CN103877144B - Application of peanut coat active component and composition comprising peanut coat active component - Google Patents
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- CN103877144B CN103877144B CN201310159719.5A CN201310159719A CN103877144B CN 103877144 B CN103877144 B CN 103877144B CN 201310159719 A CN201310159719 A CN 201310159719A CN 103877144 B CN103877144 B CN 103877144B
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Abstract
The invention discloses an application of a peanut coat active component and a composition comprising the peanut coat active component. The peanut coat active component can be used for preparing foods and medicines for preventing and treating neurodegenerative diseases. The invention also provides a composition comprising the peanut coat active component and Omega-3 series unsaturated fatty acid. The invention provides a new application of the peanut coat active component, thus offering a new thinking for preparing foods and medicines for preventing and treating neurodegenerative diseases like Alzheimer's disease. In addition, the composition also contains omega-3 series unsaturated fatty acid besides the peanut coat active component, thus better improving memory function.
Description
Technical field
The invention belongs to food, pharmaceutical product use technical field, more particularly, to a kind of application of peanut coat active component and bag
Composition containing this peanut coat active component.
Background technology
With the development of world population, the growth of elderly population is global trend, and the trouble of senile dementia
Sick rate and the incidence of disease are also gradually increased, and senile dementia is not only a difficult medical problem, and asks becoming a prominent society
Topic.At present, global patients with Alzheimer disease may reach 17,000,000, with the process of aging population, the sending out of senile dementia
Sick rate has gone up the 4th for common cause of death, is only second to cardiovascular and cerebrovascular diseases, cancer and apoplexy.Senile dementia is in China
Also it has been increasingly becoming the serious chronic disease threatening senior health and fitness.
Senile dementia refers to the dementia of senescence phase generation, including Alzheimer disease（AD, Alzheimer ' s
Dementia）, vascular dementia（Vascular dementia）, Mixed dementia（mixed dementia）And other reasonses
Caused dementia.The pathological characteristicses of Alzheimer disease show as intracerebral much starch sample senile plaque expelling（SP）, neurofibrillary tangles
（NFT）And selective neuronal and synaptic loss, various neurotransmitters especially acetylcholine degradation, correspondingly form
With amyloid beta（Aβ）, Protein tau and the big Mechanism Study field of neuron loss three.
Constantly further investigate recently as to aspects such as the nervous physiology of AD, biochemistry, pharmacology, the medicine of preventing and treating AD is opened
Send out research constantly to make progress.At present acetylcholine esterase inhibition is mainly passed through to the drug therapy of senile dementia（acChE）
To improve levels of acetylcholine in the patient, i.e. acetylcholinesteraseinhibitors inhibitors, it is clinically to be used for treating AD earliest
For a ripe class medicine.Currently for light, moderate AD patients treatment, the medicine through U.S. FDA approval listing has 4 kinds, he
Crin（tacrine）, donepezil（donepezil）, Rivastigmine（Rivastigmine）And galanthamine
（Galantamine）, it is acetylcholinesteraseinhibitors inhibitors.Other medicines such as β, gamma secretase depressant, pervone,
The brain metabolism regulators such as Nimodipine, also officials come Jilin, vitamin E etc. affects medicine of Radical Metabolism etc..
Types of drugs currently used for treatment AD various it is adaptable to different mechanism of action, they rise to a certain extent
Arrive the effect alleviating senile dementia, but either listed or in the medicine grinding, have certain toxic and side effect, current AD
Still for the persistent ailment of a radical cure of having no idea.Therefore, find new effectively treatment medicine and the method for the AD cause of disease, become and work as
The focus of modern research and difficult point.In numerous candidate drug, we have invested nerve growth factor sight.
Nerve growth factor（NGF）Belong to neurotrophic factor（NTF）The member of family, maintains the quantity of neuron and deposits
Live.NGF maintains to the normal development of nervous system and function, promotes the regeneration of injured nerve and reparation to have important function.NGF
As a kind of effective ways of possible treatment AD, but still suffer from following point at present：（1）Expensive；（2）Relative molecular mass
Greatly it is impossible to pass through blood brain barrier（Blood Brain Barrier, BBB）, only intraventricular injection, long-term treatment exists many
Technical problem.Therefore, find and intend NGF activity, and can be NGF mimics by the micromolecular compound of blood-brain barrier,
Through becoming current study hotspot.Up to the present, scientists have discovered that nearly hundred kinds of NGF mimics, can be effective
Promotion nerve cell increase, part of compounds has been completed structural modification or fully synthetic.At present, NGF mimics is existing
Compound is in three phase clinical stages.
Bread is the staff of life, and vast territory and abundant resources for China, fruits and vegetables oppress, all kinds of delicacies, very richly endowed, the existence of the mankind and multiply from
Do not begin to drink food, by diet, obtain energy and various nutrition to meet the needs of body.Therefore, exploitation tool in conventional food
The composition having medical value not only can avoid the toxic and side effect of medicine additionally it is possible to substantially reduce the research and development of medicine, production cost.
Content of the invention
The invention provides peanut coat active component is in preparation prevention, the food and medicine for the treatment of nerve degenerative diseases
Application, described peanut coat active component is prepared via a method which：Take dry peanut coat, with the mixed solution of second alcohol and water
Extraction, collects leaching liquor, is adsorbed with HP-20 resin, after washing, with the aqueous solution wash-out of ethanol, collects eluent, will wash
Described peanut coat active component is obtained, in this peanut coat active component, the percentage by weight of anthocyanidin contains after de- liquid concentration, drying
Amount is more than 50%.
The research of the present invention finds, peanut coat active component has the significant activity intending nerve growth factor, animal drugs
Experiment of science also demonstrates that peanut coat active component can improve the learning and memory function of mouse, therefore can be by peanut coat active component
As active ingredient, add pharmacy or food acceptable carrier, preparation prevention, the food for the treatment of nerve degenerative diseases and medicine
Product.
Described nerve degenerative diseases concretely senile dementia.
Described food acceptable carrier refers to the conventional carrier of field of food, such as starch, sucrose, microcrystalline cellulose
Deng filler, the adhesive such as starch slurry, hydroxypropylcellulose, gelatin, polyethylene glycol, magnesium stearate, superfine silica gel powder, polyethylene glycols
Deng wetting agent, the sorbefacient such as poly- sorb fat, lecithin, the surface-active such as poloxamer, fatty acid sorbitan, poly- sorb fat
Agent, it can in addition contain add other assistant agent such as flavouring agent, sweetener.
Described pharmaceutically acceptable carrier can refer to prior art.
Peanut coat active component is made after medicine, can be administered in a unit, method of administration can be enteron aisle
Or non-bowel.
The formulation of medicine can be solid pharmaceutical preparation, liquid preparation or semisolid dosage form, and above-mentioned various formulations can adopt existing
Production method is prepared.
Prepare during peanut coat active component it is preferred that when extracting, ethanol is 40 with the volume ratio of water：60～80：20, more
It is preferably 60：40.
Extraction liquid ratio is 2：1～1000：1, preferably 5：1～100：1, more preferably 16：1.
The temperature of extraction is 20～50 DEG C, preferably 35～45 DEG C, more preferably 40 DEG C.Suitable temperature is conducive to effectively
The dissolving of composition and diffusion, promote its leaching.
The time of extraction is 0.5～48h, preferably 1～12h.Extraction time is long to make impurity leaching content increase, and holds
It is easily caused the hydrolysis of the peanut coat active component of leaching.
The number of times of extraction is 1～5 time, preferably 2 times.
When extracting twice, when extracting for the first time, liquid ratio is 16：1, extraction time is 1h, and the time of second extraction is
0.5h.
It will be understood by those in the art that liquid ratio is the volume of leaching liquor（ml）Quality with raw material（g）Ratio.
After extraction, filtration, after can first filtrate being concentrated, use resin adsorption again, general filtrate is concentrated into 1/50 volume.
Described HP-20 resin is specifically as follows the HP-20 resin of Mitsubishi Chemical.
During wash-out, using the aqueous solution of ethanol as eluant, eluent, wherein ethanol and the volume ratio of water are 20：80～80：20, excellent
Elect 40 as：60.
For ensureing elution efficiency, during wash-out, eluant, eluent and HP-20 resin volume ratio are 1：1～10：1, preferably 2：1
～5：1, more preferably 2：1.
Concentration can be carried out using conventional method, concentrate after terminating, be generally dried to water content≤5%.
Present invention also offers a kind of composition, unsaturated containing described peanut coat active component and Omega-3 series
Aliphatic acid.
Omega-3 series unrighted acid can promote the protein metabolism of musculature, development of vision system, promotes
Cardiovascular sound, lower the risk of heart attack or apoplexy additionally it is possible to reduce cholesterol, triglycerides in blood plasma, improve high
The blood lipid level of triglycerides patient, prevention diabetes and reduction angiocardiopathy incidence；In addition, often taking Omega-3
Serial unrighted acid can not only strengthen learning ability, memory and concentration, also can alleviate pressure and make mood greatly shake
Put forth energy.
Peanut coat active component and Omega-3 series unrighted acid are mixed the improvement to memory capability more
Significantly, can be used for the food of nerve degenerative diseases and the medicines such as preparation prevention, treatment senile dementia.
Wherein, peanut coat active component can be prepared using aforesaid preparation method.
The weight ratio of peanut coat active component and Omega-3 series unrighted acid is for 1：1～5：1.
Described omega-3 series unrighted acid is DHA, clupanodonic acid, eicosapentaenoic
In acid, eicosatetraenoic acid, tetracosa carbon five olefin(e) acid, nisioic acid, parinaric acid and leukotrienes at least one
Kind.
Preferably, described omega-3 series unrighted acid is DHA.
Compared with prior art, beneficial effects of the present invention are：
（1）The invention provides a kind of new application of peanut coat active component, peanut coat active component has significantly to be intended
The activity of nerve growth factor, can play the effect of prevention and treatment senile dementia, old silly for preparation prevention, treatment
The food of the nerve degenerative diseases such as slow-witted disease and medicine provide new thinking.
（2）, with a part for daily eaten peanut as base stock, Product Safety is high, to people for peanut coat active component
Body has no toxic side effect, and raw material sources are wide, with low cost, and the preparation method of peanut coat active component is simple, quick, significantly simultaneously
Reduce research and development and production cost.
（3）In present invention offer composition in addition to containing peanut coat active component, also contain omega-3 series unsaturated
Aliphatic acid, can preferably improve memory function.All there were significant differences for the effectiveness comparison of composition and any of which.
Brief description
Fig. 1 is the microphoto on the differentiation impact of PC12 cellular neural projection for the different disposal after 48 hours；
Wherein,
A, 0.5%DMSO, negative control；
B, 40ng/mL NGF, positive control；
C, 0.3ug/mL peanut coat active component；
D, 1ug/mL peanut coat active component；
E, 3ug/mL peanut coat active component；
Fig. 2 is the impact of the nervous process differentiation rate to PC12 cell for the peanut coat active component after 24 and 48 hours；
Fig. 3 is peanut coat active component and DHA respectively or is used in mixed way the water maze test shadow to mouse
Ring result figure.
Specific embodiment
Explain the present invention with reference to specific embodiment further.
The preparation of embodiment 1 peanut coat active component
（1）By drying peanut clothing, 1000 grams contain the alcohol water mixed solvents of 60% ethanol after 40 DEG C of extractions 1 hour with 16 liters,
Filter, obtain the 1st filtrate；
（2）Add 2 liters of alcohol water mixed solvents containing 60% ethanol in residue, after extracting 30 minutes at 40 DEG C, filter, obtain
2nd filtrate；
（3）Diaion is used after 40 DEG C of reduced pressure concentrations are to 1/50 volume after filtrate twice is merged（Mitsubishi Chemical）HP-
20 five liters of resins are adsorbed, and subsequently, remove the materials such as desaccharification with 15 liters of water elutions, are entered with the aqueous solution of 10 liters of ethanol containing 40%
Row wash-out, collects eluent；
（4）By eluent after 40 DEG C of reduced pressure concentrations are to 1/50 volume, freeze-drying to water content is 5%, obtains peanut
115.8 grams of clothing active component.
The mensure of anthocyanidin content：
Quantitative analysis is carried out using Vanillin-perchloric acid.
Calibration curve：In brown test tube, 1ml catechin（catechin）Titer（0.1,0.2,0.3mg/ml）（Ac）Plus
Enter 2% vanillic aldehyde of 9.0mL（vanillin）/（HCl-MeOH）（2g vanillic aldehyde is dissolved in 100ml HCl-MeOH（1：2, v/v）
In）, with water（A0）As blank.Mixed solution measures absorbance in 500nm after being incubated 20min at 18～22 DEG C.According to upper
State method, with [（Ac）-（Ao）] value generate catechin solution calibration curve.
Peanut coat active component is tested：In brown test tube, the test solution of 1ml（The peanut coat active component of 40mg is molten
In the water of 100ml）Addition 9mL2% vanillic aldehyde/（HCl-MeOH）, mixed solution is after 18～22 DEG C of incubation 20min in 500nm
Measurement absorbance.
Each sample is surveyed three times, is calculated the content of anthocyanidin in sample according to calibration curve, thus being calculated this
In peanut coat active component in embodiment, the content of anthocyanidin is 56%.
The Bioactivity of embodiment 2 peanut coat active component
In nerve retrograde affection animal model, research finds that nerve growth factor has promotion nerve growth and nerve
Defencive function, can stop or reduce the regression of neuron, can stop the progress of senile dementia to a certain extent.Because PC12 is thin
Born of the same parents have the general features of nerve cell, and in the presence of nerve growth factor, PC12 cell can stop dividing, and grows projection, turns
Chemical conversion neuron cell.Therefore, by the use of PC12 cell as Biological Activity Identification system, screen active sample and be used for
The health food of preparation prevention and treatment senile dementia or medicine have reliability.
（1）The culture of PC12 cell
Connect 20 × 104Individual PC12 cell is inoculated in the culture medium of DMEM containing 10ml（Wherein contain 10% horse serum, 5% tire ox blood
Clearly）In culture dish, 37 DEG C, 5%CO2Incubator in culture, change a subculture two days later, after three days squamous subculture；
With PBS, cell is washed twice, add 10ml PBS in culture dish, 37 DEG C, 5%CO2Incubator in culture 10
Minute, it is then transferred to the disposable centrifuge tube of 15ml, blood counting chamber after centrifugation counts.
（2）Active testing
After cell count, in 24 porocyte culture plates, every hole is initially charged 1ml containing serum（10% horse serum, 5% tire ox blood
Clearly）DMEM culture medium, every hole accesses 2 × 104Individual cell, CO2Incubator is cultivated 24 hours；
Discard culture medium, add 1ml to contain the DMEM solution of different extracts（Without serum）, put into 37 DEG C, 5%CO2Training
Cultivate in foster case, with 1%DMSO as negative control, 40ng NGF is positive control；
Every 24 hours under inverted microscope, Continuous Observation cellular morphology changes, the nervous process differentiation rate of record cell,
Under each visual field, about 100 cells, randomly select at 3, and count mapping analysis.
Nervous process differentiation rate：Nervous process is longer than cell number and the total cell number purpose under the visual field of one times of cell space diameter
Ratio.
（3）Experimental result
A in Fig. 1：Negative control 0.5%DMSO；b：40ng/ml NGF：Positive control, in Fig. 2,0.5%DMSO is negative right
According to 40ng/mL NGF is positive control, and peanut coat active component is made into variable concentrations with 0.5%DMSO solution.Referring to Fig. 1, figure
2, peanut coat active component processes PC12 cell, peanut coat activearm after 24h, 48h under the test concentrations of 0.3,1 or 3 μ g/ml
Divide and all show significant plan Nerve Growth Factor Activity, nervous process differentiation rate is between 30～60%.
Embodiment 3 peanut coat active component and DHA are used in mixed way the impact to mouse memory function
Experiment main material：ICR mouse（8 week old hero mouse）（It is purchased from medical courses in general institute of Zhejiang Province animal center）.
Experimental implementation：Peanut coat active component（10mg/kg）With DHA（DHA）（5mg/kg）Respectively or mixed
Close gastric infusion（10mg/kg peanut coat active component+5mg/kg DHA）, each is processed and control group is 10 mouse, flower
Raw clothing active component and/or DHA（DHA）, all with the aqueous solution containing 40% glycerine, 6% fatty acid glyceride be molten
Agent control group is above-mentioned solvent.It was administered once at interval of 24 hours, the maximum volume of administration is 100 μ l, and administration number of times is 7 times..
Learning and memory and the detection-water maze test of spatial memory function.
Water maze laboratory is the Behaviors survey of test animal spatial memory capacity, and its process is divided into training and test two
Stage.Above animal was put in water maze in continuous four days, record it and climb up the learning and memory energy of the Time evaluation animal of platform
Power；Platform was removed in 5th day, record the number of times of animal travels platform and evaluate animal in the holdup time being placed with platform quadrant
Ability of learning and memory.
As shown in figure 3, compared with the control, in water maze test, it is little that the administration of peanut coat active component can significantly improve ICR
Mouse wear platform number of times, what DHA oral administration group also can significantly improve ICR mouse wears platform number of times it is most important that peanut coat is active
Component and DHA mixing administration group effect are substantially better than and are used alone peanut coat active component or DHA.
Claims (16)
1. application in preparing the health food of improving studing ability or spatial memory capacity for the peanut coat active component, institute
State peanut coat active component to be prepared via a method which：Take dry peanut coat, with the mixed solution extraction of second alcohol and water, collect
Leaching liquor, is adsorbed with HP-20 resin, after washing, with the aqueous solution wash-out of ethanol, collects eluent, eluent is concentrated,
Described peanut coat active component is obtained, in this peanut coat active component, the weight percent content of anthocyanidin is more than after drying
50%, during extraction, ethanol is 40 with the volume ratio of water:60-60:40.
2. application as claimed in claim 1 is it is characterised in that extraction liquid ratio is 2：1～1000：1.
3. it is characterised in that extraction time is 0.5～48h, extraction temperature is 20～50 for application as claimed in claim 1 or 2
℃.
4. application as claimed in claim 1 or 2 is it is characterised in that during wash-out, in the aqueous solution of ethanol, the body of ethanol and water
Long-pending ratio is 20：80～80：20.
5. application in preparation prevention, the medicine for the treatment of nerve degenerative diseases for the peanut coat active component, described peanut coat is lived
Property component is prepared via a method which：Take dry peanut coat, with the mixed solution extraction of second alcohol and water, collect leaching liquor, use
HP-20 resin is adsorbed, and after washing, with the aqueous solution of ethanol wash-out, collects eluent, eluent is concentrated, be dried after make
Obtain described peanut coat active component, in this peanut coat active component, the weight percent content of anthocyanidin is more than 50%, during extraction,
Ethanol is 40 with the volume ratio of water:60-60:40.
6. application as claimed in claim 5 is it is characterised in that extraction liquid ratio is 2：1～1000：1.
7. it is characterised in that extraction time is 0.5～48h, extraction temperature is 20～50 to the application as described in claim 5 or 6
℃.
8. the application as described in claim 5 or 6 is it is characterised in that during wash-out, in the aqueous solution of ethanol, the body of ethanol and water
Long-pending ratio is 20：80～80：20.
9. a kind of composition it is characterised in that containing as described in any one of claim 1～8 peanut coat active component and
Omega-3 series unrighted acid.
10. composition as claimed in claim 9 it is characterised in that peanut coat active component and Omega-3 series unsaturated lipid
The weight ratio of fat acid is for 1：1～5：1.
11. compositions as described in claim 9 or 10 are it is characterised in that described Omega-3 series unrighted acid is two
Dodecahexaene acid, clupanodonic acid, eicosapentaenoic acid, eicosatetraenoic acid, tetracosa carbon five olefin(e) acid, tetracosa carbon
At least one in acid, parinaric acid and leukotrienes.
12. compositions as described in claim 9 or 10 are it is characterised in that described Omega-3 series unrighted acid is two
Dodecahexaene acid.
A kind of 13. compositions containing peanut coat active component, it includes being more than 50% anthocyanidin containing weight percent content
Peanut coat active component and Omega-3 series unrighted acid, described peanut coat active component and Omega-3 series insatiable hunger
With the weight of aliphatic acid than for 1：1～5：1, wherein said peanut coat active component is by the peanut coat mixing of second alcohol and water
Solution extraction obtains.
14. compositions according to claim 13, described Omega-3 series unrighted acid is two dodecahexaenes
Acid, clupanodonic acid, eicosapentaenoic acid, eicosatetraenoic acid, two
At least one in 14 carbon 5 alkene acids, nisioic acid, parinaric acid and leukotrienes.
The composition containing peanut coat active component described in 15. claims 13 or 14 improves mankind's ability of learning and memory in preparation
Or the application in spatial memory capacity health food.
The composition containing peanut coat active component described in 16. claims 13 or 14 is in preparation prevention, treatment nervus retrogression
Application in the medicine of disease.
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| CN201310159719.5A CN103877144B (en) | 2013-05-02 | 2013-05-02 | Application of peanut coat active component and composition comprising peanut coat active component |
| JP2014043822A JP2014218493A (en) | 2013-05-02 | 2014-03-06 | Composition and food |
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| CN201310159719.5A CN103877144B (en) | 2013-05-02 | 2013-05-02 | Application of peanut coat active component and composition comprising peanut coat active component |
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| CN108285458A (en) * | 2018-02-28 | 2018-07-17 | 浙江天草生物科技股份有限公司 | The method that procyanidine A2 is extracted from peanut coat |
| CN109620857B (en) * | 2019-01-16 | 2021-05-25 | 浙江大学 | Peanut coat active component and application thereof in preparation of anti-obesity and anti-diabetic drugs |
| CN111454242B (en) * | 2020-05-14 | 2021-12-14 | 湖南华诚生物资源股份有限公司 | Method for separating multiple active ingredients from peanut coat |
| CN113796536B (en) * | 2021-09-18 | 2023-11-03 | 山东省花生研究所 | Method for enriching phenolic compounds in peanut skin, peanut skin extract prepared by method and application of peanut skin extract |
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| US20030180406A1 (en) * | 2002-03-21 | 2003-09-25 | Helmut Sies | Treatment of diseases involving defective gap junctional communication |
| US20120065172A1 (en) * | 2001-07-27 | 2012-03-15 | Fotini Sampalis | Natural marine source phospholipids comprising polyunsaturated fatty acids and their applications |
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| EP1909801B2 (en) * | 2005-05-23 | 2020-03-18 | Massachusetts Institute of Technology | Compostions containing pufa and/or uridine and methods of use thereof |
| CN101099562A (en) * | 2006-07-07 | 2008-01-09 | 赵恩厚 | Omega-3 unsaturated fatty acids appetizer oil and producing method and application thereof |
| JP2008088131A (en) * | 2006-10-04 | 2008-04-17 | Seishin Enterprise Co Ltd | Anticholesteric composition |
| JP5443015B2 (en) * | 2008-02-19 | 2014-03-19 | 株式会社岐阜セラツク製造所 | Pharmaceutical composition |
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| US20120065172A1 (en) * | 2001-07-27 | 2012-03-15 | Fotini Sampalis | Natural marine source phospholipids comprising polyunsaturated fatty acids and their applications |
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