CN109078033A - Saussurea involucrata culture inhibits protein non-enzyme glycosylation to react the application in anti-aging product - Google Patents
Saussurea involucrata culture inhibits protein non-enzyme glycosylation to react the application in anti-aging product Download PDFInfo
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Abstract
The invention discloses Saussurea involucrata cultures, and protein non-enzyme glycosylation to be inhibited to react the application in anti-aging product, the anti-aging includes that anti-wrinkle and anti-age pigment are formed, since non-enzymatic glycation will lead to human skin aging, and experiments have shown that Saussurea involucrata culture can effectively inhibit non-enzymatic glycation, therefore, illustrate that Saussurea involucrata culture can realize the anti-aging of human skin, applied to preparing in resisting age of skin product.
Description
Technical field
The present invention discloses the technical field for being related to Saussurea involucrata culture application more particularly to Saussurea involucrata culture inhibits protein non-
Application of the enzyme glycosylation in anti-aging product.
Background technique
Skin aging refers to that skin function aging damages, makes skin to the protective capacities of body, regulating power etc. subtracts
It moves back, thus skin does not adapt to the variation of internal and external environment, changing for the appearances integral status such as color, color, form, texture occurs
Become.The aging of skin is divided into: intrinsic aging and extrinsic aging.It is naturally old that intrinsic aging refers to that skin increases with the age
Change, shows as whiteness of skin, fine wrinkles, flexibility decrease, cutis laxa etc. occur；Extrinsic aging is main the reason is that day
Shine caused by light aging, show as wrinkle, cutis laxa, coarse, Beige color dermatodyschroia, telangiectasis,
Pigmented spots are formed.
Patent of invention ZL201410635060 discloses red goosefoot extract due to containing a variety of active ingredients, can effectively facilitate
The collagen secretion rate of dermal fibroblasts effectively prevents collagen saccharification and resists the injury of UV light, can resist
Skin aging.
Patent of invention 2011104395270 discloses natural plant extracts with anti-sunlight function and preparation method and answers
With lanatechead saussurea herb with flower is as one such ingredient, for sun-proof.
Therefore, there is presently no met related Saussurea involucrata culture by inhibiting protein non-enzyme glycosylation reaction in anti-skin
The report applied in terms of aging.
Summary of the invention
In consideration of it, the present disclosure provides Saussurea involucrata cultures to inhibit protein non-enzyme glycosylation reaction in anti-aging product
In application.
Technical solution provided by the invention, specifically, Saussurea involucrata culture inhibits protein non-enzyme glycosylation reaction anti-ageing
Application in old product.
It is preferred that the resisting age of skin is that anti-wrinkle and anti-age pigment are formed.
Further preferably, the Saussurea involucrata culture is the alcohol extract of Saussurea involucrata culture, it is specific the preparation method comprises the following steps:
Saussurea involucrata culture 5.0g is weighed, after being crushed, crosses the sieve that granularity is 30 mesh, 125mL volumetric concentration, which is added, is
75% ethyl alcohol, under the conditions of temperature is 65 DEG C, after ultrasonic extraction 30min, the condition for being 350 revs/min in revolving speed by extracting solution
After lower centrifugation, filter to get filtrate, it is spare；
Filter residue is added into the ethyl alcohol that 100ml volumetric concentration is 75%, under the conditions of temperature is 60 DEG C, Microwave Extraction
After 40min, extracting solution after centrifugation, is filtered to get filtrate under conditions of revolving speed is 350 revs/min；
After inciting somebody to action filtrate mixing twice, under the conditions of temperature is 70 DEG C, it is concentrated by evaporation after being 1:20 to solid-to-liquid ratio, obtains paste alcohol
Extract.
Further preferably, the product is drug or health care product.
Further preferably, the administration mode of the drug is oral, injection, infusion or cutaneous penetration.
Further preferably, the product is cosmetics and skin care item.
Further preferably, the concentration of Saussurea involucrata culture is more than or equal to 12.5mg/ml in the product.
Although saussurea involucrata has been disclosed for oxidation resistant effect, Saussurea involucrata culture of the invention is to pass through plant by Herba Saussureae Involueratae
(specific cultural method is detailed in ZL200510115902.0) that object cell culture technology obtains, color, shape and Herba Saussureae Involueratae
It is entirely different, and the content of brass in Saussurea involucrata culture, polysaccharide, chlorogenic acid, ligustrin is significantly larger than wild Tianshan Mountains
Saussurea involucrata, in addition, also without containing colchicin this small amount of toxic component in Herba Saussureae Involueratae in Saussurea involucrata culture.Therefore, saussurea involucrata
Although culture derives from Herba Saussureae Involueratae, different with Herba Saussureae Involueratae.
Non-enzymatic glycation will lead to human skin aging, and the present invention is found by experiment that: Saussurea involucrata culture can
Effectively inhibit non-enzymatic glycation, to illustrate that Saussurea involucrata culture can be realized the anti-aging of human skin, is applied to preparation
In resisting age of skin product.
Specific embodiment
Explanation is further explained to the present invention combined with specific embodiments below, but is not intended to restrict the invention
Protection scope.
Non-glycosylation (nonenzymatic glycosylation, NEG) aging theory, egg caused by non-glycosylation
The main reason for white matter crosslinking damage is aging.Non-glycosylation not only results in the crosslinking denaturation of various albumen, antioxidase
With the damage of the functions such as DNA repair enzyme, but also the reduction of energy supply will cause, the reduction of metabolic function, dysequilibrium etc.
Ageing process.Research has shown that Advanced glycation endproducts (advanced glycationend products, AGEs) are not only
Cell and function of organization can be directly affected, the generation of disease is participated in, it can also be by changing albumen in conjunction with specific receptors
Matter and cell function lead to the pathological change of body.
Non-enzymatic glycation leads to being mainly shown as skin aging: wrinkle and age pigment are formed.
1) wrinkle
There are a large amount of collagen and elastin laminin in fell skin tissue, both protein metabolisms are slow, and containing more
Lysine and hydroxylysine, this for occur NEG reaction provides material base.
The glucose of collagen and extracellular fluid occurs NEG and forms AGEs, and increases with advancing age.With
AGEs progressive increases, and collagen forms intermolecular cross-linking, not only reduces the permeability of connective tissue, also makes nutriment and gives up
The diffusion of object weakens, tissue ductility and hardness increase；It reduces the solubility of collagen and is difficult to be made by collagen enzyme hydrolysis
At skin flexibility decrease, wrinkle is generated.Since the AGEs irreversibility generated causes wrinkle constantly to be deepened and is not easy to calm down.Bullet
Property albumen and extracellular fluid glucose occur NEG formed AGEs, cause elastomer occur denaturation crosslinking, elasticity subside, skin
Skin is thinning, water content is reduced, the further atrophy of skin, crumple.
2) age pigment
Another appearance for being mainly characterized by cholesteroderma and old color spot of skin aging, this variation mainly with old color
The formation of element is related.Age pigment then refers to all ageing-related fluorescent material and its composite parts of intraor extracellular, such as eye
The aging collagen tissue etc. of yellow fluorescence, is additionally comprised in biochemical research in ball lens cataract, cytoplasm
From the fluorchrome etc. of ageing tissues.The formation of age pigment is dynamic consecutive variations process in organism, in different biologies
The all ages and classes stage of individual, different tissues position are all likely to form the age pigment with different biological natures.Age pigment
Including the intracorporal AGEs of biology, lipofuscin etc..Wherein lipofuscin is a kind of brown color pigment granule, mainly includes two aspects that one
It is lipid peroxidation, it is another, it is NEG.The starting reaction of lipid peroxidation is unsaturated fatty acid and oxygen in aerobic situation
Free radical or excessive metallic compound reaction lose a hydrogen atom, form peroxy radical, peroxy radical is subdivided solution shape
At various dimethyl carbonates (DMCs).From peroxidatic reaction of lipid and the NEG DMCs reacted and different biological molecules to be crosslinked
Or substance i.e. lipofuscin and AGEs that the mode of polymerization generates brown, has fluorescence, with the increase lipofuscin and AGEs at age
It can build up on Skin Cell and chromatophore, lead to cholesteroderma and the appearance of foxiness eventually.The product of AGEs and lipofuscin
Tire out and crosslink rear collagen and elastin laminin and forms skin splash in the accumulation of certain point.
To sum up, the advanced glycation end products that non-enzymatic glycation generates are the main reason for leading to human skin aging,
Therefore, as long as effectively inhibiting the progress of non-enzymatic glycation, the production of advanced glycation end products is prevented or is reduced, can be realized
Skin it is anti-aging.
Therefore, as long as being experimentally confirmed Saussurea involucrata culture has the function of inhibiting non-enzymatic glycation, can illustrate
It has the function of skin anti-aging, has the function of inhibiting non-enzymatic glycation, inventor to verify Saussurea involucrata culture
Devise following test.
Embodiment
It measures Saussurea involucrata culture and inhibits non-enzymatic glycation
Detection content: in pH7.4 phosphate buffer, it is with bovine serum albumin (BSA)-glucose under the conditions of 37 DEG C
Model under the conditions of excitation wavelength 370nm, slit 3nm, launch wavelength 440nm, slit 3nm, uses fluorescence after being incubated for 15 days
Spectrophotometric determination each group fluorescent value studies the alcohol of same batch various concentration and different batches various concentration Saussurea involucrata culture
Inhibiting effect of the extract to non-enzymatic glycation dead end product (AGEs).
Experimental principle: it is primarily due to BSA and the significant structural homology of human serum albumins (HSA), is chosen as external sugar
The protein of base model, D-Glucose are external glycosylated reduced sugar.It is slow in the phosphate that pH7.4 concentration is 0.2mol/L
In fliud flushing system, between the free amino of protein terminal and reduced sugar carbonyl group, by condensation, rearrangement, cracking, oxidative modification
Formed afterwards it is a series of structure is complicated and stable irreversible polymer, with fluorescence advanced glycation end products (AGEs).In reality
It tests to be added in system and there is the substance for inhibiting non-enzymatic glycation, the amount that AGEs is generated can be reduced with delayed response.
Experimental material and instrument: experiment is mentioned with all Saussurea involucrata cultures by Dalian Puruikang Biotechnology Co
For every bag of 10g dress.PH=7.4, concentration are that the phosphate-buffered salt of 0.2mol/L is configured according to States Pharmacopoeia specifications.96% cow's serum
Albumin (the magnificent Bioisystech Co., Ltd in Luoyang), sodium azide (analysis is pure), glucose (analysis is pure), volumetric concentration is
75% ethyl alcohol (Tianjin great Mao) is purchased from Dalian Fei Te company.Amino orphan (AG, positive control) is purchased from the examination of Chinese medicines group chemistry
Agent Co., Ltd.
UV-3000 double beam ultraviolet-visible spectrophotometer (Shimadzu, Japan)；F-7000 fluophotometer (Hitachi,
Japan)；PHS-3C type acidometer (Shanghai Lei Ci instrument plant)；Electronic balance, water bath with thermostatic control.
Experimental method:
(1) alcohol extract of various concentration Saussurea involucrata culture inhibits non-enzymatic glycation
Sample solution configuration: precision weighs Saussurea involucrata culture dry product (H180104) 10.0g, and after being crushed, crossing granularity is
The ethyl alcohol that 200mL volumetric concentration is 75% is added in the sieve of 30 mesh, under the conditions of temperature is 65 DEG C, after ultrasonic extraction 30min,
Extracting solution after centrifugation, is filtered to get filtrate under conditions of revolving speed is 350 revs/min, it is spare；Filter residue is added into 200ml volume
Extracting solution in revolving speed is 350 revs/min after Microwave Extraction 40min under the conditions of temperature is 60 DEG C by the ethyl alcohol that concentration is 75%
Under conditions of be centrifuged after, filter to get filtrate；After inciting somebody to action filtrate mixing twice, under the conditions of temperature is 70 DEG C, it is concentrated by evaporation to solid-liquid
After for 1:20, weighing obtains cream solid 4.010g, with the dissolution of 10mL water, is configured to 400mg/mL mother liquor.Successively it is diluted to
The sample solution of 50mg/mL, 25mg/mL, 12.5mg/mL, 6.25mg/mL, 3.125mg/mL.
Bovine serum albumin solution configuration: weighing 1g bovine serum albumin(BSA), and 200mg sodium azide mixed solution is added
It is dissolved in 7.4 phosphate buffer solution of 100mL pH.
The configuration of glucose solution: weighing glucose 7.45g and be added in 7.4 phosphate buffer solution of 100mL pH and dissolve,
74.5mg/mL solution.
Taking concentration respectively is the sample solution of 50mg/ml, 25mg/ml, 12.5mg/ml, 6.25mg/ml, 3.125mg/ml
And 1mL glucose solution and 1mL bovine serum albumin solution is added in blank group (water) 1mL, places 15 under 37 DEG C of aseptic conditions
It, 3 groups of operation repetitive.After 15 days, fluorescent spectrophotometer assay each group fluorescent value, location parameter are as follows: excitation wavelength are used
370nm, slit 3nm, launch wavelength 440nm, slit 3nm.
Measurement result see the table below:
Wherein, inhibiting rate %:I=(a-b)/a × 100% (a is blank group fluorescent value, and b is sample-adding group fluorescent value)
As seen from the above table, various concentration Saussurea involucrata culture alcohol extract is inhibited to non-enzymatic glycation, and
Increase as concentration increases its inhibiting effect.
(2) alcohol extract of different batches Saussurea involucrata culture inhibits non-enzymatic glycation
Sample solution configuration: precision weighs different batches Saussurea involucrata culture 5.0g, and after being crushed, crossing granularity is 30 purposes
The ethyl alcohol that 125mL volumetric concentration is 75% is added in sieve, under the conditions of temperature is 65 DEG C, after ultrasonic extraction 30min, will extract
Liquid after centrifugation, filters to get filtrate under conditions of revolving speed is 350 revs/min, spare；Filter residue, which is added 100ml volumetric concentration, is
75% ethyl alcohol, under the conditions of temperature is 60 DEG C, after Microwave Extraction 40min, the condition for being 350 revs/min in revolving speed by extracting solution
After lower centrifugation, filter to get filtrate；After inciting somebody to action filtrate mixing twice, under the conditions of temperature is 70 DEG C, being concentrated by evaporation to solid-to-liquid ratio is 1:
After 20, weighing obtains cream solid, is configured to the sample solution that concentration is 6.25mg/mL, 12.5mg/mL.
Other are measured according to (one) method.
For AG as positive control, concentration is 1.0 × 10-4, 2.0 × 10-4, 5.0 × 10-4, 1.0 × 10-3, 2.0 × 10- 3mol/L。
Wherein, inhibiting rate %:I=(a-b)/a × 100% (a is blank group fluorescent value, and b is sample-adding group fluorescent value)
As seen from the above table, each batch various concentration Saussurea involucrata culture alcohol extract has non-enzymatic glycation and inhibits to make
With when extract concentrations are 6.25mg/ml, inhibiting rate is higher than 60%, and when concentration is 12.5mg/m, inhibiting rate is up to
90% or more, compared with positive control AG, it is equivalent to 2.0 × 10-3The inhibiting effect of mol/L.
Separately to above-mentioned five batches, the external non-enzymatic glycation of the alcohol extract of two concentration Saussurea involucrata cultures inhibit into
Row statistics, specifically see the table below:
As seen from the above table, the alcohol extract of the Saussurea involucrata culture of same concentration different batches is to non-outside BSA-D glucose body
The inhibiting effect of enzyme glycosylation does not have significant difference.
(3) the different alcohol extracts of Saussurea involucrata culture inhibit non-enzymatic glycation
Following solution is carried out according to different alcohol extracting methods respectively to prepare and test, specific as follows:
Method 1: precision weighs part Saussurea involucrata culture dry product (H180104) 5.0g, and after being crushed, crossing granularity is 30 purposes
The ethyl alcohol that 125mL volumetric concentration is 75% is added in sieve, and under the conditions of temperature is 65 DEG C, alcohol reflux is extracted, and extracting solution is existed
After revolving speed is centrifuged under conditions of being 350 revs/min, filter to get filtrate, it is spare；It is 75% that filter residue, which is added 100ml volumetric concentration,
Ethyl alcohol in revolving speed be 350 revs/min of condition by extracting solution after alcohol reflux extracts 40min under the conditions of temperature is 60 DEG C
After lower centrifugation, filter to get filtrate；After inciting somebody to action filtrate mixing twice, under the conditions of temperature is 70 DEG C, being concentrated by evaporation to solid-to-liquid ratio is 1:
After 30, weighing obtains 3 parts of cream solids, and it is 6.25mg/mL, 12.5mg/mL that 3 parts of cream solids, which are each configured to concentration,
Sample solution.
Method 2: precision weighs part Saussurea involucrata culture dry product (H180104) 5.0g, and after being crushed, crossing granularity is 30 purposes
The ethyl alcohol that 125mL volumetric concentration is 75% is added in sieve, under the conditions of temperature is 65 DEG C, after ultrasonic extraction 30min, will extract
Liquid after centrifugation, filters to get filtrate under conditions of revolving speed is 350 revs/min, spare；Filter residue, which is added 100ml volumetric concentration, is
75% ethyl alcohol, under the conditions of temperature is 60 DEG C, after Microwave Extraction 40min, the condition for being 350 revs/min in revolving speed by extracting solution
After lower centrifugation, filter to get filtrate；；After inciting somebody to action filtrate mixing twice, under the conditions of temperature is 70 DEG C, being concentrated by evaporation to solid-to-liquid ratio is 1:
After 20, weighing obtains cream solid, is configured to the sample solution that concentration is 6.25mg/mL, 12.5mg/mL.
Method 3: precision weighs part Saussurea involucrata culture dry product (H180104) 5.0g, and after being crushed, crossing granularity is 15 purposes
The ethyl alcohol that 125mL volumetric concentration is 60% is added in sieve, under the conditions of temperature is 40 DEG C, after ultrasonic extraction 30min, will extract
Liquid after centrifugation, filters to get filtrate under conditions of revolving speed is 350 revs/min, under the conditions of temperature is 45 DEG C, is concentrated by evaporation to solid-liquid
After for 1:20, weighing obtains cream solid, is configured to the sample solution that concentration is 6.25mg/mL, 12.5mg/mL.
Other are measured according to (one) method, as a result see the table below
Wherein, inhibiting rate %:I=(a-b)/a × 100% (a is blank group fluorescent value, and b is sample-adding group fluorescent value)
As seen from the above table, inhibitory effect and extraction alcohol of the alcohol extracting thing of Saussurea involucrata culture for non-enzymatic glycation
The relevant property of concentration.
Those skilled in the art after considering the specification and implementing the invention disclosed here, will readily occur to of the invention its
Its embodiment.This application is intended to cover any variations, uses, or adaptations of the invention, these modifications, purposes or
Person's adaptive change follows general principle of the invention and including the undocumented common knowledge in the art of the present invention
Or conventional techniques.The description and examples are only to be considered as illustrative, and true scope and spirit of the invention are wanted by right
It asks and points out.
Claims (7)
1. Saussurea involucrata culture inhibits protein non-enzyme glycosylation to react the application in anti-aging product.
2. application according to claim 1, which is characterized in that the resisting age of skin is anti-wrinkle and anti-age pigment
It is formed.
3. application according to claim 1, which is characterized in that the Saussurea involucrata culture is the alcohol extract of Saussurea involucrata culture,
Its it is specific the preparation method comprises the following steps:
Saussurea involucrata culture 5.0g is weighed, after being crushed, crosses the sieve that granularity is 30 mesh, it is 75% that 125mL volumetric concentration, which is added,
Ethyl alcohol, under the conditions of temperature is 65 DEG C, after ultrasonic extraction 30min, by extracting solution under conditions of revolving speed is 350 revs/min from
After the heart, filter to get filtrate, it is spare；
Filter residue is added into the ethyl alcohol that 100ml volumetric concentration is 75%, under the conditions of temperature is 60 DEG C, after Microwave Extraction 40min,
Extracting solution after centrifugation, is filtered to get filtrate under conditions of revolving speed is 350 revs/min；
After inciting somebody to action filtrate mixing twice, under the conditions of temperature is 70 DEG C, it is concentrated by evaporation after being 1:20 to solid-to-liquid ratio, obtains paste alcohol extracting
Object.
4. application according to claim 1 or 2, which is characterized in that the product is drug or health care product.
5. application according to claim 4, which is characterized in that the administration mode of the drug be oral, injection, infusion or
Cutaneous penetration.
6. application according to claim 1 or 2, which is characterized in that the product is cosmetics and skin care item.
7. the application according to claim 4 or 6, which is characterized in that in the product concentration of Saussurea involucrata culture be greater than etc.
In 12.5mg/ml.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN111007135A (en) * | 2019-12-25 | 2020-04-14 | 广东食品药品职业学院 | Method for evaluating anti-saccharification effect of plant extract |
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