CN110652473A - Application of cedar extract - Google Patents
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- CN110652473A CN110652473A CN201810701136.3A CN201810701136A CN110652473A CN 110652473 A CN110652473 A CN 110652473A CN 201810701136 A CN201810701136 A CN 201810701136A CN 110652473 A CN110652473 A CN 110652473A
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Images
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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
- A61K8/9755—Gymnosperms [Coniferophyta]
- A61K8/9767—Pinaceae [Pine family], e.g. pine or cedar
-
- 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
-
- 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
- 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
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- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
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- Gerontology & Geriatric Medicine (AREA)
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- Medicines Containing Plant Substances (AREA)
Abstract
The invention relates to the field of daily chemicals and plant extracts, in particular to application of a cedar extract. The cedar extract has effects of improving cell repair related gene expression, such as longevity gene and keratinocyte proliferation related gene expression, improving skin cell repair ability, increasing epidermis thickness, and increasing stability of skin structure. Therefore, the cedar extract can be used for preparing skin external preparations as active ingredients for maintaining skin stability and resisting aging.
Description
Technical Field
The invention relates to the field of cosmetics, in particular to application of a cedar extract.
Background
The skin covers the whole human body surface and is in direct contact with the external environment, so that the mirror not only protects the body from being damaged by various external factors, but also reflects the health state of the whole body. The skin is composed of epidermis, dermis, and subcutaneous tissue, and contains skin appendages, abundant nerves, blood vessels, lymphatic vessels, and muscles.
The skin, like other organs, tends to age with increasing age. Whether the skin aging is caused by endogenous factors or exogenous stimuli, the skin aging is basically caused by the gradual loss of skin cells and skin constituent substances caused by the reduction of the activity of the skin cells, and finally the body function is reduced. Therefore, one important approach for anti-aging is to repair cells, improve cell metabolism, activate aging cells, and restore normal physiological functions of cells by improving self-repair ability of cells, reducing inflammatory level of cells, improving the ability of cells to secrete growth factors, and the like, thereby achieving anti-aging.
There is a gene which can improve the self-repair ability of cells and prolong the life of organisms. Such genes are involved in stress tolerance of organisms and they can enhance the self-defense and repair activities of organisms. Such genes are called longevity genes. If these genes are activated for a sufficiently long time, the health of the organism can be significantly improved and the life span can be extended. Sirtuin 2-related enzyme 1(sirtuin1, SIRT1) is one of the well-known longevity genes. Sirt1 is a nicotinamide adenine dinucleotide dependent histone and non-histone deacetylase, one of the Sirtuins family members. It is closely related to cell differentiation, aging, apoptosis and energy metabolism, and is involved in regulation and control of body longevity, energy metabolism, stress, inflammation and tumor. Studies in the skin found that Sirt1 has the effect of improving skin rejuvenation. For example, Sirt1 can protect subcutaneous collagen from photoaging caused by ultraviolet radiation; it can also lower down cell factors such as STAT3, cyclooxygenase 2, etc., and has anti-inflammatory effect; sirt1 also has skin cancer inhibiting effect. Activators of Sirt1 may play an important role in the treatment and prevention of future skin diseases. Some foreign cosmetic companies will use Sirt1 modulation as an important target for developing improved skin repair capabilities for anti-aging products.
The EGF family is a very important class of growth factors, all of which bind to EGF receptors and have the effect of regulating cell growth and differentiation and increasing cell activity. Human heparin-binding EGF-like growth factor (HBEGF) belongs to the EGF family of protein members. The growth factor plays an important role in the processes of wound healing, cardiac development and angiogenesis. In the skin, HBEGF is capable of promoting the proliferation, migration, and is considered to be an important marker of keratinocyte turnover (turnover). In the basal layer of the skin, the secretion amount of HBEGF directly affects the renewal rate of keratinocytes, so HBEGF plays an important role in the anti-aging process of the skin. Some of the well-known anti-aging actives are anti-aging by increasing the secretion of HBEGF in the basal layer of the skin.
Cedar (scientific name: Cedrus deodara (Roxb.) G.don) is a plant of genus cedar of family pinaceae. Produced in western asia, western himalayas and africa, the mediterranean coast. China has only one Himalayan cedar, which is distributed in the south of Tibet and in India and Afghanistan. Is distributed from Afghan to India with elevation 1300-. The medicinal use of cedar has long been known and can be traced back to the biblical age at the earliest. The cedar oil is added into cosmetics for beauty treatment and also used as an insect repellent. Cedar is also used by the original residents in the united states as a sanctum for medical treatment and decontamination ceremony. Cedar also effectively eliminates phlegm and resolves phlegm, and has been traditionally used to treat mucosal problems, particularly infections and blockages of the bronchi; the cedar can also be used for symptoms such as arthritis, rheumatism and the like, and can nourish the whole body; it also has excellent relieving effect on mental and mental stress, anxiety, obsessive compulsive disorder and fear.
The invention firstly provides that the cedar extract has the capability of improving the expression of genes related to skin repair capability and genes related to anti-aging, stabilizes the skin structure and is suitable for cosmetics with the effect of delaying aging.
Disclosure of Invention
The invention aims to provide application of cedar extract in preparing an external preparation for improving skin with an anti-aging effect.
The extract of Cedrus deodara can be used for preparing skin external agent, and can be used as anti-aging active ingredient and/or active ingredient for inducing skin cell repair.
Cedar extract is used as an active ingredient for improving the skin cell repair ability, increasing the epidermal thickness, or firming the basement membrane of the skin, thereby achieving the maintenance of the structural stability of the skin and/or anti-aging.
Specifically, the cedar extract can promote the proliferation of skin cells, particularly keratinocytes of epidermis, promote the repair of epidermal cells and promote the repair of skin surface layers, thereby maintaining the structural stability of the skin and having an anti-aging effect.
Thus, the cedar extract can be used as an active ingredient for increasing the expression level of a cell repair/proliferation-related gene (e.g., a longevity gene or a keratinocyte proliferation-related gene) and/or increasing the expression level of a basement membrane-related gene.
The extract of Cedrus deodara can promote the proliferation and repair of skin cells including dermal cells and keratinocyte epidermal cells, or promote the repair of skin surface layer and increase the thickness of epidermis by increasing the expression level of cell repair/proliferation related genes, such as longevity gene or keratinocyte proliferation related gene.
Preferably, the genes related to cell repair/proliferation are SIRT1 and HBEGF. SIRT1 is a longevity gene, and is a gene related to cell repair, and the up-regulation of the expression level of the longevity gene related to cell repair can promote the repair of skin cells including dermal cells and keratinocytes (epidermal cells) and enhance the repair capability of the skin cells. HBEGF is a keratinocyte proliferation related gene and encodes and generates human heparin combined with EGF-like growth factor HBEGF; improving expression level of keratinocyte proliferation related gene, promoting proliferation and migration of skin keratinocyte, improving stability of skin structure, and delaying skin aging.
The cedar extract can improve the synthesis level of human heparin combined with EGF-like growth factors by improving the expression level of HBEGF genes so as to promote the proliferation and migration of skin keratinocytes and maintain the stability of skin structures; by increasing the expression level of the SIRT1 gene, the synthesis of deacetylase is promoted to improve the repair capability of skin cells and/or increase the thickness of epidermis.
Preferably, the preparation method of the cedar extract comprises the following steps: extracting cedar with a solvent for 1-4 times, combining filtrates, removing the solvent and drying; the solvent is water, alcohol or alcohol-water mixture. Preferably, the alcohol is ethanol. Preferably, the volume concentration of the alcohol in the alcohol-water mixture is 10-95%, more preferably 40-80%; in a preferred embodiment of the present invention, the solvent is ethanol with a concentration of 70% by volume.
Preferably, the extraction method is ultrasonic extraction, the extraction time is 20-60 min each time, and the dosage ratio of the cedar to the solvent in each extraction is 1 kg: 5-20L.
Preferably, cedar needles, cedar wood or a mixture thereof are used as raw materials for extraction. Preferably cedar needles.
Preferably, the cedar is crushed, dried, extracted and crushed into particles of 20-300 meshes. More preferably, the powder is pulverized into particles of 50 to 200 mesh.
Preferably, the cedar is dried to a moisture content of less than 10%.
The invention discovers that the cedar extract has the capacity of maintaining true epidermal connection capacity, promoting proliferation of dermal cells and epidermal cells, promoting repair of epidermal cells or promoting repair of skin surface layers, increasing the thickness of epidermis and delaying skin aging, can effectively improve the expression level of longevity genes related to cell repair, such as SIRT1, and improve the expression level of genes related to skin repair and keratinocyte proliferation, such as HBEGF. Compared with the control group, the SIRT1 and HBEGF expression levels are respectively improved by 70 percent and 41 percent, and the epidermal thickness can be obviously increased, and the SIRT1 and HBEGF expression levels can be used as an efficacy additive to be added into skin external preparations such as skin care products and cosmetics to be used as anti-aging active ingredients.
The content of the cedar extract in the skin external preparation is 0.001 wt% -10 wt%. Preferably, the content of the cedar extract is 0.005 wt% to 5 wt%, more preferably 0.01 wt% to 5 wt%. In one embodiment of the invention, when the cedar extract content is 100 μ g/mL (about 0.01 wt% on a dry matter basis), the expression levels of SIRT1 and HBEGF are increased by 70% and 41%, respectively, compared to the control group, and are effective to increase the thickness of the reconstructed skin model epidermis.
The skin external preparations, such as cosmetics, are available in the forms of cleansing cream, milky lotion, cream, essence, astringent (such as emollient water, astringent), pack, makeup remover, gel, spray, bath foam, etc. The prepared skin external preparation has the effects of stabilizing skin basement membrane structure, stabilizing true epidermal junction, promoting skin cell proliferation and repair, and promoting skin surface layer repair or anti-aging.
Compared with the prior art, the invention has the beneficial effects that:
(1) the invention provides a new application of cedar extract as an effective additive and an active ingredient of skin external agents (cosmetics, skin care products and the like). The cedar extract can improve the expression level of genes related to cell repair/proliferation (such as longevity genes) and/or improve the expression level of genes related to basement membrane, can promote the proliferation of dermal cells, epidermal cells or skin surface layer, improve the repair capability of skin cells, increase the thickness of epidermis, stabilize true epidermal junction or stabilize the basement membrane of the skin. Therefore, the cedar extract can be used for preparing skin external preparations with the functions of resisting aging and maintaining the stability of skin structures, and has good application prospect.
(2) The preparation process of the cedar extract is simple, the process conditions are easy to control, and the preparation cost is low.
(3) The cedar extract has high activity, can generate obvious effect under the condition of extremely low content, is natural, safe and effective, and does not irritate the skin.
Drawings
Figure 1 shows the fast blue staining results of example 2 sections of 3D skin model with added cedar extract: a is a control group to which the cedar extract was not added, and B is an experimental group to which the cedar extract was added at 100. mu.g/mL.
FIG. 2 is a schematic diagram of the results of example 3, the cedar extract increasing SIRT1 gene expression
FIG. 3 is a graph showing the results of example 4 in which Cedrus deodara extract increases the expression of HBEGF gene involved in the proliferation of fine keratinocytes
Detailed Description
Example 1 production of cedar extract
Taking 10g of 50-200 mesh cedar (needle leaf) dry powder with water content of less than 5%, and ultrasonically extracting with 70% (volume concentration) ethanol 500W at room temperature for 2 times, wherein the ethanol dosage is 100mL each time, and the extraction time is 30min each time; the filtrates were combined and the filtrate was rotary evaporated to dryness to give about 0.9g of cedar extract.
Example 23D skin model test
Preparation of skin model
Human primary fibroblasts (NHDF) and human primary keratinocytes (NHEK) were extracted from skin tissue, respectively. Reviving human primary fibroblasts, mixing the human primary fibroblasts with collagen after culture and digestion, performing dermal phase culture for 3-4 days, digesting and inoculating human primary keratinocytes cultured by using a feeder layer to the surface of a dermal phase skin model, and sequentially performing epidermal immersion culture and gas-liquid phase culture for 18 days to obtain the 3D skin model for detection.
During the culture, the skin model was cultured with the normal medium (control, NT) or the medium containing 100. mu.g/mL of the extract of cedar according to example 1 (about 0.01 wt%, based on the dry weight of the cedar extract), respectively, from D3 after seeding fibroblasts on the scaffold to the final D18.
(II) frozen sections and quick blue staining
The 3D skin model was immersed in OCT and made into frozen blocks in liquid nitrogen. Then, the skin was sectioned in a cryomicrotome, and 5um thick 3D skin model sections were collected on a glass slide. Fixing in 70% ethanol for 90s at room temperature, dyeing in fast blue dye liquor for 120s, and washing in clear water. Dehydrating in alcohol with gradually increasing concentration, and sealing and taking picture after xylene is transparent.
The results are shown in fig. 1, a is the result of staining the untreated 3D skin model of the control group; and B is an experimental group, namely a 3D skin model dyeing result treated by the cedar extract.
As can be seen from figure 1, after 100 μ g/mL cedar extract was added, compared with the reconstructed skin model of the untreated group, the thickness of the epidermis was significantly increased, which indicates that the cedar extract can effectively increase the thickness of the epidermis of the reconstructed skin model, promote the proliferation of epidermal keratinocytes, make the arrangement of cells on the basal layer tighter, and make the differentiation of the stratum corneum more complete. The cedar extract has the functions of potentially delaying skin aging, repairing skin surface layers and promoting epidermal cell repair, and has the functions of resisting aging and improving the overall vitality and health condition of the skin.
Example 3 Cedar extract to increase expression of genes involved in skin cell repair
The skin model was prepared in the same manner as in example 2.
(II) extraction of RNA
Clipping the tissue about 2 x 3mm, and placing into a 2mL EP tube; adding 1mL of Trizol, adding 2 steel balls, homogenizing by using a tissue disruptor, and performing homogenization at 50Hz for 5 min; 9000rpm, 4 ℃ for 1min, transfer the supernatant to a new 1.5mL EP tube; adding 200uL chloroform, slightly reversing for several times, mixing uniformly, and standing for 5 minutes at room temperature; 9000rpm, 4 ℃ for 15 min; transferring the upper water phase (about 400uL) into a new 1.5ml EP tube, adding isopropanol with the same volume, mixing uniformly, and standing at room temperature for 10 min; 9000rpm 10min 4 ℃; discarding the supernatant, washing the precipitate with pre-cooled 70% anhydrous ethanol for 1 time at 8000rpm for 5min 4 deg.C; discarding the supernatant, air-drying for 5-10 min, and dissolving in 30ul DEPC water; the RNA concentration was determined spectrophotometrically.
(III) analysis of Gene expression
RNA purification and quantitative analysis of genes were performed according to the QuantiGene R2.0 Plex Assay kit, and gene expression was analyzed using the luminex platform.
In the test of determining gene expression, if the expression level of the gene is obviously increased compared with the experimental group without the added extract, it can be concluded that the cedar can induce the gene expression related to cell repair and anti-aging, and improve the stability of the structure of the skin and the efficacy of delaying aging.
As can be seen from FIG. 2, the SIRT1 gene expression was up-regulated 1.70-fold in the skin model after adding the cedar extract, indicating that the extract has the ability to improve the overall repair of cells.
Example 4 Cedar extract increases expression of keratinocyte proliferation-associated genes
The experimental procedure was the same as in example 3, and the expression of HBEGF gene in the skin model was measured, and the results are shown in FIG. 3.
As can be seen from FIG. 3, after adding Cedrus deodara extract (100. mu.g/mL), the expression of human heparin-binding EGF-like growth factor (HBEGF) in the skin model is up-regulated by 1.41 times, which shows that the extract has the efficacy of enhancing the renewal rate of keratinocytes, can promote the proliferation and migration of skin keratinocytes, improve the stability of skin structure, delay aging and have the anti-aging effect.
Claims (10)
1. Use of a cedar extract for the preparation of an external preparation for the skin, characterized in that the cedar extract is used as an active ingredient for delaying aging.
2. The use according to claim 1, wherein said cedar extract is used as an active ingredient for improving the capacity of skin cells to repair, increasing the thickness of the epidermis of the skin or maintaining the skin structure stable.
3. The use according to claim 1, wherein said cedar extract achieves anti-aging by promoting skin cell proliferation, skin cell repair, or skin top layer repair.
4. The use according to claim 1, wherein said cedar extract is used as an active ingredient for increasing the expression level of genes associated with cell repair and/or proliferation.
5. The use of claim 4, wherein the genes involved in cellular repair and/or proliferation are SIRT1 and HBEGF.
6. The use as claimed in claim 4, wherein said cedar extract promotes the proliferation and migration of skin keratinocytes by increasing the level of synthesis of human heparin binding EGF-like growth factor; by increasing the synthesis level of deacetylase, the skin cell repair capacity is increased and/or the epidermis thickness is increased.
7. The use according to any one of claims 1 to 6, wherein the cedar extract is prepared by a method comprising:
extracting the dried cedar powder with a solvent for 1-4 times, combining the filtrates, removing the solvent and drying;
the solvent is water, alcohol or alcohol-water mixture.
8. A skin external preparation having an anti-aging effect, an effect of improving the ability of repairing skin cells, and an effect of improving the stability of skin structure, characterized by containing a cedar extract.
9. The external preparation for skin as claimed in claim 8, wherein the content of the cedar extract is 0.001 to 10 wt%.
10. The external preparation for skin according to claim 8 or 9, further comprising one or more of a moisturizing active ingredient, a whitening active ingredient, a moisturizing active ingredient, an antioxidant active ingredient, an anti-wrinkle active ingredient, a spot-removing active ingredient, an acne-removing active ingredient, a sunscreen active ingredient, an acne/acne-removing active ingredient, a dandruff-removing active ingredient, an antiallergic active ingredient, or a sebaceous gland-inhibiting active ingredient.
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| JP2000154132A (en) * | 1998-11-19 | 2000-06-06 | Shiseido Co Ltd | Antiaging agent |
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| JP2000154132A (en) * | 1998-11-19 | 2000-06-06 | Shiseido Co Ltd | Antiaging agent |
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