Antarctic krill oligopeptide nutritional composition for promoting neural development and application thereof

Abstract

The invention discloses an euphausia superba oligopeptide nutritional composition for promoting neural development and application thereof, and belongs to the technical field of functional foods.The nutritional composition is prepared from, by weight, 0.01-10 parts of docosahexaenoic acid, 0.01-20 parts of arachidonic acid, 0.01-10 parts of phosphatidylserine and 0.01-80 parts of euphausia superba oligopeptide. The antarctic krill oligopeptide is obtained by treating antarctic krill, antarctic krill powder or degreased antarctic krill powder by utilizing an enzymolysis method, and the content of a peptide fragment with the molecular weight of less than 1000Da in the antarctic krill oligopeptide is more than 90%. The raw materials of the composition are easy to obtain, the safety is high, and the composition can be used for preparing products with the function of promoting neurodevelopment and can be taken as common food, health care products or medicines for a long time.

Description

Technical Field

[0001] The invention relates to the technical field of functional foods, and in particular to an Antarctic krill oligopeptide nutritional composition for promoting neural development and an application thereof. Background Art

[0002] The development of the nervous system is an important part of individual growth and development. Reasonable nutritional intake and food selection can help promote the development of the nervous system and prevent neurodegenerative diseases. As people pay more attention to health, the market for foods that promote neurodevelopmental activity (such as promoting neuron maturation) is gaining more and more attention. Food ingredients that are recognized as having neurodevelopmental activity include unsaturated fatty acids such as docosahexaenoic acid (DHA) and arachidonic acid (ARA), as well as some vitamins (vitamin D, vitamin B1, etc.) and minerals such as magnesium, iron and zinc. These food ingredients have been widely used in infant formula, health products and other fields.

[0003] Although unsaturated fatty acids such as DHA and ARA have a certain promoting effect on neural development, the effect of a single ingredient is often limited and it is difficult to meet the complex neural development needs. In addition, the extraction process of DHA from some marine organisms is complicated and the cost is high, which limits its application in large-scale production. Researchers have also developed a variety of nutritional compositions that promote neural development, such as the Chinese patent literature with publication number CN114586983A, which discloses a nutritional composition that promotes neural development, which includes 2'-fucosyllactose and osteopontin, and can further contain casein. The Chinese patent literature with publication number CN118556870A discloses a nutritional composition and its application in improving brain, nerve and cognitive development, which comprises medium-chain fatty acid triglycerides and casein phosphopeptides, medium-chain fatty acid triglycerides and casein phosphopeptides synergistically, and has the effect of improving brain, nerve and cognitive development. The Chinese patent document with publication number CN114246934A discloses a synergistic and synergistic active substance composition for protecting nerves, which is obtained by compounding walnut protein peptide and ginseng decoction, wherein the walnut protein peptide is prepared by alkaline protease and leucine aminopeptidase compound enzymolysis, the polypeptide molecular weight range is 300-5000Da, and the ginseng decoction is prepared by decoction. The Chinese patent document with publication number CN118252257A discloses a nutritional composition, including whey protein powder, casein glycomacropeptide and sialic acid, which has a synergistic effect on promoting brain development, and can not only promote the expression of sialidase in the brain, but also promote the expression of brain-derived neurotrophic factor. In these studies, the research on the combination of multiple functional components has been strengthened, and the effect of promoting neural development has been improved through the interaction between different components.

[0004] Antarctic krill is a shrimp animal of the genus Euphausiaceae of the order Euphausiaceae. It is rich in protein, has a variety of amino acids, and contains a variety of minerals and trace elements such as calcium, iron, zinc, potassium, selenium, and magnesium. Antarctic krill peptides are small molecule peptide substances extracted from Antarctic krill / Antarctic krill powder by enzymatic hydrolysis technology, and have the characteristics of small molecular weight, good water solubility, high stability, and high human absorption rate. Antarctic krill peptides have now been used in the formulation of a variety of functional compositions. For example, the Chinese patent document with publication number CN114028535A discloses the use of Antarctic krill peptides and other raw materials to prepare a composition with the effect of promoting fracture healing. The Chinese patent document with publication number CN118805857A discloses that Antarctic krill peptides and agar oligosaccharides can synergistically exert antioxidant and whitening effects, and are used to prepare antioxidant and whitening products. However, there are few reports on the research of Antarctic krill peptides in promoting neural development. Summary of the invention

[0005] The present invention provides an Antarctic krill oligopeptide nutritional composition for promoting neural development. The raw materials of the composition are easy to obtain and have high safety. The composition can be used to prepare products with the effect of promoting neural development and can be consumed for a long time as a common food, health product or medicine.

[0006] The specific technical solutions adopted are as follows: An Antarctic krill oligopeptide nutritional composition for promoting neural development comprises, by weight, 0.01-10 parts of docosahexaenoic acid (DHA), 0.01-20 parts of arachidonic acid (ARA), 0.01-10 parts of phosphatidylserine (PS) and 0.01-80 parts of Antarctic krill oligopeptide; the Antarctic krill oligopeptide is obtained by treating Antarctic krill, Antarctic krill powder or defatted Antarctic krill powder by enzymatic hydrolysis, and the content of peptide segments with a molecular weight of less than 1000Da in the Antarctic krill oligopeptide is greater than 85%.

[0007] Furthermore, the Antarctic krill oligopeptide nutritional composition comprises, by weight, 0.05-5 parts of docosahexaenoic acid, 0.05-10 parts of arachidonic acid, 0.05-5 parts of phosphatidylserine and 0.05-40 parts of Antarctic krill oligopeptide; the content of peptide segments with a molecular weight of less than 1000Da in the Antarctic krill oligopeptide is greater than 90%.

[0008] Most preferably, the Antarctic krill oligopeptide nutritional composition comprises, by weight, 1 part of docosahexaenoic acid, 2 parts of arachidonic acid, 1 part of phosphatidylserine and 8 parts of Antarctic krill oligopeptides; the content of peptides with a molecular weight of less than 1000 Da in the Antarctic krill oligopeptides is greater than 90%, the minimum limit of oligopeptide purity in marine fish oligopeptide powder in the national standard "GB / T 22729-2008" is 75%, and the purity of the Antarctic krill oligopeptides in the present invention is more than 15% higher than the minimum limit of the national standard.

[0009] Specifically, the preparation method of Antarctic krill oligopeptide includes: (1) dissolving defatted Antarctic krill powder in water to obtain a raw material solution; (2) Adjusting the pH value of the raw material solution, adding protease, and performing enzymolysis at the optimal temperature of the protease; (3) After inactivating the enzyme, filter and cool the supernatant; (4) The solution obtained in step (3) is decolorized, and further solid-liquid separation is performed to obtain a supernatant, and the supernatant is sterilized and then dried to obtain the Antarctic krill oligopeptide.

[0010] Preferably, the weight ratio of the defatted Antarctic krill powder to water is 1:8 to 1:12.

[0011] The protease includes at least one of a composite protease (commercially available), alkaline protease, neutral protease, flavor protease, papain, and bromelain, preferably a mixed protease of a composite protease (commercially available) and alkaline protease.

[0012] Based on the amount of defatted Antarctic krill powder, the amount of protease added is 0.5-3wt%, preferably 2-3wt%, and the enzymolysis time is preferably 3-6 h. Under the above conditions, the degree of enzymolysis is relatively high.

[0013] Preferably, the drying method is spray drying. Spray drying has a fast speed and helps to maintain the activity of Antarctic krill oligopeptide in the product.

[0014] The content of peptide segments with a molecular weight of less than 1000 Da in the Antarctic krill oligopeptide prepared by the method of the present invention is greater than 85%, and further greater than 90%. After the Antarctic krill oligopeptide is compounded with DHA, ARA and PS components, it is easier to be absorbed by the body on the one hand, and has a better effect of promoting neuron development on the other hand.

[0015] The present invention also provides the use of the Antarctic krill oligopeptide nutritional composition in preparing products that promote neural development.

[0016] The present invention also provides a product for promoting nerve development, the ingredients of which include the Antarctic krill oligopeptide nutritional composition.

[0017] The product for promoting neural development is a food or a medicine, and its forms include but are not limited to powders, tablets, capsules, granules or drinks, etc., and its ingredients also include pharmaceutically acceptable excipients or food-acceptable additives.

[0018] Furthermore, the product for promoting neural development is a modulated milk powder, the ingredients of which include 0.12-0.24 wt % of the Antarctic krill oligopeptide nutritional composition and 99.76-99.88 wt % of the milk-based powder.

[0019] Preferably, the milk-based powder comprises 30-40wt% whole milk powder, 20-30wt% skim milk powder, 10-20wt% desalted whey powder, 10-20wt% lactose, 3-5wt% vegetable fat powder, 1-5wt% vitamins and 1-5wt% minerals.

[0020] Experimental verification shows that the promotion of neural development specifically refers to improving the development of neuronal cells (further, mouse hippocampal neuronal cells), and improving the development of neuronal cells specifically includes at least one of the following methods: (1) Promote the growth and development of neuronal cells; (2) Increase the proliferation rate of neuronal cells; (3) Alleviate the damage caused by aluminum chloride to neuronal cells and increase the expression of microtubule-associated protein-2 (MAP-2) and axon length in neuronal cells.

[0021] Compared with the prior art, the present invention has the following beneficial effects: (1) The present invention obtains a nutritional composition capable of improving the development of neuronal cells by compounding docosahexaenoic acid, arachidonic acid, phosphatidylserine and Antarctic krill oligopeptides (peptide content with molecular weight <1000Da>90%) in specific proportions. The raw materials of the composition are easy to obtain and have high safety. The composition can promote the growth and development of neuronal cells, increase the proliferation rate of neuronal cells, alleviate the damage caused by aluminum chloride to neuronal cells, and increase the expression of MAP-2 and the length of axons in neuronal cells. The composition can be made into ordinary food, health products or medicines for long-term consumption.

[0022] (2) The combination of docosahexaenoic acid, arachidonic acid, phosphatidylserine and Antarctic krill oligopeptide has a significant synergistic effect on the growth and development of neuronal cells. Removing any active ingredient will lead to the weakening or disappearance of the proliferation ability of neuronal cells and the growth ability of axons. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 This is a statistical chart of neuronal cell proliferation after treatment with different concentrations of the three-component mixture (DHA, ARA, PS). # P <0.05,## P <0.01, indicating a significant difference compared with the control group.

[0024] Figure 2 This is a statistical chart of neuronal cell proliferation after treatment with a 100 μg / mL three-component mixture (25 μg / mL DHA, 50 μg / mL ARA, 25 μg / mL PS) and a four-component mixture with different ratios. #P <0.05,## P <0.01, indicating a significant difference compared with the 100 μg / mL three-component mixture.

[0025] Figure 3 This is a statistical chart of neuronal cell proliferation after treatment with different active ingredient combinations. ## P <0.01, indicating a significant difference compared with the control group; ** P <0.01, indicating a significant difference compared with the four-component mixture, and NS indicates no significant difference.

[0026] Figure 4 The fluorescence images and quantitative results of MAP-2 in neuronal cells after treatment with different active ingredients. A is the fluorescence image, B is the fluorescence quantitative result, and C is the average synaptic length. # P <0.05,## P <0.01, indicating a significant difference compared with the control group; + P <0.05,++ P <0.01, indicating a significant difference compared with the AlCl3 group; * P <0.05,** P <0.01, indicating a significant difference compared with the AlCl3+ DHA+ ARA+ PS+ oligopeptide group.

[0027] Figure 5 This is a statistical chart of the proliferation of neuronal cells after being treated with different concentrations of modulated milk powder. # P <0.05,## P <0.01, indicating a significant difference compared with the control group; NS indicates no significant difference.

[0028] Figure 6 The fluorescence images and quantitative results of MAP-2 in neuronal cells after being treated with different concentrations of modulated milk powder. A is the fluorescence image, B is the fluorescence quantitative result, and C is the average synaptic length. ## P <0.01, indicating a significant difference compared with the control group; + P <0.05,++ P <0.01, indicating a significant difference compared with the AlCl3 group; * P <0.05,** P <0.01, indicating a significant difference compared with the AlCl3+100μg / mL formula milk powder group 1. DETAILED DESCRIPTION

[0029] The present invention will be further described below in conjunction with the examples and accompanying drawings. It should be understood that these examples are intended only to illustrate the present invention and are not intended to limit the scope of the present invention. The operating methods in the following examples that do not specify specific conditions are usually performed under normal conditions or according to the conditions recommended by the manufacturer. The content that is not described in detail in this specification sheet belongs to the prior art known to professionals in the field. The experimental materials used in the examples, if not otherwise specified, can all be purchased by conventional biochemical reagent companies.

[0030] In the embodiment, the preparation method of Antarctic krill oligopeptide comprises the following steps: (1) dissolving defatted Antarctic krill powder in water, wherein the weight ratio of defatted Antarctic krill powder to water is 1:10, to obtain a raw material solution; (2) adjusting the pH value of the raw material solution, adding 2.5wt% protease (a mixed protease of a composite protease (commercially available) and an alkaline protease) of defatted Antarctic krill powder, and performing enzymolysis at the optimal temperature of the protease for 4 hours; (3) heating to 100°C to inactivate the enzyme for 20 minutes, filtering and cooling the supernatant; (4) further adding 15wt% activated carbon of defatted Antarctic krill powder, and decolorizing at 70°C for 1 hour; (5) after decolorization, centrifuging to separate the solid and liquid; (6) passing the obtained clear liquid through a ceramic membrane, a nanofiltration membrane, an evaporator, and an ultra-high temperature instantaneous sterilization process; (7) finally spray drying to obtain Antarctic krill oligopeptide. The test results show that the content of peptide segments with a molecular weight of <1000Da in the Antarctic krill oligopeptide is >90%.

[0031] In the following examples, docosahexaenoic acid, arachidonic acid, phosphatidylserine, milk-based powder (including whole milk powder, skim milk powder, desalted whey powder), etc. are all commercially available. The components of the milk-based powder include 35wt% whole milk powder, 25wt% skim milk powder, 15wt% desalted whey powder, 15wt% lactose, 4wt% vegetable fat powder, 3.75wt% minerals and 2.25wt% vitamins.

[0032] Example 1 (Powder) According to weight proportions, 1 part of docosahexaenoic acid (DHA, the same below), 2 parts of arachidonic acid (ARA, the same below), 1 part of phosphatidylserine (PS, the same below), 8 parts of Antarctic krill oligopeptides, and 88 parts of milk-based powder are mixed evenly in proportion and packaged to obtain a formulated milk powder.

[0033] Example 2 (Powder) According to weight, 1.5 parts of DHA, 3 parts of ARA, 1.5 parts of PS, 12 parts of Antarctic krill oligopeptide, 2 parts of fruit powder and 80 parts of oligofructose are mixed evenly in proportion and packaged to obtain a powder product.

[0034] Example 3 (solid beverage) According to weight parts, 2 parts of DHA, 4 parts of ARA, 2 parts of PS, 16 parts of Antarctic krill oligopeptides, 16 parts of xylo-oligosaccharides, 4 parts of isomaltooligosaccharides and 56 parts of skim milk powder are mixed evenly in proportion and packaged to obtain a solid beverage product.

[0035] Example 4 (granules) According to weight proportions, 3 parts of DHA, 6 parts of ARA, 6 parts of PS, 24 parts of Antarctic krill oligopeptide and 12 parts of lactose are uniformly mixed in proportion, granulated, dried and then packaged to obtain a granule product.

[0036] Example 5 (Tablets) According to weight proportions, 3.5 parts of DHA, 7 parts of ARA, 3.5 parts of PS, 28 parts of Antarctic krill oligopeptides, 2.5 parts of galacto-oligosaccharides, 52.7 parts of sorbitol, 1.8 parts of magnesium stearate and 1 part of silicon dioxide are uniformly mixed in proportion, granulated, dried and tableted to obtain a tablet product.

[0037] Example 6 (Hard Capsule)

[0038] According to weight parts, 4 parts of DHA, 8 parts of ARA, 4 parts of PS, 32 parts of Antarctic krill oligopeptide, 50 parts of starch and 2 parts of magnesium stearate are mixed evenly in proportion, and then filled with a capsule filling machine to obtain a hard capsule product.

[0039] Example 7 (microcapsules)

[0040] According to weight proportions, 5 parts of DHA, 10 parts of ARA, 5 parts of PS, 40 parts of Antarctic krill oligopeptide, 8 parts of maltodextrin and 32 parts of whey protein powder are taken; after DHA, PS, ARA and Antarctic krill oligopeptide are evenly mixed, they are then encapsulated with a mixed solution of maltodextrin and whey protein powder, dried at low temperature and then packaged to obtain a microcapsule product.

[0041] Example 8 (Soft Capsule)

[0042] According to weight parts, 3 parts of DHA, 6 parts of ARA, 3 parts of PS, 24 parts of Antarctic krill oligopeptide and 64 parts of Antarctic krill oil are mixed evenly in proportion to obtain the contents, and gelatin, glycerin and water are mixed in a ratio of 1:0.4:1 to prepare a gelatin solution, which is pressed into pills using an automatic soft capsule pill press, and then dried to obtain a soft capsule product.

[0043] Example 9 (Liquid Beverage)

[0044] According to weight parts, 3 parts of DHA, 6 parts of ARA, 3 parts of PS, 24 parts of Antarctic krill oligopeptide, 12 parts of whole milk powder, 10 parts of red date powder, 6 parts of wolfberry powder and 36 parts of purified water are evenly mixed in proportion, and packaged to obtain a formula dairy product.

[0045] Sample analysis 1 Materials and methods 1.1 In vitro culture of neuronal cells Mouse hippocampal neuron cell HT-22 cell line was purchased from Shanghai Chuanqiu Biotechnology Co., Ltd. The culture medium was a high-glucose medium containing 10% fetal bovine serum and 1% penicillin-streptomycin double antibody. The cells were cultured in an incubator at 37°C, 5% CO2 and 95% air. The cells were passaged when the cell density reached 70%~80%.

[0046] 1.2 Neuronal cell proliferation HT-22 cells were plated at 1.5 × 10 3 The cells were seeded in 96-well plates at a density of 100 μg / ml and treated after culturing for 24 h. Firstly, a concentration screening experiment was carried out. Docosahexaenoic acid (DHA), arachidonic acid (ARA), and phosphatidylserine (PS) were mixed at a mass fraction of 1:2:1 to obtain a three-component mixture. The three-component mixture was cultured in cells at concentrations of 25 μg / mL, 50 μg / mL, 100 μg / mL, 200 μg / mL, 400 μg / mL, 600 μg / mL, and 800 μg / mL, respectively (respectively recorded as three-component experimental groups 1-7). The 100 μg / mL three-component mixture contained 25 μg / mL DHA, 50 μg / mL ARA, and 25 μg / mL PS. Meanwhile, the three-component mixture of 25 μg / mL DHA, 50 μg / mL ARA, and 25 μg / mL PS was mixed with 50 μg / mL, 100 μg / mL, 200 μg / mL, 400 μg / mL, 800 μg / mL, and 1000 μg / mL PSA, respectively. Six four-component mixture experimental groups (respectively recorded as four-component experimental groups 1-6) were obtained by compounding Antarctic krill oligopeptides with a concentration of 1 μg / mL. The proliferation rate of cells was calculated after culture.

[0047] At the same time, a comparative experiment of different active ingredient combinations was set up. At the levels of 25 μg / mL DHA, 50 μg / mL ARA, 25μg / mL PS and 200μg / mL Antarctic krill oligopeptide, the three-three combinations (combination 1: DHA+ARA+ PS; combination 2: DHA+ ARA+ Antarctic krill peptide; combination 3: DHA+PS+ Antarctic krill peptide; combination 4: ARA+ PS+ Antarctic krill peptide) were used to treat cells for 24 hours, and the cell proliferation rate was calculated after culture. Finally, the cells were treated with different concentrations of modulated milk powder for 24 hours, and the cell proliferation rate was calculated after culture.

[0048] Specifically, after the culture was completed, the cells were rinsed once with PBS, and 20 μL MTT (5 g / L) and 200 μL serum-free DMEM medium were added to each well. After incubation at 37°C for 4 h, the supernatant was discarded, and 150 μL dimethyl sulfoxide was added to each well to dissolve the generated crystals, and the cells were shaken at room temperature for 20 min, and then the optical density (OD) value was measured at 490 nm.

[0049] 1.3 Immunofluorescence staining of neurons HT-22 cells were plated at 5 × 10 4 The cells were plated at a density of 1000 μmol / L in a 24-well plate containing a cell crawler (10×10 cm), and the cells were treated after culturing for 24 hours. For the comparison experiment of different active ingredient combinations, the cells were treated with 300 μmol / L AlCl3 and a four-component mixture / three-component mixture for 24 hours (the four-component mixture was the treatment level of 25 μg / mL DHA, 50 μg / mL ARA, 25 μg / mL PS and 200μg / mL Antarctic krill oligopeptide, and the three-component mixture was a three-three combination); for the concentration screening experiment of the modulated milk powder, the cells were treated with 300 μmol / L AlCl3 and different concentrations of modulated milk powder for 24 hours.

[0050] After the culture was completed, the medium was discarded, the cells were rinsed twice with PBS, and 4% PFA was added for fixation for 15 min. After the excess paraformaldehyde was washed with PBS, 0.1% TrionX-100 in PBS was added for permeabilization for 15 min. After washing with PBS, the cells were blocked at room temperature for 1 h, washed again with PBS, and the primary antibody MAP-2 (ET1602-12, Huaan, Hangzhou, China) was added and incubated overnight at 4°C. Finally, the secondary antibody was removed, and DAPI (P36981, Thermo Fisher, Massachusetts, USA) was added for sealing. The stained cell sections were photographed by fluorescence microscopy.

[0051] 1.4 Statistical analysis All experimental data were expressed as “mean ± standard error”, and statistical analysis was performed using one-way analysis of variance followed by Student-Newman-Keuls test.

[0052] 2 Experimental Results 2.1 Determination of the critical concentrations of DHA, ARA and PS for promoting neuronal cell growth In order to determine the critical concentration of DHA, ARA and PS to promote the growth of neuronal cells, the three-component composition of different concentrations was prepared with the conventional addition amount and addition ratio of DHA, ARA and PS (10 mg, 20 mg and 10 mg), and the neuronal cells were treated. After 24 hours of treatment, compared with the control group, the three-component composition of 100 μg / mL had no effect on the proliferation of neuronal cells, while the three-component composition of 200 μg / mL could significantly promote the proliferation of neuronal cells, with a proliferation rate of 113.9% ( Figure 1 ).

[0053] 2.2 Determination of the effective concentration of Antarctic krill oligopeptides in promoting neuronal cell growth In order to determine the effective concentration of Antarctic krill oligopeptides in promoting neuronal cell growth, different concentrations of krill peptides were added to a 100 μg / mL mixture of DHA, ARA and PS to prepare a four-component mixture and treat the cells. Figure 2 As shown in the figure, compared with the 100μg / mL three-component mixture, the four-component mixture containing 200μg / mL Antarctic krill oligopeptide significantly promoted the proliferation of neuronal cells, and the cell proliferation rate increased by 15%. Taking into account the cost of raw materials and the effective concentration, the relative proportions of DHA, ARA, PS and Antarctic krill oligopeptide were determined (DHA: ARA: PS: Antarctic krill oligopeptide = 1:2:1:8). In addition, this proliferation was dose-dependent within a certain range ( Figure 2 ).

[0054] 2.3 Effects of the four-component mixture on the growth and development of neuronal cells In order to determine the optimal effect of the four-component composition of DHA, ARA, PS and Antarctic krill oligopeptides in promoting the growth of neuronal cells, a four-component mixture of 25 μg / mL DHA, 50 μg / mL ARA, 25 μg / mL PS and 200μg / mL Antarctic krill oligopeptides was selected and compared with different three-component mixtures. The results showed that compared with the control, after 24 hours of treatment, the four-component mixture significantly increased the proliferation of neuronal cells, and the cell proliferation rate reached 122.9%. The three-component mixture had no effect on cell proliferation. In addition, compared with the three-component mixture, the four-component mixture significantly increased the proliferation of cells ( Figure 3A cell injury model was established by aluminum chloride (AlCl3), and MAP-2 immunofluorescence staining was performed on the neuronal cells treated with the four-component mixture and the three-component mixture. The results are shown in Figure 4 As shown in Figures A to C, AlCl3 treatment significantly reduced the expression of MAP-2 and axon length in neuronal cells, while the four-component mixture and the three-component mixture increased the expression of MAP-2 and axon length. Among them, the four-component mixture had a better effect than the three-component mixture.

[0055] 2.4 Effects of formula milk powder products on neuronal cell growth, development and microscopic morphology In order to evaluate the effects of formula milk powder products on neuronal cell growth and morphology, experiments were conducted at a ratio of DHA: ARA: PS: Antarctic krill oligopeptide = 1:2:1:8. The specific added amounts of docosahexaenoic acid DHA, arachidonic acid ARA, phosphatidylserine PS and Antarctic krill oligopeptide (10 mg, 20 mg, 10 mg and 80 mg) per 100 g of formula milk powder (the formula milk powder contains only milk-based powder in addition to the four-component mixture, and the specific components of the milk-based powder are as described above) are referred to as "formulated milk powder group 1"; the specific added amounts of docosahexaenoic acid DHA, arachidonic acid ARA, phosphatidylserine PS and Antarctic krill oligopeptide (20 mg, 40 mg, 20 mg and 160 mg) per 100 g of formula milk powder are referred to as "formulated milk powder group 2". Figure 5 As shown in the figure, compared with the control, the 200, 400, 600, 800 and 1000 μg / mL of the modified milk powder group 1 significantly increased the proliferation of cells, and the cell proliferation rates reached 109.8%, 113.5%, 114.6%, 122.6% and 117.6%, respectively; compared with the control, the 100, 200, 400, 600, 800 and 1000 μg / mL of the modified milk powder group 2 significantly increased the proliferation of cells, and the cell proliferation rates reached 110.2%, 119.2%, 119.2%, 127.6, 124.1% and 121.1%, respectively. However, compared with the concentrations of the modified milk powder group 1, the modified milk powder group 2 had no significant effect on cell proliferation. In addition, MAP-2 immunofluorescence staining was performed on cells treated with 100 and 200 μg / mL of the modified milk powder 1 group. The results showed that compared with the control group, AlCl3 treatment significantly reduced the expression of MAP-2 and axon length in neuronal cells, while 200 μg / mL of the modified milk powder 1 group increased the expression of MAP-2 and axon length in neuronal cells ( Figure 6 AC in the ).

[0056] 3 Experimental conclusions HT-22 cells are immortalized mouse hippocampal neuron cell lines, which are used to study the mechanisms of neuronal development and differentiation and are one of the more commonly used in vitro research cell lines. The MTT experiment is one of the reliable and convenient methods to test cell activity. Through the MTT experiment, this study found that a four-component mixture of 25 μg / mL DHA, 50 μg / mL ARA, 25 μg / mL PS and 200 μg / mL Antarctic krill oligopeptides significantly promoted the proliferation of HT-22 cells after 24 hours, while the three-component combination with different active ingredient configurations had no effect on cell proliferation; 200 μg / mL of the modulated milk powder group 1 significantly promoted the proliferation of HT-22 cells after 24 hours, and the effect increased with the increase of treatment concentration within a certain range. MAP2 is an important neuronal cytoskeletal protein that plays a key role in the development, formation and regeneration of the nervous system. The structure and function of neurons can be observed by staining. The results of MAP-2 immunofluorescence staining and quantitative analysis showed that the four-component mixture of 25 μg / mL DHA, 50 μg / mL LARA, 25 μg / mL PS and 200 μg / mL Antarctic krill oligopeptides and 200 μg / mL of modulated milk powder group 1 significantly improved the synaptic damage caused by AlCl3, which was beneficial to the neurodevelopment of HT-22 cells.

[0057] The embodiments described above provide a detailed description of the technical solutions of the present invention. It should be understood that the above are only specific embodiments of the present invention and are not intended to limit the present invention. Any modifications, supplements or similar substitutions made within the scope of the principles of the present invention should be included in the protection scope of the present invention.

Claims

1. An Antarctic krill oligopeptide nutritional composition for promoting neural development, characterized in that: The invention comprises, by weight, 0.01 to 10 parts of docosahexaenoic acid, 0.01 to 20 parts of arachidonic acid, 0.01 to 10 parts of phosphatidylserine and 0.01 to 80 parts of Antarctic krill oligopeptide; the Antarctic krill oligopeptide is obtained by treating Antarctic krill, Antarctic krill powder or defatted Antarctic krill powder by enzymatic hydrolysis, and the content of peptide segments with a molecular weight of less than 1000 Da in the Antarctic krill oligopeptide is greater than 85%.

2. The Antarctic krill oligopeptide nutritional composition according to claim 1, characterized in that: Calculated by weight, it comprises 0.05-5 parts of docosahexaenoic acid, 0.05-10 parts of arachidonic acid, 0.05-5 parts of phosphatidylserine and 0.05-40 parts of Antarctic krill oligopeptide; the content of peptide segments with a molecular weight of less than 1000Da in the Antarctic krill oligopeptide is greater than 90%.

3. The Antarctic krill oligopeptide nutritional composition according to claim 1, characterized in that: Calculated by weight, it comprises 1 part of docosahexaenoic acid, 2 parts of arachidonic acid, 1 part of phosphatidylserine and 8 parts of Antarctic krill oligopeptides; the content of peptide segments with a molecular weight of less than 1000Da in the Antarctic krill oligopeptides is greater than 90%.

4. The Antarctic krill oligopeptide nutritional composition according to claim 1, characterized in that: The preparation method of Antarctic krill oligopeptide comprises: (1) dissolving defatted Antarctic krill powder in water to obtain a raw material solution; (2) Adjusting the pH value of the raw material solution, adding protease, and performing enzymolysis at the optimal temperature of the protease; (3) After inactivating the enzyme, filter and cool the supernatant; (4) The solution obtained in step (3) is decolorized, and further solid-liquid separation is performed to obtain a supernatant, and the supernatant is sterilized and then dried to obtain the Antarctic krill oligopeptide.

5. Use of the Antarctic krill oligopeptide nutritional composition according to any one of claims 1 to 4 in the preparation of products that promote neural development.

6. A product for promoting neural development, characterized in that: The ingredients include the Antarctic krill oligopeptide nutritional composition according to any one of claims 1 to 4.

7. The product for promoting neural development according to claim 6, characterized in that: Product forms include powder, tablet, capsule, granule or drink.

8. The product for promoting neural development according to claim 6, characterized in that: The ingredients also include pharmaceutically acceptable excipients or food-acceptable additives.

9. The product for promoting neural development according to claim 6, characterized in that: The product for promoting neural development is a modulated milk powder, which comprises 0.12-0.24 wt % of the Antarctic krill oligopeptide nutritional composition and 99.76-99.88 wt % of the milk-based powder.

10. The product for promoting neural development according to claim 9, characterized in that: The milk-based powder includes 30-40wt% whole milk powder, 20-30wt% skim milk powder, 10-20wt% desalted whey powder, 10-20wt% lactose, 3-5wt% vegetable fat powder, 1-5wt% vitamins and 1-5wt% minerals.

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