CN106265211B - Cistanche salsa supernatant mixture, preparation method thereof and skin barrier repair effect thereof - Google Patents

Cistanche salsa supernatant mixture, preparation method thereof and skin barrier repair effect thereof Download PDF

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CN106265211B
CN106265211B CN201510297211.0A CN201510297211A CN106265211B CN 106265211 B CN106265211 B CN 106265211B CN 201510297211 A CN201510297211 A CN 201510297211A CN 106265211 B CN106265211 B CN 106265211B
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cistanche
supernatant
skin
supernatant mixture
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王滢
孙懿
范倩文
陈默
祝乐
赵亚
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Jia Hua Bio Tech Ltd Shanghai
Shanghai Jahwa United Co Ltd
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Shanghai Jahwa United Co Ltd
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Abstract

The invention discloses a method for extracting a cistanche supernatant mixture from cistanche, which comprises the following steps: decocting in water bath; concentrating, and precipitating with organic solvent; taking the supernatant, and centrifuging to obtain a cistanche salsa supernatant mixture. In some embodiments of the invention, the extraction method further comprises a step of ultrafiltration after the addition of decoction extraction. The invention also relates to a cistanche salsa supernatant mixture prepared by the method and application of the cistanche salsa supernatant mixture in cosmetics with skin barrier repair effect.

Description

Cistanche salsa supernatant mixture, preparation method thereof and skin barrier repair effect thereof
Technical Field
The invention relates to the field of natural medicinal chemistry and cosmetics, in particular to a cistanche supernatant mixture obtained by extracting in a water decoction mode and then adopting an alcohol precipitation supernatant mode. In addition, the invention also relates to the skin barrier repair effect of the cistanche salsa supernatant mixture extracted by the method, and particularly relates to the application of the cistanche salsa supernatant mixture in preparing cosmetics with the skin barrier repair effect.
Background
Skin barrier function includes physical barrier, pigment barrier, nerve barrier, immune barrier and other aspects related to skin function, and most of the skin barriers referred to in our daily life refer to the physical skin barrier. From the viewpoint of cell differentiation and tissue formation, the physical barrier function of the skin depends not only on the stratum corneum of the epidermis, but also on the entire layer structure of the epidermis; from the aspects of biochemical composition and functional role, the physical barrier structure of epidermis is closely related not only to the lipid of epidermis, but also to various proteins, water, inorganic salts and other metabolites of epidermis.
Modern skin science believes that the healthy comfort of skin appearance is inseparable from the perfection of barrier function, which is also contrary to the most important health view of "consolidating superficial body and protecting" in traditional theory. Impaired or dysfunctional skin barrier is an important root cause of the problem of disfiguring skin. The barrier function of the skin is disrupted and water in the epidermis is lost through the stratum corneum, causing the skin to become dry, thereby causing or aggravating some skin disorders. Many skin diseases such as senile skin pruritus, atopic dermatitis, psoriasis, acne, and dry skin are associated with a decrease in skin barrier function.
The skin barrier is mainly composed of a series of substances/structures such as cornification envelopes and lipid membranes, intermediate filaggrin, keratin, cornification desmosomes, lamellar bodies and horny layer keratinocyte intercellular substances, tight junctions and the like, and can prevent water loss, prevent external invasion and maintain homeostasis in an organism. A part of epidermal cells start from the basal layer and finally migrate to the skin surface through a series of growth and differentiation processes to form a completely keratinized cell mantle, which is the material basis of the barrier structure of the epidermal stratum corneum. The above process involves two main classes of proteins: namely cytokeratin (Keratin) and Keratin intermediate filament-associated protein (KI-PAP). These important intermediate silk-related proteins include, silk polymerization protein (Filaggrin), Loricrin (Loricrin), Involucrin (Involucrin), Keratinocyte Transglutaminase (TGK), and the like. The intermediate silk related protein is an important substance for forming the barrier structure of the stratum corneum of the skin, and the abnormal expression of the protein can cause a series of skin diseases characterized by the damage of the barrier function of the skin.
The integrity of the skin barrier, while also being associated with the viability of keratinocytes, facilitates the renewal of the skin cells with proliferation.
Cistanche deserticola (Cistanches Deserticola Y.C.Ma) is a perennial parasitic medicinal plant of Cistanche genus of Orobanchaceae family, and has been found to have the effects of resisting aging, enhancing human immunity, resisting fatigue and the like. The cistanche deserticola grows in extremely arid desert and hot desert, and a set of system adapting to the environment is formed under the long-term action of natural conditions. If the cistanche deserticola leaves are degenerated into scale shape, the body is rich in water to form meat quality, and a unique root suction device is provided. Meanwhile, under the stimulation of severe environment, the cistanche can generate a certain class of secondary metabolites to protect the living of organisms.
At present, most of researches on cistanche deserticola focus on the anti-aging and free radical scavenging effects of an ethanol precipitation part of an aqueous extract. The research on the supernatant part after the cistanche is subjected to water extraction and alcohol precipitation is not found. That is, the prior art does not disclose the cistanche supernatant mixture of the present invention, which is obtained by extracting the cistanche supernatant mixture in a water decoction manner and then precipitating the supernatant with alcohol. Surprisingly, after the cistanche salsa supernatant mixture acts on keratinocytes, the expression of two proteins (Involucrin and Claudin-1) which are closely related to the skin barrier in the cells is improved, and the effect of the extract on protecting the skin barrier is proved.
Disclosure of Invention
The invention aims to provide a cistanche salsa supernatant mixture with a skin barrier repair effect. The sugar content range of the cistanche salsa supernatant mixture prepared by the method is 23.37-62.60%, and the protein content range is 15.74-35.57%. In the invention, the expression conditions of two proteins (Involucrin and Claudin-1) closely related to the skin barrier in cells are mainly detected after the cistanche salsa supernatant mixture acts on keratinocytes. The research proves that the expression of the two proteins is improved, and further proves that the cistanche salsa supernatant mixture has the effect of protecting the skin barrier.
In one aspect, the invention provides a method for extracting a cistanche supernatant mixture from cistanche, which comprises the following steps:
a) decocting in water bath;
b) concentrating, and precipitating with organic solvent;
c) taking the supernatant, and centrifuging to obtain a cistanche salsa supernatant mixture.
In a preferred embodiment of the present invention, the method for extracting a cistanche supernatant mixture from cistanche further comprises a process of performing ultrafiltration after step a). In some embodiments of the invention, ultrafiltration is capable of retaining molecules with a molecular weight above 10000.
In some preferred embodiments, step a) is carried out at a temperature of 80 to 100 ℃. In some preferred embodiments, the weight ratio of cistanche salsa raw material to water in step a) is 1:20 to 1: 30.
In some preferred embodiments, the organic solvent of step b) is a lower alcohol having 1 to 4 carbon atoms. In some preferred embodiments, the organic solvent of step b) is acetone.
In another aspect of the present invention, there is provided a cistanche supernatant mixture prepared by the method of the present invention.
In some preferred embodiments, the cistanche supernatant mixture of the present invention has a sugar content ranging from 23.37% to 62.60% and a protein content ranging from 15.74% to 35.57% by weight.
In another aspect of the present invention, it is provided that the cistanche supernatant mixture prepared according to the method of the present invention can be applied to skin external preparations, health foods and medicines, and particularly, can be applied to cosmetics, and can be added in different amounts according to different types of preparations.
In another aspect of the present invention, there is provided a cosmetic having skin barrier repair effect, comprising the cistanche supernatant mixture prepared according to the method of the present invention and a cosmetically acceptable excipient. In a preferred embodiment of the invention, the cosmetic product further comprises another additive having an aging effect on skin barrier repair. In one embodiment, the cosmetic is selected from: face cleaning lotion, cosmetic water, lotion, cream, jelly and facial mask. In a preferred embodiment, the cistanche salsa supernatant mixture is present in the skin external preparation in an amount of 0.0001-20% (w/w), preferably 0.001-10% (w/w), and more preferably 0.01-5% (w/w).
The composition for external preparation for skin is a general concept of all ingredients generally used for the external part of skin, and may be, for example, a cosmetic composition or a pharmaceutical composition. The cosmetic composition may be a basic cosmetic, a face makeup cosmetic, a body cosmetic, a hair care cosmetic, etc., and the formulation thereof is not particularly limited and may be appropriately selected depending on the purpose.
The cosmetic composition also contains different cosmetically acceptable media or matrix excipients according to different formulations and purposes.
The cosmetically, dermatologically or pharmaceutically acceptable vehicle that can be used in the composition for external application to skin of the present invention is in the form of a water phase, an oil phase, a gel, a wax-in-water emulsion, an oil-in-water emulsion or a water-in-oil emulsion. The aqueous phase is a mixture of one or more water-soluble or dispersible components, which may be liquid, semi-solid, or solid at room temperature (25 ℃). The vehicle includes or may be in the form of a suspension, dispersion or solution in an aqueous or hydro-alcoholic vehicle, which may contain a thickening or gelling agent. The person skilled in the art can select suitable product forms, the components contained therein, based on the knowledge of the person skilled in the art.
The composition may comprise an aqueous phase which may contain water or a mixture of water and at least one hydrophilic organic solvent such as an alcohol, in particular a linear or branched lower monohydric alcohol containing from 2 to 5 carbon atoms, such as ethanol or propanol; polyols, such as propylene glycol, sorbitol, glycerol, panthenol or polyethylene glycols and mixtures thereof.
When the composition of the invention is in the form of an emulsion, the composition may also optionally comprise a surfactant.
The composition may also comprise film-forming polymers such as polyurethanes, polyacrylic acid homo-or copolymers, polyesters, hydrocarbon-based resins and/or silicone resins. The polymer may be dissolved or dispersed in a cosmetically acceptable vehicle and optionally combined with a plasticizer.
The compositions of the present invention may also comprise an oil phase containing oil-soluble or oil-dispersible components that are liquid at room temperature (25 ℃) and/or substances that are oily or waxy at room temperature, such as waxes, semisolids, gums, and mixtures thereof. The oil phase may also contain an organic solvent.
Typically liquid at room temperature, suitable oily substances include: hydrocarbon-based oils of animal origin, such as perhydrosqualene; hydrocarbon-based vegetable oils, such as liquid triglycerides of C4-10 fatty acids, e.g. heptanoic acid or octanoic acid triglycerides, or oils, e.g. sunflower oil, corn oil, soybean oil, grapeseed oil, castor oil, avocado oil, octanoic/decanoic acid triglycerides, jojoba oil; linear or branched hydrocarbons of mineral or synthetic origin, such as liquid paraffin and its derivatives, vaseline; synthetic esters and ethers, in particular esters of fatty alcohols, such as isopropyl myristate, 2-ethylhexyl palmitate, 2-octyldodecyl stearate, isostearyl isostearate; hydroxylated esters, such as isostearyl lactate, octyl hydroxystearate, octyl dodecyl hydroxystearate, heptanoates, octanoates and decanoates of fatty alcohols; polyol esters such as propylene glycol dicaprylate, neopentyl glycol diheptanoate, diethylene glycol diisononanoate, and pentaerythritol esters; c12-26-containing fatty alcohols, such as octyldodecanol, 2-butyloctanol, 2-hexyldecanol, 2-undecylpentadecanol, oleyl alcohol; fluoro and/or fluorosilicone oils based in part on hydrocarbons, silicone oils, volatile or non-volatile linear or cyclic polymethylsiloxanes which are liquid or semi-solid at room temperature, such as cyclic polydimethylsiloxanes and polydimethylsiloxanes, optionally containing phenyl groups, such as phenyltrimethicones, silicones and mixtures thereof.
The composition of the present invention may further comprise any component commonly used in the cosmetic field. These components include preservatives, aqueous phase thickeners (extract biopolymers, synthetic polymers) and fatty phase thickeners, fragrances, hydrophilic and lipophilic active agents and mixtures thereof.
The compositions of the invention may also comprise an additional particulate phase, which may be a pigment and/or a pearlescent agent and/or a filler used in cosmetic compositions.
Pigments may be present in the composition, suitable inorganic pigments include titanium oxide, zirconium oxide and cerium oxide as well as zinc oxide, iron oxide and ferric blue; suitable organic pigments include barium, strontium, calcium and aluminum lakes and carbon black.
Pearling agents may be present in the composition, suitable pearling agents include mica coated with titanium oxide, iron oxide or natural pigments.
Fillers may be present in the composition, suitable fillers include talc, silica, zinc stearate, mica, kaolin, nylon powder, polyethylene powder, teflon, starch, boron nitride, copolymer microspheres, such as silicone resin microbeads.
The oil phase of the compositions of the present invention may comprise one or more waxes, gums or mixtures thereof. Waxes include hydrocarbon-based waxes, fluoro waxes, and/or silicone waxes, and may be derived from vegetable, mineral, animal, and/or synthetic sources. Suitable waxes include beeswax, carnauba wax, candelilla wax, paraffin wax, microcrystalline wax, ozokerite; synthetic waxes include polyethylene waxes, silicone waxes containing C16-45. Gums are generally polydimethylsiloxanes or sodium carboxymethylcellulose or extracts, and semisolid substances are generally hydrocarbon-based compounds, such as lanolin and its derivatives.
The compositions of the present invention may be formulated into any suitable product form. Such product forms include, but are not limited to, aerosol sprays, creams, lotions, solids, liquids, dispersions, foams, gels, lotions, mousses, ointments, powders, patches, pomades, solutions, hand pump sprays, sticks, masks and towelettes. The compositions of the present invention may be conveniently used to prepare or as cosmetic, dermatological or pharmaceutical topical products by various methods well known in the art.
The composition for external skin preparations of the present invention may include one or more of the following ingredients: anti-allergic agents, antimicrobial agents, antioxidants, chelating agents, colorant depigmenting agents, emollients, emulsifiers, exfoliants, film formers, fragrances, humectants, insect repellents, lubricants, pharmaceutically active agents, moisturizers, light stabilizers, preservatives, skin protectants, skin penetration enhancers, sunscreens, stabilizers, surfactants, thickeners, viscosity modifiers, vitamins, or any combination thereof.
Drawings
FIG. 1 is a flow chart of the extraction process of the cistanche salsa supernatant mixture of the present invention.
FIG. 2 shows the results of the effect of the mixture of cistanche salsa supernatants of the present invention on the viability of epidermal cells.
FIG. 3 shows the effect of the mixture of cistanche salsa supernatant of the present invention on the expression of the protein associated with skin barrier repair.
Detailed Description
The invention is further illustrated below with reference to specific examples. It is to be understood, however, that these examples are illustrative only and are not to be construed as limiting the scope of the present invention. Test methods in which specific conditions are not specified in the following examples are generally carried out under conventional conditions or under conditions recommended by the manufacturer. All percentages and parts are by weight unless otherwise indicated.
The instrument used in the examples of the present invention is as follows:
SHB-III circulating water type multipurpose vacuum pump: zhengzhou great wall science, Industrial and trade, Inc.
Model 85-2 constant temperature magnetic stirrer: shanghai si le instruments ltd.
DHG-9070A type electric heating constant-temperature air-blast drying oven: shanghai, a constant technology, Inc.
N-1100D-WD vacuum rotary evaporator: japan EYELA Corp.
UV260 visible spectrophotometer: shimadzu, japan.
GL-21M low temperature high speed centrifuge: shanghai centrifuge research institute, Inc.
Labconco large freeze dryer: labconco, USA.
Edwards mini freeze dryer: edwards, UK.
Full wavelength double plate multifunctional instrument (microplate reader): novostar of BMG Labtech in USA
A cell culture box: U.S. Thermo serial II
The following examples 1 to 6 describe the preparation method of the cistanche salsa supernatant mixture according to the present invention and examine the preparation process. The cistanche deserticola is produced in Gansu or Xinjiang.
Example 1
Collecting Cistanchis herba bulb 500g, extracting at 80 deg.C in water bath at a ratio of 1:20, concentrating the extract to 3-5L, ultrafiltering, collecting the trapped fluid with molecular weight above about 10000, concentrating, adding organic solvent (lower alcohol such as methanol and ethanol, acetone), stirring for precipitation, standing, centrifuging at 4 deg.C overnight, and centrifuging to obtain supernatant.
Example 2
Collecting Cistanchis herba bulb 500g, extracting at 80 deg.C in water bath at a ratio of 1:30, concentrating the extract to 3-5L, ultrafiltering, collecting the trapped fluid with molecular weight above about 10000, concentrating, adding organic solvent (lower alcohol such as methanol and ethanol, and acetone), stirring for precipitation, standing, centrifuging at 4 deg.C overnight, and centrifuging to obtain supernatant.
Example 3
Collecting Cistanchis herba bulb 500g, extracting at 80 deg.C in water bath at a ratio of 1:50, concentrating the extract to 3-5L, ultrafiltering, collecting the trapped fluid with molecular weight above about 10000, concentrating, adding organic solvent (lower alcohol such as methanol and ethanol, and acetone), stirring for precipitation, standing, centrifuging at 4 deg.C overnight, and centrifuging to obtain supernatant.
Example 4
Collecting Cistanchis herba bulb 500g, extracting at 100 deg.C in water bath at a ratio of 1:20, concentrating to 3-5L, ultrafiltering, collecting retentate with molecular weight of about 10000 or more, concentrating, adding organic solvent (lower alcohol such as methanol and ethanol, and acetone), stirring for precipitation, standing, centrifuging at 4 deg.C overnight, and centrifuging to obtain supernatant.
Example 5
Collecting Cistanchis herba bulb 500g, extracting at 100 deg.C in water bath at a ratio of 1:30, concentrating to 3-5L, ultrafiltering, collecting retentate with molecular weight of about 10000 or more, concentrating, adding organic solvent (lower alcohol such as methanol and ethanol, and acetone), stirring for precipitation, standing, centrifuging at 4 deg.C overnight, and centrifuging to obtain supernatant.
Example 6
Collecting Cistanchis herba bulb 500g, extracting at 100 deg.C in water bath with the ratio of Cistanchis herba material to water added at each time of 1:50, concentrating the extract to 3-5L, ultrafiltering, collecting trapped fluid with molecular weight above about 10000, concentrating, adding organic solvent (lower alcohol such as methanol and ethanol, and acetone), stirring for precipitation, standing, precipitating, centrifuging, standing at 4 deg.C overnight, and centrifuging to obtain supernatant.
Example 7: determination of sugar content
The sugar content of the cistanche supernatant mixtures prepared in examples 1 to 6 was measured by the phenol-sulfuric acid method.
The principle of the phenol-sulfuric acid method is that a sugar sample to be measured is hydrolyzed into monosaccharide under the condition of strong acid, the monosaccharide is dehydrated in concentrated sulfuric acid to generate furfural, and then the furfural and phenol form a yellow compound, and the yellow compound can be measured by utilizing a colorimetric method.
Reagent: phenol (analytically pure) is steamed again to prepare 50g/L (5%, w/v) aqueous solution; concentrated sulfuric acid (analytically pure, d 1.84 g/cm)3) (ii) a Standard solution (150. mu.g/mL): 7.5mg of glucose which had been dried was taken, dissolved in water, and dissolved to 50 mL. The sample solution to be tested is prepared into a solution with the solid content of 1 percent and diluted by 50 times.
The concentrations of the upper samples are respectively 0,30,60,90,120 and 150 mu g/ml during the measurementAdding 0.2mL of solution, 0.4mL of 5% phenol, adding 2.0mL of concentrated sulfuric acid after shaking, standing at room temperature for 30min, and determining OD with blank tube (0#) as control490
Taking 0.2mL sample, determining OD according to the same method of standard solution490
The results are shown in the following table:
TABLE 1
Figure BDA0000730529580000091
Example 8: protein content determination (Kjeldahl method)
The protein content of the cistanche supernatant mixtures prepared in examples 1 to 6 was measured. The protein in the sample is decomposed under the condition of catalytic heating, and the generated ammonia is combined with sulfuric acid to generate the sulfuric acid. Alkalifying and distilling to free ammonia, absorbing with boric acid, titrating with sulfuric acid or hydrochloric acid standard titration solution, instantly obtaining nitrogen content according to acid consumption, and multiplying by corresponding conversion coefficient to obtain protein content.
The results are shown in the following table:
TABLE 2
Figure BDA0000730529580000092
By comparing and analyzing the sugar content and the protein content in tables 1-2, and considering the operability and cost economy of industrial production, we chose the extract as the subject with the extraction temperature of 100 ℃ and the material ratio of 1:20, and examined the skin barrier repair effect.
Therefore, the supernatant mixture of cistanche deserticola prepared in example 4 was taken in the following examples to prepare a crude drug concentration sample of 1g/ml with a solid content of 11.38%.
In the work efficiency evaluation, samples were prepared with a mother liquor concentration of 1%, i.e. 10 mg/ml.
Example 9: skin barrier effect of cistanche salsa supernatant mixture
(1) Effect of cistanche salsa supernatant mixture on epidermal cell vitality
Mu.l of cell culture medium containing 0.2% growth factor was added to each well of a 96-well plate, and 150. mu.l of fibroblast suspension was added to the plate using a discharge gun. The cell inoculation amount is 8500 cells/hole, and the cells are placed at 37 ℃ and 5% CO2Culturing in an incubator for 24 h. Sucking 150 μ l of culture medium, adding 150 μ l/well of culture medium containing Cistanchis herba supernatant mixture to make final concentration of 0.1mg/ml and 0.3mg/ml, and adding 150 μ l/well of culture medium containing Cistanchis herba precipitate to make final concentration of 0.02mg/ml and 0.1 mg/ml. After the culture is continued for 48h, the culture medium is replaced again according to the method described previously, after 24h, the culture medium is sucked out, after being washed by PBS, 125 mu l of 0.2mg/ml XTT reaction liquid (containing 0.1% PMS and Sigma) is added, and after the culture is continued for 2h, the absorbance is measured at 450nm on a microplate reader.
The experimental results are shown in fig. 2, and the cistanche salsa supernatant mixture respectively increases the epidermal cell viability by 7.06% and 13.34% under the conditions of the concentrations of 0.1mg/ml and 0.3 mg/ml. And the cistanche salsa precipitation part reduces the activity of epidermal cells by 15.71 percent and 19.37 percent respectively under the conditions of the concentration of 0.02mg/ml and 0.1 mg/ml. From the results, the cistanche sediment part has certain influence on the cell state, so in the following experiment for investigating the influence on skin-related protein, the cistanche supernatant mixture is mainly detected.
(2) Effect of cistanche salsa supernatant mixture on expression of barrier-associated proteins of heel skin
Mu.l of cell culture medium was added to each well of 8-well plates, and then 500. mu.l of keratinocyte suspension was added thereto. The cell inoculum size was 1.83X 104 cells/well, and the cells were incubated at 37 ℃ in a 5% CO2 incubator for 24 h. Sucking out the culture medium, and adding 1ml of culture medium containing Cistanchis herba supernatant mixture to make the final concentration 0.1 mg/ml. The drug concentration has been tested for cytotoxicity, and has no toxic effect on keratinocytes at this cell density. 5% CO at 37 ℃2The incubator is used for 72 h. After fixing the cells with acetone, the cells in each well were treated with monoclonal antibody (primary antibody) for 12 hours and washed away, followed by staining with a secondary antibody (sigma) containing a fluorescent group for 2 hours. After washing, the nuclei were finally stained with DAPI solution, washed off, and mounted.
In the invention, the expression conditions of two proteins (Involucrin and Claudin-1) closely related to the skin barrier in cells are mainly detected after the cistanche salsa supernatant mixture acts on keratinocytes, and the skin barrier protection effect of the extract is proved. Involucrin (Involucrin) is a key protein formed by a cutin envelope in the redifferentiation process of a keratinocyte, and the cutin envelope is a tough outer membrane formed after cell membranes in the cutin layer are widely crosslinked, is the basis of an epidermis defense barrier function and ensures that the skin is not damaged by external stimuli, so that the Involucrin is very important for maintaining the normal barrier function of the skin; claudin-1, a tight junction protein, is the main protein strand skeleton that constitutes intercellular tight junction, has important effects in maintaining the barrier function and barrier function specific to tight junction, regulating and controlling the exchange of water and inorganic salts in the skin, preventing other substances from passing through, and maintaining the internal steady state of the skin.
The experimental results are shown in fig. 3, and the cistanche salsa supernatant mixture can obviously improve the expression of involucrin and tight junction protein closely related to the skin barrier function in keratinocytes under the condition of 0.1mg/ml (non-toxic concentration verified by XTT experiment): involucrin and Claudin content.
The mixture of the supernatant of cistanche deserticola prepared in examples 1-6 was used for the preparation of skin external preparations. The skin external preparation is preferably a cosmetic composition such as a lotion, essence, cream, etc. The weight percentage of the sealwort pectin polysaccharide in the skin external preparation is 0.0001-20% (w/w), the preferable weight percentage is 0.001-10% (w/w), and the more preferable weight percentage is 0.01-5% (w/w).
The following are examples of the specific use of the cistanche supernatant mixture in skin external preparations, and the formulation and preparation method of these preparations. In the tables, "-" indicates no addition.
The cistanche supernatant mixtures used in the following examples 10 to 20 were all the cistanche supernatant mixtures prepared in example 4, and the crude drug concentration thereof was 1 g/ml; the units of the components in examples 3-13 are weight percentages.
Example 10: preparation of face cream
Figure BDA0000730529580000111
Figure BDA0000730529580000121
Example 11: preparation of the emulsion
Figure BDA0000730529580000122
Example 12: preparation of jelly
Figure BDA0000730529580000131
Example 13: preparation of astringent
Figure BDA0000730529580000132
Example 14: preparation of essence
Figure BDA0000730529580000133
Figure BDA0000730529580000141
Example 15: preparation of facial mask
Figure BDA0000730529580000142
Example 16: preparation of eye cream
Figure BDA0000730529580000151
Example 17: preparation of an aerosol (cleaning foam)
Figure BDA0000730529580000152
Example 18: preparation of the spray
Figure BDA0000730529580000161
Example 19: preparation of shower gel
Figure BDA0000730529580000162
Example 20: preparation of facial cleanser
Figure BDA0000730529580000163
Figure BDA0000730529580000171
The cistanche supernatant mixtures described in the above examples 10-20 were subjected to ① stability tests, and the color and morphology of each material remained stable when the material was placed at-20 deg.C, 4 deg.C, room temperature, 48 deg.C and circulated (four temperature conditions were changed continuously) for 12 weeks, and ② skin safety tests confirmed that the mixture was not irritating to the skin, did not cause erythema, desquamation, stinging pain, burning sensation, and other adverse reactions, and could be used with ease.

Claims (8)

1. Use of a cistanche supernatant mixture for the preparation of a product for skin barrier repair, wherein the cistanche supernatant mixture is prepared by the following method:
a) after water extraction, ultrafiltration is carried out;
b) concentrating, and precipitating with organic solvent;
c) taking the supernatant, centrifuging to obtain a cistanche salsa supernatant mixture,
wherein the content of sugar in the cistanche salsa supernatant mixture is 23.37-62.60%, the content of protein is 15.74-35.57%,
wherein the ultrafiltration has a cut-off of molecules having a molecular weight of 10000 or more, and the product is a composition for external use for skin.
2. Use according to claim 1, wherein step a) is carried out at a temperature of 80-100 ℃.
3. The use of claim 1, wherein the weight ratio of cistanche tubulosa raw material to water in step a) is 1:20 to 1: 30.
4. Use according to claim 1, wherein the organic solvent precipitation of step b) is carried out using an organic solvent selected from the group consisting of: methanol, ethanol, acetone.
5. The use of claim 1, comprising increasing the expression of involucrin and claudin within keratinocytes.
6. The use of claim 1, wherein the product is a skin external composition, and the weight percentage of the cistanche supernatant mixture in the skin external composition is 0.0001-20 wt%.
7. The use of claim 1, wherein the cistanche supernatant mixture is present in the skin external composition in an amount of 0.001 to 10 wt%.
8. The use of claim 1, wherein the cistanche supernatant mixture is present in the skin external composition in an amount of 0.01 to 5 wt%.
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