CN115463064A - Saussurea involucrate callus extract and preparation method and application thereof - Google Patents
Saussurea involucrate callus extract and preparation method and application thereof Download PDFInfo
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- CN115463064A CN115463064A CN202211276641.0A CN202211276641A CN115463064A CN 115463064 A CN115463064 A CN 115463064A CN 202211276641 A CN202211276641 A CN 202211276641A CN 115463064 A CN115463064 A CN 115463064A
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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]
-
- 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
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/19—Preparation or pretreatment of starting material involving fermentation using yeast, bacteria or both; enzymatic treatment
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Dermatology (AREA)
- Mycology (AREA)
- Rheumatology (AREA)
- Biotechnology (AREA)
- Botany (AREA)
- Microbiology (AREA)
- Gerontology & Geriatric Medicine (AREA)
- Birds (AREA)
- Epidemiology (AREA)
- Pain & Pain Management (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Cosmetics (AREA)
Abstract
The invention provides a saussurea involucrate callus extract as well as a preparation method and application thereof, belonging to the technical field of plant tissue extraction. The saussurea involucrate callus extract provided by the invention is prepared by taking saussurea involucrate callus cell powder as a raw material, performing alcohol pretreatment and performing enzymolysis, has a remarkable inhibition effect on DPPH and ABTS free radicals, has an obvious inhibition effect on secretion of inflammatory factors of a HaCaT cell model induced by UVB, can inhibit pigmentation after inflammation on a B16-F10 melanoma cell model stimulated by PGE-2, and has a whitening effect.
Description
Technical Field
The invention belongs to the technical field of plant tissue extraction, and relates to a preparation method and application of a saussurea involucrate callus extract.
Background
Saussurea involucrate (Saussurea involuccata) is a perennial high mountain herbaceous plant of the genus Sphingopsis in the family of Compositae, and the overground part of the Saussurea involucrate is a commonly used medicinal material in Uygur nationality in China. The method for obtaining the saussurea involucrate cells containing a large amount of bioactive substances by using a plant cell culture technology is an effective way for solving the problem of wild saussurea involucrate resources, and has the advantages of no land occupation, no influence of natural environment, manual control of production and the like, so that the method is widely concerned by academia and industry.
The herba Saussureae Involueratae contains flavonoids, polysaccharides and hyaluronic acid, and has effects of relieving pain, resisting inflammation, regulating cardiovascular system, inhibiting cancer cell, terminating pregnancy, resisting oxidation, resisting aging, resisting radiation, relieving spasm, lowering blood pressure, relieving asthma and enhancing immunity. Research results show that the saussurea involucrate culture has antirheumatic, anti-inflammatory and analgesic effects; the saussurea involucrate culture has the functions of inhibiting nonspecific immunity and cellular immunity and enhancing humoral immunity, namely has the function of immunoregulation; the water-soluble polysaccharide of the saussurea involucrate culture has obvious scavenging effect on oxygen free radicals and hydroxyl free radicals, and is a good antioxidant for scavenging the free radicals; the saussurea involucrate culture has radioprotective effect. The saussurea involucrate culture can be applied to functional foods (solid beverage, tablet candy, high-grade beverage, health wine and the like), health-care foods (tablets, capsules, medicinal granules, oral liquid, health-care wine and the like), daily chemicals (sun cream, freckle removing cream, beauty cream, essence, toothpaste and the like) and medicines (oral preparation, medicinal liquor, rheumatism pain relieving plaster, gynecological protection pad and the like).
Saussurea involucrate is widely used in hospitals in clinic, and the saussurea involucrate is widely developed and applied by adopting Chinese and western combined medicines and various treatment modes in hospitals. The results of the method for treating the patients with diabetic peripheral neuropathy by combining long needle deep puncture, electric needle and snow lotus acupoint injection show that the blood viscosity can be reduced, the blood flow variable can be improved, the effective perfusion of microcirculation can be increased, the blood and oxygen supply of peripheral nerve tissues can be increased, the disturbance of glycolipid metabolism of the patients can be partially corrected, and the nerve conduction velocity of the patients can be well improved. The saussurea involucrate injection and lidocaine planetary ganglion block are used for treating migraine to obtain satisfactory effect, and the mechanism of the injection is related to the functions of activating blood circulation, dredging channels, easing pain, resisting inflammation and regulating vegetative nerve of saussurea involucrate besides the function of lidocaine. The saussurea involucrate injection is respectively injected at the Quchi point and the Ashi point at the pressure pain point, has obvious curative effect on migraine, and has short treatment course, no hormone and high thorough cure rate. The acupoint injection greatly enhances the stimulation to the acupoints, promotes local metabolism, increases the blood flow speed and flow at the focus part, improves the blood circulation, and thus has the effects of eliminating inflammation and relieving pain.
Chinese invention patent CN113855620A discloses an anti-inflammatory and/or whitening composition and a preparation method and application thereof, wherein the composition is obtained by taking saussurea involucrate cell culture and schisandra chinensis as raw materials, extracting by hot reflux and alcohol extraction to obtain an extract, and compounding with resveratrol; the composition provided by the invention is applied to a cosmetic formula through reasonable proportioning and synergistic enhancement of whitening, anti-inflammatory and antibacterial peptide regulation and control effects, and can significantly enhance the anti-inflammatory and whitening effects. Chinese invention patent CN102335216B discloses an extract from saussurea involucrate, which can be used as a melanin generation inhibitor after being separated by a chromatographic column, and also discloses the application of the extract in the fields of medicines, cosmetics and the like.
The research aims to develop the application of the saussurea involucrate in the field of cosmetics by extracting and preparing callus of the saussurea involucrate and researching the effects of relieving, resisting inflammation and whitening.
Disclosure of Invention
In order to solve the technical problems, the invention provides saussurea involucrate callus as well as a preparation method and application thereof, and the saussurea involucrate callus is developed by establishing an effective extraction method, researching the effects of oxidation resistance, inflammation resistance and whitening and developing the application of the saussurea involucrate callus in the field of cosmetics.
In order to achieve the purpose, the invention adopts the following technical scheme:
on one hand, the invention requests to protect the saussurea involucrate callus extract, which is prepared by taking the saussurea involucrate callus cell powder as a raw material through alcohol pretreatment, enzymolysis and thermal reflux.
Preferably, the alcohol is 1, 3-butanediol, and the concentration of the alcohol is 10% -70%.
Further preferably, the alcohol is 1, 3-butanediol at a concentration of 30%.
Preferably, the enzymes are cellulases and pectinases.
Further preferably, the addition amount of the cellulase is 0.5 to 2.5 percent of the mass of the saussurea involucrate callus cell powder; the addition amount of the pectinase is 0.5-2.5% of the mass of the saussurea involucrate callus cell powder.
Still further preferably, the addition amount of the cellulase is 1 percent of the mass of the saussurea involucrate callus cell powder; the addition amount of the pectinase is 1 percent of the mass of the saussurea involucrate callus cell powder.
Preferably, the conditions of the enzymolysis are as follows: the temperature is 40-60 ℃; the pH is 4-5; the time is 1.5-2.5h.
Further preferably, the enzymolysis conditions are as follows: the temperature is 50 ℃; the pH was 4.5; the time is 2h.
Preferably, the feed-liquid ratio of the saussurea involucrate callus cell powder to the alcohol is 1.
Further preferably, the feed-liquid ratio of the saussurea involucrate callus cell powder to the alcohol is 1.
Preferably, the conditions of the hot reflux are as follows: the extraction temperature is 80-120 deg.C, and the extraction time is 2-3h.
Further preferably, the conditions of the hot reflux are as follows: the extraction temperature is 100 ℃, and the extraction time is 2.5h.
On the other hand, the invention also provides a preparation method of the saussurea involucrate callus extract, which comprises the following steps:
(1) Accurately weighing saussurea involucrate callus cell powder, adding alcohol, adjusting the material-liquid ratio of the saussurea involucrate callus cell powder to the extraction solvent, and pretreating to obtain a treatment solution;
(2) Adding enzyme into the treated liquid for enzymolysis to obtain an enzymolysis liquid;
(3) And (3) performing thermal reflux extraction on the enzymolysis liquid, cooling, performing fine filtration, and collecting filtrate to obtain the saussurea involucrate callus extract.
In another aspect, the invention also provides an application of the saussurea involucrate callus extract or the saussurea involucrate callus extract obtained by the preparation method in preparing antioxidant, anti-inflammatory and/or whitening cosmetics.
In another aspect, the invention also provides an antioxidant cosmetic, which comprises the saussurea involucrate callus extract or the saussurea involucrate callus extract obtained by the preparation method.
In another aspect, the invention also provides an anti-inflammatory cosmetic, which comprises the saussurea involucrate callus extract or the saussurea involucrate callus extract obtained by the preparation method.
In another aspect, the invention also provides a whitening cosmetic, which comprises the saussurea involucrate callus extract or the saussurea involucrate callus extract obtained by the preparation method.
Compared with the prior art, the invention has the following beneficial effects:
1. the callus tissue extract of the saussurea involucrate is prepared by a multi-step method in a synergistic manner, the cell permeability is improved by butanediol pretreatment and the synergistic effect of cellulase and pectinase, so that the active ingredients in the saussurea involucrate can be effectively extracted, and the flavone and polyphenol content of the callus tissue extract of the saussurea involucrate is improved;
2. the saussurea involucrate callus extract extracted by the method has good DPPH and ABTS free radical scavenging effect and strong oxidation resistance;
3. the saussurea involucrate callus extract extracted by the invention has good anti-inflammatory effect, can effectively inhibit the expression of COX-2 and inhibit the secretion of inflammatory factors PGE-2 and IL-1 beta in cells;
4. the saussurea involucrate callus extract extracted by the invention has good whitening effect, can obviously inhibit pigmentation after inflammation and inhibit tyrosinase activity.
Drawings
FIG. 1 shows the contents of flavone and polyphenol in the callus extracts of saussurea involucrate obtained by extraction in examples 1-5 and comparative examples 1-4;
FIG. 2 is a data chart of DPPH free radical scavenging experiment by saussurea involucrate callus extract;
FIG. 3 is a data diagram of the experiment of eliminating ABTS free radicals by the callus extract of saussurea involucrate.
Detailed Description
The following non-limiting examples are presented to enable those of ordinary skill in the art to more fully understand the present invention and are not intended to limit the invention in any way. The following is merely an exemplary illustration of the scope of the invention as claimed, and various changes and modifications of the invention of the present application may be made by those skilled in the art based on the disclosure, which also fall within the scope of the invention as claimed.
The present invention will be further described below by way of specific examples. The various chemicals used in the examples of the present invention were obtained by conventional commercial routes unless otherwise specified.
In the following examples, saussurea involucrate callus cell powder was purchased from Ansaibo Biotech, inc.; 1, 3-butanediol was purchased from OXEA, U.S.A.; cellulase was purchased from Biotopped under the cat # C6270; pectinase was purchased from Biotopped under the accession number P6280; DPPH free radical ladder loving (Shanghai) chemical industry development limited company, the goods number is D4313; ABTS was purchased from Biotopped under the product number S016; the PGE-2 kit is purchased from Shanghai Weiao biotechnology, inc., and has a product number of EM0340; the IL-1 beta kit is purchased from Weiao biotechnology limited Shanghai, and has a product number of EH0205; the BCA protein assay kit is purchased from Shanghai Weiao Biotechnology Limited, and has the product number of WB0123; mouse tyrosinase (TyR) ELISA kits were purchased from Jianglai organisms under the accession number JL20441.
Example 1
Saussurea involucrate callus extract and preparation thereof
Accurately weighing 5g of herba Saussureae Involueratae callus cell powder, adding 250mL of 30%1, 3-butanediol; adding 1.0% of cellulase and 1.0% of pectinase after alcohol treatment, adjusting the pH value to 4.5 by using citric acid and NaOH at the temperature of 50 ℃, and performing enzymolysis for 2 hours; performing enzymolysis, extracting at 100 deg.C under reflux for 2.5 hr, cooling, fine filtering with 0.45 μm filter plate, and collecting filtrate to obtain herba Saussureae Involueratae callus extract.
Example 2
Accurately weighing 5g of herba Saussureae Involueratae callus cell powder, adding 250mL of 10%1, 3-butanediol; adding 1.0% of cellulase and 1.0% of pectinase after alcohol treatment, adjusting the pH value to 4.5 by using citric acid and NaOH at the temperature of 50 ℃, and performing enzymolysis for 2 hours; performing enzymolysis, extracting at 100 deg.C under reflux for 2.5 hr, cooling, fine filtering with 0.45 μm filter plate, and collecting filtrate to obtain herba Saussureae Involueratae callus extract.
Example 3
Accurately weighing 5g of herba Saussureae Involueratae callus cell powder, adding 250mL of 70%1, 3-butanediol; adding 1.0% of cellulase and 1.0% of pectinase after alcohol treatment, adjusting the pH value to 4.5 by using citric acid and NaOH at the temperature of 50 ℃, and performing enzymolysis for 2 hours; performing enzymolysis, performing heat reflux extraction at 100 deg.C for 2.5h, cooling, fine filtering with 0.45 μm filter plate, and collecting filtrate to obtain callus extract of herba Saussureae Involueratae.
Example 4
Accurately weighing 5g of herba Saussureae Involueratae callus cell powder, adding 250mL of 30%1, 3-butanediol; adding 1.3% of cellulase and 0.7% of pectinase after alcohol treatment, adjusting the pH value to 4 by using citric acid and NaOH at the temperature of 40 ℃, and performing enzymolysis for 1.5h; performing enzymolysis, extracting at 80 deg.C under reflux for 2 hr, cooling, fine filtering with 0.45 μm filter plate, and collecting filtrate to obtain callus extract of herba Saussureae Involueratae.
Example 5
Accurately weighing 5g of herba Saussureae Involueratae callus cell powder, adding 250mL of 30%1, 3-butanediol; adding 1.5% of cellulase and 0.5% of pectinase after alcohol treatment, adjusting the pH value to 5 by using citric acid and NaOH at the temperature of 60 ℃, and performing enzymolysis for 2.5h; performing enzymolysis, performing heat reflux extraction at 120 deg.C for 3 hr, cooling, fine filtering with 0.45 μm filter plate, and collecting filtrate to obtain callus extract of herba Saussureae Involueratae.
Comparative example 1
Accurately weighing 5g of herba Saussureae Involueratae callus cell powder, adding 250mL of 30%1, 3-butanediol; extracting at 100 deg.C under reflux for 2.5 hr, cooling, fine filtering with 0.45 μm filter plate, and collecting filtrate to obtain extract.
Comparative example 2
Accurately weighing 5g of herba Saussureae Involueratae callus cell powder, adding 250mL of 30%1, 3-butanediol; adding 1.0% of cellulase and 1.0% of pectinase after alcohol treatment, adjusting the pH value to 4.5 by using citric acid and NaOH at the temperature of 50 ℃, and carrying out enzymolysis pretreatment for 2 hours; after enzymolysis, fine-filtering with a 0.45 μm filter plate, and collecting the filtrate to obtain the extract.
Comparative example 3
Accurately weighing 5g of herba Saussureae Involueratae callus cell powder, adding 250mL of 90%1, 3-butanediol; adding 1.0% of cellulase and 1.0% of pectinase after alcohol treatment, adjusting the pH value to 4.5 by using citric acid and NaOH at the temperature of 50 ℃, and performing enzymolysis for 2 hours; performing enzymolysis, extracting at 100 deg.C under reflux for 2.5 hr, cooling, fine filtering with 0.45 μm filter plate, and collecting filtrate to obtain herba Saussureae Involueratae callus extract.
Comparative example 4
Drying herba Saussureae Involueratae, accurately weighing 5g of herba Saussureae Involueratae powder, adding 250mL of 1, 3-butanediol; adding 1.0% of cellulase and 1.0% of pectinase after alcohol treatment, adjusting the pH value to 4.5 by using citric acid and NaOH at the temperature of 50 ℃, and performing enzymolysis for 2 hours; performing enzymolysis, performing heat reflux extraction at 100 deg.C for 2.5h, cooling, fine filtering with 0.45 μm filter plate, and collecting filtrate to obtain callus extract of herba Saussureae Involueratae.
In the examples and the comparative examples, the contents of flavone and polyphenol in the examples 1 to 5 are obviously higher than those in the comparative examples 1 to 4, and the specific results are shown in figure 1.
Effect test
Preparing a solution to be detected: the callus extracts of saussurea involucrate obtained in examples 1-5 and comparative examples 1-4 are prepared into solutions to be tested with the concentrations of 0.5%, 1.0%, 2.0% and 5.0% by using absolute ethyl alcohol.
1. Inhibiting activity on DPPH free radical
(1) Preparing a DPPH ethanol solution:
weighing 20mg of DPPH, adding absolute ethyl alcohol to dissolve, fixing the volume in a 250mL volumetric flask, and preparing the DPPH concentration into 2 multiplied by 10 -4 mol/L; can be stored at 0-4 deg.C in dark place, and can be used after being prepared, and is effective within 4h. The positive control adopts vitamin C with the concentration of 0.5mg/mL.
(2) The reagents are added according to the table 1, and after the reaction is carried out for 30min in the dark, the absorbance values of tubes A, B and C are measured at 517 nm.
TABLE 1
| Number of | DPPH solution | Solvent(s) | Liquid to be tested | Total volume |
| A | 1mL | - | 1mL | 2mL |
| B | 1mL | 1mL | - | 2mL |
| C | - | 1mL | 1mL | 2mL |
(3) DPPH free radical inhibition rate calculation formula: DPPH inhibition (%) = (B + C-se:Sub>A)/B.
The results of the experiment are shown in FIG. 2.
2. ABTS free radical inhibition assay
(1) Preparing an ABTS aqueous solution: precisely weighing 0.03841g of ABTS, dissolving and dissolving in 10mL of water;
(2)K 2 S 2 O 8 preparing an aqueous solution: accurately weighing 0.0662g K 2 S 2 O 8 Dissolving and fixing in 100mL of water;
(3) Mother liquor preparation: mixing aqueous ABTS solution and K 2 S 2 O 8 Mixing the aqueous solution in equal volume, and reacting at low temperature in a dark place for 12-16h;
(4) ABTS working solution: the mother liquor is diluted by absolute ethyl alcohol until the OD value at 734nm is 0.7 +/-0.02, and the preparation is prepared as before.
(5) Vitamin C with concentration of 0.5mg/mL is selected as positive control, and is stored at 0-4 deg.C in dark place and is ready for use;
(6) The specific experimental steps are as follows:
adding the reagents according to the table 2, reacting for 30min in the dark at room temperature, and measuring the light absorption value of the reagents at 734nm by using a microplate reader
TABLE 2
| Number of | ABTS working solution | Solvent(s) | Liquid to be tested |
| Experimental group (A) | 0.8mL | - | 0.2mL |
| Blank group (A0) | 0.8mL | 0.2mL | - |
(7) ABTS free radical inhibition rate calculation formula: ABTS inhibition (%) = [ (A0-a)/A0 ] × 100%.
The results of the experiment are shown in FIG. 3.
3. Inhibition effect on UVB-induced HaCaT cell model inflammatory factor
HaCaT cells were cultured at 3X 10 5 cells/mL were seeded in 6-well plates, blank, model and sample sets were set at 37 ℃ with 5% CO 2 After 24h incubation under the conditions, UVB (radiation dose of 100 mJ/cm) was applied to the model group and the sample group 2 ) Stimulating HaCaT cells, adding herba Saussureae Involueratae callus extract into sample group, placing all group cells in incubator at 37 deg.C and 5% CO 2 Culturing for 24h in the environment, collecting supernatant, and detecting the expression quantity of PGE-2 and IL-1 beta by using an ELISA kit. Specific results are shown in Table 4.
4. Whitening efficacy test for PGE-2 induced B16-F10 melanocytes
(1) Preparation of a sample: weighing 2mg of PGE-2, dissolving in 1mL of DMSO, and then diluting with a DMEM medium to a concentration of 20 mug/mL; the saussurea involucrate callus extract is prepared to 2.0 percent concentration by serum-free culture medium.
(2) Collecting B16-F10 cell solution at 5X 10 5 cells/well Density were seeded in 24-well plates, charged with 5% CO 2 Incubate at 37 ℃ for 24h.
(3) After the cells are attached to the wall, the culture medium is discarded, 500. Mu.L of culture medium is added to each well of the blank group and the model group, and 500. Mu.L of sample solution diluted by the serum-containing culture medium is added to each well of the sample group.
(4) After 4 hours in the incubator, 5. Mu.L of 2mg/mL PGE-2 solution was added to each well of the model group and the sample group. At 5% of CO 2 After incubation at 37 ℃ for 36h, cells were collected. The cells were divided into two groups, one group for melanin content determination and the other group for tyrosinase content determination.
(5) Determination of melanin content
Cells for melanin content determination were centrifuged at 1000rpm for 5min, and the supernatant was discarded. 300uL of 1mol/L NaOH lysate (containing 10% DMSO) was added, taken out after 30min in a 80 ℃ water bath, and allowed to stand for 30min, after which the OD value was measured at a wavelength of 475 nm. Preparing PMSF cell lysate, adding 200 mu L of lysate into each tube of cells, vortexing for 10 minutes, centrifuging for 10 minutes at 1000rcf 4 ℃, taking supernatant, adding 25 mu L of cleaved protein into a 96-well plate by using a BCA kit according to the kit specification, adding 200 mu L of BCA reagent, reacting for 30 minutes at 37 ℃, and measuring the OD value at 562 nm. Total protein concentration was calculated from the standard.
Melanin change amount = (sample group OD value/protein concentration)/(model group OD value/protein concentration).
(6) Tyrosinase content determination
The mouse tyrosinase (TyR) ELISA kit is used, the OD value is measured at 450nm according to the kit instruction operation, and the tyrosinase content is calculated according to the standard curve.
Tyrosinase content rate of change = (sample group tyrosinase content/sample group total protein concentration)/(blank group tyrosinase content/blank group total protein concentration). Specific results are shown in Table 4.
Results and analysis of the experiments
TABLE 4
Note: all experiments were repeated 3 times and the results are expressed as Mean ± standard deviation (Mean ± SD).
As can be seen from fig. 2 to 3, the saussurea involucrate callus extracts extracted in examples 1 to 5 have the effect of promoting DPPH and ABTS free radical scavenging, and compared with comparative examples 1 to 4, the saussurea involucrate callus extracts extracted in examples 1 to 5 have more excellent free radical scavenging rate, which indicates that the saussurea involucrate callus extracts provided by the application can promote the scavenging of oxygen radicals and have antioxidant effect.
By detecting the contents of PGE-2 and IL-1 beta in a UVB-induced HaCaT cell model high-expression inflammatory factor model, it can be seen from Table 4 that the extracts obtained in examples 1-5 can remarkably inhibit the secretion of inflammatory factors PGE-2 and IL-1 beta. The results show that the saussurea involucrate callus extract has the effects of relieving and resisting inflammation.
PGE-2 is adopted to induce B16-F10 melanocytes, and the result of detection of melanin content (see table 4) shows that the saussurea involucrate callus extract obtained in examples 1-5 can obviously inhibit the secretion of melanin and tyrosine kinase and can inhibit the expression activity of tyrosine kinase compared with comparative examples 1-4. The results show that the saussurea involucrate callus extract has certain effect of inhibiting pigmentation after inflammation and has certain whitening effect.
Finally, it should be noted that the above-mentioned contents are only used for illustrating the technical solutions of the present invention, and do not limit the protection scope of the present invention, and those skilled in the art can make simple modifications or equivalent substitutions on the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention.
Claims (14)
1. The saussurea involucrate callus extract is characterized in that saussurea involucrate callus cell powder is used as a raw material and is prepared by carrying out alcohol pretreatment, enzymolysis and thermal reflux.
2. The saussurea involucrate callus extract as claimed in claim 1, wherein the alcohol is 1, 3-butanediol, and the mass concentration is 10-70%.
3. The saussurea involucrate callus extract as claimed in claim 1, wherein the enzyme is cellulase and pectinase.
4. The saussurea involucrate callus extract as claimed in claim 3, wherein the addition amount of the cellulase is 0.5-2.5% of the mass of the saussurea involucrate callus cell powder; the addition amount of the pectinase is 0.5-2.5% of the mass of the saussurea involucrate callus cell powder.
5. The saussurea involucrate callus extract as claimed in claim 4, wherein the addition amount of the cellulase is 1% of the mass of the saussurea involucrate callus cell powder; the addition amount of the pectinase is 1 percent of the mass of the saussurea involucrate callus cell powder.
6. The saussurea involucrate callus extract as claimed in claim 1, wherein the conditions of enzymolysis are: the temperature is 40-60 ℃; the pH is 4-5; the time is 1.5-2.5h.
7. The saussurea involucrate callus extract as claimed in claim 1, wherein the feed-liquid ratio of the saussurea involucrate callus cell powder to alcohol is 1.
8. The saussurea involucrate callus extract as claimed in claim 7, wherein the ratio of the saussurea involucrate callus cell powder to alcohol is 1.
9. The saussurea involucrate callus extract as set forth in claim 1, wherein the conditions of the thermal refluxing are: the extraction temperature is 80-120 deg.C, and the extraction time is 2-3h.
10. A method for preparing saussurea involucrate callus extract as claimed in claim 1, comprising the steps of:
(1) Accurately weighing herba saussureae involucratae callus cell powder, adding alcohol, and pretreating to obtain treatment solution;
(2) Adding enzyme into the treatment solution for enzymolysis to obtain an enzymolysis solution;
(3) And (3) performing thermal reflux extraction on the enzymolysis liquid, cooling, performing fine filtration, and collecting filtrate to obtain the saussurea involucrate callus extract.
11. Use of the saussurea involucrate callus extract of any one of claims 1 to 9 or the saussurea involucrate callus extract obtained by the preparation method of claim 10 in preparing antioxidant, anti-inflammatory and/or whitening cosmetics.
12. An antioxidant cosmetic comprising the saussurea involucrate callus extract as set forth in any one of claims 1 to 9 or the saussurea involucrate callus extract obtained by the production method as set forth in claim 10.
13. An anti-inflammatory cosmetic comprising the saussurea involucrate callus extract according to any one of claims 1 to 9 or the saussurea involucrate callus extract obtained by the production method according to claim 10.
14. A whitening cosmetic, characterized by comprising the saussurea involucrate callus extract of any one of claims 1 to 9 or the saussurea involucrate callus extract obtained by the production method of claim 10.
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