CN104435366A - Composition containing herbal extract and makeup material composition - Google Patents
Composition containing herbal extract and makeup material composition Download PDFInfo
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- CN104435366A CN104435366A CN201410598149.4A CN201410598149A CN104435366A CN 104435366 A CN104435366 A CN 104435366A CN 201410598149 A CN201410598149 A CN 201410598149A CN 104435366 A CN104435366 A CN 104435366A
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Abstract
The invention discloses a composition containing a herbal extract and a makeup material composition. The composition containing the herbal extract comprises 5-20% of a guava leaf extract, 5-20% of an angelica sinensis whole grass extract, 5-35% of a tea extract and a lotus leaf extract by mass, totaling 100%. The makeup material composition is the composition containing the herbal extract. The composition which contains mixed extracts such as the lotus leaf extract, the tea extract, the guava leaf extract and the angelica sinensis whole grass extract can inhibit reproductive growth of propionibacterium acnes which causes acnes of skins and inflammatory response, has a very good improving effect on skin problem, promotes blood circulation of the skins and maintains lubricative and clean skins, and is free of irritation and side effects and high in safety when being used.
Description
Technical field
The present invention relates to a kind of compositions containing medicinal herbal extract, be specifically related to the compositions, particularly cosmetic combination that can improve the skin problem such as acne, skin allergy.Its extract mixtures using Folium Nelumbinis extract, Folium Camelliae sinensis extract, Folium Psidii Guajavae extract and Radix Angelicae Sinensis herb extract to mix by a certain percentage all presents excellent effect as medicinal herbal extract in antibacterial, antioxidation, antiinflammatory and skin sedation etc.
Background technology
Acne be most people at the generable pleomorphic skin disease of all age brackets, mainly embody with the symptom such as acne, pustule, pimple, tuberosity on the positions such as face, breast, shoulder.The concrete occurrence cause of acne can't get across at present completely, but be commonly considered as dyskeratosis in excessive sebaceous gland sebum secretion, hormonal imbalance, inherited genetic factors, hair follicle, the many factors such as amount reproduction and inflammatory reaction of propionibacterium acnes (Propionibacterium acnes) causes, especially, by stimulate in the androgen of sebaceous gland and hair follicle decompose sebum generate the propionibacterium acnes of free fatty induce the exacerbate inflammation caused to be considered to most important factor.
Main pathogens-the propionibacterium acnes of acne is that the anaerobic existed in skin is everlasting bacterium, is the microorganism of the close fatty grown in skin follicle.Propionibacterium acnes can be hydrolyzed in the sebum formed by androgen or other factors after triacylglycerol, and secretion lipase (lipase) and chemotactic factor generate free fatty and the disease that causes inflammation.
The orientation treatment of this acne is mainly to relax the improper keratinization of hair follicle cell, and cortex secretion normalization, suppresses to carry out for the purpose of propionibacterium acnes excessive multiplication and minimizing inflammatory reaction etc.
Current is benzene peroxide and BHA (beta-hydroxy acid) to the more effective prescription of propionibacterium acnes.But benzene peroxide generally belongs to pharmaceuticals composition, cannot use in cosmetics, and BHA in cosmetics, content can not more than 0.5%, therefore effect is also general.And benzene peroxide has stronger zest, also the multiple side effect such as erythema, edema, xeroderma can be caused.
So exploitation is a kind of based on natural goods, and can has and become research and development focus with the new medical material had no side effect of the equal drug effect of benzene peroxide.
Relate to " Dermatologic preparation composition containing green tea chitting piece extract " in No. 2013-011542nd, Korean Patent Publication No., about the compositions containing effective ingredient green tea (Camellia sinensis) seed extract, be specially skin care, aging resistance, anti-wrinkle, improve skin elasticity, whitening, moisturizing, the prevention of the xerosis cutis diseases such as allergy and improvement, sebum regulates, antiinflammatory, the prevention of the skin problems such as acne and improvement, skin-color improves, remove cutin, prevention depilation, the Dermatologic preparation composition containing green tea chitting piece extract of the effect such as anti-dandruff.
Summary of the invention
The object of the invention is to provide a kind of compositions containing medicinal herbal extract for relaxing acne containing mixed extracts such as Folium Nelumbinis extract, Folium Camelliae sinensis extract, Folium Psidii Guajavae extract and Radix Angelicae Sinensis herb extract.The present invention all to have excellent effect containing the compositions of medicinal herbal extract relaxing in acne, antioxidation, antiinflammatory, skin sedation etc., and is suitable for human-body safety.
Compositions of the present invention not only has the effect relaxing acne, also all has excellent effect to the aspect such as antibacterial, antioxidation, antiinflammatory, skin sedation, can improve irritated grade for skin problem.The present composition also can be used as effective ingredient in functional cosmetic combination.
The means of technical solution problem
For relaxing a kind of compositions containing medicinal herbal extract of acne, comprise Folium Psidii Guajavae extract 5-20%, Radix Angelicae Sinensis herb extract 5-20%, Folium Camelliae sinensis extract 5-35% and Folium Nelumbinis extract by mass percentage, each constituent mass percentage ratio sum is 100%.
Preferably, described Folium Psidii Guajavae extract is Folium Psidii Guajavae ethanol extraction.
Preferably, described Radix Angelicae Sinensis herb extract is Radix Angelicae Sinensis herb ethanol extraction.
Preferably, described Folium Camelliae sinensis extract is Folium Camelliae sinensis ethanol extraction.
Preferably, described Folium Nelumbinis extract is Folium Nelumbinis ethanol extraction; More preferably, described Folium Nelumbinis extract be by the aqueous solution of described Folium Nelumbinis ethanol extraction with an organic solvent multistage extract and separate after extract and separate composition in the extract that obtains.
Wherein, the described organic solvent adopted in described multistage extract and separate is preferably one or more in hexane, chloroform, ethyl acetate and butanols.
Wherein, preferably, in time adopting multiple organic solvent in described multistage extract and separate, variant organic solvent uses successively.More preferably, in time adopting multiple organic solvent in described multistage extract and separate, hexane, chloroform, ethyl acetate and butanols is used to carry out multistage extract and separate successively.
The present invention also provides a kind of cosmetic combination, and it comprises the above-mentioned compositions containing medicinal herbal extract.
In described cosmetic combination, the content of the described compositions containing medicinal herbal extract is preferably 1000-10000ppm, and being more preferably 1500-8000ppm, is 1800-4500ppm best.
Particular content of the present invention
The present invention relates to a kind of by used in combination by a certain percentage to Folium Nelumbinis, Folium Camelliae sinensis, Folium Psidii Guajavae and Radix Angelicae Sinensis herb extract, for the compositions in cosmetic preparation.Described compositions all has excellent effect to the aspect such as antibacterial, antioxidation, antiinflammatory, skin sedation, and has the effects such as mitigation, calmness, improvement to the such as skin problem such as acne.
A kind of compositions containing medicinal herbal extract for relaxing acne of the present invention, comprises Folium Psidii Guajavae extract 5-20%, Radix Angelicae Sinensis herb extract 5-20%, Folium Camelliae sinensis extract 5-35% and Folium Nelumbinis extract by mass percentage; Each constituent mass percentage ratio sum is 100%.
Get the mixture of single extract in described Folium Psidii Guajavae extract, Radix Angelicae Sinensis herb extract, Folium Camelliae sinensis extract and Folium Nelumbinis extract or two or more extracts, carry out antibacterial activity, antioxidation, anti-inflammatory respectively, improve the detection of the aspects such as blood circulation, present inventor find said extracted thing to antibacterial, antiinflammatory, antioxidation, improve blood circulation all there is excellent effect, and can be used as and improve skin health state and cosmetic combination safely and effectively.Present inventor in order to obtain relaxing the aspect such as acne, skin sedation, antioxidation, antiinflammatory and all there is excellent effect and to human-body safety and be applicable to multiple dosage form material for the purpose of, finally described Folium Psidii Guajavae extract, Radix Angelicae Sinensis herb extract, Folium Camelliae sinensis extract and Folium Nelumbinis extract are mixed to get the present composition by proper proportion after having carried out large quantity research.
In the present invention, described Folium Psidii Guajavae refers to the leaf of Fructus psidii guajavae immaturus (Psidium guajava), described Psidium is in the dicotyledon of Myrtales Myrtaceae, original producton location is american torrid zone area and South Asia region, mainly be distributed in subtropical zone, in Korea S mainly in the growth of Jizhou Island area.Folium Psidii Guajavae contains the mineral compositions such as abundant polyphenol, vitamin A, B, C and ferrum, is commonly referred to as " king of vitamin ", all has remarkable effect for various allergic skin problem and anti-aging aspect.Folium Psidii Guajavae contains more carotenoid (Carotenoid) composition and the polyphenol component such as guaijaverin (Guaijaverin), long-living glycosides (Amritoside), therefore can inflammation-inhibiting skin problem lipid peroxide of increasing there is excellent antioxidant effect when occurring, can also inflammation-inhibiting trigger cell-mastocyte (Mast cell) retting conditions phenomenon and eliminate pruritis, can also suppress the synthesis of histamine discharge and to inflammation relax have better effects.Folium Psidii Guajavae not only has antiinflammatory, antioxidant effect also containing abundant vitamin A, contributes to pore refining, reduces wrinkle and effectively can remove the flaws such as brown macules, can improve skin condition on the whole.And as " king of vitamin " also containing abundant vitamin B, moisture of skin can be made to increase, prevent cell senescence.
Described Folium Psidii Guajavae extract is Folium Psidii Guajavae ethanol extraction.Particularly, described Folium Psidii Guajavae extract is the Folium Psidii Guajavae extract obtained by this area general extraction methods extraction Folium Psidii Guajavae.Described Folium Psidii Guajavae extract is preferably prepared by following method: extract after Folium Psidii Guajavae drying sample 100g adds the ethanol of concentration 30%, at 60 DEG C 4 hours, filter, at 55 DEG C concentrated at 3 hours, 60 DEG C dry 48 hours, to obtain final product; But Folium Psidii Guajavae extract of the present invention is not limited in the Folium Psidii Guajavae extract that the method obtains.
In the present invention, described Radix Angelicae Sinensis herb refers to the herb of Radix Angelicae Sinensis (Angelica gigas Nakai).Radix Angelicae Sinensis is belong to umbelliferous perennial herb as resource plant, and the medicinal herbs be always considered to a kind of property of medicine moderate notoxic, having no side effect, are especially widely used in gynaecopathia, is extraordinary medical material for women.Show according to current research, Radix Angelicae Sinensis main component decursin (Decursin) can suppress the activity of androgen (androgen can activate sebaceous gland function and promote that sebum generates).Further, decursin can stop 5-α-dehydrogenation testosterone to be combined with androgen receptor, can promote that receptor protein decomposes, and has excellent anti-androgenic effect.With the washing bath containing Radix Angelicae Sinensis, can make that skin becomes moist, blood circulation promoting, calming nerves, wound healing accelerates and reduces pain.Therefore, ancient times, geishas used the water face-washing containing Radix Angelicae Sinensis to wash hair, gentry in New Year's Eve with the washing bath of boiling Radix Angelicae Sinensis.Record that " Radix Angelicae Sinensis can treat the heavily disease such as cough, infertility, malignant boil, lump, sympotoms caused by cold factors, and wound can be made better to scab in " the integrated side of township's medicine ".
Described Radix Angelicae Sinensis herb extract is preferably Radix Angelicae Sinensis herb ethanol extraction.Particularly, described Radix Angelicae Sinensis herb extract is the Radix Angelicae Sinensis herb extract obtained by this area general extraction methods extraction Radix Angelicae Sinensis herb.Described Radix Angelicae Sinensis herb extract is preferably prepared by following method: after Radix Angelicae Sinensis herb drying sample 100g adds concentration 30% ethanol, extract at 60 DEG C 4 hours, filter, at 55 DEG C concentrated at 3 hours, 60 DEG C dry 48 hours, to obtain final product; But Radix Angelicae Sinensis herb extract of the present invention is not limited in the Radix Angelicae Sinensis herb extract that the method obtains.
In the present invention, described Folium Camelliae sinensis refers to the leaf of Camellia sinensis (Camellia sinensis), described Camellia sinensis belongs to the perennial seed plant of Theaceae, is considered to have the pharmacological properties in the human body such as antibacterial, antioxidation, anticancer, antiallergic, check melanin, suppression skin aging.Especially, current research shows, the epigallocatechin gallate (EGCG) (EGCG:Epigallocatechin-3-gallate) of Folium Camelliae sinensis main component polyphenol system can effectively suppress acne to generate the excessive secretion of main cause-sebum, also there is strong antibacterial action, more than 50% is reached for propionibacterium acnes suppression ratio.EGCG effectively can suppress the secretion of hair follicle cutin being crossed to the influential interleukin of keratinization (IL-1 α) simultaneously, proves that EGCG also has drug effect for the another kind of pathogenic factor of acne.
Described Folium Camelliae sinensis extract is preferably Folium Camelliae sinensis ethanol extraction.Particularly, described Radix Angelicae Sinensis Folium Camelliae sinensis extract is the Folium Camelliae sinensis extract obtained by this area general extraction methods extraction Folium Camelliae sinensis.Described Folium Camelliae sinensis extract is preferably prepared by following method: after tea-drying sample 100g adds concentration 30% ethanol, extract at 60 DEG C 4 hours, filter, at 55 DEG C concentrated at 3 hours, 60 DEG C dry 48 hours, to obtain final product; But Folium Camelliae sinensis extract of the present invention is not limited in the Folium Camelliae sinensis extract that the method obtains.
Described Folium Nelumbinis refers to the leaf of plant Flos Nelumbinis (Nelumbo nucifera).Flos Nelumbinis is the perennial water plant belonging to Nelumbonaceae, is extensively thought the effect having and purify water.In order to food of guaranteeing the quality better when ancient times, monks went on a long journey, can wrap up grain with Folium Nelumbinis and make grain without deterioration, Folium Nelumbinis serves the effect of natural antiseptic agent.Generally believe that the reason that Folium Nelumbinis can play purification and antibacterial action serves antibacterial action containing abundant alkaloid in Folium Nelumbinis.Folium Nelumbinis contains more nuciferine (nuciferine), nefering (neferine), cupreol (β-sitosterol), nuciferine (roemerine), nornuciferine (nornuciferine), the alkaloid components such as armepavine (armepavine).Alkaloid component is the immunizing composition of plant for self-protection, especially in Folium Nelumbinis, makes Folium Nelumbinis in sewage, keep clean containing a lot of alkaloid.Record Folium Nelumbinis character in " Dong-eui-bo-gam " gentle, when using with the green tea belonging to cool property, both coordinated can play better effect simultaneously.
Described Folium Nelumbinis extract refers to Folium Nelumbinis ethanol extraction.Described Folium Nelumbinis extract is the Folium Nelumbinis extract obtained by this area general extraction methods extraction Folium Nelumbinis.The extract of described Folium Nelumbinis extract preferably for obtaining in extract and separate composition with an organic solvent after multistage extract and separate of the aqueous solution of described Folium Nelumbinis ethanol extraction.Wherein, the described organic solvent adopted in described multistage extract and separate is preferably one or more in hexane, chloroform, ethyl acetate and butanols.
Wherein, preferably, in time adopting multiple organic solvent in described multistage extract and separate, variant organic solvent uses successively.More preferably, in time adopting multiple organic solvent in described multistage extract and separate, hexane, chloroform, ethyl acetate and butanols is used to carry out multistage extract and separate successively.
That is, as shown in Figure 1, described Folium Nelumbinis extract is prepared by following method best: after Folium Nelumbinis drying sample 100g adds concentration 70% ethanol, extracts 6 hours at 80 DEG C, filters; The extracting solution mixing obtained after 2 times are extracted and after extracting for 3 times, concentrated, use the different solvent of polarity to carry out the extract that multistage extract and separate obtains every section; Namely, after concentrated extract dilute with water, after separatory funnel using successively hexane, chloroform, ethyl acetate, fourth alcohol and water respectively carry out 3 extract and separate, use rotary vacuum evaporator (rotatory vacuum evaporator) ethanol evaporation, carry out lyophilization again; But Folium Nelumbinis extract of the present invention is not limited in the Folium Nelumbinis extract that the method obtains.
A kind of acne mitigation cosmetic combination containing medicinal herbal extract provided by the invention, it comprises the above-mentioned compositions containing medicinal herbal extract.
In described cosmetic preparation combination, the content of the described compositions containing medicinal herbal extract is preferably 1000-10000ppm, and being more preferably 1500-8000ppm, is 1800-4500ppm best.
On the basis meeting this area general knowledge, above-mentioned each optimum condition, can combination in any, obtains the preferred embodiments of the invention.
Agents useful for same of the present invention and raw material are all commercially.
Positive progressive effect of the present invention is: the compositions that the present invention contains the mixed extracts such as Folium Nelumbinis extract, Folium Camelliae sinensis extract, Folium Psidii Guajavae extract and Radix Angelicae Sinensis herb extract can suppress to cause the flourish of the propionibacterium acnes of skin acne, can inflammation-inhibiting reaction have skin problem and well improve effect, can cutaneous circulation be promoted and keep skin lubrication clean, and skin is not stimulated and side effect, during use, safety is high.
Accompanying drawing explanation
Fig. 1 is the extraction schematic flow sheet used in a preferred version of Folium Nelumbinis extract of the present invention.
Fig. 2 is the DPPH removal effect schematic diagram of Folium Camelliae sinensis extract of the present invention.
Fig. 3 is the hydroxyl radical free radical removal effect schematic diagram of Folium Camelliae sinensis extract of the present invention.
Fig. 4 is the nitric oxide production inhibition schematic diagram of Folium Psidii Guajavae extract of the present invention.
Fig. 5 is the antihistamine effect schematic diagram of Folium Psidii Guajavae extract of the present invention.
Fig. 6 is that Radix Angelicae Sinensis herb extract of the present invention is to intercellular adhesion protein ICAM-1 expression inhibiting effect schematic diagram.
Fig. 7 is that Radix Angelicae Sinensis herb extract of the present invention is to intercellular adhesion protein VCAM-1 expression inhibiting effect schematic diagram.
Fig. 8 is the expression in vivo inhibition schematic diagram of Radix Angelicae Sinensis herb extract of the present invention to inflammation inducible enzyme COX-2 (Cycloxygenase).
Fig. 9 is the expression in vivo inhibition schematic diagram of Radix Angelicae Sinensis herb extract of the present invention to inflammation inducible enzyme 5-LO (5 lipoxygenase).
Figure 10 is the cytotoxicity experiment result schematic diagram of the compositions containing medicinal herbal extract of a preferred embodiment of the present invention.
Figure 11 is the antihistamine effect schematic diagram of the compositions containing medicinal herbal extract of a preferred embodiment of the present invention.
Detailed description of the invention
Mode below by embodiment further illustrates the present invention, but does not therefore limit the present invention among described scope of embodiments.The experimental technique of unreceipted actual conditions in the following example, conventionally and condition, or selects according to catalogue.
the minimal inhibitory concentration (MIC) of acne being brought out to bacterium detects
The extract distilled water diluting of Folium Nelumbinis extract and each organic solvent thereof is obtained each Folium Nelumbinis extract sample of variable concentrations, in each sample, inoculate 10
7the propionibacterium acnes (P.acnes) of cfu/ml and aurococcus (Staphylococcus aureus:S.aureus), then 37 DEG C, cultivate 16 hours under 150rpm condition, measure bacteriostasis rate, and extract concentrations bacteriostasis rate being reached the sample of more than 99% carries out record as MIC value, result is as table 1.
Table 1
| Folium Nelumbinis extract | Propionibacterium acnes | Aurococcus |
| Hexane extraction separator | 8000ppm | 4000ppm |
| Chloroform extraction separator | 4000ppm | 2000ppm |
| Extraction into ethyl acetate separator | 2000ppm | 1000ppm |
| Butanol, before immunoassay separator | 4000ppm | 1000ppm |
| Last aqueous phase extract | 8000ppm | 4000ppm |
As table 1, known through detecting, in multistage extract and separate, the extraction into ethyl acetate separator effect of Folium Nelumbinis ethanol extraction is best, secondly be chloroform extraction separator and butanol, before immunoassay separator, then be hexane extraction separator, the last aqueous phase extract after hexane, chloroform, ethyl acetate, the extraction of butanols multistage and the Folium Nelumbinis ethanol extraction before multistage extraction have the minimal inhibitory concentration of equal extent.
dPPH removal effect determination test
In order to verify the antioxidant effect of Folium Camelliae sinensis extract, carry out DPPH removal effect determination test.Highly stable free radical (free radical): 2, the bitter diazanyl (DPPH:2 of 2-diphenyl-1-, 2-Diphenyl-1-picrylhydrazyl, Missouri, USA sigma) can be yellow diphenyl bitterness hydrazides (diphenylpicrylhydrazine) from purple variable color because of Hydrogen Proton free radical scavenger, this test utilizes this phenomenon to determine removal free radical activity.Show the removal effect of the Folium Camelliae sinensis extract of variant concentration in Fig. 2, and can find out that the concentration increase free radical removal effect along with Folium Camelliae sinensis extract also can increase, also demonstrate Folium Camelliae sinensis extract further and there is free radical removal effect.
hydroxyl radical free radical removal effect is tested
Hydroxyl radical free radical (OH in DRI (2-deoxy-D-ribose)
-) can because of Fenton's reaction degeneration, the method being changed to thiobarbituric acid reaction thing by the thiobarbituricacidα-sodium (2-thiobarbituric acid:TBA) measured wherein verifies the hydroxyl radical free radical removal effect of Folium Camelliae sinensis extract.Be specially the FeCl by adding 500 μ l Folium Camelliae sinensis extract solution, 0.75mmol/L in the 21mmol/L DRI of 200 μ l
3the H of EDTA 200 μ l, 7.5mmol/L of 200ml, 0.75mmol/L
2o
2ascorbic acid (ascorbic acid) the 200 μ l of 200 μ l and 0.75mmol/L, reacts 1 hour at 37 DEG C.Add mixed liquor (both the volume ratio 1:1v/v) 1.5ml of 0.5%TBA and 10%TCA (trichloroacetic acid) again, 100 DEG C of reactions after 15 minutes, calculate hydroxyl radical free radical clearance effect data according to following formula after detecting the absorbance on 532nm, and the removal effect data of each for Folium Camelliae sinensis extract concentration are presented on the chart of Fig. 3.As shown in Figure 3, along with the concentration of Folium Camelliae sinensis extract increases, free radical removal effect also increases, and demonstrates Folium Camelliae sinensis extract further and has hydroxyl radical free radical removal effect.
Clearance (%)=[absorbance of (absorbance of the absorbance-experimental group of matched group)/matched group] × 100
lPS produces NO growing amount Inhibition test
In order to verify that Folium Psidii Guajavae extract has antiphlogistic effects, the inhibition done for inflammation evocating substance-nitric oxide (nitric oxide:NO) is tested, and inhibition data are weighed particular by nitrite (nitrite) concentration in the culture fluid generated when utilizing Griess reaction to measure lipopolysaccharide (lipopolysaccharide:LPS) in process mouse macrophage.First, on 96-well culture plate, 1 × 10 is injected with containing hyclone (Fetal bovineserum:FBS) and antibiotic DMEM culture medium (Dulbecco's Modified Eagle's Medium)
4after the mouse macrophage Raw264.7 of cell/well, at 37 DEG C, 5%CO
2cultivate 24 hours in thermostat under condition.After 24 hours, after original culture medium is removed, reinject not containing hyclone and antibiotic new DMEM culture medium, process each sample.After 1 hour, after processing the LPS of 1 μ g/ml, cultivate 24 hours again.The nitric oxide formed in the training period measures by Griess reagent the NO contained in cell culture medium
2 -concentration is weighed, and is specially after being mixed by cell culture upper liquid 50 μ l and Griess reagent 50 μ l and reacts 10 minutes on 96-application of sample orifice plate, then use microplate reader (ELISA reader) to measure the absorbance of 540nm.Nitric oxide clearance (%) utilizes calculating formula: clearance (%)=(1-adds the absorbance of absorbance/the do not add matched group of sample of the reaction group of sample) × 100 calculate, and the PBS in Fig. 4 is phosphate buffered saline (PBS) (Phosphatebuffered saline).As shown in Figure 4, after LPS process, contrast with matched group (phosphate buffered saline (PBS)), find that nitric oxide production concentration reduces, checking Folium Psidii Guajavae extract can effectively reduce nitric oxide production generation further.
histamine ionization rate measures
In order to verify antiinflammatory, the antihistamine effect of Folium Psidii Guajavae extract, determine the histamine ionization rate of mastocyte.In order to get mastocyte from experimental mouse (SD Mus), after the lethal experimental mouse of ether, at tyrode (137mmol/LNaCl, 2.7mmol/L KCL, the 1mmol/LCaCl of experimental mouse Intraperitoneal injection 10ml
22H
2o, 0.4mmol/L NaH
2pO
4, 12mmol/L NaHCO
3, 5.5mmol/L glucose) after, massage stomach wall 2 minutes gently, after open abdomen, get abdominal cavity washing liquid.Remove upper liquid after this abdominal cavity washing liquid is separated 5 minutes with 3000rpm centrifugation, wash 2 times again with the tyrode not containing bovine serum albumin (BSA).The mastocyte suspension obtained by tyrode settling flux is with 1 × 10
6cell/ml amount injects each well respectively, after stablizing 10 minutes at 37 DEG C, add histamine again to dissociate the functional composition 250ml of immunomodulator compounds 48/80 (compound 48/80: the condensation polymer of N-methyl-p-ethamine and formaldehyde: Missouri, USA Sigma-Aldrich product) 250ml, on demand concentration dilution and tyrode, allotment, to 2.5ml, is cultivated 20 minutes.After stopping the free reaction of histamine at 4 DEG C, be separated upper liquid with 3000rpm centrifugalize after 5 minutes.Get upper liquid 10 μ l, S-adenosine (methyl-
14c) tris-glycine buffer (Tris-glycine buffer:pH 6.8) the 40 μ l of methionine 5 μ l, histamine methyl transferase (histamine N-methyltransferase) 5 μ l, 300mmol/L reacts after 90 minutes together with distilled water 43.5 μ l at 37 DEG C, add 3mol perchloric acid 30ul, cessation reaction.In order in and perchloric acid add 10molNaOH20ul again.Then with the extraction of 1ml toluene isoamyl alcohol, after the cocktail solution flickerization of the 700 μ l upper liquid 3ml obtained, CPM (counts per minute) value is determined with beta-counter (β-counter).Utilize histamine standard curve to calculate histamine amount, then represent with the percentage ratio of histamine total amount.Histamine ionization rate (%)=(experimental group histamine free amount/total histamine free amount) × 100.The experimental result about antihistaminic of inducing because of threshing inductive substance (compound 48/80) in experimental mouse peritoneal cavity mast cells as shown in Figure 5.In Fig. 5, sham-operation (sham) group does not use compound 48/80 process, and negative control group [(-) control] is the compound 48/80 adding 1ug/ml.And compound 48/80 is the reagent as promoting mastocyte threshing (degranulation) and histamine release.As shown in Figure 5, when adding Folium Psidii Guajavae extract, HFC obviously reduces.
fibrin (fibrin) lytic activity
This experiment is the experiment of the fibrinolytic activity in order to measure Radix Angelicae Sinensis herb extract.By Fibrinogen (fibrinogen) with 0.5% concentration be dissolved in 50mmmol/L sodium phosphate buffer agent (sodiumphosphate buffer:pH 7.4), the sodium phosphate buffer agent solution 0.1ml dissolving 100NIH/ml thrombin is added after getting this solution 5ml, add agarose (agarose) the solution 5ml of concentration 1% again, join the culture dish of diameter 9cm after mix homogeneously, under normal temperature condition, solidify 1 hour.The each sample adding variable concentrations after the fibrin solidified-agarose colloid digs out the aperture of diameter 3mm processes.Use fibrinolysin (plasmin) as positive controls, experimental result is as shown in table 2.
Table 2
Can as seen from Table 2, along with the concentration of Radix Angelicae Sinensis herb extract increases, dissolved area also significantly increases in proportion.
intercellular adhesion protein (ICAM-1) expression inhibiting test (ICC)
In order to measure by intercellular adhesion protein ICAM-1 expression inhibiting effect in the Human umbilical vein endothelial cells of TNF-alpha active (Human umbilical veinendothelial cell), carry out immunocytochemical method.Experimental result shows, cell in normal Human umbilical vein endothelial cells group (phosphate buffer processed group) seldom has the expression of ICAM-1, but more there is the cell of more than 70% to have the expression of ICAM-1 (as in Fig. 6 by the groups of cells (TNF-α processed group) of TNF-alpha active, cell so that dark color represents), after Radix Angelicae Sinensis extract process, the expression of ICAM-1 all has minimizing in various degree according to different concentration for the treatment of.In Fig. 7, Radix Angelicae Sinensis extract refers to Radix Angelicae Sinensis herb extract.
intercellular adhesion protein (VCAM-1) expression inhibiting test (ICC)
In order to measure by intercellular adhesion protein VCAM-1 expression inhibiting effect in the Human umbilical vein endothelial cells of TNF-alpha active (Human umbilical veinendothelial cell), carry out immunocytochemical method.Experimental result shows, cell in normal Human umbilical vein endothelial cells group (phosphate buffer processed group) seldom has VCAM-1 to express, but about there is the cell of more than 70% to have the expression of VCAM-1 (as in Fig. 7 by the groups of cells (TNF-α processed group) of TNF-alpha active, cell so that dark color represents), after Radix Angelicae Sinensis extract process, the expression of VCAM-1 all has minimizing in various degree according to different concentration for the treatment of.In Fig. 7, Radix Angelicae Sinensis extract refers to Radix Angelicae Sinensis herb extract.
expression inhibiting test (ICC) of vascular inflammation inducible enzyme (COX-2)
In order to measure LPS process inflammatory cell (RAW 264.7cell) model in the expression in vivo inhibition of inflammation inducible enzyme COX-2 (Cycloxygenase), carried out immunocytochemical method.Experimental result shows, cell in normal cell group (phosphate buffer process) seldom has the expression of COX-2, but bringing out in the groups of cells (LPS process) of inflammation about has the cell of more than 50% to have the expression of COX-2 (as in Fig. 8, cell so that dark color represents), after the Radix Angelicae Sinensis extract process of 200ppm concentration, the expression of COX-2 could not be suppressed, but effectively can suppress the expression of COX-2 with the Radix Angelicae Sinensis extract of 1000ppm concentration.In Fig. 8, Radix Angelicae Sinensis extract refers to Radix Angelicae Sinensis herb extract.
expression inhibiting test (ICC) of vascular inflammation inducible enzyme (5-LO)
In order to measure LPS process inflammatory cell (RAW 264.7cell) model in the expression in vivo inhibition of inflammation inducible enzyme 5-LO (5 lipoxygenase), carried out immunocytochemical method.Experimental result shows, cell in normal cell group (phosphate buffer process) seldom has the expression of 5-LO, but bringing out in the groups of cells (LPS process) of inflammation about has the cell of more than 50% to have the expression of 5-LO (as in Fig. 9, cell so that dark color represents), after the Radix Angelicae Sinensis extract process of 200ppm concentration, the expression of 5-LO could not be suppressed, but effectively can suppress the expression of 5-LO with the Radix Angelicae Sinensis extract of 1000ppm concentration.In Fig. 9, Radix Angelicae Sinensis extract refers to Radix Angelicae Sinensis herb extract.
By as above testing the drug effect verifying each medicinal herbal extract, below provide the medicinal herbal extract compositions being used for relaxing acne be mixed to get by above-mentioned each medicinal herbal extract.Medicinal herbal extract compositions has the doubly excellent drug effect of 2-10 compared with single extract in antibacterial, antioxidation, antiinflammatory, skin sedation etc., is used in cosmetic preparation, can effectively improves the skin problems such as acne, skin allergy, erythema by this compositions.
Embodiment 1-48
Number ratios shown in above-mentioned Folium Nelumbinis extract, Folium Camelliae sinensis extract, Folium Psidii Guajavae extract and Radix Angelicae Sinensis herb extract according to the form below 3,4 is mixed to get the compositions containing medicinal herbal extract of embodiment 1-48.
Table 3
Table 4
effect example 1: by the experiment effect data of the compositions of the extract of different proportion mixing
Carry out the effect experiment such as antibacterial, antioxidation, antiinflammatory, skin irritation mitigation to embodiment 1-48 containing the compositions of medicinal herbal extract, result as shown in Table 5,6.
Table 5
| Embodiment is numbered | Antibacterial effect | Antioxidant effect | Antiphlogistic effects | Skin irritation alleviation effects |
| 1 | ++++ | + | + | - |
| 2 | ++ | ++++ | ++ | ++ |
| 3 | + | ++ | +++ | +++ |
| 4 | + | + | ++ | +++ |
| 5 | +++ | ++ | ++ | ++ |
| 6 | ++ | ++ | +++ | ++ |
| 7 | ++ | ++ | +++ | ++ |
| 8 | ++++ | +++ | ++ | ++ |
| 9 | ++++ | ++ | ++ | ++ |
| 10 | ++ | + | ++ | ++ |
| 11 | ++ | +++ | ++ | ++ |
| 12 | ++ | ++ | ++ | ++ |
| 13 | ++ | +++ | ++ | ++ |
| 14 | ++ | ++ | +++ | +++ |
| 15 | ++ | ++ | +++ | +++ |
| 16 | ++ | ++ | +++ | +++ |
| 17 | ++ | ++ | ++ | +++ |
| 18 | ++ | + | ++ | +++ |
| 19 | +++ | ++ | ++ | +++ |
| 20 | +++ | ++ | ++ | ++ |
| 21 | +++ | +++ | ++ | ++ |
| 22 | ++ | ++ | +++ | +++ |
| 23 | ++ | + | +++ | +++ |
| 24 | ++ | ++ | ++ | +++ |
| 25 | ++ | + | +++ | ++ |
Table 6
As above, shown in table 5,6, the drug effect of the extractive composition of all embodiments is all than each single medicinal herbal extract good drug efficacy, and wherein the extractive composition drug effect of embodiment 41 is best, is the extractive composition of embodiment 48 subsequently.The extractive composition of embodiment 41 is Folium Nelumbinis extracts, Folium Camelliae sinensis extract, Folium Psidii Guajavae extract and Radix Angelicae Sinensis herb extract form by the mass ratio of 2:1:0.5:0.5, and the extractive composition of embodiment 48 is Folium Nelumbinis extracts, Folium Camelliae sinensis extract, Folium Psidii Guajavae extract and Radix Angelicae Sinensis herb extract form by the mass ratio of 2.5:0.5:0.5:0.5.Can confirm according to as above content, the extractive composition of these two embodiments has optimal drug effect.
effect example 2: the present invention containing medicinal herbal extract compositions for acne bring out bacterium minimum press down
bacteria concentration (MIC) detects
In order to measure extract mixtures (i.e. Folium Nelumbinis extract, Folium Camelliae sinensis extract, Folium Psidii Guajavae extract and Radix Angelicae Sinensis herb extract form by the mass ratio of 2:1:0.5:0.5 the extract mixtures) antibacterial effect of above-described embodiment 41, after the extract mixtures distilled water diluting of above-described embodiment 41 is obtained the sample of variable concentrations, in each sample, inoculate 10
7the acne of cfu/ml brings out bacterium: propionibacterium acnes (P.acnes) and aurococcus (Staphylococcus aureus:S.aureus), then 37 DEG C, cultivate 16 hours under 150rpm condition, measure bacteriostasis rate, and concentration bacteriostasis rate being reached the extract mixtures of the sample of more than 99% carries out record as MIC value, result is as table 7.
Table 7
| / | Propionibacterium acnes | Aurococcus |
| Extract mixtures MIC value | 4000ppm | 2000ppm |
effect example 3: the present invention detects containing the histamine ionization rate of the compositions of medicinal herbal extract
In order to verify antiinflammatory, the antihistamine effect of the extract mixtures (i.e. Folium Nelumbinis extract, Folium Camelliae sinensis extract, Folium Psidii Guajavae extract and Radix Angelicae Sinensis herb extract form by the mass ratio of 2:1:0.5:0.5 extract mixtures) of the above embodiment of the present invention 41, determine the histamine ionization rate of mastocyte.In order to get mastocyte from experimental mouse (SD Mus), after the lethal experimental mouse of ether, at tyrode (137mmol/LNaCl, 2.7mmol/L KCL, the 1mmol/L CaCl of experimental mouse Intraperitoneal injection 10ml
22H
2o, 0.4mmol/L NaH
2pO
4, 12mmol/L NaHCO
3, 5.5mmol/L glucose) after, massage stomach wall 2 minutes gently, after open abdomen, get abdominal cavity washing liquid.Remove upper liquid after this abdominal cavity washing liquid is separated 5 minutes with 3000rpm centrifugation, wash 2 times again with the tyrode not containing bovine serum albumin (BSA).The mastocyte suspension obtained by tyrode settling flux is with 1 × 10
6cell/ml amount injects each well respectively, after stablizing 10 minutes at 37 DEG C, add histamine again to dissociate the above-mentioned composition 250ml of immunomodulator compounds 48/80 (compound 48/80: the condensation polymer of N-methyl-p-ethamine and formaldehyde: Missouri, USA Sigma-Aldrich product) 250ml, on demand concentration dilution and tyrode, allotment, to 2.5ml, is cultivated 20 minutes.After stopping the free reaction of histamine at 4 DEG C, be separated upper liquid with 3000rpm centrifugalize after 5 minutes.Get upper liquid 10 μ l, S-adenosine (methyl-
14c) tris-glycine buffer (Tris-glycine buffer:pH6.8) the 40 μ l of methionine 5 μ l, histamine methyl transferase (histamineN-methyltransferase) 5 μ l, 300mmol/L reacts after 90 minutes together with distilled water 43.5 μ l at 37 DEG C, add 3mol perchloric acid 30ul, cessation reaction.In order in and perchloric acid add 10molNaOH20ul again.Then with the extraction of 1ml toluene isoamyl alcohol, after the cocktail solution flickerization of the 700 μ l upper liquid 3ml obtained, CPM (count per minute) value is determined with beta-counter (β-counter).Utilize histamine standard curve to calculate histamine amount, then represent with the percentage ratio of histamine total amount.Histamine ionization rate (%)=(experimental group histamine free amount/total histamine free amount) × 100.Following Figure 11 of inhibition test result that the histamine produced by threshing evocating substance (compound 48/80) in experimental mouse peritoneal cavity mast cells is free.
effect example 4: the present invention tests containing the compositions cytotoxicity (MTT) of medicinal herbal extract
Human safety detection is carried out to the extract mixtures of the above embodiment of the present invention 41 (i.e. Folium Nelumbinis extract, Folium Camelliae sinensis extract, Folium Psidii Guajavae extract and Radix Angelicae Sinensis herb extract form by the mass ratio of 2:1:0.5:0.5 extract mixtures).This experiment is by cultivating Mus fibroblast: 3T3 cell carries out cytotoxicity (MTT) and tests.After process 3T3 cell sample, after serum-free (serum free) state cultured cell, MTT analytic process is utilized to detect the cytotoxic effect (cytotoxiceffect) of cell.The full name of MTT is 3-(4,5-dimethylthiazole-2)-2,5-diphenyltetrazolium bromide bromine salt, a kind of flaxen substrate, succinate dehydrogenase in living cells mitochondrion can make MTT be reduced to water-insoluble bluish violet crystallization first a ceremonial jade-ladle, used in libation (Formazan) and be deposited in cell, and dead cell is without this function, therefore living cells quantity can be measured according to the growing amount of first a ceremonial jade-ladle, used in libation (Formazan).Experimental result as shown in Figure 10, can be clear and definite, under 1000ppm concentration, also do not have cytotoxicity.
effect example 5: the present invention (closes containing the skin irritation test of the compositions of medicinal herbal extract
patch is tested)
By extract mixtures (the i.e. Folium Nelumbinis extract of the above embodiment of the present invention 41, Folium Camelliae sinensis extract, the extract mixtures that Folium Psidii Guajavae extract and Radix Angelicae Sinensis herb extract form by the mass ratio of 2:1:0.5:0.5) be diluted to concentration 0.5% respectively with distilled water, 2.5%, speckle sheet is made after the solution of 5%, select 20 health adult men and women as study subject, stick at arm position and state each sample speckle sheet, speckle sheet is removed after spending 24 hours, stimulation degree is calculated by mathematical expression after judging skin condition change according to following criterion naked eyes, result of calculation is as shown in table 8.
Criterion:
-: there is no the phenomena such as erythema
+-:, with surrounding ratio, reddens a little
+: with surrounding ratio, significantly redden
++: with surrounding ratio, seriously redden and have swelling
Stimulation degree=[(± value) * 1]+[(+) value * 2]+[(++) value * 3]/tested number
Table 8
As table 8, can determine that extract mixtures of the present invention is without any skin irritation.
effect example 6: the present invention tests for the improvement of acne containing the compositions of medicinal herbal extract
Use the extractive composition of the embodiment of the present invention 41, prepare astringent preparation according to the formula in lower form.Select 17-29 year after skin of face has 20 of acne experimenters to be divided into 2 groups at random, allow one group sooner or later to use above-mentioned astringent preparation 30 days, allow another matched group use matched group preparation 1 containing extractive composition.Then according to the suggestion of experimenter, use following evaluation criterion to judge that acne improves degree.Result is as shown in table 9.
Astringent pharmaceutical formulation
Criterion
-: worsen
+-: is not improved, and does not also worsen
+: slightly improve
++: improve Be very effective
Table 9
| Time | Astringent preparation | Matched group preparation 1 |
| 1 week | +- | +- |
| 2 weeks | + | +- |
| 3 weeks | + | +- |
| 4 weeks | ++ | +- |
As shown in table 9, can confirm that the astringent preparation using extractive composition of the present invention to prepare has better acne to improve effect compared with matched group preparation 1, and extractive composition of the present invention does not cause any skin irritation, erythema or pruritus.
effect example 7: the blood circulation promoting of extract mixtures of the present invention also improves skin-color effect
detect
Use the extractive composition of the embodiment of the present invention 41, prepare skin breast according to the formula in following table.Respectively select 25 women as experimenter, after above-mentioned skin breast and the matched group preparation 2 not containing extractive composition of the present invention are spread upon experimenter's face, have detected skin-color and improve effect.Now skin-color improves effect assessment is by draw after naked eyes and by outcome record on table 10 according to department of dermatologry hospital Senior Expert.
Skin breast formula
Table 10
| Skin breast | Matched group preparation 2 | |
| Effective | 21 people | 5 people |
| Invalid | 4 people | 20 people |
| Improvement rate (%) | 84.0 | 20.0 |
As shown in table 10, can confirm that the skin breast using extractive composition of the present invention to prepare has better blood circulation promoting effect and improves the effect of skin-color compared with matched group preparation 2.
iNDUSTRIAL APPLICABILITY
In the raw material that the present invention can be used for the skin preparations for extenal use such as cosmetics and product manufacturing industry.
Although the foregoing describe the specific embodiment of the present invention; but those skilled in the art is to be understood that; these only illustrate; protection scope of the present invention is defined by the appended claims; those skilled in the art is under the prerequisite not deviating from principle of the present invention and essence; can do place's various changes or modifications to these embodiments, but these change and amendment all falls into protection scope of the present invention.
Claims (10)
1., containing a compositions for medicinal herbal extract, comprise Folium Psidii Guajavae extract 5-20%, Radix Angelicae Sinensis herb extract 5-20%, Folium Camelliae sinensis extract 5-35% and Folium Nelumbinis extract by mass percentage, each constituent mass percentage ratio sum is 100%.
2. compositions as claimed in claim 1, it is characterized in that, described Folium Psidii Guajavae extract is Folium Psidii Guajavae ethanol extraction.
3. compositions as claimed in claim 1, it is characterized in that, described Radix Angelicae Sinensis herb extract is Radix Angelicae Sinensis herb ethanol extraction.
4. compositions as claimed in claim 1, it is characterized in that, described Folium Camelliae sinensis extract is Folium Camelliae sinensis ethanol extraction.
5. compositions as claimed in claim 1, it is characterized in that, described Folium Nelumbinis extract is Folium Nelumbinis ethanol extraction.
6. compositions as claimed in claim 5, is characterized in that, described Folium Nelumbinis extract be by the aqueous solution of described Folium Nelumbinis ethanol extraction with an organic solvent multistage extract and separate after extract and separate composition in the extract that obtains.
7. compositions as claimed in claim 6, is characterized in that, the described organic solvent adopted in described multistage extract and separate is one or more in hexane, chloroform, ethyl acetate and butanols.
8. compositions as claimed in claim 7, it is characterized in that, in time adopting multiple organic solvent in described multistage extract and separate, variant organic solvent uses successively.
9. compositions as claimed in claim 8, is characterized in that, in time adopting multiple organic solvent in described multistage extract and separate, use hexane, chloroform, ethyl acetate and butanols to carry out multistage extract and separate successively.
10. a cosmetic combination, it comprises the compositions as described in any one of claim 1-9; The content of described compositions is preferably 1000-10000ppm, is more preferably 1500-8000ppm, is 1800-4500ppm best.
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| CN105434210A (en) * | 2015-06-03 | 2016-03-30 | 马南行 | Multifunctional self-foaming type body mousse composition and preparation method thereof |
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