CN108354919A - A kind of unsaturated fatty-acid compositions and its application for improving anti-oxidation function - Google Patents
A kind of unsaturated fatty-acid compositions and its application for improving anti-oxidation function Download PDFInfo
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- CN108354919A CN108354919A CN201711470124.6A CN201711470124A CN108354919A CN 108354919 A CN108354919 A CN 108354919A CN 201711470124 A CN201711470124 A CN 201711470124A CN 108354919 A CN108354919 A CN 108354919A
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Abstract
The present invention relates to a kind of unsaturated fatty-acid compositions for improving anti-oxidation function, it is characterised in that：EPA:The molar ratio of DHA is 2:1.The invention further relates to a kind of health foods for improving anti-oxidation function, it is characterised in that：Containing unsaturated fatty-acid compositions described in claim 1, in the unsaturated fatty-acid compositions, EPA:The molar ratio of DHA is 2:1.The present invention combines by adjusting ratio EPA and DHA, the antioxidant effect being optimal, and maximally efficient ratio combination is therefrom determined.And ratio formula can be optimal by manually reconciling, to provide product function effect.
Description
Technical field
Unsaturated fatty-acid compositions and its application that the present invention relates to a kind of for improving anti-oxidation function relate in turn
And health food, food nutrition, drug field.
Background technology
Epidemiological survey has proven to the diet ω -3PUFAs of the low incidence and intake high-content of AD (senile dementia),
Mainly eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) (Sydenham et al., 2012) are related.ω -3 long-chains
Component part of the polyunsaturated fatty acid (ω -3PUFAs) as neuron membrane, is essential nutriment, to human body device
Official and its function play a major role.They participate in inflammation and immunological process and hormone control.In addition, they participate in the hair of brain
It educates and function.
Numerous studies have proven to oxidative stress and can result in damage and the dysfunction of neuron with long-term chronic inflammation, are
Depression or the inducement of AD morbidities.
Several evidences show that the presence of inflammatory cytokine in AD increases, such as TNF-α level increases in AD patients serums
It is enclosed with presence of the interleukin-6 mRNA in APPsw transgenic mice Tg2576 hippocampus and cortex and in cerebral cortex
Around the increase of the microglia cell of the activation of senile plaque.
The microglia cell of activation can not only generate proinflammatory cytokine, but also can generate free radicals an oxidation
Nitrogen and superoxide anion.These groups induce neurodegeneration event similar with AD together with TNF-α secretion object.Studies have shown that
A β in SH-SY5Y cells25-35Toxicity and ROS and NO releases and the enhancing of oxidative damage it is related, it is quick to have raised redox
The transcription factor of sense such as NF- κ B, this is oxidation and an important factor for inflammatory reaction in AD.
In previous studies, it was recently reported that neuroinflamation can reduce the level of neurotrophic factor such as NGF and BDNF.Thing
In reality, it is reported in the concentration of neurotrophic factor and function of receptors increased and decreased in AD patient.Neurotrophy system participates in
With neure growth, survival and the relevant many physiology courses of plasticity.It also reported cholinergic neuron nerve to occur in AD
Dysfunction.
ω -3PUFAs are related to the structure and function that cell membrane phospholipid is constituted in brain, and are considered the lifting in cognitive process
It acts on.Secondly, it has been found that ω -3PUFAs have anti-oxidant and anti-inflammatory effect.Especially in the brain of aging, this spy
Property potentially contributes to the protection of neuron and prevents cell death.
Currently, fish oil or linseed oil comprising unsaturated fatty acid have been widely used in health food.However facing
In bed and experiment, the effect of different unsaturated fatty acids is not consistent：There is good anti-inflammatory, the anti-oxidant, neuroprotection of display,
There is invalid report.These different effects may be related with its separate sources insatiable hunger aliphatic acid used, or with its source difference institute
The ratio difference containing DHA with EPA leads to the difference of result.
Invention content
The purpose of the present invention is to provide a kind of unsaturated fatty-acid compositions for improving anti-oxidation function, including
EPA and DHA, it is characterised in that：EPA:The molar ratio of DHA is 2:1.
The present invention also aims to provide a kind of health food for improving anti-oxidation function, it is characterised in that：Contain
The unsaturated fatty-acid compositions having the right described in requirement 1, in the unsaturated fatty-acid compositions, EPA:The molar ratio of DHA
It is 2:1.
The present invention also provides application of the unsaturated fatty-acid compositions in the drug for improving anti-oxidation function.
The present invention provides application of the unsaturated fatty-acid compositions in the food for improving anti-oxidation function.
The present inventor has studied the anti-oxidant, anti-inflammatory and neural of different proportion DHA and EPA ratio combination by systematic comparison
Protecting effect therefrom determines maximally efficient ratio combination.And ratio formula can be optimal by manually reconciling, to carry
For product function effect.
Description of the drawings
Fig. 1 shows to prepare the flow chart of the unsaturated fatty-acid compositions for improving anti-oxidation function.
Fig. 2 a show the influence that EPA, DHA of various concentration cell viabilities beta induced to A reduce.
Fig. 2 b show the influence that EPA, DHA and its different proportion cell viability beta induced to A reduce.
Fig. 2 c show the protective effect of EPA, DHA and its different proportion cellular damage beta induced to A.
Fig. 3 a show the influence of EPA, DHA and its different proportion the ROS level variation beta induced to A.
Fig. 3 b show the influence of EPA, DHA and its different proportion the NO level variation beta induced to A.
Fig. 3 c show the influence of EPA, DHA and its different proportion the GSH level variation beta induced to A.
Fig. 4 a show that EPA, DHA and its different proportion the TNF-α mRNA beta induced on A express increased influence.
Fig. 4 b show EPA, DHA and its different proportion on the horizontal increased influence of the TNF-α that A is beta induced.
Fig. 5 a show the influence of EPA, DHA and its different proportion the NGF mRNA expression beta induced to A.
Figure 5b shows that the influences of BDNF mRNA expression EPA, DHA and its different proportion beta induced to A.
Fig. 5 c show the influence of EPA, DHA and its different proportion to the NGF levels that A is beta induced.
Fig. 5 d show the influence of EPA, DHA and its different proportion to the BDNF levels that A is beta induced.
Fig. 6 a show EPA, DHA and its different proportion Bax beta induced to A：The influence of Bcl-2 ratios.
Fig. 6 b show the influence of EPA, DHA and its different proportion the Caspase-3 protein expression beta induced to A.
Specific implementation mode
Below in conjunction with the specific embodiment of the invention, technical scheme of the present invention is verified, the embodiment verified
Only section Example of the invention.Based on the embodiments of the present invention, this field researcher is not making any creation
Property labour under the premise of the other embodiment that is obtained, shall fall within the protection scope of the present invention.
In the unsaturated fatty-acid compositions for improving anti-oxidation function of the present invention, EPA:The molar ratio of DHA is 2:
1。
EPA, DHA wherein in composition are commercially available sterling, and the preparation flow of composition is as shown in Figure 1：It surveys respectively first
The influence for trying the cell viability of the AD cell models of EPA and DHA the SH-SY5Y cell beta induced to A of various concentration, to really
Surely shield the dose concentration of EPA and DHA, then EPA and DHA combined treatments cell in varing proportions again, detects SH-
The cell viability of SY5Y cells, Antioxidant Indexes：ROS, NO, GHA, anti-inflammatory index：TNF-α, anti-apoptotic index Bc1, Bax,
Caspase-3, so that it is determined that EPA：The molar ratio of DHA is 2:Antioxidant effect is optimal when 1, only need to manually allocate EPA and DHA
To required ratio, it is uniformly mixed using conventional method and obtains the composition.
It is appreciated that unsaturated fatty-acid compositions of the present invention, can individually eat, can also be added as health food
It is applied to mixed edible in other food or in preparing anti-oxidation medicine.
The health food for improving anti-oxidation function of the present invention, contains the unsaturation described in the first aspect of the present invention
Aliphatic acid composition, in the unsaturated fatty-acid compositions, EPA:The molar ratio of DHA is 2:1.
The health food can be the form of ordinary food, can also use tablet, capsule, pill, lyophilized form, or
Any suitable administration form of person.
Health food in the scope of the invention may include the additive below one or more：Preservative, solubilizer, stabilization
Agent, wetting agent, emulsifier, sweetener, colorant, flavoring agent, deodorant, buffer solution, coating agent, antioxidant, suspending agent,
Auxiliary agent, excipient and diluent.
The present invention also provides application of the unsaturated fatty-acid compositions in the drug for improving anti-oxidation function.
The unsaturated fatty-acid compositions containing EPA and DHA can be used in pharmaceutical composition so that rich in drug
Containing EPA and DHA.
The convenient form of the pharmaceutical composition can be tablet for oral administration, pill, capsule, syrup
Agent, pulvis or granule；Sterile parenteral for parenterai administration or subcutaneous solution, suspension or the bolt for rectally
Agent, all these is all well known in the art.
Specific dosage level and dose frequency can be variation for any specific patient, will depend on more
Kind of factor, including the metabolic stability of the activity of used specific compound, this compound activity and duration, the age,
Weight, general health, gender, diet, the mode of administration and time, the rate of excretion, the combining of drug, specific disease
The severity of disease and the treatment individually carried out.The medicament and/or pharmaceutical composition of the present invention can be according to daily 1~10 time
Administration, such as once or twice daily.For oral and parenteral administration patient, the dosage level of medicament can be single dose
Amount or separated dosage.
The present invention provides application of the unsaturated fatty-acid compositions in the food for improving anti-oxidation function.
The composition of the present invention can be used in food industry so that food (such as grain food, dairy products, soya-bean oil, beans
Slurry) in be rich in EPA and DHA.
The cereal foods can have different shape or appearance forrns.For example, stick, cake can be made into
Or macaroon or its can also be oatmeal shape, sheet or rodlike.It can individually eat or with dairy products such as milk, acid
Milk or cottage cheese etc. are eaten together.
It should be understood that essence can also be added in food, such as honey, fruit, chocolate, caramel, nut, apricot
Benevolence, yoghurt flavours or combinations thereof.
Current market sales of unsaturated fat acid product largely comes from deep sea fish oil, also has from plant, various
The extraction of algae, ingredient is different, and especially there are notable differences in DHA and EPA ratios.Such as it is common from gold in the market
Fish oil DHA in prosperous fish or dog salmon：EPA is close to 1:2；And the unsaturated fatty acid in seaweed then contains higher DHA ratios
Example, this leads to health effect, and there are bigger differences.And so far, it there is no to DHA and EPA ratios group in unsaturated fatty acid
It closes in anti-oxidant, anti-inflammatory, the research in terms of neuroprotection, it is unclear between the variation of the two ratio and effect, therefore also without most
Ratio of greater inequality example formula and product.
The present inventor has studied the anti-oxidant, anti-inflammatory and neural of different proportion DHA and EPA ratio combination by systematic comparison
Protecting effect therefrom determines maximally efficient ratio combination.And ratio formula can be optimal by manually reconciling, to carry
For product function effect.
A β are by amyloid precusor protein (APP) through β-and the protein hydrolysate of gamma-secretase under pathological conditions.A
β25-35It is a kind of synthetic peptide of the overall length A β corresponding to 25-35 amino acid, beta sheet structure having the same, and keep overall length
Aβ1-42Complete toxicity.The peptide shows to form the rapid aggregation property of stable fibrinogen, and has god immediately in dissolving
Through toxicity.The present invention uses A β25-35The SH-SY5Y cells of differentiation are damaged as AD models, more different EPA/DHA ratios pair
A β in SH-SY5Y cells25-35The potential neuroprotection of the neurotoxicity of induction.
Currently, without any experiment respectively to EPA, DHA or a certain proportion of combinations carry out research and in same experiments
More different ω -3PUFAs.The present invention studies the combination of individual EPA and DHA and its different proportion to A β25-35The AD of induction
The effect of cell model.Cell viability is measured, to compare EPA, the combination of DHA or its various ratio is to A β25-35The AD of induction
The influence of the neurotoxicity of cell model and their potential synergistic effect.In addition, measure oxidative stress, proinflammatory cytokines because
The level of son and neurotrophic factor, to analyze EPA, DHA or combinations thereof may be beneficial to the cell and molecular mechanism of AD.And
The EPA/DHA of different proportion adjusts the ability of the expression of apoptosis-associated genes.
In the present invention, " FAs " refers to the EPA of various concentration, DHA or the combination of its different proportion.
Embodiment
Human neuroblastoma cell system SH-SY5Y cells all-trans retinoic acid (RA) is handled 7-8 days and is divided completely
Turn to human neure like cell.In the last day of differentiation, cell starts for testing.In A β25-35The SH- of the differentiation of induction
EPA and DHA is tested in the AD cell models of SY5Y cells in 6,12,25,50,100 μM of influences to cell viability.Wherein occur
Slight but significant decaying A β25-35The point that the cell viability of induction reduces is selected as the optimal dose of EPA and DHA and culture continues
Time.Then carry out following seven groups of researchs：(i) (culture medium) is compareed, (ii) A β25-35(add A β25-35Culture medium), (iii) A β
+ EPA (is pre-processed with EPA, then uses A β25-35Processing), (iv) A β+DHA (are pre-processed with DHA, then use A β25-35Processing) and
(v-vii) A β+EPA+DHA are (respectively with 2:1,1:1 and 1:2 EPA+DHA (totally 25 μM) processing, then uses A β25-35Processing).
Add A β25-35Before, cell EPA, DHA, combination thereof or control solvent are pre-processed 12 hours.A β are added25-35(eventually
A concentration of 20 μM) after, cell is incubated 24 hours again.Then, the cell viability in SH-SY5Y cells, oxidative stress, inflammatory are studied
Cytokine TNF-α, neurotrophic factor and Apoptosis.
In the present embodiment, as reagent, using as follows：>Eicosapentaenoic acid (the EPA of 99% purity；20：5, n-3) and
Docosahexaenoic acid (DHA；22：6, n-3) sodium salt comes from Sigma-Aldrich companies.In the medium by FAs dissolvings,
It is divided into aliquot under nitrogen flowing, and is preserved at -80 DEG C until using.
In the present embodiment, SH-SY5Y comes from ATCC (CRL-2266, Lot.61983120).Cell is being contained 10%
Fetal calf serum (FBS,Canada) and 1% Pen .- Strep DMEM/F12 culture mediums (Add and takes
Ventilation 75-cm greatly)2It is cultivated in culture bottle.It is dense with end in the DMEM/F12 containing 3%FBS (culture medium is replaced for every 2 days)
Degree is divided into complete human neure for 7-8 days for 10 μM RA (Sigma Aldrich, Canada) processing SH-SY5Y cells
Like cell.
Use 3- (4,5- dimethylthiazoles-the 2) -2,5- diphenyltetrazolium bromides for measuring cell proliferation rate and cell viability
Bromide (MTT) measures cell viability.By cell inoculation in 96 orifice plates, 90 μ L cell suspending liquids are added into each hole.
After experiment process, MTT (ATCC) is used to detect cell viability according to the manufacturer's instructions.It uses microplate reader (BioTek, USA)
Optical density is measured in 570nm.The absorbance of control group is considered as the 100% of cell viability.
As detected by mtt assay, A β25-35It lives in the cell of 20 μM of 24 hours SH-SY5Y cells for significantly reducing differentiation
Power (p<0.01, Fig. 2 a).However, significantly reducing A β with (6-100 μM) pretreatment of the EPA of various concentration or DHA25-35With agent
Measure the reduction of cell viability caused by dependence mode, and 6 in EPA, 12 (p>0.05), 25 (p<0.05), 50 μM of (p<
And 100 μM of (p 0.01)<0.05) 6 (p, or in DHA<0.05), 12-50 (p<And 100 μM of (p 0.01)<0.05) (Fig. 2 a).
Compared with EPA, the effect of DHA seems more stronger than effects of the EPA under same dose.On the basis of these results, subsequent
SH-SY5Y cells carry out oxidative stress, in the measurement of inflammation and apoptosis, we select 25 μM handled as different proportion in EPA
With the accumulated dose of DHA combinations.
It since MTT measurement is sensitive to cell quantity, is influenced, you must use another by cell Proliferation and cell viability
A kind of detection method confirmation result.It is thin in culture medium to assess using CytoTox-96 assay kits (Promega, Canada)
Total release of cytoplasm lactic dehydrogenase (LDH), this is the result of cell integrity damage.The measurement is based on from 2-P- (iodobenzenes
Base) -3- (p-nitrophenyl) -5- phenyltetrazoles chloride (INT, tetrazolium salt) arrives the coupling enzymatic conversion of formazan product, and enzyme is anti-
It should be catalyzed from release in cell and the diaphorase in measuring substrate mixture by LDH.By microplate reader at 490nm
Read absorbance.Every group of mean light absorbency is normalized to the percentage of control value.
In order to test different EPA/DHA ratios in neuroprotection have influence in various degree it is assumed that test below
It is middle to be combined using following different FAs：EPA/DHA is 2:1, EPA/DHA 1:1 or EPA/DHA is 1:2.
Fig. 2 b's the results show that with A β25-35Group is compared, in all proportions test, different EPA/DHA ratios significantly (p
<0.05) cell viability is improved.Protect SH-SY5Y cells from A β25-35The FAs effect of the neurotoxicity of induction is：EPA<2：
1EPA/DHA<DHA≤1：1EPA/DHA<1：2EPA/DHA.
Measurement (it is the index of cell death), which is discharged, by LDH further demonstrates different proportion EPA/DHA to A β25-35
The protective effect (Fig. 2 c) of the SH-SY5Y cellular damages of induction.In conjunction with said determination as a result, we can obtain knot for certain
By, EPA, DHA and combinations thereof can to varying degrees effective protection differentiation SH-SY5Y cells from A β25-35Induction
Cellular damage, the EPA of best raising cell viability：The ratio of DHA is 1：2EPA/DHA.
In the present embodiment, oxidative stress and Antioxidation reaction are measured, e.g., SH-SY5Y cells are inoculated in 96 orifice plates, to
200 μ L cell suspending liquids are added in each hole.After experiment process, with ROS kits in fluorecyte (Sigma Aldrich)
The level of ROS in quantization cell.Use fluorescence microplate reader (Reader Synergy HT, BioTek Instruments, U.S.
State), with lex=650/lem=675nm fluorescence intensities.According to the manufacturer's instructions, pass through Griess reagent systems
(Promega, Canada) measures cell intracellular nitric oxide (NO) yield.Absorbance is measured at 540nm using microplate reader.
By SH-SY5Y cell inoculations in 96 orifice plates, and 2mL cell suspending liquids are added into each hole.In experiment process
Afterwards, according to the manufacturer's instructions, GSH concentration is measured with glutathione assay kit (Sigma Aldrich).Fluorescence intensity
It is measured with fluorimeter reader, excitation wavelength 390nm, launch wavelength 478nm.
Fig. 3 a illustrate A β25-35ROS fluorescence (p is dramatically increased than control group<0.01).When with A β25-35When group compares, individually
(p is significantly reduced using the ROS fluorescence of the combination of EPA and DHA under EPA and all test ratios<0.05).However, in DHA groups
ROS fluorescence does not find significant difference.Protect SH-SY5Y cells from A β25-35The FAs effect of increased ROS fluorescence is induced to be：
DHA<1：2EPA/DHA<EPA<1：1EPA/DHA≤2:1EPA/DHA.
As shown in Figure 3b, when individually with A β25-35When handling cell, observe that nitrate levels dramatically increase about 40.29%
(p<0.05).However, with A β25-35Group is compared, and the nitrate levels of EPA processing are used alone slightly but do not significantly reduce (p<
0.05).With A β25-35Group compares, and DHA and the processing of all EPA/DHA processing groups, the equal horizontal significance difference of nitrate-free is used alone
It is different.
As shown in Figure 3c, A β25-35The GSH contents of damaging cells significantly (p<0.05) it reduces, with A β25-35Group is compared, and is owned
The EPA/DHA of ratio test significantly (p<0.05) increase GSH contents.Protect SH-SY5Y cells from A β25-35What is induced is anti-oxidant
GSH reduce FAs effect be：1:2EPA/DHA≤DHA<1:1EPA/DHA<EPA≤2:1EPA/DHA.
By the SH-SY5Y cell inoculations of differentiation in six orifice plates, and 2mL cell suspending liquids are added into each hole, experiment
After, harvest cell.GoScriptTM Reverse Transcriptase (a) are used using RNA extraction method
(Promega, Canada) synthesizes complementary DNA (cDNA) from RNA.2 μ gRNA are for the first chain cDNA synthesis.The nucleotide of primer
Nucleotide database and Primer Premier 6.0 of the sequence from NCBI.Spy in NCBI- nucleotide-BLAST
After opposite sex verification, the synthesis of primer is carried out by Invitrogen companies.Use Quantitect SYBR Greenmaster
Mix (Qiagen) prepares PCR reactions, and is carried out using Real Time PCR Detection System (Bio-Rad, the U.S.) CFX96TM real-time systems
PCR reacts.PCR processes are as follows：95 DEG C initial to be incubated 5 minutes to activate Hot-Star-Taq archaeal dna polymerases, then 94 DEG C 15
Second (denaturation), 59 DEG C 30 seconds (annealing) and 72 DEG C 30 seconds (extensions).After 38 cycles, a melting curve is produced, is used
In the specificity and homogeneity that measure primer.The rna expression of gene expression dose house-keeping gene beta-actin is (relatively fixed
Amount) and △ △ CT calibration standards.
By SH-SY5Y cell inoculations in 6 orifice plates, and 2mL cell suspending liquids are added into each hole.Through experiment process
Afterwards, it collects cell and is centrifuged 10 minutes with 10.000g, RIPA buffer solutions (RIPA, Thermo Scientific) is used in combination to crack.
By ultrasonic wave Assisted Cleavage, lysate is centrifuged 10 minutes at 4 DEG C with 10000g.Supernatant is collected, is added after boiling and contains 20-
The aliquot of 40 μ g proteins, and detached in electrophoretic buffer 60 minutes with 100V on 10%SDS-PAGE gels.Fortune
It, will be on Protein transfer to polyvinylidene fluoride (PVDF) film after row gel.Then by trace in Tris buffer solutions-polysorbas20
(TBST) washing 5 minutes in, then close (TBST and 5% alipoidic milk power) 1 hour at 20 DEG C.After closing, washed with TBST
Trace 5 minutes, is incubated with, including be used for actin, NGF, BDNF, TrkA, TrkB, TNF-α, Bcl-2, Bax with primary antibody
With the rabbit source polyclonal antibody of Caspase-3 (Abcam), 4 DEG C overnight, are then added secondary antibody, peroxidase at 20 DEG C
(HRP) 1 hour the anti-rabbit IgG combined.Trace is washed three times in TBS.Use ClarityTMWestern ECL substrates try
Agent box (Bio-rad, Canada) is with Image LabTMThe ChemiDoc of software (Bio-rad, Canada)TMIn MP systems
Detect immunoreactivity band.By the beta-actin for being normalized to detect again on same film, then with control
The percentage calculation of group, quantitative all target proteins.
Compared with the control group, after 4h incubations, by giving A β25-35, TNF-α mRNA expression dramatically increases (p<0.05,
Fig. 4 a), but with A β25-35Increase (the P of protein expression can not be found before being incubated 12 hours<0.01).However, and A
β25-35Group is compared, and has individually been restored to A β with the EPA and DHA being used in combination25-35Handle the aobvious of the TNF-α mRNA expression of reaction
Writing reduces.(Fig. 4 a) is in addition, 1：The effect of 1EPA/DHA is with obvious effects more more effective than individual EPA or DHA, these evidence tables
Bright 1:1EPA/DHA can play anti-inflammatory agent potential synergistic effect.
It is similar to TNF-α gene, A β25-35Processing occurs apparent ngf gene expression variation for 4 hours.Compared with the control group,
Aβ25-35Damaging cells NGF mRNA expression is apparent to lower (p<0.01, Fig. 5 a).Meanwhile A β25-35Damaging cells BDNF genes
(p was also lowered in mRNA expression at 4 hours<0.01, Fig. 5 b).Compared with the control, A β25-35In damaging cells, when being incubated at 12 hours
It was found that the protein expression of NGF significantly reduces (p<0.01, Fig. 5 c), and bdnf protein expression dramatically increases (p<0.01, Fig. 5 d).With
EPA, DHA and its pretreatment of various ratios can weaken A β to some extent25-35NGF mRNA and the protein expression (figure of induction
5a, c), BDNF mRNA expression (Fig. 5 b) and bdnf protein expression variation (Fig. 5 d).
Different from the gene of front, TrkA and TrkB are giving A β25-35It is affected after 24 hours.Aβ25-35It is substantially reduced
TrkA protein expressions (p<0.01).EPA, DHA and its processing of various ratios cannot significantly change A β25-35Effect to receptor, is removed
2：1EPA/DHA significantly increases this variation (Fig. 5 a).Individually giving A β25-35Cell in, it has been found that TrkB protein
Distant increase (p<0.05).2：1 and 1：2EPA/DHA pretreatments can part significantly reverse A β25-35The TrkB of induction is expressed
Change (equal p<0.05, Fig. 5 b), compared with A β groups, other groups handled with EPA and/or DHA are without significant change.
In A β25-35Different incubation times (4,8,12 and 24 hours) under test cdna Bax, Bcl-2 and Caspase-3 egg
White expression.Bcl-2 is with A β25-35When being incubated for 24 hours, protein expression is by A β25-35It is strong to reduce, by with EPA, DHA and its various
The pretreatment of ratio dramatically increases (p in various degree<0.01).About Bax, do not find significantly to become after being incubated at 4~24 hours
Change.Bax in cell：The ratio of Bcl-2 is individually giving A β25-35Dramatically increase (p<0.01).However, EPA, DHA and its various
Ratio processing can differently be obviously reduced this effect, and by Bax：Bcl-2 ratios are reduced to different degrees of controlled level
(in addition to EPA p<0.05, all p<0.01, Fig. 6 a)
For caspase-3 mRNA, A β25-35Dramatically increase its protein expression (p<0.01), EPA, DHA and its various ratios
Example processing can weaken A β to some extent25-35Effect (in addition to 1：1EPA/DHA p<0.05, all p<0.01, Fig. 6 b).
To sum up, 1:2EPA/DHA shows most effective to cell viability reduction；2:1EPA/DHA is sent out in test group
Wave the most effective ratio of antioxidation.For antiphlogistic effects, 1:1EPA/DHA is the optimal proportion in all test ratios；
When being related to Anti-G value, the combination of pure DHA ratio EPA and any other ratio is more effective.Based on these as a result, the knot obtained
By for EPA, DHA and its various ratios differently adjust A in SH-SY5Y cells by differently inhibiting inflammation and oxidative stress
β25-35The neurotoxicity of induction adjusts neurotrophic factor level, to weaken Neuron Apoptosis.
Claims (4)
1. a kind of unsaturated fatty-acid compositions for improving anti-oxidation function, including EPA and DHA, it is characterised in that：EPA:
The molar ratio of DHA is 2:1.
2. a kind of health food for improving anti-oxidation function, it is characterised in that：Contain unsaturated lipid described in claim 1
Fat acid composition, in the unsaturated fatty-acid compositions, EPA:The molar ratio of DHA is 2:1.
3. a kind of application of unsaturated fatty-acid compositions described in claim 1 in the drug for improving anti-oxidation function.
4. a kind of application of unsaturated fatty-acid compositions described in claim 1 in the food for improving anti-oxidation function.
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