CN113855620B - Composition containing saussurea involucrata cell culture and preparation method and application thereof - Google Patents
Composition containing saussurea involucrata cell culture and preparation method and application thereof Download PDFInfo
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/99—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from microorganisms other than algae or fungi, e.g. protozoa or bacteria
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/33—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
- A61K8/34—Alcohols
- A61K8/347—Phenols
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics 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
- A61K8/9783—Angiosperms [Magnoliophyta]
- A61K8/9789—Magnoliopsida [dicotyledons]
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q17/00—Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
- A61Q17/005—Antimicrobial preparations
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/08—Anti-ageing preparations
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/40—Chemical, physico-chemical or functional or structural properties of particular ingredients
- A61K2800/59—Mixtures
- A61K2800/592—Mixtures of compounds complementing their respective functions
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/40—Chemical, physico-chemical or functional or structural properties of particular ingredients
- A61K2800/59—Mixtures
- A61K2800/592—Mixtures of compounds complementing their respective functions
- A61K2800/5922—At least two compounds being classified in the same subclass of A61K8/18
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/74—Biological properties of particular ingredients
- A61K2800/78—Enzyme modulators, e.g. Enzyme agonists
- A61K2800/782—Enzyme inhibitors; Enzyme antagonists
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- Y—GENERAL 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
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The invention discloses an anti-inflammatory and/or whitening composition, and a preparation method and application thereof, and relates to the field of cosmetics. The raw materials of the anti-inflammatory and/or whitening composition comprise: the mass ratio of the saussurea involucrata cell culture to the schisandra chinensis is 1:3-4:1; the preparation method of the composition comprises the following steps: s1, mixing saussurea involucrata cell cultures, schisandra chinensis and 1, 3-butanediol, and carrying out hot reflux extraction; s2, finely filtering and then compounding resveratrol to obtain the composition; the application is the application of the composition in preparing anti-inflammatory and/or whitening products. The composition provided by the invention has the effects of synergistically enhancing whitening, anti-inflammatory and antibacterial peptide regulation through reasonable proportion, can obviously enhance the anti-inflammatory and whitening effects when applied to a cosmetic formula, is prepared from natural medicinal plants clinically verified for many years, has high biological safety, is environment-friendly, and has a wide application prospect.
Description
Technical Field
The invention relates to the field of cosmetics, in particular to a composition containing saussurea involucrata cell culture, a preparation method thereof and application thereof in cosmetics.
Background
From a medical perspective, the inflammatory response is the primary response of human tissue to the stimulation of various damaging factors, primarily the human defenses. The causes of skin inflammation mainly include the following aspects: 1. external environmental stimulus: the health condition of the skin is affected by various factors such as the external environment. The skin is irritated by the common supercooled or overheated environment, strong wind and sand, and pollutants, dust, pollen and the like in the air. 2. Stimulating components in skin care products: such as alcohol, perfume, preservative, etc., and the active ingredients in the skin care product are also part of the stimulating ingredient. The active ingredient has biplanarity in cosmetics, and moderate concentration of the active ingredient can help to improve skin condition, and unsuitable concentration can cause skin inflammation to occur due to intolerance of skin. 3. Skin oil extraction: the grease is secreted vigorously, so that sebaceous gland ducts are easy to be blocked, abnormal proliferation of bacterial groups such as propionibacterium acnes, staphylococcus aureus, malassezia and the like is easy to be caused, facial skin infection is caused, and skin problems such as closed acne and acne are easy to be induced. 4. Ultraviolet ray stimulation: the ultraviolet short wave UVB can directly cause cells such as skin keratinocytes to release inflammatory factors, so that the skin is reddish and heated in a short time. At the same time, the ultraviolet rays can also stimulate the activation of melanocytes, which leads to skin pigmentation. In addition, ultraviolet rays can accelerate skin aging and influence normal metabolism of skin.
Because some components in the natural plants have the effects of anti-inflammation, antioxidation and the like, the natural plant components have mild skin effect, small irritation and high safety, so the natural plant components have wide application prospect in cosmetics.
Saussurea involucrata: sex taste: bitter, slightly bitter and warm; the main functions are as follows: clearing away heat and toxic materials, dispelling pathogenic wind and dampness, relieving swelling and pain. Can be used for treating head injury, gynecological diseases, rheumatoid arthritis, apoplexy, and alpine reaction, and for topical application. The saussurea involucrata is grown on the mountain grass slope, cliff and ice rock seam with the altitude of 4000 m, and has the capability of absorbing ultraviolet rays due to the fact that the saussurea involucrata grows in a special environment near the snow mountain and receives extremely strong ultraviolet radiation. It is a rare and precious Chinese herbal medicine special for Xinjiang. Saussurea involucrata is the only saussurea involucrata plant listed in the book of Chinese plant Red book. The saussurea involucrata cell culture is prepared by selecting isolated tissue of saussurea involucrata of Compositae, taking callus formed by dedifferentiation as a secondary seed, and performing secondary culture under certain conditions to obtain mauve agglomerate particles.
Fructus Schisandrae chinensis is a dry mature fruit of Schisandra chinensis (Schisandra chinensis (Turcz.) Baill) belonging to Schisandra of Magnoliaceae, which is a well-known traditional Chinese medicine and also plays a role in modern medicine. Modern medical research shows that the schisandra chinensis contains various functional components such as schisandra chinensis B, triterpenes, lignans, polysaccharides and the like, and has the activities of reducing blood sugar, resisting oxidation, protecting liver and the like. Patent CN200910069813.5 discloses the application of schisandra chinensis extract in preparing sun-proof skin care products, which proves that schisandra chinensis extract has the effect of absorbing ultraviolet rays and can reduce inflammatory reaction caused by the ultraviolet rays.
The inflammatory reaction is a stress reaction of the skin to external stimulus, and the inflammation possibly aggravates skin pigmentation, accelerates skin aging and the like, so that the plant anti-inflammatory extract with high safety, environmental friendliness, remarkable anti-inflammation and definite action mechanism is developed, and the plant anti-inflammatory extract has a wide application market.
Disclosure of Invention
Aiming at the problems of poor safety, low effect and the like of anti-inflammatory raw materials in the prior art in the field of cosmetics, the invention provides an anti-inflammatory composition containing saussurea involucrata cell cultures, a preparation method thereof and application thereof in the field of cosmetics. The anti-inflammatory composition is compounded by carefully selecting saussurea involucrata cell cultures and shizandra berry, can synergistically play the roles of enhancing anti-inflammatory, regulating and controlling antibacterial peptide, resisting oxidation and whitening, so that the anti-inflammatory composition is applied to cosmetics, and the product effect is improved.
In order to achieve the above purpose, the technical scheme adopted by the invention is as follows:
in one aspect, the invention discloses an anti-inflammatory composition, which comprises the following traditional Chinese medicine raw materials: saussurea involucrata cell culture and schisandra chinensis.
Preferably, the mass ratio of the saussurea involucrata cell culture to the schisandra chinensis extract is 1:3-4:1, and further preferably, the mass ratio is 1:1.
Preferably, the above composition further comprises resveratrol.
Preferably, the resveratrol is 0.1% -0.3% by mass.
Wherein, the saussurea involucrata cell culture, the schisandra chinensis and the resveratrol in the composition are screened by a cell experiment to obtain a plurality of plant extracts, and all three extracts are determined to have the functions of inhibiting inflammatory factor prostaglandin E2 (PGE-2) and regulating and controlling antibacterial peptide LL-37.
Saussurea involucrata, also known as Saussurea involucrata, belongs to a plant of saussura (Saussurea) of Compositae, can independently survive under severe natural conditions of high mountain, and is closely related to the formed special adaptation mechanism and active ingredients. Research shows that the active components of saussurea involucrata mainly comprise polysaccharides, flavonoids, alkaloids, sterols, volatile oil and the like, and the active components have the biological functions of resisting inflammation, resisting radiation, scavenging free radicals, resisting bacteria, resisting viruses and the like. And excessive inflammatory factors are activated in the skin after UVB irradiation, so that the active ingredients of the snow lotus have a great effect in resisting skin injury caused by UVB irradiation. The content of active ingredients of the saussurea involucrata culture is compared with that of natural plant extracts, and the main functional active substances are higher than a plurality of times. The total flavone is 4 times higher and the total protein is nearly 2 times higher. Compared with wild saussurea involucrata, the saussurea involucrata culture has higher content of main active ingredients.
Fructus Schisandrae chinensis is a dry mature fruit of Schisandra chinensis (Schisandra chinensis (Turcz.) Baill) belonging to Schisandra of Magnoliaceae, which is a well-known traditional Chinese medicine and also plays a role in modern medicine. Modern medical research shows that the schisandra chinensis contains various functional components such as schisandra chinensis B, triterpenes, lignans, polysaccharides and the like, and has the activities of reducing blood sugar, resisting oxidation, protecting liver and the like.
Resveratrol is a plant polyphenol, also known as stilbene trisol. In recent years, attention has been paid to wide biological activities, and a great deal of research has been conducted to confirm that resveratrol has functions of anti-oxidative stress, anti-inflammatory, antibacterial, regulating fat deposition, etc.
On the other hand, the invention also discloses a preparation method of the composition, which comprises the following steps:
s1, mixing saussurea involucrata cell cultures, schisandra chinensis and 1, 3-butanediol, performing hot reflux extraction, performing fine filtration, and collecting filtrate to obtain an extracting solution;
s2, adding resveratrol into the extract obtained in the step S1 to obtain the composition.
Preferably, the volume percentage of the 1, 3-butanediol solution in step S1 is 10-50%, preferably 30%.
Preferably, in the step S1, the feed liquid ratio of the saussurea involucrata and the schisandra chinensis to the 1, 3-butanediol is 1:40-60, preferably 1:50.
in yet another aspect, the present invention provides the use of the above composition for the preparation of an anti-inflammatory, whitening, antioxidant and/or antibacterial peptide modulating product.
In yet another aspect, the present invention provides an anti-inflammatory, whitening, antioxidant and/or antimicrobial peptide-modulating product comprising the above composition.
Preferably, the product is a cosmetic product.
Preferably, the cosmetic product includes, but is not limited to, a cream, an emulsion, an essence, a barrier cream, a BB cream, a foundation, a shampoo, a conditioner, or a toner for skin care.
In yet another aspect, the present invention provides the use of the above composition for the preparation of anti-inflammatory, whitening, antioxidant and/or modulating antimicrobial peptides.
Compared with the prior art, the invention has the following beneficial effects:
1. the raw materials of the anti-inflammatory composition disclosed by the invention are natural medicinal plant components, and are natural medicinal plants which are clinically verified for many years, so that the anti-inflammatory composition is high in biological safety and environment-friendly.
2. After a large number of natural medicinal plant extracts are screened through a cell experiment, the invention determines that the two natural medicinal plant components of saussurea involucrata and schisandra chinensis have the functions of inhibiting inflammatory factor prostaglandin E2 (PGE-2) and regulating antibacterial peptide LL-37, and through reasonable proportion, the functions of anti-inflammatory, regulating antibacterial peptide, anti-oxidation and whitening can be synergistically enhanced, and the effects of anti-inflammatory, regulating antibacterial peptide, anti-oxidation and whitening are superior to the effects of independently anti-inflammatory, regulating antibacterial peptide, anti-oxidation and whitening.
3. After a large number of natural medicinal plant extracts are screened through a cell experiment, the invention also determines that resveratrol has the functions of inhibiting inflammatory factors prostaglandin E2 (PGE-2) and regulating antibacterial peptide LL-37, and the resveratrol and the composition are reasonably compounded, so that the effects of anti-inflammatory, regulating antibacterial peptide, anti-oxidation and whitening can be further synergistically exerted.
Drawings
FIG. 1 shows the effect of examples 1-5, comparative examples 1-4 on PGE-2 secretion.
FIG. 2 shows the regulation of the antibacterial peptide LL-37 of examples 1-5, comparative examples 1-4.
FIG. 3 shows the effect of examples 6-10, comparative examples 5-8 on PGE-2 secretion.
FIG. 4 shows the regulation of the antibacterial peptide LL-37 in examples 6-10, and comparative examples 5-8.
Detailed Description
Other advantages and effects of the present invention will become apparent to those skilled in the art from the following disclosure, which describes the embodiments of the present invention with reference to specific examples. The invention may be practiced or carried out in other embodiments that depart from the specific details, and the details of the present description may be modified or varied from the spirit and scope of the present invention.
Before the embodiments of the invention are explained in further detail, it is to be understood that the invention is not limited in its scope to the particular embodiments described below; it is also to be understood that the terminology used in the examples of the invention is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the invention.
Where numerical ranges are provided in the examples, it is understood that unless otherwise stated herein, both endpoints of each numerical range and any number between the two endpoints are significant both in the numerical range. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.
It should be noted that the raw materials used in the present invention are all common commercial products, and the sources thereof are not particularly limited, and some raw materials and the sources of the detection kit are shown in table 1.
TABLE 1
| Raw materials | Manufacturer' s | Goods number |
| Schisandra chinensis | Beijing Tongren Tang | Without any means for |
| Saussurea involucrata cell culture | An Saibo | Without any means for |
| Resveratrol | Biotopped | 501-36-09 |
| PGE-2ELISA kit | Biotechwell | EH0424 |
| LL-37ELISA kit | CUSABIO | B04030098 |
| Radix stephaniae tetrandrae extract | Zhejiang lake Jiamei biochemicals Co.Ltd | 20210722 |
| L-tyrosine | BEIJING J&K SCIENTIFIC Ltd. | 185061 |
| Mushroom tyrosinase | JSENB | 9002-10-2 |
| Vitamin C | BEIJING J&K SCIENTIFIC Ltd. | 231406 |
Basic embodiment
An anti-inflammatory and/or whitening composition comprising the following raw materials: saussurea involucrata cell culture and schisandra chinensis;
preferably, the mass ratio of the saussurea involucrata cell culture to the schisandra chinensis in the composition is 1:3-4:1, and more preferably, the mass ratio is 1:1.
The invention also discloses a preparation method of the anti-inflammatory composition, which comprises the following steps:
s1, mixing saussurea involucrata cell cultures, schisandra chinensis and 1, 3-butanediol, performing hot reflux extraction, performing fine filtration, and collecting filtrate to obtain an extracting solution;
s2, adding resveratrol into the extract obtained in the step S1 to obtain the composition.
Examples 1-5 an anti-inflammatory composition and method of making same
The raw materials and amounts of the anti-inflammatory compositions described in examples 1-5 are shown in Table 2
TABLE 2
The preparation method comprises the following steps: the method comprises the following steps:
s1, weighing saussurea involucrata cell cultures and schisandra chinensis, and mixing the saussurea involucrata and schisandra chinensis according to a feed liquid ratio: 30%1, 3-butanediol = 1:50, 30%1, 3-butanediol was added;
s2, carrying out thermal reflux for 2.5h at 100 ℃, cooling, and carrying out suction filtration by using a 0.45 mu m filter plate to obtain the composition.
Comparative example 1
This comparative example differs from example 2 in that: the raw materials of the anti-inflammatory composition comprise 10g of saussurea involucrata cell culture, excluding schisandra chinensis.
Comparative example 2
This comparative example differs from example 2 in that: the raw materials of the anti-inflammatory composition comprise 10g of schisandra chinensis, excluding saussurea involucrata cell cultures.
Comparative example 3
This comparative example differs from example 1 in that: the raw materials of the anti-inflammatory composition comprise 2g of saussurea involucrata cell culture and 8g of schisandra chinensis.
Comparative example 4
This comparative example differs from example 3 in that: the raw materials of the anti-inflammatory composition comprise 8.75g of saussurea involucrata cell culture and 1.25g of schisandra chinensis.
Evaluation of Effect
1. The effect of the composition on PGE-2 secretion and the ability to modulate the antibacterial peptide LL-37 were examined.
(1) The detection method comprises the following steps:
s1, respectively filtering the samples of the examples 1-5 and the comparative examples 1-4, and preparing the samples into samples with concentrations of 5%, 2.5%, 1% and 0.5% respectively by using a MEM culture medium;
s2, inoculating: according to 6X 10 5 Cell/well seeding Density cells were seeded into 6-well plates, incubator (37 ℃,5% CO) 2 ) Incubating overnight;
s3, UVB irradiation: when the cell plating rate in the 6-hole plate reaches 80% -90%, the culture solution is discarded, 1mL of PBS solution is added into each hole, the cells are washed, and the washing is repeated for 2 times. Then adding a proper amount of PBS to cover cells, and performing UVB radiation with the radiation dose of 100mJ/cm 2 ；
S4, drug administration: setting a blank group, a UVB group and a sample group in an experiment, wherein the blank group is not irradiated by UVB, and is accessed into 2mL of culture medium; after UVB irradiation, 2mL of culture medium is inoculated into the UVB group; after UVB irradiation, the medicine group is inoculated with 2mL of sample culture medium with different concentrations, 37 ℃ and 5% CO 2 Culturing in an incubator for 24 hours;
s5, collecting supernatant: after culturing for 24 hours, collecting cell culture supernatant in an EP tube, and placing the EP tube in a refrigerator at the temperature of-80 ℃ for freezing preservation;
s6, detecting the content of PGE-2: detecting the cell sample according to the PGE-2ELISA kit operating instructions;
s7, LL-37 content detection: detecting the cell sample according to the LL-37ELISA kit protocol;
s8, result statistical analysis: test results were plotted using GraphPad Prism Program software, and the sample groups were compared to the negative control groups, using SPSS statistical analysis, p <0.05 (x) indicated significant differences, and p <0.01 (x) indicated very significant differences.
(2) Detection result: the results are shown in tables 3-4.
TABLE 3 detection of effects on PGE-2 secretion
TABLE 4 detection results of regulatory antibacterial peptide LL-37
Antibacterial peptides can be divided into various types according to their structures, with Cathelicidin and descensin being the main types, and antibacterial peptide LL-37 being the only antibacterial peptide of the Cathelicidin family that is present in human skin. It not only has bactericidal effect, but also plays a key role in the release of inflammatory factors in immunomodulation and the occurrence of inflammatory diseases. LL-37 has been shown to inhibit inflammatory responses by inhibiting the release of the pro-inflammatory cytokine PGE-2, etc., against increases in NF- κB protein expression and transcriptional activity by UVB-irradiated HaCaT cells. UVB radiation causes cells to generate inflammatory reaction, the expression of the antibacterial peptide LL-37 is increased, and the release of inflammatory factors PGE-2 and the like is inhibited, so that the inflammatory reaction is inhibited, and after the inflammatory reaction is reduced, the expression of the antibacterial peptide LL-37 is reduced and regulated to an equilibrium state, so that the regulation of the LL-37 has an important influence on the inhibition of the inflammatory reaction.
2. The antioxidant efficacy of the composition was tested.
(1) The detection method comprises the following steps:
s1, sample preparation: weighing 20mg DPPH, adding absolute ethyl alcohol to dissolve and fixing volume in 250mL volumetric flask, wherein the concentration of DPPH solution is 2×10 -4 The mol/L is preserved in dark at 0-4 ℃ and is effective within 4 hours after being prepared for use; the positive control is vitamin C with the vitamin concentration of 1mg/mL; examples 1-5 and comparative examples 1-4 were formulated with absolute ethanol at 6 concentrations of 100%, 50%, 10%, 5%, 1% and 0.5%, respectively, as samples to be tested.
S2, taking 1mL of to-be-detected liquid and 1mL of DPPH solution, and uniformly mixing (A pipe); mixing 1mL of absolute ethyl alcohol with 1mL of DPPH solution uniformly (B pipe); mixing 1mL of absolute ethanol with 1mL of the solution to be tested (C tube) (shown in Table 5); after 30min of reaction, the absorbance of A, B, C tubes was measured at 517nm using an enzyme-labeled instrument (infinite M200 PRO, TECAN).
TABLE 5
S3, data processing: clearance (%) = [ (b+c) -a ]/B.
(2) Detection result: the results are shown in Table 6.
TABLE 6 detection results of DPPH clearance
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3. The whitening efficacy of the composition was tested.
(1) The detection method comprises the following steps:
s1, sample preparation: 45.30mg of L-tyrosine is weighed and dissolved in 50mL of PBS buffer solution to prepare 5mM L-tyrosine solution, and the solution is preserved at 4 ℃ for standby; completely dissolving 25kU of mushroom tyrosinase powder in 2mL of precooled sterile PBS solution, subpackaging 50 mu L of the solution in each tube, and storing the solution in-20 ℃, when in use, taking one tube of 50 mu L of mushroom tyrosinase solution, adding 2.45mL of PBS buffer solution, diluting the solution to 2.5mL for use, storing the solution at 4 ℃ for later use, and finishing the use within 24 hours; the positive control is vitamin C with the concentration of 1mg/mL; examples 1-5 and comparative examples 1-4 were formulated with PBS solution at 5 concentrations of 0.5%, 1%, 5%, 10% and 100%, respectively, as samples to be tested.
S2, preheating the mushroom tyrosinase, the prepared sample to be tested, PBS and L-tyrosine solution in a shaking table at 37 ℃ for 10min; taking 20 mu L of a sample solution to be detected and PBS solvent, respectively adding the sample solution and PBS solvent into a 96-well culture plate, adding 160 mu L of L-tyrosine (0.5 mM), uniformly mixing, and then preserving heat in a 37 ℃ incubator (KYC-100B, shanghai Fuma laboratory equipment Co., ltd.); then 20 mu L (final concentration of 25U/mL) of preheated 250U/mL mushroom tyrosinase solution at 37 ℃ is added, the mixture is uniformly mixed (the reagent preparation ratio is shown in table 7), and an enzyme-labeled instrument (infinite M200 PRO, TECAN) is immediately used for measuring absorbance values A0 and B0 at 490 nm; after the measurement, the reaction was carried out in a constant temperature incubator at 37℃for 30 minutes, and absorbance values A30 and B30 at 490nm were measured by using a microplate reader.
TABLE 7
| Reagent(s) | A (sample group) | B (solvent group) |
| L-tyrosine solution (mu L) | 160 | 160 |
| Sample to be tested or PBS solvent (μL) | 20 | 20 |
| Mushroom tyrosinase solution (mu L) | 20 | 20 |
S3, data processing:
(3) Detection result: the results are shown in Table 8.
TABLE 8 tyrosinase inhibition assay results
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The detection results show that the embodiment 1-5 of the mass ratio of the saussurea involucrata cell culture to the schisandra fruit in the protection scope of the invention can obviously inhibit the secretion of PGE-2, and simultaneously regulate and control the content of the antibacterial peptide LL-37 to restore the antibacterial peptide LL-37 to an equilibrium state, and the anti-inflammatory and antioxidant and whitening capacities of the antibacterial peptide LL-37 are better than those of the saussurea involucrata cell culture composition and the schisandra fruit composition in the comparative example 1-2, so that the two natural medicinal plant raw materials are compounded according to a certain proportion, and the anti-inflammatory and antioxidant and whitening capacities of the antibacterial peptide LL-37 are synergistically enhanced. The detection results of comparative examples 1 to 5 and comparative examples 3 to 4 prove that the anti-inflammatory composition with the mass ratio of the saussurea involucrata cell culture to the schisandra chinensis which is not in the protection scope of the invention cannot exert the synergistic effect.
Examples 6-10 an anti-inflammatory composition and method of making same
The raw materials and amounts of the anti-inflammatory compositions described in examples 6-10 are shown in Table 9
TABLE 9
| Example 6 | Example 7 | Implementation of the embodimentsExample 8 | Example 9 | Example 10 | |
| Saussurea involucrata cell culture/g | 5 | 5 | 5 | 5 | 5 |
| Schisandra chinensis/g | 5 | 5 | 5 | 5 | 5 |
| Resveratrol/% | 0.1 | 0.15 | 0.2 | 0.25 | 0.3 |
The preparation method comprises the following steps: the method comprises the following steps:
s1, weighing saussurea involucrata cell cultures and schisandra chinensis, and mixing the saussurea involucrata and schisandra chinensis according to a feed liquid ratio: 30%1, 3-butanediol = 1:50, 30%1, 3-butanediol was added;
s2, carrying out thermal reflux for 2.5 hours at the temperature of 100 ℃, cooling, and carrying out suction filtration by using a 0.45 mu m filter plate;
s3, compounding 0.2% resveratrol to obtain the composition.
Comparative example 5
This comparative example differs from example 8 in that: the raw materials of the anti-inflammatory composition comprise 10g of saussurea involucrata cell culture, excluding schisandra chinensis.
Comparative example 6
This comparative example differs from example 8 in that: the raw materials of the anti-inflammatory composition comprise 10g of schisandra chinensis, excluding saussurea involucrata cell cultures.
Comparative example 7
This comparative example differs from example 8 in that: the raw materials of the anti-inflammatory composition comprise 5g of paeonia lactiflora, excluding saussurea involucrata cell cultures.
Comparative example 8
This comparative example differs from example 8 in that: the raw materials of the anti-inflammatory composition comprise 5g of baikal skullcap root, excluding shizandra berry.
Evaluation of Effect
1. The effect of the composition on PGE-2 secretion and the ability to modulate the antibacterial peptide LL-37 were examined.
(1) Detection method
Consistent with the detection method described above.
(2) Detection result: the results are shown in tables 10-11
TABLE 10 detection of effects on PGE-2 secretion
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TABLE 11 detection results of regulatory antibacterial peptide LL-37
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2. The antioxidant efficacy of the composition was tested.
(1) Detection method
Consistent with the detection method described above.
(2) Detection result: the results are shown in Table 12.
TABLE 12 detection results of DPPH clearance
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3. The whitening efficacy of the composition was tested.
(1) The detection method comprises the following steps:
consistent with the detection method
(2) Detection result: the results are shown in Table 13.
TABLE 13 tyrosinase inhibition assay results
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The detection results show that the mass ratio of the saussurea involucrata cell culture to the schisandra chinensis to the resveratrol in examples 6-10 can obviously inhibit the secretion of PGE-2 within the protection scope of the invention, meanwhile, the content of the antibacterial peptide LL-37 is regulated and controlled to restore the balance state, and the effects of resisting inflammation and regulating the antibacterial peptide, the effects of resisting oxidation and whitening are better than those of the composition compounded by the saussurea involucrata cell culture and the schisandra chinensis, and the detection results of comparative examples 6-10 and example 2 prove that the three natural medicinal plant raw materials are compounded according to a certain proportion, so that the anti-inflammatory and antibacterial peptide regulating capacity, the antioxidant and the whitening capacity of the saussurea involucrata cell culture and the schisandra chinensis can be synergistically enhanced. The results of comparative examples 8 and 7 to 8 show that the anti-inflammatory, antioxidant and whitening effects are remarkably reduced even if one of the raw materials is replaced with the other natural medicinal plant raw material having the same weight part and anti-inflammatory and antibacterial effects, and the results of comparative examples 6 to 10 and 5 to 6 show that the saussurea involucrata cell culture, schisandra chinensis and resveratrol with specific mass ratios can exert a synergistic effect.
Finally, it should be noted that the above description is only for illustrating the technical solution of the present invention, and not for limiting the scope of the present invention, and that the simple modification and equivalent substitution of the technical solution of the present invention can be made by those skilled in the art without departing from the spirit and scope of the technical solution of the present invention.
Claims (7)
1. An anti-inflammatory composition, characterized in that it comprises the following raw materials: saussurea involucrata cell culture and schisandra chinensis; the mass ratio of the saussurea involucrata cell culture to the schisandra chinensis is 1:3-4:1;
the preparation method of the composition comprises the following steps:
s1, taking saussurea involucrata cell cultures and schisandra chinensis according to a feed liquid ratio: 30%1, 3-butanediol = 1:50, 30%1, 3-butanediol was added;
s2, carrying out heat reflux on the extracting solution obtained in the step S1 at 100 ℃ for 2.5 hours, cooling, and carrying out suction filtration by using a 0.45 mu m filter plate to obtain the composition.
2. The composition of claim 1, wherein the composition further comprises resveratrol; the resveratrol is 0.2 percent by mass percent;
the preparation method of the composition comprises the following steps:
s1, weighing saussurea involucrata cell cultures and schisandra chinensis, and mixing the saussurea involucrata and schisandra chinensis according to a feed liquid ratio: 30%,1, 3-butanediol = 1:50, 30%1, 3-butanediol was added: s2, carrying out thermal reflux for 2.5 hours at the temperature of 100 ℃, cooling, and carrying out suction filtration by using a 0.45 mu m filter plate;
s3, compounding 0.2% resveratrol to obtain the composition.
3. Use of a composition according to any one of claims 1-2 for the preparation of anti-inflammatory, whitening, antioxidant cosmetics.
4. Use of a composition according to any one of claims 1-2 for the preparation of a cosmetic for modulating an antibacterial peptide.
5. An anti-inflammatory, whitening, anti-oxidant cosmetic comprising the composition of any one of claims 1-2.
6. A cosmetic for the modulation of antimicrobial peptides, characterized in that it comprises a composition according to any one of claims 1-2.
7. The cosmetic according to any one of claims 5 or 6, wherein the cosmetic is a cream, essence, barrier cream, barrier milk, BB cream, foundation or toner for skin care.
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| CN115607492A (en) * | 2022-10-18 | 2023-01-17 | 安赛搏(重庆)生物技术有限公司 | Radix ceratostigmae callus extract and preparation method and application thereof |
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| CN117180164B (en) * | 2023-11-08 | 2024-01-30 | 北京尧景基因技术有限公司 | Extraction method of saussurea involucrata purple callus vesicles and application of saussurea involucrata purple callus vesicles in preparation of antioxidant or skin whitening products |
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