CN103877144A - Application of peanut coat active component and composition comprising peanut coat active component - Google Patents

Application of peanut coat active component and composition comprising peanut coat active component Download PDF

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Publication number
CN103877144A
CN103877144A CN201310159719.5A CN201310159719A CN103877144A CN 103877144 A CN103877144 A CN 103877144A CN 201310159719 A CN201310159719 A CN 201310159719A CN 103877144 A CN103877144 A CN 103877144A
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active component
arachidis hypogaeae
acid
testa arachidis
application
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CN103877144B (en
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戚建华
森大辅
向兰
木村雄辉
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HANGZHOU NAIQIRUI BIO-PHARMACEUTICAL TECHNOLOGY Co Ltd
Gifu Shellac Manufacturing Co Ltd
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HANGZHOU NAIQIRUI BIO-PHARMACEUTICAL TECHNOLOGY Co Ltd
Gifu Shellac Manufacturing Co Ltd
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Priority to JP2014043822A priority patent/JP2014218493A/en
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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

The application of Testa arachidis hypogaeae active component and the compositions that comprises this Testa arachidis hypogaeae active component
Technical field
The invention belongs to food, medicine purposes technical field, the compositions that relates in particular to a kind of application of Testa arachidis hypogaeae active component and comprise this Testa arachidis hypogaeae active component.
Background technology
Along with the development of world population, the growth of aging population is global trend, and the prevalence of alzheimer disease and sickness rate also increase gradually, and alzheimer disease is not only a difficult medical problem, and will become an outstanding social problem.At present, global patients with Alzheimer disease may reach 1,700 ten thousand, and along with the process of aged tendency of population, the sickness rate of alzheimer disease has risen to the 4th of common cause of death, is only second to cardiovascular and cerebrovascular disease, cancer and apoplexy.Alzheimer disease also becomes serious threat senior health and fitness's pertinacious disease gradually in China.
Alzheimer disease refers to the dementia that the geratic period occurs, and comprises Alzheimer (AD, Alzheimer ' s Dementia), vascular dementia (Vascular dementia), dementia due to Mixed dementia (mixed dementia) and other reasons.The pathological characteristics of Alzheimer shows as much starch sample senile plaque (SP) in brain, neurofibrillary tangles (NFT) and selective neuronal and synapse disappearance, various neurotransmitters is acetylcholine degradation especially, has correspondingly formed with amyloid beta (A β), Protein tau and neuron loss three large Mechanism Study fields.
Along with the aspects such as the nervous physiology to AD, biochemistry, pharmacology are constantly furtherd investigate, the drug development of control AD constantly makes progress in recent years.At present mainly to improve the levels of acetylcholine in patient body by acetylcholine esterase inhibition (acChE) to the Drug therapy of senile dementia, i.e. acetylcholinesteraseinhibitors inhibitors, it is to be used for the treatment of clinically an AD the most ripe class medicine the earliest.At present, for treatment light, moderate AD patients, medicine through U.S. FDA approval listing has 4 kinds, and this bright (Rivastigmine) and galantamine (Galantamine) of tacrine (tacrine), donepezil (donepezil), profit, is acetylcholinesteraseinhibitors inhibitors.Other medicine is as β, gamma secretase depressant, the brain metabolism regulators such as vincamine, nimodipine, and also officials come Jilin, vitamin E etc. affects the medicine etc. of Radical Metabolism.
The types of drugs that is used for the treatment of at present AD is various, is applicable to different mechanism of action, and they to a certain extent, play the effect of alleviating senile dementia, but no matter be go on the market or at the medicine grinding, have certain toxic and side effects, AD is still the persistent ailment of the radical cure of having no idea at present.Therefore, find new effective medicine and method for the AD cause of disease, become focus and the difficult point of current research.In numerous candidate drug, we have invested nerve growth factor sight.
Nerve growth factor (NGF) belongs to the member of neurotrophic factor (NTF) family, maintains neuronic quantity and survival.NGF maintains neural normal development and function, promotes regeneration and the reparation of injured nerve to have important function.NGF is as a kind of effective ways that may treat AD, but still has at present following point: (1) is expensive; (2) relative molecular mass is large, can not see through blood-brain barrier (Blood Brain Barrier, BBB), intracerebral ventricle injection only, and there are many technical problems in long-term treatment.Therefore, find and intend NGF activity, and can be NGF mimics by the micromolecular compound of blood brain barrier, become current study hotspot.Up to the present, scientists has had been found that nearly hundred kinds of NGF mimics, can effectively promote neurocyte to increase, and part of compounds has completed structural modification or complete synthesis.At present, the existing compound of NGF mimics is in three phase clinical stages.
Bread is the staff of life, and vast territory and abundant resources in China, the fruit and vegerable flesh of fish, and all kinds of delicacies, very richly endowed, the mankind's existence and procreation be unable to do without diet, by diet, obtain energy and various nutrition to meet the needs of body.Therefore, the composition that exploitation has a medical value in daily food not only can be avoided the toxic and side effects of medicine, can also greatly reduce research and development, the production cost of medicine.
Summary of the invention
The invention provides the application of Testa arachidis hypogaeae active component in food and the medicine of preparation prevention, treatment neurodegenerative diseases, described Testa arachidis hypogaeae active component is prepared by the following method: get dry Testa arachidis hypogaeae, with the mixed solution lixiviate of second alcohol and water, collect lixiviating solution, adsorb with HP-20 resin, after washing, with the aqueous solution eluting of ethanol, collect eluent, after concentrated eluent, dry, make described Testa arachidis hypogaeae active component, in this Testa arachidis hypogaeae active component, the weight percent content of anthocyanidin is greater than 50%.
Research of the present invention is found, Testa arachidis hypogaeae active component has the activity of significant plan nerve growth factor, animal pharmacology experiment also proves that Testa arachidis hypogaeae active component can improve the learning and memory function of mice, therefore can be using Testa arachidis hypogaeae active component as effective ingredient, add pharmacy or food acceptable carrier, food and the medicine of preparation prevention, treatment neurodegenerative diseases.
Described neurodegenerative diseases specifically can be alzheimer disease.
Described food acceptable carrier refers to the carrier of field of food routine, the filleies such as such as starch, sucrose, microcrystalline Cellulose, the binding agents such as starch slurry, hyprolose, gelatin, Polyethylene Glycol, the wetting agents such as magnesium stearate, micropowder silica gel, polyethylene glycols, the absorption enhancers such as poly-Pyrusussuriensis fat, lecithin, the surfactants such as poloxamer, smooth, the poly-Pyrusussuriensis fat of fatty acid Pyrusussuriensis, can also add other adjuvant such as flavouring agent, sweeting agent in addition.
Described pharmaceutically acceptable carrier can be with reference to prior art.
Testa arachidis hypogaeae active component is made after medicine, can be with unit dosage form administration, route of administration can be intestinal or non-intestinal.
The dosage form of medicine can be solid preparation, liquid preparation or semisolid dosage form, and above-mentioned various dosage forms can adopt existing production method to be prepared.
While preparing Testa arachidis hypogaeae active component, preferred, when lixiviate, the volume ratio of ethanol and water is 40:60~80:20, more preferably 60:40.
Lixiviate liquid ratio is 2:1~1000:1, is preferably 5:1~100:1, more preferably 16:1.
The temperature of lixiviate is 20~50 DEG C, is preferably 35~45 DEG C, more preferably 40 DEG C.Suitable temperature is conducive to dissolving and the diffusion of effective ingredient, promotes its leaching.
The time of lixiviate is 0.5~48h, is preferably 1~12h.The long meeting of extraction time increases impurity leaching amount, and easily causes the hydrolysis of the Testa arachidis hypogaeae active component having leached.
The number of times of lixiviate is 1~5 time, is preferably 2 times.
When lixiviate twice, when lixiviate for the first time, liquid ratio is 16:1, and extraction time is 1h, and the time of lixiviate is for the second time 0.5h.
It will be understood by those in the art that liquid ratio is the ratio of the volume (ml) of lixiviating solution and the quality (g) of raw material.
After lixiviate, filtration, after can first filtrate being concentrated, use resin absorption, general filtrate is concentrated into 1/50 volume again.
Described HP-20 resin is specifically as follows the HP-20 resin of Mitsubishi Chemical.
When eluting, using the aqueous solution of ethanol as eluant, wherein the volume ratio of ethanol and water is 20:80~80:20, is preferably 40:60.
For ensureing elution efficiency, when eluting, eluant and HP-20 resin volume ratio are 1:1~10:1, are preferably 2:1~5:1, more preferably 2:1.
Concentrate and can adopt conventional method to carry out, after concentrated end, be conventionally dried to water content≤5%.
The present invention also provides a kind of compositions, contains described Testa arachidis hypogaeae active component and Omega-3 series unsaturated fatty acid.
Omega-3 series unsaturated fatty acid can promote protein metabolism, the development of vision system of muscular tissue, promote cardiovascular sound, lower the risk of heart attack or apoplexy, can also reduce cholesterol in blood plasma, triglyceride, improve high triglyceride patient's blood lipid level, prevent diabetes and reduction cardiovascular disease incidence rate; In addition, often take Omega-3 series unsaturated fatty acid and can not only strengthen learning capacity, memory and absorbed power, also can alleviate pressure and make emotion greatly rouse oneself.
By Testa arachidis hypogaeae active component and Omega-3 series unsaturated fatty acid mix to memory ability to improve effect more remarkable, can be used for food and the medicine of the neurodegenerative diseases such as preparation prevention, treatment senile dementia.
Wherein, Testa arachidis hypogaeae active component can adopt aforesaid preparation method to be prepared.
The weight ratio of Testa arachidis hypogaeae active component and Omega-3 series unsaturated fatty acid is 1:1~5:1.
Described omega-3 series unsaturated fatty acid is at least one in docosahexenoic acid, clupanodonic acid, eicosapentaenoic acid, eicosatetraenoic acid, the acid of tetracosa carbon pentaene, nisioic acid, parinaric acid and linolenic acid.
Preferably, described omega-3 series unsaturated fatty acid is docosahexenoic acid.
Compared with prior art, beneficial effect of the present invention is:
(1) the invention provides the new purposes of one of Testa arachidis hypogaeae active component, Testa arachidis hypogaeae active component has the activity of significant plan nerve growth factor, can bring into play the effect of prevention and treatment senile dementia, for the food of the neurodegenerative diseases such as preparation prevention, treatment senile dementia and drug provision new thinking.
(2) Testa arachidis hypogaeae active component is taking a part for the daily Semen arachidis hypogaeae of being eaten as base stock, and Product Safety is high, and human body is had no side effect, and raw material sources are wide, with low cost, the preparation method of Testa arachidis hypogaeae active component is simple, quick simultaneously, greatly reduces research and development and production cost.
(3) the invention provides in compositions except containing Testa arachidis hypogaeae active component, also contain omega-3 series unsaturated fatty acid, can better improve memory function.More all there were significant differences with any effect wherein for compositions.
Brief description of the drawings
Fig. 1 is the microphotograph of different disposal on PC12 cellular neural projection differentiation impact after 48 hours;
Wherein,
A, 0.5%DMSO, negative control;
B, 40ng/mL NGF, positive control;
C, 0.3ug/mL Testa arachidis hypogaeae active component;
D, 1ug/mL Testa arachidis hypogaeae active component;
E, 3ug/mL Testa arachidis hypogaeae active component;
Fig. 2 is the impact of the nervous process differentiation rate of Testa arachidis hypogaeae active component on PC12 cell after 24 and 48 hours;
Fig. 3 is that Testa arachidis hypogaeae active component and docosahexenoic acid are distinguished or mixing use affects result figure to the water maze test of mice.
Detailed description of the invention
Further explain the present invention below in conjunction with specific embodiment.
The preparation of embodiment 1 Testa arachidis hypogaeae active component
(1) 1000 grams, drying peanut clothing is extracted after 1 hour at 40 DEG C with 16 liters of alcohol water mixed solvents containing 60% ethanol, filter, obtain the 1st time filtrate;
(2) in residue, add 2 liters of alcohol water mixed solvents containing 60% ethanol, extract after 30 minutes at 40 DEG C, filter, obtain the 2nd time filtrate;
(3) will twice filtrate after the volume of 40 DEG C of concentrating under reduced pressure to 1/50, use Diaion(Mitsubishi Chemical after merging) five liters of HP-20 resins adsorb, subsequently, except the material such as desaccharide, carry out eluting, collection eluent with 10 liters of aqueous solutions containing 40% ethanol with 15 premium on currency eluting;
(4) by eluent after the volume of 40 DEG C of concentrating under reduced pressure to 1/50, lyophilization to water content is 5%, obtains 115.8 grams of Testa arachidis hypogaeae active components.
The mensuration of anthocyanidin content:
Adopt vanillin-salt acid system to carry out quantitative analysis.
Standard curve: in brown test tube, 1ml catechin (catechin) titer (0.1,0.2,0.3mg/ml) (Ac) add 9.0mL 2% vanillin (vanillin)/(HCl-MeOH) (2g vanillin is dissolved in 100ml HCl-MeOH(1:2, v/v) in), using water (A0) as blank.After hatching 20min at 18~22 DEG C, mixed solution measures absorbance at 500nm.According to said method, generate the standard curve of catechin solution by [(Ac)-(Ao)] value.
Testa arachidis hypogaeae active component test: in brown test tube, the test solution (the Testa arachidis hypogaeae active component of 40mg is dissolved in the water of 100ml) of 1ml adds 9mL2% vanillin/(HCl-MeOH), and mixed solution is measured absorbance at 500nm at 18~22 DEG C after hatching 20min.
Each sample is surveyed three times, calculates the content of anthocyanidin in sample according to standard curve, is 56% thereby calculate in the present embodiment the content of anthocyanidin in Testa arachidis hypogaeae active component.
The Bioactivity of embodiment 2 Testa arachidis hypogaeae active components
In nerve retrograde affection animal model, research finds that nerve growth factor has the nerve growth of promotion and neuroprotective function, can stop or reduce neuronic regression, can stop to a certain extent the progress of senile dementia.Because PC12 cell has the general features of neurocyte, under the effect of nerve growth factor, PC12 cell can stop division, grows projection, changes into neuron cell.Therefore, utilize PC12 cell as Biological Activity Identification system, the activated sample of screening tool has reliability for the preparation of health food or the medicine of prevention and treatment alzheimer disease.
(1) cultivation of PC12 cell
Connect 20 × 10 4individual PC12 cell is inoculated in containing in 10ml DMEM culture medium (wherein containing 10% horse serum, 5% hyclone) culture dish, and 37 DEG C, 5%CO 2incubator in cultivate, change two days later a subculture, after three days successive transfer culture;
Cell is washed to twice with PBS, then add 10ml PBS in culture dish, 37 DEG C, 5%CO 2incubator in cultivate 10 minutes, then transfer to the disposable centrifuge tube of 15ml, on centrifugal rear blood counting chamber, count.
(2) active testing
After cell counting, in 24 porocyte culture plates, every hole first adds 1ml to contain the DMEM culture medium of serum (10% horse serum, 5% hyclone), and every hole accesses 2 × 10 4individual cell, CO 2incubator is cultivated 24 hours;
Discard culture medium, add 1ml to contain the DMEM solution (not containing serum) of different extracts, put into 37 DEG C, 5%CO 2incubator in cultivate, with the negative contrast of 1%DMSO, the positive contrast of 40ng NGF;
Under inverted microscope, every 24 hours, Continuous Observation cellular morphology changed, and recorded the nervous process differentiation rate of cell, and under each visual field, approximately 100 cells, choose at random 3 places, and add up mapping analysis.
Nervous process differentiation rate: nervous process is longer than the ratio of total cell number under the cell number of one times of cell space diameter and the visual field.
(3) experimental result
A in Fig. 1: negative control 0.5%DMSO; B:40ng/ml NGF: positive control, in Fig. 2, the negative contrast of 0.5%DMSO, the positive contrast of 40ng/mL NGF, Testa arachidis hypogaeae active component is made into variable concentrations with 0.5%DMSO solution.Referring to Fig. 1, Fig. 2, Testa arachidis hypogaeae active component is processed PC12 cell under the test concentrations of 0.3,1 or 3 μ g/ml, and after 24h, 48h, Testa arachidis hypogaeae active component all demonstrates significant plan Nerve Growth Factor Activity, and nervous process differentiation rate is between 30~60%.
Embodiment 3 Testa arachidis hypogaeae active components are mixed the impact using Mus memory function with DHA
Experiment main material: ICR mice (male Mus in 8 week age) (being purchased from medical courses in general institute of Zhejiang Province animal center).
Experimental implementation: Testa arachidis hypogaeae active component (10mg/kg) and docosahexenoic acid (DHA) are (5mg/kg) respectively or mix gastric infusion (10mg/kg Testa arachidis hypogaeae active component+5mg/kg DHA), each processing and matched group are 10 mices, Testa arachidis hypogaeae active component and/or docosahexenoic acid (DHA), all taking the aqueous solution containing 40% glycerol, 6% fatty acid glyceride as solvent control group is as above-mentioned solvent.At interval of administration in 24 hours 1 time, the maximum volume of administration is 100 μ l, and administration number of times is 7 times.。
Detection-the water maze test of learning and memory and spatial memory function.
Water maze laboratory is behavioristics's experiment of test animal spatial memory capacity, and its process is divided into two stages of training and testing.Animal was put into water maze in continuous four days above, record it and climb up the ability of learning and memory of the Time evaluation animal of platform; Platform was removed in the 5th day, record the number of times of animal spanning platform and evaluate the ability of learning and memory of animal in the holdup time that is placed with platform quadrant.
As shown in Figure 3, compared with the control, in water maze test, what the administration of Testa arachidis hypogaeae active component can significantly improve ICR mice wears platform number of times, what DHA oral administration group also can significantly improve ICR mice wears platform number of times, the most important thing is that Testa arachidis hypogaeae active component and DHA mixing administration group successful are better than using separately Testa arachidis hypogaeae active component or DHA.

Claims (9)

1. the application of Testa arachidis hypogaeae active component in food and the medicine of preparation prevention, treatment neurodegenerative diseases, described Testa arachidis hypogaeae active component is prepared by the following method: get dry Testa arachidis hypogaeae, with the mixed solution lixiviate of second alcohol and water, collect lixiviating solution, adsorb with HP-20 resin, after washing, with the aqueous solution eluting of ethanol, collect eluent, after concentrated eluent, dry, make described Testa arachidis hypogaeae active component, in this Testa arachidis hypogaeae active component, the weight percent content of anthocyanidin is greater than 50%.
2. application as claimed in claim 1, is characterized in that, when lixiviate, the volume ratio of ethanol and water is 40:60~80:20.
3. application as claimed in claim 1, is characterized in that, lixiviate liquid ratio is 2:1~1000:1.
4. application as claimed in claim 1, is characterized in that, extraction time is 0.5~48h, and extraction temperature is 20~50 DEG C.
5. application as claimed in claim 1, is characterized in that, when eluting, in the aqueous solution of ethanol, the volume ratio of ethanol and water is 20:80~80:20.
6. a compositions, is characterized in that, contains just like the Testa arachidis hypogaeae active component described in claim 1~5 any one and Omega-3 series unsaturated fatty acid.
7. compositions as claimed in claim 6, is characterized in that, the weight ratio of Testa arachidis hypogaeae active component and Omega-3 series unsaturated fatty acid is 1:1~5:1.
8. compositions as claimed in claim 6, it is characterized in that, described omega-3 series unsaturated fatty acid is at least one in docosahexenoic acid, clupanodonic acid, eicosapentaenoic acid, eicosatetraenoic acid, the acid of tetracosa carbon pentaene, nisioic acid, parinaric acid and linolenic acid.
9. compositions as claimed in claim 6, is characterized in that, described omega-3 series unsaturated fatty acid is docosahexenoic acid.
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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
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