CN109276493A - Longan flower extract with anti-aging, whitening, anti-allergic and cell repairing functions - Google Patents
Longan flower extract with anti-aging, whitening, anti-allergic and cell repairing functions Download PDFInfo
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- CN109276493A CN109276493A CN201810751433.9A CN201810751433A CN109276493A CN 109276493 A CN109276493 A CN 109276493A CN 201810751433 A CN201810751433 A CN 201810751433A CN 109276493 A CN109276493 A CN 109276493A
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- 229910001424 calcium ion Inorganic materials 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 239000000084 colloidal system Substances 0.000 description 1
- 238000003927 comet assay Methods 0.000 description 1
- 231100000170 comet assay Toxicity 0.000 description 1
- 239000013065 commercial product Substances 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 231100000135 cytotoxicity Toxicity 0.000 description 1
- 230000003013 cytotoxicity Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 230000003111 delayed effect Effects 0.000 description 1
- 235000015872 dietary supplement Nutrition 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 235000014103 egg white Nutrition 0.000 description 1
- 210000000969 egg white Anatomy 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 210000001339 epidermal cell Anatomy 0.000 description 1
- DZYNKLUGCOSVKS-UHFFFAOYSA-N epigallocatechin Natural products OC1Cc2cc(O)cc(O)c2OC1c3cc(O)c(O)c(O)c3 DZYNKLUGCOSVKS-UHFFFAOYSA-N 0.000 description 1
- 229940030275 epigallocatechin gallate Drugs 0.000 description 1
- ZMMJGEGLRURXTF-UHFFFAOYSA-N ethidium bromide Chemical compound [Br-].C12=CC(N)=CC=C2C2=CC=C(N)C=C2[N+](CC)=C1C1=CC=CC=C1 ZMMJGEGLRURXTF-UHFFFAOYSA-N 0.000 description 1
- 229910001447 ferric ion Inorganic materials 0.000 description 1
- 239000003337 fertilizer Substances 0.000 description 1
- 239000012894 fetal calf serum Substances 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 210000002950 fibroblast Anatomy 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 229930003944 flavone Natural products 0.000 description 1
- 235000011949 flavones Nutrition 0.000 description 1
- 238000011010 flushing procedure Methods 0.000 description 1
- 235000013373 food additive Nutrition 0.000 description 1
- 239000002778 food additive Substances 0.000 description 1
- 238000013467 fragmentation Methods 0.000 description 1
- 238000006062 fragmentation reaction Methods 0.000 description 1
- 239000003292 glue Substances 0.000 description 1
- 239000004220 glutamic acid Substances 0.000 description 1
- 235000013922 glutamic acid Nutrition 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 229960000890 hydrocortisone Drugs 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 230000002757 inflammatory effect Effects 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 230000000977 initiatory effect Effects 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 230000033001 locomotion Effects 0.000 description 1
- 229910001629 magnesium chloride Inorganic materials 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 230000000116 mitigating effect Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
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- 239000002773 nucleotide Substances 0.000 description 1
- 125000003729 nucleotide group Chemical group 0.000 description 1
- 230000020520 nucleotide-excision repair Effects 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 238000012856 packing Methods 0.000 description 1
- 230000000149 penetrating effect Effects 0.000 description 1
- 239000002953 phosphate buffered saline Substances 0.000 description 1
- 239000000419 plant extract Substances 0.000 description 1
- 150000008442 polyphenolic compounds Chemical class 0.000 description 1
- 235000013824 polyphenols Nutrition 0.000 description 1
- 239000001103 potassium chloride Substances 0.000 description 1
- 235000011164 potassium chloride Nutrition 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 238000010791 quenching Methods 0.000 description 1
- 230000000171 quenching effect Effects 0.000 description 1
- 239000002516 radical scavenger Substances 0.000 description 1
- 238000010992 reflux Methods 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 230000009758 senescence Effects 0.000 description 1
- UQDJGEHQDNVPGU-UHFFFAOYSA-N serine phosphoethanolamine Chemical compound [NH3+]CCOP([O-])(=O)OCC([NH3+])C([O-])=O UQDJGEHQDNVPGU-UHFFFAOYSA-N 0.000 description 1
- 230000009759 skin aging Effects 0.000 description 1
- 230000037394 skin elasticity Effects 0.000 description 1
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 238000007447 staining method Methods 0.000 description 1
- 210000000130 stem cell Anatomy 0.000 description 1
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Classifications
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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]
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/77—Sapindaceae (Soapberry family), e.g. lychee or soapberry
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/16—Emollients or protectives, e.g. against radiation
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/18—Antioxidants, e.g. antiradicals
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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
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/08—Antiallergic agents
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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
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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
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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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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- 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
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/805—Corresponding aspects not provided for by any of codes A61K2800/81 - A61K2800/95
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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
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/84—Products or compounds obtained by lyophilisation, freeze-drying
Abstract
The invention relates to a longan flower extract with anti-aging, whitening, anti-allergic and cell repairing functions, which is characterized in that an effective dose of longan flower extract is applied to skin cells, and the longan flower extract has the effects of eliminating DPPH and ABTS + free radical, reducing power, chelating iron ions and inhibiting lipid peroxide; inhibiting tyrosinase activity; scavenging Nitric Oxide (NO) free radicals, and inhibiting lipoxygenase-1(LOX-1) activity; inhibiting the capacity of the hyaluronidase, improving the content of hyaluronic acid of the skin, promoting the wound repair and reducing the generation of wrinkles; inhibiting Matrix Metalloproteinases (MMPs) activity, and reducing collagen degradation; promoting cell repair and improving anti-allergic ability.
Description
Technical field
The present invention about a kind of longan extracts application, refer in particular to using longan extracts as cosmetics,
The additive of the external skin products such as skin care products, to reach anti-aging, anti-oxidant, anti-inflammatory, inhibit melanin, crease-resistant, antiallergy
And the purpose of the machining functions such as cell repair.
Background technique
Modern Asians increasingly improves white-skinned colour of skin demand, and the attention also the same for health, therefore
How to propose that a kind of light spot mode of health is a development priority in fact for medical exterior-applied article industry.Traditional plant is protected for beauty
It supports, mostly some inrigorating qi and promoting blood circulations, the temperature compensation drug for adjusting constitution also have a little list, compound to be used directly for skin-whitening, remove
The purposes such as acne, anti-inflammatory.But mechanism analysis and its matching in medical exterior-applied article for the effective component in plant extract
Side's design application is still the emphasis that domestic medical exterior-applied article industry needs active development.Therefore as can exploitation traditional plant becomes low
Dosage, dynamical light spot product, it will help promote medical exterior-applied article scientific and technological industry.
Have now been found that the problem of be mostly be by vitamin C (Vitamin C), to benzene inside existing whitening product
Diphenol (Hydroquinone), azalaic acid (Acelaic acid) and Arbutin (Arbutin) come the effect of reaching light spot, though
These right substances truly have significant effect for light spot, however, the effect of some light spot agent is directly to destroy melanocyte,
Therefore it will cause the poisoning of cell.Therefore inhibiting the safety of melanin production and antioxidant content to function concentration is value
It must pay attention to.The present invention be extract natural plants longan extracts have remove DPPH and ABTS+ free radical ability,
Reducing power, chelates ferric ions and anti-lipid peroxidation object efficiency；Inhibit Tyrosinase activity；Remove nitric oxide (NO) freely
Base inhibits lipoxygenase lipoxygenase-1 (LOX-1) activity.
Most of document all points out that flos longan has effects that anti-oxidant and anti-inflammatory, but rarer inhibition melanin intercrescence
At and the active correlative study repaired of DNA, do not have its whitening and the discovery of anti-aging function, and a beauty that commercial product is contained yet
Bai Chengfen is for example: vitamin C (Vitamin C), hydroquinone (Hydroquinone), azalaic acid (Acelaic acid) and
Arbutin (Arbutin), though having remarkable efficacy, the effect of some whitening agents is directly to destroy melanocyte, therefore
It will cause the poisoning of cell.
Inventor be still have when actual implementation use in view of above-mentioned existing whitening, anti-aging product composition it is more
Place's missing, this is improved, and grind accordingly in tireless spirit by its practical experience for enriching professional knowledge and many years
Create the present invention.
Summary of the invention
Flos longan contains a large amount of Polyphenols and flavone compound, has good oxidation resistance, and has drop blood
The effect of rouge and inhibition lipid synthesis.However pertinent literature discuss it is rare, therefore the present invention i.e. with longan extracts into
A series of anti-oxidant, DNA of row are repaired, are inhibited melanin GCMS computer and anti-inflammatory Activity Assessment.
Main purpose of the present invention is to provide a kind of application of longan extracts, is referred to flos longan (Dimocarpus
Longan flower, DLF) water extracts and ethanolic extract is used as with Scavenging ability and inhibits Tyrosinase active
Cosmetics additive；Whereby, longan extracts can be used as external skin product with the skin aging for preventing or mitigating individual
Phenomenon reaches anti-aging, anti-oxidant, anti-inflammatory, inhibits the machining functions such as melanin, crease-resistant, antiallergy and cell maintenance.
In order to reach above-mentioned implementation purpose, a kind of application of longan extracts of the present invention, is by the dragon of effective dose
Dim eyesight extract is applied to Skin Cell, has and removes free radical, anti-lipid peroxidation object efficiency and rouge oxygen is inhibited to close
Effect that is anti-oxidant, inhibiting melanin production, anti-inflammatory and cell repair can be improved in enzyme (lipoxygenase-1, LOX-1) activity
Fruit.
Preferably, the longan extracts include that water extracts DLFW and ethanolic extract DLFE.
Preferably, which, which has, removes or reduces ability of the free radical to prevent cell senescence.
Preferably, the free radical are DPPH (2,2-diphenyl-1-picrylhydrazyl) free radical or ABTS (2,2-
Azino-bis-3-ethylbenzothiazoline-6-sulfonic acid) free radical.
Preferably, the longan extracts are as whitening or oxidation resistant cosmetic constituent, food additives or doctor
Medicine constituent.
Preferably, the longan extracts have the ability for inhibiting UV to irradiate caused chafing reaction or as skins
Antiphlogistic.
Preferably, cosmetic constituent or medical component of the longan extracts as anti-inflammatory.
There is the correlation factor for adjusting p53 and NER repair system to be irradiated to repair by UV for preferably, the longan extracts
The ability of the damage of caused DNA.
Preferably, which has the activity for inhibiting extracellular mushroom Tyrosinase, and then reduces melanin
The ability of generation.
Preferably, the activity which can inhibit NO and LOX-1 achieve the purpose that inhibit inflammatory response.
Preferably, the longan extracts have the ability for inhibiting sodium hyaluronate enzyme, improve skin sodium hyaluronate content, promote wound
Mouth is repaired, and is reduced wrinkle and is generated.
Preferably, which, which has, inhibits matrix metalloproteinase (MMPs) activity, reduces collagen drop
The ability of solution.
Preferably, the longan extracts have the ability for promoting cell repair.
Preferably, which, which has, improves antianaphylactic ability.
Detailed description of the invention
Fig. 1 is cell toxicant rationality test chart of the present invention.
Fig. 2 is Scavenging ability test chart of the present invention.
Fig. 3 is whitening effect test figure of the present invention.
Fig. 4 is anti-inflammatory Test Drawing of the present invention.
Fig. 5 is DNA protection test figure of the present invention.
Fig. 6 is that DNA of the present invention protects comparative diagram.
Fig. 7 is the crease-resistant Test Drawing of the present invention.
Fig. 8 is that the present invention inhibits sodium hyaluronate enzyme ability test figure.
Fig. 9 is the wound-healing abilities Test Drawing of cell of the present invention.
Figure 10 is UVB cell damage Test Drawing of the present invention.
Figure 11 is antiallergic ability test figure of the present invention.
Specific embodiment
Advantage in the purpose of the present invention and its structure function will cooperate specific real according to structure shown in following drawing
It applies example to be explained, so that juror can have the present invention deeper into and specifically understand.
Firstly, the present inventor is separated by flos longan is made water extraction DLFW, ethanolic extract DLFE.For example, water extracts
DLFW then can obtain water-soluble filtrate drying to mix flos longan with water to obtain water-soluble filtrate, and mix
Temperature is preferably 80 to 90 DEG C；Ethanolic extract DLFE can be to mix flos longan to obtain ethyl alcohol dissolubility filtrate with ethyl alcohol,
Then the drying of ethyl alcohol dissolubility filtrate is obtained.Weight ratio between flos longan and water is preferably 1:8 to 1:10, more preferably
For 1:9；And the weight ratio between flos longan and ethyl alcohol is preferably 1:4 to 1:6, is more preferably 1:5.In addition, in order to allow water to extract
DLFW can completely be contacted with ethanolic extract DLFE with biological cell, and water extracts DLFW and ethanolic extract DLFE can be with mixed solution
Form acts on biological cell.For example, water extraction DLFW or ethanolic extract DLFE can first be dissolved in solvent (such as secondary water,
DMSO or PBS) in, then biological cell acted on resulting mixed solution.The water extraction DLFW and ethanolic extract of above-mentioned material
DLFE has reduction because of H2O2HaCaT Cellular Oxidation pressure caused by stimulation improves glutathion inside cell
(glutathione, GSH) content, and raising antioxidative enzyme: superoxide dismutase (superoxide dismutase,
SOD), the table of catalase (catalase) and glutathione peroxidase (glutathione peroxidase, GPx)
Now measure.
Longan extracts (water extracts DLFW, ethanolic extract DLFE), which have, inhibits intracellular Tyrosinase
(tyrosinase) activity and the ability of reduction Melanin productions.It is tested with immunofluorescent test and Western blot, longan
After extracts (water extracts DLFW, ethanolic extract DLFE) act on cell, it was demonstrated that have and inhibit melanin GCMS computer in
Recipient MC1R (melanocortin-1receptor) and MITF (microphthalmia-associated
Transcription factor) performance, and influence Tyrosinase, TRP-2 (tyrosinase-related proteins-
2) and the performance of TRP-1 (tyrosinase-relatedproteins-1), and then inhibit melanin production.
In anti-inflammatory ability test, longan extracts have inhibit intracellular inflammatory factor (TNF-α, IL-1 β and
IL-6 the ability of content), and there is the p38, IL-6, IL-1 β and TNF-α protein expression for reducing inflammation introductory path.It is dynamic
Object tests SKH-1 mouse after UVB irradiates with immunohistochemistry staining method (immunohistochemistry, IHC) and west
The test of Fang Modian method confirms that longan extracts have the p38, IL-6, IL-1 β and TNF-α protein for reducing inflammation introductory path
Performance.
In DNA is repaired and protective capability is tested, longan extracts, which have, adjusts Nucleotide Sequence Analysis
The correlation factor of (nucleotideexcision repair, NER) system is irradiated the damage of caused DNA to repair by UV, with
And it adjusts the correlation factor of p53 and NER repair system and caused DNA damage is irradiated by UV to repair.
By following specific embodiments, can further prove the present invention can practical application range, but be not intended to any
Form limits the scope of the invention.
Embodiment 1: the water extraction of flos longan
Flos longan is bought from peasant association of the township Nantou County Zhong Liao.After flos longan is smash essence, 100 grams are taken to smash extraction object and 900 grams
Water mixes in double ground round-bottomed flasks.Then, merging heating, which is wrapped and mixed in a manner of being heated to reflux at 80 to 90 DEG C, smashes extraction
Object with water about 3 hours.It is after time reaches, double ground round-bottomed flasks are cooling, then with cloth funnel and aspiration pump to mixture mistake
Filter, and obtain clear filtrate.Finally, filtrate is first concentrated under reduced pressure, the filtrate after freeze concentration at -80 DEG C about one day, after
It is placed in Dewar bottle and is freeze-dried that (extraction yield about 7.2%, 100 cron dim eyesights are available to obtain final product water extraction object DLFW
7.2 grams of water extract object DLFW).It should be noted that water extraction object DLFW first must be dissolved in secondary water before carrying out subsequent Bioexperiment
Or in DMSO equal solvent, if therefore the concentration that is hereafter mentioned or similar indicating without especially defining the weight phase for referring both to water extraction object DLFW
Extract the concentration of the total volume of object DLFW and solvent for water.
Embodiment 2: the ethyl alcohol extraction of flos longan
Flos longan is bought from peasant association of the township Nantou County Zhong Liao.After flos longan is smash essence, 100 grams of extraction object of smashing is taken to be soaked in 500
Gram 95% ethyl alcohol up to three days.Then, mixture is filtered with cloth funnel and aspiration pump, and obtains clear filtrate.In addition, mistake
The filter residue obtained after filter is soaked in 500 grams of 95% ethyl alcohol up to after three days, with cloth funnel and aspiration pump to mixing herein again
Object filtering, and obtain clear filtrate herein.Finally, filtrate is first concentrated under reduced pressure after resulting filtrate mixing twice, then
Filtrate at -80 DEG C after freeze concentration about one day is placed in Dewar bottle and is freeze-dried to obtain the extraction of final product ethyl alcohol
Object DLFE (6.0 grams of ethanolic extract DLFE can be obtained in extraction yield about 6.0%, 100 cron dim eyesights).It should be noted that after progress
Before continuous Bioexperiment, first ethanolic extract DLFE must be dissolved in secondary water or DMSO equal solvent, therefore hereafter mentioned
If concentration similar is indicated without especially defining the weight for referring both to ethanolic extract DLFE relative to ethanolic extract DLFE and solvent
Total volume concentration.
Embodiment 3: cell toxicant rationality test
Effect in view of existing whitening agent or dermal melanin inhibitor will cause cytotoxicity, therefore the present inventor exists
After obtaining this longan extracts, the cell poisoning test experiments of this compound are first carried out, it is dense using 100,250 and 500 μ g/mL
After the longan extracts of degree act on HaCaT cell 24 hours, is tested using MTT and measure its survival degree.Its result such as Fig. 1
Shown, the water extraction DLFW and ethanolic extract DLFE of flos longan is not toxic in cell, with commercially available common ingredient ascorbic acid
(ascorbic acid, AA) is compared to more secure.
Embodiment 4: Scavenging ability test
The effect of water extraction DLFW, the ethanolic extract DLFE of this experiment discussion flos longan remove free radical, using DPPH and
Two kinds of free radicals of ABTS+ test its Scavenging activity, and result is as shown in Fig. 2, ethanolic extract DLFE is removing DPPH free radical
About just reach 50% clearance rate in test in 45 μ g/mL, and ethanolic extract DLFE removes the ability of two kinds of free radicals more
Extract DLFW better than water, longan extracts have good inhibiting rate for two kinds of free radicals as the result is shown.
Embodiment 5: whitening effect test
Tyrosinase is ferment important in melanin GCMS computer approach, and the water of flos longan is extracted DLFW and ethanolic extract
DLFE carries out extracellular mushroom Tyrosinase activity test assessment with various concentration 100,250 and 500 μ g/mL.Its result such as Fig. 3
It is shown, by flos longan water extract DLFW and ethanolic extract DLFE effect after, inhibit extracellular mushroom Tyrosinase activity with
Concentration increases and improves, and ethanolic extract DLFE (EC50Extract DLFW (EC better than water for the inhibiting rate of 410 μ g/mL)50For 624 μ
G/mL), it was demonstrated that longan extracts have the active ability for inhibiting extracellular mushroom Tyrosinase.
Embodiment 6: anti-inflammatory test
NO is to be primarily involved in one of medium of inflammatory response, and LOX-1 is the most common ferment for causing inflammatory response, by
The water extraction DLFW and ethanolic extract DLFE anti-inflammatory effect of this test assessment flos longan.Its result is as shown in figure 4, flos longan
Water extracts DLFW and ethanolic extract DLFE, has the ability of good removing NO free radical and inhibits LOX-1 activity, it was demonstrated that flos longan
Water extraction DLFW and ethanolic extract DLFE can via inhibit NO and LOX-1 activity come reach inhibit inflammatory response activity,
Has the effect of anti-inflammatory.
Embodiment 7:DNA protection test
UV exposure or physiological metabolism will increase the damage that oxidative pressure causes DNA.This test is with pUC119DNA
With H2O2(1mM) is induced and with UVB (20mJ/cm2) irradiation, cause DNA damage, assesses the water extraction DLFW and ethyl alcohol extraction of flos longan
Take whether object DLFE there is protection DNA to avoid the DNA damage caused by irradiating because of oxidative pressure and UVB.Referring to figure 5.,
PUC119DNA was supercoil body structure (Supercoiled form, SC-form) originally, through H2O2(1mM) induction and UVB (20mJ/
cm2) it will cause DNA break after irradiation, form line style (Linear form, L-form) and style of opening (Open form, O-
Form DNA fragmentation).It is continuous referring to Fig. 6, when the ethanolic extract DLFE (100,250 and 500 μ g/mL) of various concentration is added again
With H2O2After induction and UVB irradiation, ethanolic extract DLFE energy effective protection DNA, and protection effect with the increase of concentration as the result is shown
Fruit is better, it was demonstrated that flos longan have protection DNA from being irradiated because of oxidative pressure and UVB caused by damage.
Embodiment 8: crease-resistant test
Collagen is primarily present in skin histology in the connective tissue of skin corium, is the highest albumen of people's in-vivo content
Matter has very strong stretching force and can support skin corium overall structure, and collagen is also the main constituents of extracellular matrix,
It keeps skin elasticity and there is the ability repaired, and collagen can be lost with factors such as aging and environment, produce skin
The problems such as raw wrinkle and osteoporosis.Its matrix metalloproteinase (Matrix metalloproteinases, MMPs) is glue
Former protease, MMP-2 and MMP-9 mainly decompose typeⅡ Collagen.To adjust light aging important factor, fiber
Mother cell can be caused matrix metalloproteinase largely to show by UV exposure, and then degradation extracellular matrix (ECM) makes dermal tissue
Collagen degradation.
The ethanolic extract DLFE (100,250 and 500 μ g/mL) of flos longan is acted on Hs68 with various concentration by this test
After cultivating 72 hours in cell, collagen content is measured.Whether the ethanolic extract DLFE of the test assessment flos longan whereby
With the activity for inhibiting MMPs.Its result as shown in fig. 7, through flos longan ethanolic extract DLFE effect after, with control group
(Control, also referred to as control group) is compared, and collagen content increases as concentration improves, therefore can deduce flos longan
Ethanolic extract DLFE has the activity for inhibiting MMP-9 and MMP-2, and can effectively increase fibroblast and generate collagen egg
White content.
Embodiment 9: inhibit sodium hyaluronate enzyme ability test
Sodium hyaluronate is present in the skin corium in human skin tissue and connective tissue, reduces the generation of wrinkle, and has
The effect of promoting wound to repair, and sodium hyaluronate enzyme is a kind of ferment that sodium hyaluronate can be made to degrade, and disintegrates the structure of sodium hyaluronate, is lost
Lose original function.Inhibit the ability of sodium hyaluronate enzyme by the ethanolic extract DLFE of this test assessment flos longan.
By the ethanolic extract DLFE (500 μ g/mL) and standard items epigallocatechin of flos longan
(Epigallocatechin gallate, EGCG) is separately added into sodium hyaluronate enzyme (3000unit/mL) effect 18 hours, as a result
As shown in figure 8, the position after the ethanolic extract DLFE of flos longan effect compares the deeply obvious many of color with control group,
Display DLFE can be effectively prevented sodium hyaluronate and be decomposed by sodium hyaluronate enzyme, go out DLFE pairs of ethanolic extract of flos longan through quantitative analysis
The inhibiting rate of sodium hyaluronate enzymatic activity is 82%.As a result, it was confirmed that the ethanolic extract DLFE of flos longan, which has, inhibits sodium hyaluronate enzyme activity
The ability of property.
Embodiment 10: the wound-healing abilities test of cell
Whether there is the ability for promoting cell repair with wound healing (wound healing) test assessment DLFE.First
After acting on HaCaT cell 4 hours with longan extracts DLFE (500 μ g/mL), culture solution is drained, with UVB (50mJ/cm2)
Irradiation adds DLFE effect, after culture 4 hours, with the mobile variation of micro- sem observation cell.
As a result as shown in figure 9, the cell of control group and simple irradiation UVB are without too big mobile variation, especially merely
The cell for irradiating UVB, has almost no change for from 0 to 48 hour.Cell after longan extracts DLFE effect was at 12 hours
Have the tendency that afterwards significantly to centre movement, by 48 hours without gap.As a result it reconfirms, longan extracts
DLFE has the ability for promoting cell repair.
The test of embodiment 11:UVB cell damage
Comet Assay is to detect DNA in single cell with colloid electrophoresis and with the dyeing of EtBr progress nucleus and shine through UVB
Degree of injury after penetrating.After acting on HaCaT cell 4 hours with DLFE (500 μ g/mL), culture solution is drained, with UVB (50mJ/
cm2) irradiation, longan extracts DLFE effect is added, after culture 4 hours, with the degree of injury of micro- sem observation cell.
The results are shown in Figure 10, the cell irradiated by UVB merely, and DNA is impaired and has apparent trailing phenomenon, 1000 thin
In born of the same parents, there is impaired situation there are about the DNA of 50% cell.Cell after being acted on by DLFE, in 500 μ g/mL of high concentration
Under effect, in 1000 cells, there is impaired situation there are about the DNA of 8% cell.Judge that there is DLFE protection DNA to exempt from whereby
The injury caused by UVB irradiation.
CPD is generated photoproduct after DNA is irradiated by UVB, and the results are shown in Figure 10, through UVB (50mJ/cm2) irradiation
The fluorescence performance amount of cell afterwards, CPD increases, and after DLFE (500 μ g/mL) effect, hence it is evident that reduce UVB photoproduct
It generates, reconfirms the DNA damage caused by DLFE has protection DNA and reduces because of UVB irradiation.
Embodiment 12: antiallergic ability test
Figure 11 is please referred to the β of Calcimycin A23187 or antigen induction-hexosaminidase degranulation test: with Calcimycin
The degree of A23187 and antigen induction RBL-2H3 mast cell degranulation are by β-hexosaminidase release test, via following side
The method of formula improvement measures: inoculation RBL-2H3 mast cell (2 × 104Cells/well) in 96 hole micro-plates and RBL-2H3
Mast cell (3 × 104Cells/well) in 48 hole micro-plates, Calcimycin A23187 Induction experiments and antigen are supplied respectively to
Used in Induction experiments, the sample of various concentration is added in cell, stands 20 hours；Use enlightening cortisol (10nM) as positive control
Processed group.In antigen induction experiment, cell is first with anti-dinitrophenol IgE monoclonal antibody (anti-dinitrophenol
IgE, anti-DNP IgE) (5 μ g/mL) passive high quick Tai Luodeshi buffer (135mM at least 2 hours, cell preheated
Sodium chloride, 5mM potassium chloride, 1.8mM calcium chloride, 1.0mM magnesium chloride, 5.6mM glucose, 20mM 4- ethoxy croak piperazine ethanesulfonic acid,
PH 7.4) after cleaning down, distinctly with the calcium ion carrier A 23187 of Tai Luodeshi buffer (Calcimycin, 1 μM) or
Antigen dinitrobenzene-fetal calf serum conjugate (100ng/mL) stimulates one hour.It, will for measurement β-hexosaminidase release total amount
Unprovoked cell is cracked with 0.5% Triton X-100 solution, or without handling the release for making cell spontaneity
β-hexosaminidase；1 μM of poly- nitrogen-acetylglucosamine (antigen) of packing supernatant (50 μ L) and equivalent is formulated in 0.1M citric acid
In salt buffer (pH 4.5), as release β-hexosaminidase substrate.After cultivating 1 hour at 37 DEG C, the end of 100 μ L is added
Only buffer (0.1M Na2/NaHCO3, pH 10.0) and quenching reaction.By micro-plate analyzer (Multiskan Ascent,
Thermo Scientific) it is set in 405nm wavelength, brightness is inhaled in measurement；β-hexosaminidase release suppression percentage is experiment
Group is calculated divided by the percentage of controlling value (untreated cell)；It is 0.5% to the maximum tolerance dose of dimethyl sulfoxide, and institute
There is experiment in triplicate.
The direct degranulation test of β-hexosaminidase of sample induction: in RBL-2H3 mast cell, by the β-of sample initiation
The burst size of glycuronidase is determined with improved β-hexosaminidase release experiment；Briefly, by RBL-2H3 fertilizer
Maxicell (4 × 104Cells/well it) is inoculated in 48 hole micro-plates, stands 10 hours after sample is added.Tai Luodeshi is delayed
5.6mM glucose, 2mg/mL bovine serum albumin(BSA) (BSA) and 2mM glutamic acid is added to prepare used in sample and cell in fliud flushing.50
The supernatant of μ L is transferred to 96 hole micro-plates, measures β-hexosaminidase burst size, Calcimycin A23187 according to foregoing manner
(1 μM) as positive control group, all experiments are carried out more than in triplicate.
In conclusion the invention demonstrates that the water extraction DLFW and ethanolic extract DLFE of flos longan has Scavenger of ROS and suppression
Melanin production processed, and the protection of the DNA with skin epidermal cells, repair ability and the generation with inhibition inflammatory response,
With the potentiality for developing into whitening, anti-oxidant and anti-inflammatory skin care products or food supplement, and really can be by above-mentioned
Before disclosed embodiment reaches desired use effect, and the present invention is not also disclosed in application, really comply fully with specially
The regulation and requirement of sharp method.Therefore the application of patent of invention is proposed in accordance with the law, it earnestly asks and gives examination, and grant quasi patent, then true feeling moral is just.
Above-mentioned taken off diagram and explanation, only presently preferred embodiments of the present invention, non-is to limit the scope of protection of the present invention；
Generally it is familiar with the personnel of this technology, institute's diagnostic categories under this invention, made other equivalent change or modifications should all regard
Not depart from design scope of the invention.
Claims (10)
1. a kind of longan extracts are used to prepare the purposes of the composition of anti-aging, whitening, antiallergy and cell maintenance, described
Composition is to be applied to Skin Cell with the longan extracts containing effective dose, to remove free radical, anti-lipid peroxidation
Object efficiency and inhibition lipoxygenase activity, and then improve anti-oxidant, inhibition melanin production, anti-inflammatory and cell repair.
2. purposes as described in claim 1, wherein the flos longan extract includes that water extracts DLFW and ethanolic extract DLFE.
3. purposes as described in claim 1, wherein the flos longan extract, which has, removes or reduce free radical to prevent carefully
The ability of born of the same parents' aging.
4. purposes as described in claim 1, wherein the free radical is DPPH free radical or ABTS free radical.
5. purposes as described in claim 1, wherein the flos longan extract has the phase for adjusting p53 and NER repair system
The factor is closed to repair the ability for the damage for irradiating caused DNA by UV.
6. purposes as described in claim 1, wherein the flos longan extract, which has, inhibits extracellular mushroom Tyrosinase
Activity, and then reduce the ability of melanin production.
7. purposes as described in claim 1, wherein the activity that the flos longan extract can inhibit NO and LOX-1 reaches inhibition
The purpose of inflammatory response.
8. purposes as described in claim 1 improves skin wherein the flos longan extract has the ability for inhibiting sodium hyaluronate enzyme
Skin sodium hyaluronate content promotes wound reparation, reduces wrinkle and generates.
9. purposes as described in claim 1, wherein the flos longan extract also has inhibition matrix metal proteinase activity,
Reduce the ability of collagen degradation.
10. purposes as described in claim 1, wherein the flos longan extract, which has, improves antianaphylactic ability.
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| CN107137307A (en) * | 2017-05-09 | 2017-09-08 | 江西中医药大学 | A kind of anti-oxidant lightening compositions containing opc |
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| TWI803146B (en) * | 2022-01-11 | 2023-05-21 | 捷康生技有限公司 | Use of longan flower extract for increasing melatonin in individuals |
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