Hair dye on the domestic and international market has experienced a series of variation, have hair dyed to semi-permanent hair dyeing up to present permanent hair dyeing from provisional, in all these hair dyes, all added the bleach of hair, promptly elder generation decomposes the oxidation of melanin in the hair and makes hair decolouring back so that colouring once more.
In realizing process of the present invention, the inventor finds that there is following problem at least in prior art: existing hair dye can have in various degree infringement to hair, and along with the metabolic of people constantly carries out, newly-generated hair still is a primary colors, especially for the people of containing poliosis in the hair, fundamentally solve the problem that makes the hair blackening, so from angle safely and effectively, the natural goods of market demand replace existing hair dye, thereby reach black hair truly.
Summary of the invention
The purpose of the embodiment of the invention is the defective at above-mentioned prior art, provides a kind of Chinese medicine black hair that can pitch-black hair through proved recipe.
Another purpose of the embodiment of the invention provides the pharmaceutical composition that the above-mentioned Chinese medicine composition that contains effective dose and pharmaceutical carrier are formed.
Another purpose of the embodiment of the invention provides this preparation of drug combination method.
The technical scheme taked of the present invention is to achieve these goals:
A kind of Chinese medicine black hair comprises following parts by weight of Chinese traditional medicine compositions through proved recipe:
1～40 part of Cacumen Platycladi, 1～40 part of Herba Ecliptae, 1～30 part of Fructus Ligustri Lucidi, 1～30 part in Fructus Mori, 1～30 part of Radix Scutellariae, 1～30 part of Herba Menthae.
The invention provides optimized technical scheme is: a kind of Chinese medicine black hair comprises following parts by weight of Chinese traditional medicine compositions through proved recipe:
10～30 parts of Cacumen Platycladis, 10～30 parts of Herba Ecliptaes, 5～20 parts of Fructus Ligustri Lucidi, 5～20 parts in Fructus Mori, 5～20 parts of Radix Scutellariaes, 5～20 parts of Herba Menthaes.
The invention provides preferred technical scheme is: a kind of Chinese medicine black hair comprises following parts by weight of Chinese traditional medicine compositions through proved recipe: 20 parts of Cacumen Platycladis, 20 parts of Herba Ecliptaes, 10 parts of Fructus Ligustri Lucidi, 10 parts in Fructus Mori, 10 parts of Radix Scutellariaes, 10 parts of Herba Menthaes.
The present invention also provides a kind of pharmaceutical composition, contains the above-mentioned Chinese medicine composition and the pharmaceutical carrier of effective dose.
Preparation of drug combination method of the present invention may further comprise the steps:
The preparation method of lotion: get Cacumen Platycladi, Herba Ecliptae, Fructus Ligustri Lucidi, Fructus Mori, Radix Scutellariae and Herba Menthae and mix, after giving the distilled water merceration 20-40min that adding mixing medical material gross weight 5-7 doubly measures in the mixing medical material, heating, boil the back and keep boiling 40min-80min, filter, get filtrate, give to add in the medicinal residues and mix the distilled water that medical material gross weight 3-5 doubly measures, boil the back and keep boiling 20-40min, filter, merge filtrate twice, leave standstill 36-60h, get supernatant,, promptly get lotion with citric acid adjust pH 6.0-6.5.
The preparation method of liniment/tincture: get Cacumen Platycladi, Herba Ecliptae, Fructus Ligustri Lucidi, Fructus Mori, Radix Scutellariae and Herba Menthae and mix, adding medical material gross weight 5-7 doubly measures 75% ethanol in the medical material to mixing, airtight immersion 20-40min under the room temperature, twice of 85 ℃ of heating and refluxing extraction, after boiling for the first time, keep boiling 20-60min, filtration back medicinal residues reuse mixing medical material gross weight 3-5 doubly measures 75% ethanol and boils for the second time, keep boiling 20-40min, filter, merge filtrate twice, leave standstill 36-60h, get supernatant with citric acid adjust pH 6.0-6.5, promptly get liniment/tincture.
The beneficial effect of the embodiment of the invention is: the present invention is primarily aimed at heat in blood and contains partially, prematured white hair person due to the Rhizoma Imperatae mistake is supported, its effect mainly is by nourishing the liver and kidney, cooling blood for hemostasis, removing damp to relieve itching, the hair follicle that gos deep into hair improves the activity of tryrosinase, promote the propagation of melanoma cells, produce more melanin, thereby reach pitch-black hair.The present invention adopts Chinese medicine compound, and each Chinese medicine Synergistic of distinguishing the flavor of is compared with single medicinal material, has multisystem, many target spots and brings into play the characteristics of comprehensive drug action at many levels, fundamentally solves the problem that hair color bleaches.
The medicinal function of each medicine of the present invention and effect:
Cacumen Platycladi: bitter and puckery flavor, little suffering.Cooling blood for hemostasis, growing and blacking hair is used for the heat in blood alopecia, early whitening of beard and hair." Japan hanako materia medica " carries: " burn extracting juice, be coated with head, black profit hair on the temple "; Compendium of Material Medica: " black profit hair on the temple ".
Herba Ecliptae: sweet in the mouth, acid, one-tenth, cold in nature.Liver-nourishing kidney-boosting, cooling blood for hemostasis, crow must hair color.Compendium of Material Medica " black beard and hair, kidney tonifying the moon "; " Bencao Jingshu " " Eclipta prostrata is apt to removing heat from blood.The white person of beard and hair ... then beard and hair blackening, and tooth is also therefore and admittedly.So the grass that bleaches at all times, when being victory now.”
Fructus Ligustri Lucidi: property is sweet, bitter, flat, the function tonification Liver and kidney, and black hair makes eye bright.Compendium of Material Medica: " reinforcing YIN-essence, strong waist knee joint bleach and send out, and make eye bright ".Can breed hair follicle cell, promote hair growth, strengthen tyrosinase activity, increase melanic generation, be used for the treatment of early whitening of beard and hair.
Fructus Mori: sweet in flavor and cold in property.The liver and the kidney tonifying, nourishing blood to promote the production of body fluid is used for early whitening of beard and hair." book on Chinese herbal medicine is looked for the truth ": " heat extraction yin nourishing ... crow palpus hair color ".
Radix Scutellariae: property hardship, cold, the function clearing away heat-damp and expelling toxic material, bacteria growing inhibiting, removing scurf and relieving itching has the effect of stronger anti-chlosma; Experimentation shows that Radix Scutellariae extract has stronger activation to tryrosinase.
Herba Menthae: the nature and flavor suffering, cold has dispelling wind-heat, refresh oneself, the effect of rash.Withered to head pruritus, hair, oils and fats too much, seborrheic alopecia and tinea capitis etc. have the improvement effect.Modern study shows that Folium Menthae extract has the effect that strengthens tyrosinase activity.
The specific embodiment
The invention will be further described below in conjunction with specific embodiment, but not as a limitation of the invention.
Embodiment 1
Take by weighing raw material (g) according to following weight:
Cacumen Platycladi 20g, Herba Ecliptae 20g, Fructus Ligustri Lucidi 10g, Fructus Mori 10g, Radix Scutellariae 10g, Herba Menthae 10g.
Chinese medicine black hair of the present invention is through the preparation method of the lotion of proved recipe:
Get Cacumen Platycladi 20g, Herba Ecliptae 20g, Fructus Ligustri Lucidi 10g, Fructus Mori 10g, Radix Scutellariae 10g and Herba Menthae 10g and mix, after mixing the distilled water merceration 30min that adds 6 times of amounts of mixing medical material gross weight in the medical material, heating, boil the back and keep boiling 1h, filter, give to add the distilled water that mixes 4 times of amounts of medical material gross weight in the medicinal residues, boil the back and keep boiling 30min, filter, merge filtrate twice, leave standstill 48h, get supernatant, with citric acid adjust pH 6.0-6.5, promptly get lotion.
Using method: after user is cleaned hair, evenly water with well-done medicine soup hair is all soaked, the massaging scalp while watering, after 15 minutes, the reuse towel wraps up, and is clean with clear water after 3 hours.
Embodiment 2
Take by weighing raw material (g) according to following weight:
Cacumen Platycladi 10g, Herba Ecliptae 25g, Fructus Ligustri Lucidi 15g, Fructus Mori 5g, Radix Scutellariae 15g, Herba Menthae 5g.
Chinese medicine black hair of the present invention is through the preparation method of the lotion of proved recipe:
Get what Cacumen Platycladi 10g, Herba Ecliptae 25g, Fructus Ligustri Lucidi 15g, Fructus Mori 5g, Radix Scutellariae 15g and Herba Menthae 5g and mix, after mixing the distilled water merceration 20min that adds 5 times of amounts of mixing medical material gross weight in the medical material, heating, boil the back and keep boiling 40min, filter, give to add the distilled water that mixes 3 times of amounts of medical material gross weight in the medicinal residues, boil the back and keep boiling 20min, filter, merge filtrate twice, leave standstill 36h, get supernatant, with citric acid adjust pH 6.0-6.5, promptly get lotion.
Using method such as embodiment 1.
Embodiment 3
Take by weighing raw material (g) according to following weight:
Cacumen Platycladi 35g, Herba Ecliptae 10g, Fructus Ligustri Lucidi 1g, Fructus Mori 25g, Radix Scutellariae 30g, Herba Menthae 1g.
Chinese medicine black hair of the present invention is through the preparation method of the lotion of proved recipe:
Get Cacumen Platycladi 35g, Herba Ecliptae 10g, Fructus Ligustri Lucidi 1g, Fructus Mori 25g, Radix Scutellariae 30g and Herba Menthae 1g and mix, after mixing the distilled water merceration 40min that adds 7 times of amounts of mixing medical material gross weight in the medical material, heating, boil the back and keep boiling 80min, filter, give to add the distilled water that mixes 5 times of amounts of medical material gross weight in the medicinal residues, boil the back and keep boiling 40min, filter, merge filtrate twice, leave standstill 60h, get supernatant, with citric acid adjust pH 6.0-6.5, promptly get lotion.
Using method such as embodiment 1.
Embodiment 4
Take by weighing raw material (g) according to following weight:
Cacumen Platycladi 30g, Herba Ecliptae 15g, Fructus Ligustri Lucidi 20g, Fructus Mori 20g, Radix Scutellariae 5g, Herba Menthae 15g.
Chinese medicine black hair of the present invention is through the preparation method of the liniment/tincture of proved recipe:
Getting Cacumen Platycladi 30g, Herba Ecliptae 15g, Fructus Ligustri Lucidi 20g, Fructus Mori 20g, Radix Scutellariae 5g and Herba Menthae 15g mixes, give to mix and add 6 times of amounts of medical material gross weight, 75% ethanol in the medical material, airtight immersion 30min under the room temperature, twice of 85 ℃ of heating and refluxing extraction, after boiling for the first time, keep boiling 40min, after 4 times of amounts of medicinal residues reuse mixing medical material gross weight, 75% ethanol boils after filtering, keep boiling 30min, filter, merge filtrate twice, leave standstill 48h, get supernatant,, promptly get liniment/tincture with citric acid adjust pH 6.0-6.5.
Using method: medicinal liquid is squirted hair, rub while spraying every day 1-2 time, each position 3-5 of reuse finger massage head minute, with comb combing hair repeatedly, make the telangiectasis under the scalp thereafter, blood flow increasing helps medicinal liquid infiltration root of hair.
Embodiment 5
Take by weighing raw material (g) according to following weight:
Cacumen Platycladi 15g, Herba Ecliptae 10g, Fructus Ligustri Lucidi 5g, Fructus Mori 15g, Radix Scutellariae 20g, Herba Menthae 20g.
Chinese medicine black hair of the present invention is through the preparation method of the liniment/tincture of proved recipe:
Getting Cacumen Platycladi 15g, Herba Ecliptae 10g, Fructus Ligustri Lucidi 5g, Fructus Mori 15g, Radix Scutellariae 20g and Herba Menthae 20g mixes, give to mix and add 5 times of amounts of medical material gross weight, 75% ethanol in the medical material, airtight immersion 20min under the room temperature, twice of 85 ℃ of heating and refluxing extraction, after boiling for the first time, keep boiling 20min, after 3 times of amounts of medicinal residues reuse mixing medical material gross weight, 75% ethanol boils after filtering, keep boiling 20min, filter, merge filtrate twice, leave standstill 36h, get supernatant,, promptly get liniment/tincture with citric acid adjust pH 6.0-6.5.
Using method such as embodiment 4.
Embodiment 6
Take by weighing raw material (g) according to following weight:
Cacumen Platycladi 1g, Herba Ecliptae 2g, Fructus Ligustri Lucidi 30g, Fructus Mori 30g, Radix Scutellariae 1g, Herba Menthae 30g.
Chinese medicine black hair of the present invention is through the preparation method of the liniment/tincture of proved recipe:
Getting Cacumen Platycladi 1g, Herba Ecliptae 2g, Fructus Ligustri Lucidi 30g, Fructus Mori 30g, Radix Scutellariae 1g and Herba Menthae 30g mixes, give to mix and add 7 times of amounts of medical material gross weight, 75% ethanol in the medical material, airtight immersion 40min under the room temperature, twice of 85 ℃ of heating and refluxing extraction, after boiling for the first time, keep boiling 1h, after 5 times of amounts of medicinal residues reuse mixing medical material gross weight, 75% ethanol boils after filtering, keep boiling 40min, filter, merge filtrate twice, leave standstill 60h, get supernatant with citric acid adjust pH 6.0-6.5, promptly get liniment/tincture.
Using method such as embodiment 4.
The present invention has following pharmacological action: the Chinese medicine black hair has the effect that activates tryrosinase, promotes cell proliferation and melanin formation promoting through proved recipe, above pharmacological action is used to prevent and treat poliosis for it is clinical provide experimental basis.
The pharmacodynamic study that the Chinese medicine black hair of the present invention preparation is carried out through proved recipe water extract (hereinafter to be referred as water extract) and ethanol extraction (hereinafter to be referred as ethanol extract) below, its method and result are as follows:
One, to the influence of tyrosinase activity
1 material and method
1.1 the Chinese medicine black hair is through the prescription of proved recipe
Cacumen Platycladi 20g, Fructus Ligustri Lucidi 10g, Herba Ecliptae 20g, Fructus Mori 10g, Radix Scutellariae 10g, Herba Menthae 10g
1.2 the preparation of Chinese medicine black hair experience formula extraction
1.2.1 the Chinese medicine black hair is through the preparation of proved recipe water extract
The Chinese medicine black hair is behind 6 times of amount distilled water of proved recipe medication material gross weight merceration 30min, and heating is after boiling, after keeping boiling 1h, filter, after the distilled water that adds 4 times of amounts of medical material total amount again boils, keep boiling 30min, filter, merge filtrate twice, boil off solvent and become paste, be dissolved in pH6.8,0.05mol/L in the phosphate buffer, be diluted to concentration and be 1g/ml, standby.
1.2.2 the Chinese medicine black hair is through the preparation of proved recipe ethanol extract
The Chinese medicine black hair is airtight immersion 30min under 6 times of amounts of proved recipe medication material gross weight, 75% ethanol room temperature, and 85 ℃ of heating and refluxing extraction twice are after boiling for the first time, keep boiling 40min, after the filtration, after 4 times of amounts of medicinal residues reuse medical material gross weight, 75% ethanol boils, keep boiling 30min, filter, merge secondary filtrate, fling to ethanol and become paste, add the 5mL dimethyl sulfoxine, reuse pH6.8,0.05mol/L phosphate buffer (PBS) are diluted to concentration and are 1g/ml, and be standby.
1.3 reagent and instrument
Reagent: tryrosinase, levodopa derive from sigma company; Sodium hydrogen phosphate (Tianjin Da Mao chemical reagent factory, lot number: 20090721); (Guangzhou newly becomes the chemical plant to potassium dihydrogen phosphate, lot number: 20010526).
Instrument: ELX800 type microplate reader (U.S. BIOTEK company); SW-CJ-IF type superclean bench (Wujiang Correspondent cleaning equipment factory); BT224S type electronic analytical balance (Sartorius company); 110S electric-heated thermostatic water bath (SHIPING company).
1.4 the configuration of reagent
1.4.3PBS the configuration of solution
First liquid: 0.05mol/L Na
2HPO
4Solution takes by weighing sodium hydrogen phosphate 7.099g, and adding distil water is to 1000ml.
Second liquid: 0.05mol/L KH
2PO
4Solution takes by weighing potassium dihydrogen phosphate 6.803g, and adding distil water is to 1000ml.
First, second liquid is divided in the brown bottle, preserves in 4 ℃ of refrigerators, time spent first, second two liquid mix by same ratio, can obtain the PBS liquid of pH=6.8.
1.4.1 the preparation of tryrosinase solution
Accurately take by weighing the 25ku tryrosinase, be dissolved among the 125ml PBS, i.e. enzyme activity unit 200u/ml, jolting is dissolved standby.
1.4.2 the preparation of levodopa solution
The 1h precision takes by weighing levodopa 20mg before the measuring, be dissolved among the 50mlPBS, i.e. and concentration 0.4mg/ml, jolting, fully the dissolving back is standby.
1.4.4 be subjected to the preparation of reagent thing
2 Chinese medicine black hair that with concentration are 1g/ml are high, medium and low three dose concentrations with freshly prepared PBS solution dilution respectively through the proved recipe concentrated solution, are respectively 1mg/ml, 0.5mg/ml, 0.25mg/ml.
2 experimental techniques
The Chinese medicine black hair of the variable concentrations of above configuration is mixed with tryrosinase 50ul through proved recipe diluent 100ul, behind 36.8 ℃ of thermostat water bath incubation 5min, add 50ul L-dopa and continue incubation 2min, immediately in microplate reader 490nm place's photometry density OD value.Each sample repeated measure 6 times is averaged, and calculates the activity ratio of tryrosinase again.
The tyrosinase activity activity ratio is pressed formula and is calculated: activity ratio %=[(C-D)-(A-B)]/(A-B) * 100
In the formula: A=A475-medicine+enzyme
B=A475-medicine-enzyme
C=A475+ medicine+enzyme
D=A475+ medicine-enzyme
Wherein,
The A value is the OD value of tyrosine solution+50ul 0.4%L-dopa solution of 100ul medicinal liquid+50ul 500u/ml.
The B value is the OD value of 100ul medicinal liquid+50ulPBS solution+50ul 0.4%L-dopa solution.
The C value is the OD value of tyrosine solution+50ul 0.4%L-dopa solution of 100ul PBS+50ul 500u/ml.
The D value is the OD value of 100ul PBS+50ul PBS solution+50ul 0.4%L-dopa solution.
3 experimental results
Table 1 result shows that the Chinese medicine black hair all has the effect that activates tryrosinase through proved recipe water extract and ethanol extract, and each Chinese prescription is along with the increase of concentration, and tyrosinase activity increases gradually, presents dose-dependence.Wherein, activity ratio is the highest be water extract when the 1mg/ml level, the activity ratio of tryrosinase is reached 32.43%.
Table 1 Chinese medicine black hair is through the influence to tryrosinase of proved recipe water extract and ethanol extract
Two, to B16 melanoma cells propagation and melanogenic influence
1 material and method
1.1 main agents and instrument
The source of cell strain: from Shanghai cell research institute of the Chinese Academy of Sciences.
Reagent: DMEM high glucose medium (U.S. Hyclone company); Hyclone (U.S. Hyclone company); Trypsin U.S. Sigma company); Tetramethyl azo tetrazolium bromide (MTT, Amersco company); Dimethyl sulfoxide (1Guanghua Chemical Plant Co., Ltd., Guangdong).
Instrument: ELX800 type microplate reader (U.S. BIOTEK company); BH2-RFL-T2 type inverted microscope (OLYMPUS company); 311 type CO
2Incubator (Thermo company); SW-CJ-IF type superclean bench (Wujiang Correspondent cleaning equipment factory); 3K18 type high speed tabletop centrifuge (U.S. Sigma company); BT224S type electronic analytical balance (Sartorius company); 110S type electric-heated thermostatic water bath (SHIPING company).
1.2 the Chinese medicine black hair is through the prescription of proved recipe
Cacumen Platycladi 20g, Fructus Ligustri Lucidi 10g, Herba Ecliptae 20g, Fructus Mori 10g, Radix Scutellariae 10g, Herba Menthae 10g.
1.3 the preparation of Chinese medicine black hair experience formula extraction
1.3.1 the preparation of water extract
After the Chinese medicine black hair adds the distilled water merceration 30min of 6 times of amounts of medical material gross weight through proved recipe, after the heated and boiled, keep seething with excitement behind the 1h, filter, after the distilled water that adds 4 times of amounts of medical material total amount again boils, keep boiling 30min, filter, merge filtrate twice, be condensed into extractum, use pH6.8, the 0.05mol/L phosphate buffer dissolves and is diluted to concentration is 1g/ml, and 4 ℃ of preservations are standby.
1.3.2 the preparation of alcohol extract
The Chinese medicine black hair is airtight immersion 30min under 6 times of amounts of proved recipe medication material gross weight, 75% ethanol room temperature, and 85 ℃ of heating and refluxing extraction twice are after boiling for the first time, keep boiling 40min, after the filtration, after 4 times of amounts of medicinal residues reuse medical material total amount, 75% ethanol secondary boils, keep boiling 30min, filter, merge filtrate twice, fling to ethanol and become paste, reuse pH6.8,0.05mol/L phosphate buffer (PBS) dissolved dilution to concentration is 1g/ml, 4 ℃ of preservations are standby.
2 experimental techniques
2.1 the cultivation of mice B16 melanotic tumor cell
Mouse black-in tumor cell B16 cell strain, (contain the 100U/ml penicillin with the high sugared DMEM culture medium that contains 10%PBS, when the 100g/ml streptomycin) being cultured to nearly fusion state, with 0.25% trypsin and 0.02%EDTA digestion, centrifugal, with above-mentioned culture medium re-suspended cell, the adjustment cell density is 1 * 105/ml, plants 96 orifice plates (being used for cell proliferation test) and 6 orifice plates (being used for melanin content measures) respectively.Behind the cell inoculation 24h, change liquid, measure after continuing to hatch 48h (cell proliferation test) and 72h (melanin content mensuration) with the fresh culture that adds medicine.
2.2 tried preparation of thing solution and experiment grouping
Water extract and alcohol extract are respectively with serum-free DMEM culture medium dilution 100mg/ml, filtration sterilization, be 10mg/ml with the coubling dilution dilution again, 5mg/ml, 2.5mg/ml, 1.25mg/ml, 0.625mg/ml, 0.3125mg/ml, 0.15625mg/ml, 0.078125mg/ml the blank group is to contain the culture medium (pure water or ethanol) of being tried the thing coordinative solvent.
2.3MTT method detects cell proliferation
The B16 cell inoculation of getting place's exponential phase is in 96 orifice plates, and (inoculum density is 1 * 10 to every hole 100l
4/ hole).Behind the 24h, inhale and go culture medium, handle the fresh culture (10mg/ml of factor to contain variable concentrations, 5mg/ml, 2.5mg/ml, 1.25mg/ml, 0.625mg/ml, 0.1325mg/ml, 0.0613mg/ml, 0.0306mg/ml) change liquid, establish not dosing blank, after continuing to cultivate 48h, culture medium is gone in suction, with the PBS washing, adds fresh culture, every hole adds 5g/LMTT solution 10l, put incubator and cultivate 4h, inhale and go culture medium, every hole adds 150lDMSO, shake well is measured every hole 490nm light absorption value with microplate reader.Each experiment repeats 4 times.
2.4 the mensuration of melanin content
In 6 orifice plates, (inoculum density is 1 * 10 to every hole 1000l with the B16 cell inoculation of exponential phase
5/ ml).Behind the 24h, inhale and go culture medium, handle the fresh culture (5mg/ml of factor to contain variable concentrations, 2.5mg/ml, 1.25mg/ml, 0.625mg/ml, 0.3125mg/ml) changing liquid, every hole 2000l establishes not dosing blank, after continuing to cultivate 72h, culture medium is gone in suction, with PBS washing, trypsinization, stop digestion, collect Digestive system.Digestive system changes the 1.5mlEP pipe respectively in each hole, and is centrifugal, removes supernatant, adds 10% trichloroacetic acid, centrifugal, removes supernatant, adds 1NNaOH solution, and 80 ℃ of dissolvings extremely fully of water-bath change ELISA Plate over to, measure every hole 490nm light absorption value with microplate reader immediately.Each experiment repeats 4 times.Melanocyte appreciation rate (A medicine group-A blank group)/A blank group * 100%.
2.5 statistical method
The data of surveying with all using
The one way analysis of variance check is adopted in expression, and statistic processes uses the SPSS13.0 statistical software to finish.
3 experimental results
3.1 the mensuration of mice B16 melanoma cells proliferation activity
Table 2 result shows, with the blank group relatively, the effect (P＜0.05) that the Chinese medicine black hair has obvious promotion melanocyte propagation through the proved recipe water extract when concentration (0.15625,0.3125,0.625,1.25mg/ml), concentration 0.625mg/ml appreciation rate is the highest; Concentration on cell proliferation when (5,10mg/ml) has the obvious suppression effect; Concentration 2.5mg/ml compares no difference of science of statistics (P＞0.05) with the blank group; The Chinese medicine black hair has obvious facilitation (P＜0.05) to melanocyte propagation through the proved recipe ethanol extract when concentration (0.15625,0.3125mg/ml), and appreciation rate is higher during concentration 0.3125mg/ml; Concentration shows as the effect of obvious inhibition cell proliferation when (0.625,1.25,5,10mg/ml), concentration 10mg/ml suppression ratio is the highest; Concentration 0.078125mg/ml and blank group no difference of science of statistics (P＞0.05).
Table 2 Chinese medicine black hair is through the influence to B16 melanoma cells proliferation activity of proved recipe water extract and ethanol extract
Annotate: * represents to compare with the blank group, and there is obvious facilitation P＜0.05; # represents to compare with the blank group, and there is obvious inhibitory action P＜0.05.
3.2 the mensuration of melanin content in the mice B16 melanoma cells
Table 3 result shows, with the blank group relatively, the Chinese medicine black hair more all has significant difference through each concentration of proved recipe water extract and blank group, shows as obviously the increasing of melanin content (P＜0.05), melanin content was the highest when concentration was 1.25mg/ml.The Chinese medicine black hair is all variant through each concentration group of proved recipe ethanol extract and the comparative statistics of blank group, shows as obviously the increasing of melanin content (P＜0.05), and melanin content was the highest when concentration was 2.5mg/ml.
Table 3 Chinese medicine black hair through proved recipe water extract and ethanol extract to the synthetic influence of melanoma cells melanocyte (
The A value)
Annotate: * represents to compare with the blank group, and there is obvious facilitation P＜0.05; # represents to compare with the blank group, and there is obvious inhibitory action P＜0.05.
Three, conclusion
By originally experimental studies have found that: the Chinese medicine black hair all has the effect that activates tryrosinase, promotes cell proliferation and melanin formation promoting through proved recipe water extract and ethanol extract.
Above-described embodiment, the present invention specific embodiment a kind of more preferably just, the common variation that those skilled in the art carries out in the technical solution of the present invention scope and replacing all should be included in protection scope of the present invention.
Claims (6)
1. a Chinese medicine black hair comprises following parts by weight of Chinese traditional medicine compositions through proved recipe:
1～40 part of Cacumen Platycladi, 1～40 part of Herba Ecliptae, 1～30 part of Fructus Ligustri Lucidi, 1～30 part in Fructus Mori, 1～30 part of Radix Scutellariae, 1～30 part of Herba Menthae.
2. Chinese medicine black hair according to claim 1 is characterized in that through proved recipe: comprise following parts by weight of Chinese traditional medicine compositions:
10～30 parts of Cacumen Platycladis, 10～30 parts of Herba Ecliptaes, 5～20 parts of Fructus Ligustri Lucidi, 5～20 parts in Fructus Mori, 5～20 parts of Radix Scutellariaes, 5～20 parts of Herba Menthaes.
3. Chinese medicine black hair according to claim 1 is characterized in that through proved recipe: comprise following parts by weight of Chinese traditional medicine compositions: 20 parts of Cacumen Platycladis, 20 parts of Herba Ecliptaes, 10 parts of Fructus Ligustri Lucidi, 10 parts in Fructus Mori, 10 parts of Radix Scutellariaes, 10 parts of Herba Menthaes.
4. a pharmaceutical composition is characterized in that, contains each described Chinese medicine composition and pharmaceutical carrier in the claim 1～3 of effective dose.
5. the described preparation of drug combination method of claim 4 may further comprise the steps:
The preparation method of lotion: get Cacumen Platycladi, Herba Ecliptae, Fructus Ligustri Lucidi, Fructus Mori, Radix Scutellariae and Herba Menthae and mix, after giving the distilled water merceration 20-40min that adding mixing medical material gross weight 5-7 doubly measures in the mixing medical material, heating, boil the back and keep boiling 40min-80min, filter, get filtrate, give to add in the medicinal residues and mix the distilled water that medical material gross weight 3-5 doubly measures, boil the back and keep boiling 20-40min, filter, merge filtrate twice, leave standstill 36-60h, get supernatant,, promptly get lotion with citric acid adjust pH 6.0-6.5.
6. the described preparation of drug combination method of claim 4 may further comprise the steps:
The preparation method of liniment/tincture: get Cacumen Platycladi, Herba Ecliptae, Fructus Ligustri Lucidi, Fructus Mori, Radix Scutellariae and Herba Menthae and mix, adding medical material gross weight 5-7 doubly measures 75% ethanol in the medical material to mixing, airtight immersion 20-40min under the room temperature, twice of 85 ℃ of heating and refluxing extraction, after boiling for the first time, keep boiling 20-60min, filtration back medicinal residues reuse mixing medical material gross weight 3-5 doubly measures 75% ethanol and boils for the second time, keep boiling 20-40min, filter, merge filtrate twice, leave standstill 36-60h, get supernatant with citric acid adjust pH 6.0-6.5, promptly get liniment/tincture.
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| CN102526221A (en) * | 2012-01-18 | 2012-07-04 | 王秀峰 | Pharmaceutical formulation for preventing grey hair and hair loss |
| WO2015006897A1 (en) * | 2013-07-15 | 2015-01-22 | Tian Yi Er | Formula of hair-darkening chinese medicine preparation and preparation method thereof |
| CN105486647A (en) * | 2015-09-29 | 2016-04-13 | 吉林农业大学 | Extraction and performance determination for hair-blackening active composition in cacumen platycladi |
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| CN102526221B (en) * | 2012-01-18 | 2013-09-11 | 王秀峰 | Pharmaceutical formulation for preventing grey hair and hair loss |
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| CN113209264A (en) * | 2021-06-01 | 2021-08-06 | 山东中医药大学 | Traditional Chinese medicine composition for reducing grey hair and promoting growth of black hair and preparation method thereof |
| CN113318026A (en) * | 2021-06-15 | 2021-08-31 | 福州膳柏健康产业有限公司 | Formula of traditional Chinese medicine hair care liquid for enhancing tyrosinase activity |
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