CN115025027B - Biological factor eyelash nourishing liquid - Google Patents

Biological factor eyelash nourishing liquid Download PDF

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CN115025027B
CN115025027B CN202210672094.1A CN202210672094A CN115025027B CN 115025027 B CN115025027 B CN 115025027B CN 202210672094 A CN202210672094 A CN 202210672094A CN 115025027 B CN115025027 B CN 115025027B
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extract
hair
astragalus
azone
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CN115025027A (en
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罗彬相
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Shanghai Youya Pharmaceutical Technology Co ltd
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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00Cosmetics or similar toiletry preparations
    • A61K8/18Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/97Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
    • A61K8/9783Angiosperms [Magnoliophyta]
    • A61K8/9794Liliopsida [monocotyledons]
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01HNEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H4/00Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00Cosmetics or similar toiletry preparations
    • A61K8/18Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/67Vitamins
    • A61K8/678Tocopherol, i.e. vitamin E
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00Cosmetics or similar toiletry preparations
    • A61K8/18Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/97Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
    • A61K8/9783Angiosperms [Magnoliophyta]
    • A61K8/9789Magnoliopsida [dicotyledons]
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61QSPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q1/00Make-up preparations; Body powders; Preparations for removing make-up
    • A61Q1/02Preparations containing skin colorants, e.g. pigments
    • A61Q1/10Preparations containing skin colorants, e.g. pigments for eyes, e.g. eyeliner, mascara
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61QSPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q7/00Preparations for affecting hair growth
    • YGENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02ATECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

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  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
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  • Engineering & Computer Science (AREA)
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  • Developmental Biology & Embryology (AREA)
  • Mycology (AREA)
  • Microbiology (AREA)
  • Cell Biology (AREA)
  • Dermatology (AREA)
  • Environmental Sciences (AREA)
  • Cosmetics (AREA)
  • Medicines Containing Plant Substances (AREA)

Abstract

The invention discloses a biological factor eyelash nourishing liquid, which comprises astragalus extract, dried ginger extract, vitamin E, glycerol, azone, propylene glycol, sodium hydroxide and lactic acid. The agrobacterium rhizogenes containing FGF-10 gene Ri plasmid is used for infecting the astragalus explant, so that FGF-10 protein can be produced, the activity of germinal cells and the environment of hair follicles are improved, the activity and content of hair follicle stimulating growth factors are improved, the regeneration of hair mother cells and hair papilla cells is stimulated, and the hair regeneration speed can be improved by combining the drug property of astragalus root. The gingerol and Jiang Xitong contained in the dried ginger can promote blood circulation, increase nutrition of hair follicle, stimulate hair follicle and promote hair regeneration. The azone and propylene glycol can produce synergistic effect, act on the lipid bilayer of cells to increase the fluidity of the bilayer, promote the diffusion of FGF protein and medicine between cells, and thus increase the permeation of effective substances. The invention has the characteristics of safety and quick response.

Description

Biological factor eyelash nourishing liquid
Technical Field
The invention relates to the technical field of cosmetics, in particular to a biological factor eyelash nourishing liquid.
Background
Women during pregnancy and lactation have alopecia and eyelash drop. The fetus develops during pregnancy, the demand for the mother is large, and in the process, the deficiency of trace elements is easy to occur, and the conditions of the body are reduced, endocrine disturbance is caused, so that eyelash is fallen. During lactation, the nutrition demand is gradually increased along with the growth of children, and breast feeding can cause malnutrition of the mother and hair or eyelashes to fall off.
The eyelash is shed to seriously influence the image, extra pressure can be brought to females, bad emotion during pregnancy has bad influence on child development, females during lactation can be more fidgety, and postpartum depression risk is increased. However, the administration of the liquid for women during pregnancy and lactation requires a certain care, and thus a liquid for nourishing eyelashes, which can promote the rapid growth of eyelashes, is needed. Alopecia is more studied in the market, but it is less studied for eyelash shedding during pregnancy and lactation.
Therefore, a biological factor eyelash nourishing liquid with safe design and quick effect and a preparation method thereof are necessary.
Disclosure of Invention
The invention aims to provide a biological factor eyelash nourishing liquid and a preparation method thereof, which are used for solving the problems in the background technology.
In order to solve the technical problems, the invention provides the following technical scheme: a biological factor eyelash nourishing liquid comprises radix astragali extract, rhizoma Zingiberis extract, vitamin E, glycerol, azone, propylene glycol, sodium hydroxide, and lactic acid.
According to the technical scheme, the biological factor eyelash nourishing liquid comprises, by mass, 3-5 parts of astragalus extract, 1-3 parts of dried ginger extract, 0.5-2 parts of vitamin E, 2-8 parts of glycerol, 0.8-3.5 parts of azone, 0.4-3 parts of propylene glycol, 0.1-0.6 part of sodium hydroxide and 0.5-4 parts of lactic acid.
According to the technical scheme, the biological factor eyelash nourishing liquid comprises, by mass, 5 parts of astragalus extract, 1.5 parts of dried ginger extract, 0.5 part of vitamin E, 6 parts of glycerin, 1.2 parts of azone, 1.5 parts of propylene glycol, 0.3 part of sodium hydroxide and 1.6 parts of lactic acid.
A biological factor eyelash nourishing liquid and a preparation method thereof comprise the following specific steps:
step one: preparation of astragalus extract: cleaning radix astragali hairy root, grinding with liquid nitrogen in a mortar, adding deionized water, stirring, standing for 3-5 hr, centrifuging at 8000-12000rpm for 20-40min, and collecting supernatant to obtain radix astragali extract;
step two: preparing dried ginger extract: soaking rhizoma Zingiberis powder in 90% ethanol for 24 hr, slowly percolating at a rate of 1-3 ml per minute, collecting 850ml of primary percolate, storing in a container, percolating until the percolate is colorless, aroma and pungency of rhizoma Zingiberis are light, collecting percolate, concentrating to soft extract below 60deg.C, adding primary percolate, mixing, filtering, standing, clarifying, and filtering to obtain rhizoma Zingiberis recens fluid extract;
step three: grinding 10% glycerol, adding purified water, swelling, adding radix astragali extract, rhizoma Zingiberis extract, propylene glycol, azone, propylene glycol, VE, and lactic acid, stirring, and adjusting pH to 5.5-6.5 with sodium hydroxide.
According to the technical scheme, the astragalus extract is extracted from hairy roots of explants, and the explants can be roots, stems and leaves.
According to the technical scheme, the hairy roots of the explants are infected with the astragalus explants by agrobacterium rhizogenes: taking stem as explant, pH of culture medium is 5-6, sucrose concentration is 2-5%, age of the explant is 30-35D, ultrasonic treatment time is 4s, pre-culture and co-culture are 2-3D, infection time is 5-10min, and exogenous hormone is added to 0.1mg/mLNAA.
According to the technical scheme, the agrobacterium rhizogenes is one of A4, 15834, 1025 and 1000 strains.
According to the technical scheme, the Ri plasmid of the agrobacterium rhizogenes contains FGF-10 genes, the FGF10 genes are amplified by PCR, and the PCR is connected to the Ri plasmid after double digestion.
According to the technical scheme, the astragalus hairy roots are subjected to PCR detection: according to the designed pair of FGF-10 gene primers, the same band as the gene primer appears in the band (627 bp), while the untransformed astragalus explant does not amplify the band, proving that the Agrobacterium Ri plasmid has been integrated into the genome of hairy roots.
According to the technical scheme, the hairy roots of the astragalus membranaceus are transferred into a liquid culture medium for expansion proliferation culture: and (3) culturing for 25-35D by using an MS liquid culture medium, wherein the concentration of sucrose is 30g/L, and JA of 30 mu mol/L is added.
Astragalus mongholicus is a leguminous Chinese medicinal material. The main functions are as follows: tonify qi, strengthen superficies, expel pus, promote urination, promote granulation and promote hair growth.
The FGF10 has wide clinical application, mainly comprises the functions of promoting injury repair (such as burn, cornea injury, lung injury and the like), has a certain effect on facial red blood filaments, and can promote stem cell proliferation and differentiation. Especially, FGF10 can improve the activity of germinal cells and the environment of hair follicles, increase the activity and content of the growth factors for promoting hair follicles, and stimulate the regeneration of hair mother cells and hair papilla cells, thereby achieving the effect of promoting hair regeneration.
The radix astragali explant is infected by agrobacterium rhizogenes containing FGF-10 gene Ri plasmid, FGF-10 protein can be produced in the radix astragali explant, and the prepared radix astragali extract pair can promote the rapid growth of hair and promote the regeneration speed by combining the drug property of radix astragali.
Gingerol and Jiang Xitong contained in rhizoma Zingiberis can dilate blood vessel and promote blood circulation, and they are all easily dissolved in ethanol. The ginger fluid extract is prepared from ethanol and has the functions of dilating subcutaneous blood vessels, promoting blood circulation, increasing nutrition of hair follicles, stimulating hair follicles and promoting hair regeneration.
Azone (Azone) is an excellent percutaneous absorption enhancer. The preparation method is mainly applied to the lipid bilayer of cells to increase the fluidity of the bilayer, reduce the phase transition temperature and promote the diffusion of FGF protein and medicine among cells, thereby increasing the permeation of the medicine. Azone has transdermal promoting effect on many lipophilic and hydrophilic drugs, and can produce synergistic effect with Propylene Glycol (PG). Azone has optimal transdermal enhancement effect especially for low concentration medicine, and has low side effect. Therefore, azone and PG are selected as the composite transdermal enhancer.
Since the extract of zingiber officinale is lipophilic, and glycerol has thickening, moisturizing and lubricating effects and can be used as a carrier solvent, PG is commonly used as a solvent, a latent solvent, a wetting agent, a humectant and a preservative in medicaments, and has a penetration promoting effect, glycerol and PG are added to increase the dissolution and uniformly disperse the medicaments.
VE has high boiling point, is stable to heat, is easily dissolved in ethanol, is nontoxic, has obvious skin aging resisting effect, and can improve skin roughness, scales, smoothness, wrinkles and the like, so VE is selected as an antioxidant.
Compared with the prior art, the invention has the following beneficial effects: in the present invention, the number of the components,
(1) The agrobacterium rhizogenes containing FGF-10 gene Ri plasmid is used for infecting the astragalus explant, so that FGF-10 protein can be generated in the astragalus explant, the activity of germinal cells and the environment of hair follicles are improved, the activity and the content of the hair follicle stimulating growth factors are improved, and the regeneration of hair mother cells and hair papilla cells is stimulated, so that the purposes of promoting hair regeneration and improving the hair regeneration speed can be achieved by combining the drug property of the astragalus;
(2) Gingerol substances such as gingerol and Jiang Xitong contained in the dried ginger can dilate blood vessels and promote blood circulation, and are all easily dissolved in ethanol, and ginger fluid extract is prepared by ethanol, and mainly has the effects of dilating subcutaneous blood vessels, promoting blood circulation, increasing hair follicle nutrition, stimulating hair follicles and promoting hair regeneration;
(3) Azone is an excellent percutaneous absorption promoter, and Propylene Glycol (PG) can produce synergistic effect when being combined, and mainly acts on a cell lipid bilayer to increase the fluidity of the bilayer, reduce the phase transition temperature, and promote the diffusion of FGF protein and medicines through cells, thereby increasing the permeation of effective substances.
Detailed Description
The following description of the technical solutions in the embodiments of the present invention will be clear and complete, and it is obvious that the described embodiments are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
The invention provides the technical scheme that: a biological factor eyelash nourishing liquid comprises radix astragali extract, rhizoma Zingiberis extract, vitamin E, glycerol, azone, propylene glycol, sodium hydroxide, and lactic acid.
According to the technical scheme, the biological factor eyelash nourishing liquid comprises, by mass, 3-5 parts of astragalus extract, 1-3 parts of dried ginger extract, 0.5-2 parts of vitamin E, 2-8 parts of glycerol, 0.8-3.5 parts of azone, 0.4-3 parts of propylene glycol, 0.1-0.6 part of sodium hydroxide and 0.5-4 parts of lactic acid.
According to the technical scheme, the biological factor eyelash nourishing liquid comprises, by mass, 5 parts of astragalus extract, 1.5 parts of dried ginger extract, 0.5 part of vitamin E, 6 parts of glycerin, 1.2 parts of azone, 1.5 parts of propylene glycol, 0.3 part of sodium hydroxide and 1.6 parts of lactic acid.
A biological factor eyelash nourishing liquid and a preparation method thereof comprise the following specific steps:
step one: preparation of astragalus extract: cleaning radix astragali hairy root, grinding with liquid nitrogen in a mortar, adding deionized water, stirring, standing for 3-5 hr, centrifuging at 8000-12000rpm for 20-40min, and collecting supernatant to obtain radix astragali extract;
step two: preparing dried ginger extract: soaking rhizoma Zingiberis powder in 90% ethanol for 24 hr, slowly percolating at a rate of 1-3 ml per minute, collecting 850ml of primary percolate, storing in a container, percolating until the percolate is colorless, aroma and pungency of rhizoma Zingiberis are light, collecting percolate, concentrating to soft extract below 60deg.C, adding primary percolate, mixing, filtering, standing, clarifying, and filtering to obtain rhizoma Zingiberis recens fluid extract;
step three: grinding 10% glycerol, adding purified water, swelling, adding radix astragali extract, rhizoma Zingiberis extract, propylene glycol, azone, propylene glycol, VE, and lactic acid, stirring, and adjusting pH to 5.5-6.5 with sodium hydroxide.
According to the technical scheme, the astragalus extract is extracted from hairy roots of explants, and the explants can be roots, stems and leaves.
According to the technical scheme, the hairy roots of the explants are infected with the astragalus explants by agrobacterium rhizogenes: taking stem as explant, pH of culture medium is 5-6, sucrose concentration is 2-5%, age of the explant is 30-35D, ultrasonic treatment time is 4s, pre-culture and co-culture are 2-3D, infection time is 5-10min, and exogenous hormone is added to 0.1mg/mLNAA.
According to the technical scheme, the agrobacterium rhizogenes is one of A4, 15834, 1025 and 1000 strains.
According to the technical scheme, the Ri plasmid of the agrobacterium rhizogenes contains FGF-10 genes, the FGF10 genes are amplified by PCR, and the PCR is connected to the Ri plasmid after double digestion.
According to the technical scheme, the astragalus hairy roots are subjected to PCR detection: according to the designed pair of FGF-10 gene primers, the same band as the gene primer appears in the band (627 bp), while the untransformed astragalus explant does not amplify the band, proving that the Agrobacterium Ri plasmid has been integrated into the genome of hairy roots.
According to the technical scheme, the hairy roots of the astragalus membranaceus are transferred into a liquid culture medium for expansion proliferation culture: and (3) culturing for 25-35D by using an MS liquid culture medium, wherein the concentration of sucrose is 30g/L, and JA of 30 mu mol/L is added.
Case 1:
a biological factor eyelash nourishing liquid comprises radix astragali extract, rhizoma Zingiberis extract, vitamin E, glycerol, azone, propylene glycol, sodium hydroxide, and lactic acid.
According to the mass ratio, astragalus extract 5 parts, dried ginger extract 1.5 parts, vitamin E0.5 parts, glycerin 6 parts, azone 1.2 parts, propylene glycol 1.5 parts, sodium hydroxide 0.3 part and lactic acid 1.6 parts.
A biological factor eyelash nourishing liquid and a preparation method thereof comprise the following specific steps:
step one: preparation of astragalus extract: cleaning hairy roots of radix astragali, grinding with liquid nitrogen in a mortar, adding deionized water, stirring, standing for 4 hr, centrifuging at 10000rpm for 30min, and collecting supernatant to obtain radix astragali extract;
step two: preparing dried ginger extract: soaking rhizoma Zingiberis powder in 90% ethanol for 24 hr, slowly percolating at a rate of 2ml per minute, collecting primary percolate 850ml, storing in a container, continuously percolating until the percolate is colorless, aroma and pungency of rhizoma Zingiberis are light, collecting percolate, concentrating at 60deg.C below to soft extract, adding primary percolate, mixing, filtering, standing, clarifying, and filtering to obtain rhizoma Zingiberis recens fluid extract;
step three: grinding 10% glycerol, adding purified water, swelling, adding radix astragali extract, rhizoma Zingiberis extract, propylene glycol, azone, propylene glycol, VE, and lactic acid, stirring, and adjusting pH to 5.5-6.5 with sodium hydroxide.
According to the technical scheme, the astragalus extract is extracted from hairy roots of explants, and the roots are selected as the explants.
According to the technical scheme, the hairy roots of the explants are infected with the astragalus explants by agrobacterium rhizogenes: taking the stem as an explant, wherein the pH value of the culture medium is 5-6, the concentration of sucrose is 3%, the age of the explant is 30-35D, the ultrasonic treatment time is 4s, the pre-culture and co-culture are carried out for 2-3D, the infection time is 10min, and 0.1mg/mLNAA exogenous hormone is added.
According to the technical scheme, the agrobacterium rhizogenes is an A4 strain.
According to the technical scheme, the Ri plasmid of the agrobacterium rhizogenes contains FGF-10 genes, the FGF10 genes are amplified by PCR, and the PCR is connected to the Ri plasmid after double digestion.
According to the technical scheme, the astragalus hairy roots are subjected to PCR detection: according to the designed pair of FGF-10 gene primers, the same band as the gene primer appears in the band (627 bp), while the untransformed astragalus explant does not amplify the band, proving that the Agrobacterium Ri plasmid has been integrated into the genome of hairy roots.
According to the technical scheme, the hairy roots of the astragalus membranaceus are transferred into a liquid culture medium for expansion proliferation culture: and (3) culturing 30D by using an MS liquid culture medium, wherein the concentration of sucrose is 30g/L, and JA of 30 mu mol/L is added.
Case 2:
a biological factor eyelash nourishing liquid comprises radix astragali extract, rhizoma Zingiberis extract, vitamin E, glycerol, azone, propylene glycol, sodium hydroxide, and lactic acid.
According to the technical scheme, the biological factor eyelash nourishing liquid comprises, by mass, 4 parts of astragalus extract, 3 parts of dried ginger extract, 0.8 part of vitamin E, 4 parts of glycerol, 0.8 part of azone, 0.9 part of propylene glycol, 0.2 part of sodium hydroxide and 2 parts of lactic acid.
A biological factor eyelash nourishing liquid and a preparation method thereof comprise the following specific steps:
step one: preparation of astragalus extract: cleaning hairy roots of radix astragali, grinding with liquid nitrogen in a mortar, adding deionized water, stirring, standing for 5 hr, centrifuging at 8000rpm for 40min, and collecting supernatant to obtain radix astragali extract;
step two: preparing dried ginger extract: soaking rhizoma Zingiberis powder in 90% ethanol for 24 hr, slowly percolating at a rate of 1-3 ml per minute, collecting 850ml of primary percolate, storing in a container, percolating until the percolate is colorless, aroma and pungency of rhizoma Zingiberis are light, collecting percolate, concentrating to soft extract below 60deg.C, adding primary percolate, mixing, filtering, standing, clarifying, and filtering to obtain rhizoma Zingiberis recens fluid extract;
step three: grinding 10% glycerol, adding purified water, swelling, adding radix astragali extract, rhizoma Zingiberis extract, propylene glycol, azone, propylene glycol, VE, and lactic acid, stirring, and adjusting pH to 5.5-6.5 with sodium hydroxide.
According to the technical scheme, the astragalus extract is extracted from hairy roots of explants, and the explants can be roots, stems and leaves.
According to the technical scheme, the hairy roots of the explants are infected with the astragalus explants by agrobacterium rhizogenes: taking the stem as an explant, wherein the pH value of the culture medium is 5-6, the sucrose concentration is 4%, the age of the explant is 30-35D, the ultrasonic treatment time is 4s, the pre-culture and co-culture are carried out for 3D, the infection time is 10min, and 0.1mg/mLNAA exogenous hormone is added.
According to the technical scheme, the agrobacterium rhizogenes is 1025 strains.
According to the technical scheme, the Ri plasmid of the agrobacterium rhizogenes contains FGF-10 genes, the FGF10 genes are amplified by PCR, and the PCR is connected to the Ri plasmid after double digestion.
According to the technical scheme, the astragalus hairy roots are subjected to PCR detection: according to the designed pair of FGF-10 gene primers, the same band as the gene primer appears in the band (627 bp), while the untransformed astragalus explant does not amplify the band, proving that the Agrobacterium Ri plasmid has been integrated into the genome of hairy roots.
According to the technical scheme, the hairy roots of the astragalus membranaceus are transferred into a liquid culture medium for expansion proliferation culture: and (3) culturing 35D by using an MS liquid culture medium, wherein the concentration of sucrose is 30g/L, and JA of 30 mu mol/L is added.
In order to detect the effect of the biological factor eyelash nourishing liquid, the in vitro test is specially carried out:
1. skin irritation test
The skin irritation test of applying eyelash nourishing liquid for multiple times is adopted. Taking 5 rabbits, and adding 10% sodium sulfideDehairing on both sides of the spine, each side area being about 40cm 2 After 24 hours, the left dehairing area of the rabbit was coated with about 1g of eyelash tonic solution, the right dehairing area was coated with about 1g of purified water 1 time a day for 7 days, the medicines were washed off with warm water after the last application of the medicines for 24 hours, the irritation reactions of the skin of the rabbit after 1 hour, 24 hours, 48 hours and 72 hours after the residual eyelash tonic solution was removed were observed, and the scoring was performed with reference to table 1:
TABLE 1 evaluation index of skin irritation test
Each rabbit was scored according to the scoring principle of table 1, averaged and the results are shown in table 2:
table 2 skin irritation test results (n=5)
From the experimental results, the eyelash tonic liquid has slight irritation, and the skin is free from edema after application.
2. Eyelash nourishing liquid external transdermal experiment
The treated rat skin, with the horny layer facing upwards, is fixed on Franz diffusion cell of a transdermal apparatus, and eyelash nourishing liquid 2g (for eliminating air bubbles) is uniformly coated on one side of the horny layer, and the effective diffusion area is 2.48cm 2 7ml of physiological saline containing 20% PEG400 was added to the receiving tank as a receiving solution. The diffusion cell is placed in a constant temperature water bath with the temperature of 37+/-1 ℃ and the stirring speed of 300r/min, 1ml is sampled at 1, 2, 4, 6, 8, 10, 12 and 24 hours respectively, and the physiological saline containing 20 percent PEG400 with the same temperature and the same volume is supplemented. Filtering the obtained sample by using a microporous filter membrane with the size of 0.45 mu m, taking 20 mu l of continuous filtrate for sample injection, measuring the content, and calculating the accumulated transmission amount of a certain time unit area according to the following formula:
Qs={Cn*Vo+V}/S
wherein, qs: accumulated permeation quantity per unit area at the nth sampling; cn: the concentration in the receiving liquid at the nth sampling; vo: a diffusion cell volume; v: sampling volume; s: diffusion area.
TABLE 3 cumulative permeation per unit area
The visual analysis method test results are used for analysis, and the skin permeability of the eyelash nourishing liquid is good. Azone and propylene glycol play a major role in promoting penetration of eyelash nourishing liquid in the percutaneous penetration process.
3. Test of Effect of eyelash nourishment liquid on growth of body hair in dehairing zone of mouse
Collecting 20 Kunming mice, cutting back hair, applying depilatory (10% NaS) to the back of the mice for about 2-3 min, cleaning with warm water, cleaning with the back of the mice smooth, and removing hair with a depilating area of about 3×3cm 2 . The next day mice were randomly divided into 2 groups of 10 mice each, each with blank, mascara solution applied to the depilatory area 2 times daily, 0.2 ml/each for 15 consecutive days. Taking 10 longest hairs from the same position of each mouse dehairing area on 16 days, measuring the lengths of the longest hairs by using a microscopic micrometer under a microscope, and calculating the average value of the hair lengths of all groups of mice by using the average lengths of the 10 hairs to represent the hair length of each mouse; the mice were then sacrificed by cervical removal, the fur from the back dehairing area was cut off, a round piece of fur was taken from the same position in each mouse dehairing area with a 14mm punch, all hairs on the piece were scraped off with a scalpel, and the weights were measured on a ten-thousandth balance to calculate the average hair weights for each group.
TABLE 4 Effect of eyelash tonic on body hair length and weight in the depilatory region of mice
The experimental results are shown in Table 4, and the hair length and hair weight per body surface area of the mascara-fed mice are increased to different extents as compared to the blank matrix group, and the mascara-fed mice are longer and denser than the body hair of the blank matrix group, indicating that the mascara-fed can promote regeneration of the body hair of the chemically dehaired mice.
It is noted that relational terms such as first and second, and the like are used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions. Moreover, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus.
Finally, it should be noted that: the foregoing description is only a preferred embodiment of the present invention, and the present invention is not limited thereto, but it is to be understood that modifications and equivalents of some of the technical features described in the foregoing embodiments may be made by those skilled in the art, although the present invention has been described in detail with reference to the foregoing embodiments. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims (1)

1. A biological factor eyelash nourishing liquid is characterized in that: comprises radix astragali extract, rhizoma Zingiberis extract, vitamin E, glycerol, azone, propylene glycol, sodium hydroxide, and lactic acid;
the astragalus extract is extracted from hairy roots of explants, wherein the explants are roots, stems and leaves;
the hairy roots of the explants are infected with the astragalus explants by agrobacterium rhizogenes: taking a stem segment as an explant, wherein the pH value of a culture medium is 5-6, the concentration of sucrose is 4%, the age of the explant is 30-35D, the ultrasonic treatment time is 4s, the pre-culture and co-culture are carried out for 3D, the infection time is 10min, and 0.1mg/mLNAA exogenous hormone is added;
the agrobacterium rhizogenes is 1025 strains;
the Ri plasmid of the agrobacterium rhizogenes contains FGF-10 genes, the FGF10 genes are amplified by PCR, and the agrobacterium rhizogenes is connected to the Ri plasmid after double enzyme digestion;
according to the mass ratio, 4 parts of astragalus extract, 3 parts of dried ginger extract, 0.8 part of vitamin E, 4 parts of glycerin, 0.8 part of azone, 0.9 part of propylene glycol, 0.2 part of sodium hydroxide and 2 parts of lactic acid;
the preparation method of the biological factor eyelash nourishing liquid comprises the following specific steps:
step one: preparation of astragalus extract: cleaning hairy roots of radix astragali, grinding with liquid nitrogen in a mortar, adding deionized water, stirring, standing for 5 hr, centrifuging at 8000rpm for 40min, and collecting supernatant to obtain radix astragali extract;
step two: preparing dried ginger extract: soaking rhizoma Zingiberis powder in 90% ethanol for 24 hr, slowly percolating at a rate of 1-3 ml per minute, collecting 850ml of primary percolate, storing in a container, percolating until the percolate is colorless, aroma and pungency of rhizoma Zingiberis are light, collecting percolate, concentrating to soft extract below 60deg.C, adding primary percolate, mixing, filtering, standing, clarifying, and filtering to obtain rhizoma Zingiberis extract;
step three: grinding 10% glycerol, adding purified water, swelling, adding radix astragali extract, rhizoma Zingiberis extract, propylene glycol, azone, vitamin E, and lactic acid, stirring, and adjusting pH to 5.5-6.5 with sodium hydroxide.
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