CN108354919A - A kind of unsaturated fatty-acid compositions and its application for improving anti-oxidation function - Google Patents

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

A kind of unsaturated fatty acid composition and its application for improving antioxidant function

technical field

The invention relates to an unsaturated fatty acid composition for improving anti-oxidation function and application thereof, and further relates to the fields of health food, food nutrition and medicine.

Background technique

Epidemiological surveys have confirmed that the low incidence of AD (Alzheimer's disease) is associated with a high intake of dietary ω-3PUFAs, mainly eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) (Sydenham et al., 2012) related. ω-3 long-chain polyunsaturated fatty acids (ω-3PUFAs), as components of neuronal membranes, are essential nutrients and play a major role in human organs and their functions. They are involved in inflammatory and immunological processes and hormone regulation. Furthermore, they are involved in the development and function of the brain.

A large number of studies have confirmed that oxidative stress and long-term chronic inflammation can cause neuronal damage and dysfunction, and are the inducements of depression or AD.

Several lines of evidence showed increased presence of inflammatory cytokines in AD, such as elevated levels of TNF-α in the serum of AD patients and the presence of interleukin-6 mRNA in the hippocampus and cortex of APPsw transgenic mice Tg2576 and surrounding senile plaques in the cerebral cortex. Increase in activated microglia.

Activated microglia can produce not only pro-inflammatory cytokines, but also free radicals nitric oxide and superoxide anions. Together with TNF-α secretion, these groups induce neurodegenerative events similar to AD. Studies have shown that _Aβ25–35 toxicity in SH-SY5Y cells is associated with ROS and NO release and enhanced oxidative damage, which upregulates redox-sensitive transcription factors such as NF-κB, which are important for oxidative and inflammatory responses in AD. factor.

In previous studies, it was reported that neuroinflammation can reduce the levels of neurotrophic factors such as NGF and BDNF. Indeed, increased and decreased neurotrophic factor concentrations and receptor functions have been reported in AD patients. The neurotrophic system is involved in many physiological processes related to neuronal growth, survival and plasticity. Dysfunction in neurogenesis of cholinergic neurons has also been reported in AD.

Omega-3 PUFAs are involved in the structure and function of cell membrane phospholipid composition in the brain and are thought to play an important role in cognitive processes. Second, omega-3 PUFAs have been found to have antioxidant and anti-inflammatory effects. Especially in the aging brain, this property may contribute to the protection of neurons and prevent cell death.

Currently, fish oil or linseed oil containing unsaturated fatty acids has been widely used in health food. However, in clinical and experimental studies, the effects of different unsaturated fatty acids are not consistent: some have shown good anti-inflammatory, anti-oxidant, and neuroprotective effects, and some have reported ineffectiveness. These different effects may be related to the different sources of unsaturated fatty acids used, or the different ratios of DHA and EPA contained in different sources lead to differences in results.

Contents of the invention

The object of the present invention is to provide an unsaturated fatty acid composition for improving antioxidant function, comprising EPA and DHA, characterized in that the molar ratio of EPA:DHA is 2:1.

The purpose of the present invention is also to provide a health food for improving antioxidant function, characterized in that: containing the unsaturated fatty acid composition described in claim 1, in the unsaturated fatty acid composition, the mole of EPA:DHA The ratio is 2:1.

The invention also provides the application of the unsaturated fatty acid composition in medicines for improving anti-oxidation function.

The invention provides the application of the unsaturated fatty acid composition in food for improving anti-oxidation function.

The present inventors studied the antioxidant, anti-inflammatory and neuroprotective effects of different ratio combinations of DHA and EPA through systematic comparison, and determined the most effective ratio combination. And it can achieve the optimal ratio formula through manual blending, so as to provide product functional effect.

Description of drawings

Fig. 1 shows a flow chart for preparing an unsaturated fatty acid composition for improving antioxidant function.

Figure 2a shows the effects of different concentrations of EPA and DHA on the reduction of cell viability induced by Aβ.

Figure 2b shows the effects of EPA, DHA and their different ratios on Aβ-induced decrease in cell viability.

Figure 2c shows the protective effects of EPA, DHA and their different ratios on Aβ-induced cell damage.

Figure 3a shows the effects of EPA, DHA and their different ratios on the changes of ROS levels induced by Aβ.

Figure 3b shows the effects of EPA, DHA and their different ratios on the change of NO level induced by Aβ.

Figure 3c shows the effects of EPA, DHA and their different ratios on the changes of GSH levels induced by Aβ.

Figure 4a shows the effects of EPA, DHA and their different ratios on the increase of TNF-α mRNA expression induced by Aβ.

Figure 4b shows the effect of EPA, DHA and their different ratios on the increase of TNF-α level induced by Aβ.

Figure 5a shows the effects of EPA, DHA and their different ratios on Aβ-induced NGF mRNA expression.

Figure 5b shows the effects of EPA, DHA and their different ratios on Aβ-induced BDNF mRNA expression.

Figure 5c shows the effects of EPA, DHA and their different ratios on Aβ-induced NGF levels.

Figure 5d shows the effects of EPA, DHA and their different ratios on Aβ-induced BDNF levels.

Figure 6a shows the effect of EPA, DHA and their different ratios on the Bax:Bcl-2 ratio induced by Aβ.

Figure 6b shows the effects of EPA, DHA and their different ratios on the expression of Caspase-3 protein induced by Aβ.

Detailed ways

In the following, the technical solutions of the present invention will be verified in combination with specific embodiments of the present invention, and the verified embodiments are only part of the embodiments of the present invention. Based on the embodiments of the present invention, other embodiments obtained by researchers in the field without any creative effort all belong to the protection scope of the present invention.

In the unsaturated fatty acid composition for improving antioxidant function of the present invention, the molar ratio of EPA:DHA is 2:1.

The EPA and DHA in the composition are commercially available pure products, and the preparation process of the composition is shown in Figure 1: first, the effects of different concentrations of EPA and DHA on the cell viability of the AD cell model of SH-SY5Y cells induced by Aβ were tested respectively. influence, so as to determine the dose concentration of EPA and DHA that play a protective role, and then treat the cells with different ratios of EPA and DHA, and detect the cell viability of SH-SY5Y cells, antioxidant indicators: ROS, NO, GHA, anti-inflammatory indicators: TNF -α, anti-apoptosis indicators Bc1, Bax, and Caspase-3, so as to determine the best anti-oxidation effect when the molar ratio of EPA:DHA is 2:1, only need to manually adjust EPA and DHA to the required ratio, and use the general method Mix evenly to obtain the composition.

It can be understood that the unsaturated fatty acid composition of the present invention can be eaten alone as a health food, or added to other foods for mixed consumption or used in the preparation of antioxidant drugs.

The health food for improving antioxidant function of the present invention contains the unsaturated fatty acid composition described in the first aspect of the present invention, and in the unsaturated fatty acid composition, the molar ratio of EPA:DHA is 2:1.

The health food can be in the form of common food, or in tablet, capsule, pill, freeze-dried form, or any suitable administration form.

Health food within the scope of the present invention may include one or more of the following additives: preservatives, solubilizers, stabilizers, wetting agents, emulsifiers, sweeteners, coloring agents, flavoring agents, deodorants, buffer solutions , coating agents, antioxidants, suspending agents, adjuvants, excipients and diluents.

The invention also provides the application of the unsaturated fatty acid composition in medicines for improving anti-oxidation function.

The unsaturated fatty acid composition containing EPA and DHA can be used in a pharmaceutical composition to enrich the medicine with EPA and DHA.

The convenient form of the pharmaceutical composition may be tablets, pills, capsules, syrups, powders or granules for oral administration; sterile parenteral or subcutaneous solutions for parenteral administration, Suspensions or suppositories for rectal administration, all of which are well known in the art.

The specific dosage level and dosage frequency may vary for any particular patient and will depend on many factors including the activity of the particular compound being used, the metabolic stability and duration of the activity of the compound, age, body weight , general health, sex, diet, mode and timing of administration, rate of excretion, combination of drugs, severity of a particular condition, and treatments administered individually. The medicament and/or pharmaceutical composition of the present invention may be administered according to a regimen of 1 to 10 times a day, such as once or twice a day. For oral and parenteral administration to patients, the daily dosage level of the medicament may be a single dose or divided doses.

The invention provides the application of the unsaturated fatty acid composition in food for improving anti-oxidation function.

The composition of the present invention can be used in the food industry to enrich foods (eg cereals, dairy products, soybean oil, soybean milk) with EPA and DHA.

The cereals may have different shapes or appearances. For example, it can be made into sticks, cakes or macaroons, or it can be in the form of cereal, flakes or bars. It can be eaten alone or with dairy products such as milk, yogurt or cottage cheese.

It should be understood that flavors such as honey, fruit, chocolate, caramel, nut, almond, yogurt flavors or combinations thereof may also be added to the food.

Most of the unsaturated fatty acid products currently on the market come from deep-sea fish oil, and some are extracted from plants and various algae. The components are different, especially in the ratio of DHA and EPA. For example, the DHA:EPA ratio of fish oil from Jinchang fish or salmon commonly found on the market is close to 1:2; while the unsaturated fatty acids from seaweed contain a higher ratio of DHA, which leads to a large difference in their health effects. So far, there is no research on the ratio combination of DHA and EPA in unsaturated fatty acids in terms of anti-oxidation, anti-inflammation, and neuroprotection. It is not clear about the change and effect of the ratio of the two, so there is no optimal ratio formula and product. .

The present inventors studied the antioxidant, anti-inflammatory and neuroprotective effects of different ratio combinations of DHA and EPA through systematic comparison, and determined the most effective ratio combination. And it can achieve the optimal ratio formula through manual blending, so as to provide product functional effect.

Aβ is the proteolysis product of amyloid precursor protein (APP) by β- and γ-secretase under pathological conditions. Aβ _25-35 is a synthetic peptide corresponding to full-length Aβ of 25-35 amino acids, has the same β-sheet structure, and maintains the full toxicity of full-length Aβ _1-42 . The peptide exhibits rapid aggregation properties forming stable fibrils and is immediately neurotoxic upon dissolution. The present invention uses Aβ _25-35 -damaged differentiated SH-SY5Y cells as an AD model, and compares the potential neuroprotective effects of different EPA/DHA ratios on Aβ _25-35- induced neurotoxicity in SH-SY5Y cells.

At present, there are no experiments that have separately studied EPA, DHA, or a combination of certain ratios and compared different omega-3 PUFAs in the same experiment. The present invention studies the effects of individual EPA and DHA and their combinations in different ratios on the AD cell model induced by Aβ _25-35 . Cell viability was measured to compare the effects of EPA, DHA, or combinations thereof in various ratios on Aβ _25-35- induced neurotoxicity in AD cell models, as well as their potential synergistic effects. In addition, the levels of oxidative stress, pro-inflammatory cytokines and neurotrophic factors were measured to analyze the cellular and molecular mechanisms by which EPA, DHA or their combination may benefit AD. As well as the ability of different ratios of EPA/DHA to regulate the expression of apoptosis-related genes.

In the present invention, "FAs" refer to different concentrations of EPA, DHA or combinations thereof in different ratios.

Example

Human neuroblastoma cell line SH-SY5Y cells were treated with all-trans retinoic acid (RA) for 7-8 days to completely differentiate into human neuron-like cells. On the last day of differentiation, cells were started for experiments. The effects of EPA and DHA at 6, 12, 25, 50, 100 μM on cell viability were tested in the AD cell model of Aβ _25-35 induced differentiated SH-SY5Y cells. The point at which a slight but significant attenuation of the _Aβ25-35 -induced decrease in cell viability occurred was chosen as the optimal dose and culture duration of EPA and DHA. The following seven sets of studies were then performed: (i) control (medium), (ii) Aβ _25-35 (medium plus Aβ _25-35 ), (iii) Aβ+EPA (pretreated with EPA, followed by Aβ _{25 -35} treatment), (iv) Aβ+DHA (pretreated with DHA, then treated with Aβ _25-35 ) and (v-vii) Aβ+EPA+DHA (treated with 2:1, 1:1 and 1:2, respectively EPA+DHA (total 25 μM) treatment, followed by treatment with Aβ _25-35 ). Cells were pretreated with EPA, DHA, their combination or control vehicle for 12 hours before adding Aβ _25-35 . After the addition of Aβ _25-35 (20 μM final concentration), the cells were incubated for an additional 24 hours. Then, cell viability, oxidative stress, inflammatory cytokine TNF-α, neurotrophic factors and apoptosis in SH-SY5Y cells were investigated.

In this example, as reagents, the following were used: >99% pure eicosapentaenoic acid (EPA; 20:5, n-3) and docosahexaenoic acid (DHA; 22:6, n-3) 3) The sodium salt was from Sigma-Aldrich Company. FAs were dissolved in culture medium, aliquoted under nitrogen flow, and stored at -80 °C until use.

In this example, SH-SY5Y is from ATCC (CRL-2266, Lot. 61983120). Cells were incubated with 10% fetal bovine serum (FBS, Canada) and DMEM/F12 medium with 1% penicillin-streptomycin ( Canada) in ventilated 75-cm ² flasks. SH-SY5Y cells were differentiated into fully human neuron-like cells by treating them with RA (Sigma Aldrich, Canada) at a final concentration of 10 μM in DMEM/F12 containing 3% FBS (medium changed every 2 days) for 7–8 days cell.

Cell viability was measured using 3-(4,5-dimethylthiazole-2)-2,5-diphenyltetrazolium bromide (MTT), which measures cell proliferation rate and cell viability. Cells were seeded in 96-well plates, and 90 μL of cell suspension was added to each well. After experimental treatments, cell viability was measured with MTT (ATCC) according to the manufacturer's instructions. Optical density was measured at 570 nm using a microplate reader (BioTek, USA). The absorbance of the control group was considered as 100% of cell viability.

Aβ _25-35 at 20 μM for 24 hours significantly decreased the cell viability of differentiated SH-SY5Y cells as detected by the MTT assay (p<0.01, Figure 2a). However, pretreatment with different concentrations of EPA or DHA (6-100 μM) significantly attenuated the decrease in cell viability induced by Aβ _25-35 in a dose-dependent manner, and in EPA 6,12 (p>0.05), 25 (p<0.05), 50 μM (p<0.01) and 100 μM (p<0.05), or 6 (p<0.05), 12-50 (p<0.01) and 100 μM (p<0.05) in DHA (Fig. 2a). Compared to EPA, the effect of DHA appears to be stronger than that of EPA at the same dose. On the basis of these results, we chose 25 μM as the total dose of EPA and DHA combination in different ratio treatments in subsequent assays of oxidative stress, inflammation and apoptosis in SH-SY5Y cells.

Because the MTT assay is sensitive to cell number, which is influenced by cell proliferation and cell viability, results must be confirmed with another assay. The CytoTox-96 assay kit (Promega, Canada) was used to assess the total release of cytoplasmic lactate dehydrogenase (LDH) in the medium as a result of impairment of cell integrity. The assay is based on the coupled enzymatic conversion of 2-P-(iodophenyl)-3-(p-nitrophenyl)-5-phenyltetrazolium chloride (INT, a tetrazolium salt) to the formazan product, The enzymatic reaction is catalyzed by the release of LDH from the cells and diaphorase in the assay substrate mixture. Absorbance was read at 490 nm by a microplate reader. The mean absorbance of each group was normalized to the percentage of the control value.

To test the hypothesis that different EPA/DHA ratios have different effects on neuroprotection, different combinations of FAs were used in the following tests: EPA/DHA 2:1, EPA/DHA 1:1 or EPA/DHA 1 :2.

The results in Figure 2b show that different EPA/DHA ratios significantly (p<0.05) increased cell viability in all ratio tests compared to the Aβ _25-35 group. The potency of FAs protecting SH-SY5Y cells from _Aβ25-35 -induced neurotoxicity was: EPA<2:1EPA/DHA<DHA≤1:1EPA/DHA<1:2EPA/DHA.

The protective effect of different ratios of EPA/DHA on _Aβ25-35- induced SH-SY5Y cell injury was further confirmed by LDH release assay, which is an indicator of cell death (Fig. 2c). Combining the above assay results, we can definitely conclude that EPA, DHA and their combination can all effectively protect differentiated SH-SY5Y cells from _Aβ25-35 -induced cell damage to varying degrees, and optimally improve cell viability The EPA:DHA ratio is 1:2 EPA/DHA.

In this example, oxidative stress and antioxidant response were measured, for example, SH-SY5Y cells were seeded in a 96-well plate, and 200 μL of cell suspension was added to each well. After experimental treatment, the level of intracellular ROS was quantified with the fluorescent intracellular ROS kit (Sigma Aldrich). Fluorescence intensity was detected at lex=650/lem=675 nm using a fluorescent microplate reader (Reader Synergy HT, BioTek Instruments, USA). Intracellular nitric oxide (NO) production was measured by the Griess reagent system (Promega, Canada) according to the manufacturer's instructions. Absorbance was measured at 540 nm using a microplate reader.

Seed SH-SY5Y cells in a 96-well plate, and add 2 mL of cell suspension to each well. After experimental treatments, GSH concentrations were measured with a Glutathione Assay Kit (Sigma Aldrich) according to the manufacturer's instructions. Fluorescence intensity was measured with a fluorometer reader with an excitation wavelength of 390 nm and an emission wavelength of 478 nm.

Figure 3a illustrates that Aβ _25-35 significantly increased ROS fluorescence compared to the control group (p<0.01). ROS fluorescence was significantly reduced (p<0.05) for EPA alone and the combination of EPA and DHA at all ratios tested when compared to the Aβ _25-35 group. However, no significant difference was found in ROS fluorescence in the DHA group. The potency of FAs protecting SH-SY5Y cells from _Aβ25-35 -induced increased ROS fluorescence was: DHA<1:2EPA/DHA<EPA<1:1EPA/DHA≤2:1EPA/DHA.

As shown in Figure 3b, when cells were treated with Aβ _25-35 alone, a significant increase in nitrate levels of about 40.29% (p<0.05) was observed. However, nitrate levels were slightly but not significantly decreased (p<0.05) with EPA alone compared to the Aβ _25-35 group. There were no significant differences in nitrate levels between DHA alone and all EPA/DHA treatments compared to the Aβ _25-35 group.

As shown in Figure 3c, the GSH content of Aβ _25-35 injured cells was significantly (p < 0.05) decreased, and EPA/DHA for all ratios tested significantly (p < 0.05) increased GSH content compared with the Aβ _25-35 group. The potency of FAs protecting SH-SY5Y cells from _Aβ25-35 -induced decrease in anti-oxidative GSH was: 1:2EPA/DHA≤DHA<1:1EPA/DHA<EPA≤2:1EPA/DHA.

Differentiated SH-SY5Y cells were seeded in a six-well plate, and 2 mL of cell suspension was added to each well. After the experiment, the cells were harvested. Complementary DNA (cDNA) was synthesized from RNA using the RNA extraction method using GoScript(TM) Reverse Transcriptase (a) (Promega, Canada). 2 μg RNA was used for first-strand cDNA synthesis. The nucleotide sequences of the primers were obtained from NCBI's Nucleotide database and Primer Premier 6.0. Primer synthesis was performed by Invitrogen after specificity verification in NCBI-nucleotide-BLAST. Quantitect SYBR Greenmastermix (Qiagen) was used to prepare PCR reactions, and real-time PCR detection system (Bio-Rad, USA) CFX96TM real-time system was used for PCR reactions. The PCR procedure was as follows: initial incubation at 95°C for 5 minutes to activate the Hot-Star-Taq DNA polymerase, followed by 94°C for 15 seconds (denaturation), 59°C for 30 seconds (annealing), and 72°C for 30 seconds (extension). After 38 cycles, a melting curve was generated and used to determine primer specificity and identity. Gene expression levels were normalized with RNA expression (relative quantification) of the housekeeping gene β-actin with ΔΔCT correction.

Seed SH-SY5Y cells in a 6-well plate, and add 2 mL of cell suspension to each well. After experimental treatment, cells were harvested and centrifuged at 10.000 g for 10 min and lysed with RIPA buffer (RIPA, Thermo Scientific). Lysis was assisted by ultrasound, and the lysates were centrifuged at 10,000 g for 10 min at 4°C. Supernatants were collected, boiled and aliquots containing 20–40 μg protein were added and separated on a 10% SDS-PAGE gel at 100 V in electrophoresis buffer for 60 min. After running the gel, the proteins were transferred to a polyvinylidene fluoride (PVDF) membrane. The blots were then washed in Tris buffer-Tween 20 (TBST) for 5 minutes and then blocked (TBST and 5% non-fat dry milk) for 1 hour at 20°C. After blocking, blots were washed with TBST for 5 min and incubated with primary antibodies including rabbit sources for Actin, NGF, BDNF, TrkA, TrkB, TNF-α, Bcl-2, Bax and Caspase-3 (Abcam) Polyclonal antibody, overnight at 4°C, followed by secondary antibody, peroxidase (HRP)-conjugated anti-rabbit IgG, for 1 hour at 20°C. Blots were washed three times in TBS. Immunoreactive bands were detected on a ChemiDoc ^™ MP system with Image Lab ^™ software (Bio-rad, Canada) using the Clarity ^™ Western ECL Substrate Kit (Bio-rad, Canada). All target proteins were quantified by normalizing to β-actin reprobed on the same membrane and then calculated as a percentage of the control.

Compared with the control group, TNF-α mRNA expression was significantly increased by administration of Aβ _25-35 as early as after 4 h incubation (p<0.05, Fig. 4a), but no change in protein expression could be found before incubation with Aβ _25-35 for 12 h increased (P<0.01). However, EPA and DHA alone and in combination restored the significant decrease in TNF-α mRNA expression in response to Aβ _25-35 treatment compared to the Aβ _25-35 group. (Fig. 4a) Furthermore, the effect of 1:1 EPA/DHA was significantly more potent than that of EPA or DHA alone, evidence that 1:1 EPA/DHA could exert a potential synergistic effect on anti-inflammatory agents.

Similar to TNF-α gene, Aβ _25-35 treatment for 4 hours showed obvious changes in NGF gene expression. Compared with the control group, the expression of NGF mRNA in Aβ _25-35 injured cells was significantly down-regulated (p<0.01, Figure 5a). At the same time, the expression of BDNF gene mRNA in Aβ _25-35 injured cells was also down-regulated at 4 hours (p<0.01, Figure 5b). Compared with the control, in the Aβ _25-35 -injured cells, the protein expression of NGF was found to be significantly decreased (p<0.01, Fig. 5c) and the protein expression of BDNF was significantly increased (p<0.01, Fig. 5d) at 12 hours of incubation. Pretreatment with EPA, DHA and their various ratios could attenuate _Aβ25-35 -induced changes in NGF mRNA and protein expression (Fig. 5a, c), BDNF mRNA expression (Fig. 5b) and BDNF protein expression (Fig. 5d) to varying degrees .

Unlike the preceding genes, TrkA and TrkB were affected 24 hours after Aβ _25-35 administration. Aβ _25-35 significantly decreased the expression of TrkA protein (p<0.01). Treatment with EPA, DHA and their various ratios could not significantly change the effect of Aβ _25-35 on the receptor, except that 2:1 EPA/DHA significantly increased this change (Fig. 5a). A less pronounced increase in TrkB protein was also found in cells administered Aβ _25-35 alone (p<0.05). 2:1 and 1:2 EPA/DHA pretreatment partially and significantly reversed the Aβ _25-35- induced TrkB expression changes (all p<0.05, Fig. 5b), compared with the Aβ group, the other groups treated with EPA and/or DHA No significant changes.

Genes Bax, Bcl-2 and Caspase-3 protein expression were tested at different incubation times (4, 8, 12 and 24 hours) of Aβ _25-35 . When Bcl-2 was incubated with Aβ _25-35 for 24 hours, the protein expression was strongly reduced by Aβ _25-35 , and increased significantly by pretreatment with EPA, DHA and their various ratios (p<0.01). Regarding Bax, no significant changes were found after 4-24 hours of incubation. The ratio of Bax:Bcl-2 in cells was significantly increased after Aβ _25-35 alone (p<0.01). However, EPA, DHA, and their various ratio treatments could significantly attenuate this effect differently and reduce the Bax:Bcl-2 ratio to varying degrees of control (all p<0.01 except EPA p<0.05, Fig. 6a)

For caspase-3, Aβ _25-35 significantly increased its protein expression (p<0.01), while EPA, DHA and their various ratio treatments could weaken the effect of Aβ _25-35 to varying degrees (except 1:1 EPA /DHA p<0.05, all p<0.01, Figure 6b).

In conclusion, 1:2 EPA/DHA was shown to be the most effective in reducing cell viability; 2:1 EPA/DHA was the most effective ratio to exert antioxidant effects in the test group. For anti-inflammatory effect, 1:1 EPA/DHA was the best ratio of all tested ratios; pure DHA was more effective than the combination of EPA and any other ratio when it came to anti-apoptotic effect. Based on these results, it was concluded that EPA, DHA and their various ratios differentially modulate _Aβ25-35- induced neurotoxicity in SH-SY5Y cells, modulating neurotrophic factor levels by differentially suppressing inflammation and oxygen stress , thereby reducing neuronal apoptosis.

Claims (4)

1. An unsaturated fatty acid composition for improving antioxidant function, comprising EPA and DHA, is characterized in that: the mol ratio of EPA:DHA is 2:1. 2. A health food for improving antioxidant function, characterized in that: containing the unsaturated fatty acid composition according to claim 1, in the unsaturated fatty acid composition, the mol ratio of EPA:DHA is 2:1 . 3. the application of a kind of unsaturated fatty acid composition described in claim 1 in the medicine that improves antioxidant function. 4. the application of a kind of unsaturated fatty acid composition described in claim 1 in the food that improves antioxidant function.