RU2741495C2 - Linoleic acid or alpha-linolenic acid for use in reducing cognitive disorders/early neurogenesis induced by early life stress - Google Patents

Abstract

FIELD: food industry.

SUBSTANCE: group of inventions relates to the use of linoleic acid (LA) and alpha-linolenic acid (ALA) for preparing a composition, including LA and ALA in weight ratio LA/ALA in range of 0.1–12, for prevention of cognitive function reduction induced by early life human stress, where the human being is an unborn fetus, an infant or toddler, and to prevent the early life (ES) induced cell hippocampus decline in a human, where a human being is an unborn fetus, an infant or toddler.

EFFECT: group of inventions provides a composition comprising linoleic acid (LA) and alpha-linolenic acid (ALA) in a low weight ratio of LA/ALA, which helps to prevent cognitive function reduction induced by stress in early life, also helps to reduce early stage induced neurogenesis, and also promotes the survival of hippocampal cells.

15 cl, 1 dwg, 3 tbl, 2 ex

Description

FIELD OF THE INVENTION

The present invention relates to the field of nutritional compositions that have a beneficial effect in reducing stress-induced disorders in early life, for example, nutritional compositions that have a beneficial effect on cognitive impairments induced by stress in the early period of life, in particular stress in early life period of life experienced by a person.

BACKGROUND OF THE INVENTION

Early life stress is known to negatively affect cognitive performance throughout a patient's life. Cognitive decline induced by stress early in life cannot be cured or can only be cured to a limited extent. During early life, the developing human brain is highly sensitive to modulations of environmental influences, and hence any adaptation of structure and function is programmed by such early environmental influences for the rest of life. A direct association between stress in early life and cognitive impairment is supported by clinical evidence. Such an association is described by Lucassen et al. in Trends in Neurosciences , 2013, 36, 621-631.

The hippocampus is one of the very few areas of the brain that retains the ability to create new neurons throughout adulthood. This process is called adult neurogenesis. Newly formed neurons are essential for the functioning of the hippocampus and are involved in specific aspects of hippocampal-dependent learning and memory. Naninck et al. in an article entitled "Chronic early life stress alters developmental and adult neurogenesis and impairs cognitive function in mice" in Hippocampus (2015) vol 25 (3): 309-328, describes that early life (ES) stress induces changes in neurogenesis and also reduces the survival of neurons formed in adulthood, and that altered levels of neurogenesis are functionally important for cognitive impairment in adulthood. In short, stress early in life disrupts cognitive function and hippocampal neurogenesis in adulthood, also called "adult neurogenesis." The authors hope that reaching an understanding of the underpinnings of such changes induced by early life stress will prove to be very important for mental health and should provide the basis for future therapeutic interventions.

SUMMARY OF THE INVENTION

During early life, the development of the human brain is highly sensitive to changes under the influence of the environment and any adaptation of structure and function is therefore programmed by such early environmental influences during the rest of life. The present invention provides a method for providing a pathway for influencing brain structures and functions through nutritional interventions to alleviate disturbances due to early life stress.

The inventors have surprisingly found that administration of a nutritional composition comprising linoleic acid (LA) and alpha-linolenic acid (ALA) at a low LA / ALA weight ratio is effective in preventing stress-induced cognitive decline in early life. Mice that were stressed early in life and fed with the nutritional compositions of the invention outperformed mice that were subjected to the same stress but fed with a nutritional composition with a higher LA / ALA weight ratio in the object recognition test, object placement tests and the water test. Morris maze, conducted in adulthood, which has shown a significant preventive effect on cognitive decline. Therefore, unexpectedly, through dietary intervention, the negative effects of stress early in life can be prevented.

Also, the inventors have surprisingly found that administration of a nutritional composition comprising linoleic acid (LA) and alpha-linolenic acid (ALA) in a low LA / ALA weight ratio is effective in preventing the decrease in neurogenesis induced by stress in early life (ES). It was found that dietary intervention with a nutritional composition comprising a low LA / ALA weight ratio restored neurogenesis in adulthood after exposure to stress in early life. Mice that were stressed early in life and fed with the nutritional composition of the invention demonstrated unexpected survival of hippocampal cells in adulthood compared to mice that were exposed to the same stress but fed with a nutritional composition with higher LA / ALA, demonstrating reliable preventive effect in relation to the reduction of neurogenesis. Therefore, unexpectedly through dietary intervention, the negative effects of stress early in life can be prevented. The use of a low nutritional mass ratio of LA / ALA in the early period of life protects against stress-induced disorders in the early period of life, in particular, protects against a decrease in neurogenesis.

DETAILED DESCRIPTION OF THE INVENTION

Therefore, the present invention relates to a method for preventing stress-induced cognitive decline in early life in humans, comprising administering to the patient a nutritional composition comprising linoleic acid (LA) and alpha-linolenic acid (ALA) in an LA / ALA weight ratio in the range of 0, 1-12. In one embodiment, the method for preventing early life stress-induced cognitive decline is a non-medical method.

In other words, the invention relates to a nutritional composition comprising linoleic acid (LA) and alpha-linolenic acid (ALA) in an LA / ALA weight ratio in the range of 0.1-12 for use in the prevention of stress-induced cognitive decline in the early period of life. in a person.

The invention can also be formulated as the use of linoleic acid (LA) and alpha-linolenic acid (ALA) in the preparation of a nutritional composition comprising LA and ALA in an LA / ALA weight ratio in the range of 0.1-12 for the prevention of cognitive decline induced by stress in the early period of human life.

The invention can also be formulated as the use of a nutritional composition comprising linoleic acid (LA) and alpha-linolenic acid (ALA) in a LA / ALA weight ratio in the range of 0.1-12 for the prevention of early stress-induced cognitive decline life of a person.

The present invention also relates to a method for preventing a decrease in early life stress induced neurogenesis (ES) in a human, comprising administering to a human a nutritional composition comprising linoleic acid (LA) and alpha-linolenic acid (ALA) in an LA / ALA weight ratio in the range 0.1-12. In one embodiment of the invention, the method for preventing the decrease in neurogenesis induced by stress in early life (ES) is a non-medical method.

Alternatively, the present invention relates to a method for restoring neurogenesis in an adult in a human after stress in early life, comprising administering to a human a nutritional composition comprising linoleic acid (LA) and alpha-linolenic acid (ALA) in an LA / ALA weight ratio in the range of 0, 1-12. In one embodiment, the method of restoring neurogenesis in adulthood is a non-medical method.

In other words, the invention relates to a nutritional composition comprising linoleic acid (LA) and alpha-linolenic acid (ALA) in an LA / ALA weight ratio in the range of 0.1-12 for use in the prevention of a decrease in neurogenesis induced by stress in early life (ES ) in humans.

In other words, the invention relates to a nutritional composition comprising linoleic acid (LA) and alpha-linolenic acid (ALA) in an LA / ALA weight ratio in the range of 0.1-12, for use in the restoration of neurogenesis in humans in adulthood after early stress. period of life.

The invention can also be described as the use of linoleic acid (LA) and alpha-linolenic acid (ALA) in the preparation of a nutritional composition comprising LA and ALA in an LA / ALA weight ratio in the range of 0.1-12 for the prevention of stress-induced reduction in neurogenesis in early life (ES) in humans.

The invention can also be presented as the use of linoleic acid (LA) and alpha-linolenic acid (ALA) in the preparation of a nutritional composition comprising LA and ALA in an LA / ALA weight ratio in the range of 0.1-12 to restore neurogenesis in humans after stress in early life.

The invention can also be presented as the use of a nutritional composition comprising linoleic acid (LA) and alpha-linolenic acid (ALA) in a LA / ALA weight ratio in the range of 0.1-12 for the prevention of stress-induced decrease in neurogenesis in the early period of life (ES) in humans.

The invention may also be described as the use of a nutritional composition comprising linoleic acid (LA) and alpha-linolenic acid (ALA) in an LA / ALA weight ratio in the range of 0.1-12 to restore neurogenesis in an adult human after stress early period of life.

In a preferred embodiment of the invention, prevention of the decrease in neurogenesis induced by stress in early life (ES) occurs by prevention of decreased survival of hippocampal cells in humans.

In one aspect, the present invention relates to a method for preventing a decrease in hippocampal cell survival induced by stress in early life (ES) in a human, comprising administering to a human a nutritional composition comprising linoleic acid (LA) and alpha-linolenic acid (ALA) in a weight ratio LA / ALA in the range of 0.1-12. In one embodiment, the method comprises preventing early life stress (ES) stress-induced reduction in hippocampal cell survival (ES) is a non-medical method.

In other words, the invention relates to a nutritional composition comprising linoleic acid (LA) and alpha-linolenic acid (ALA) in an LA / ALA weight ratio in the range of 0.1-12 for use in the prevention of stress-induced reduction in hippocampal cell survival in the early period. life (ES) in humans.

The invention can also be described as the use of linoleic acid (LA) and alpha-linolenic acid (ALA) in the preparation of a nutritional composition comprising LA and ALA in an LA / ALA weight ratio in the range of 0.1-12 for the prevention of decreased survival of hippocampal cells stress induced early life (ES) in humans.

The invention can also be described as the use of a nutritional composition comprising linoleic acid (LA) and alpha-linolenic acid (ALA) in an LA / ALA weight ratio in the range of 0.1-12 to prevent early stress-induced decrease in hippocampal cell survival. period of life (ES) in humans.

All that has been said below for the process according to the invention applies equally in the application according to the invention and to the composition for use according to the invention and vice versa.

Nutritional composition

The nutritional composition to be administered according to the method of the invention comprises linoleic acid (LA) and alpha-linolenic acid (ALA) in an LA / ALA weight ratio in the range of 0.1-12. Such a composition is referred to as a "nutritional composition according to the invention" or a "nutritional composition".

The nutritional composition of the invention can take any form of nutrition suitable for early administration to a human patient. The nutritional composition according to the invention can be a food additive, for example, as an additive to a regular diet, a fortification mixture to be added to a regular diet, a balanced diet, or an infant food suitable for feeding infants (for example, a first level milk formula or a second level milk formula ). Preferably, the nutritional composition is suitable for feeding infants and / or preschool children. In a preferred embodiment, the nutritional composition is selected from a first level infant formula, a second level infant formula, a third level infant formula and a nutritional supplement for pregnant women, most preferably the nutritional composition is a first level infant formula.

Depending on the form of the nutritional composition, it can be a balanced meal or supplement. Preferably, the nutritional composition is a balanced diet. It is preferred that the composition comprises a lipid component, a digestible carbohydrate component and a protein component. It may additionally contain ingredients such as dietary fiber, minerals, vitamins, organic acids, nucleotides and flavorings. The lipid component preferably provides 2.9-6 g lipid per 100 kcal, the protein component preferably provides 1.8 to 5.5 g per 100 kcal, respectively 1.8 to 2.5 g per 100 kcal and the digestible carbohydrate component is preferably provides from 9 to 14 g per 100 kcal, composition. Total calories are defined as the sum of calories derived from protein, lipids, digestible carbohydrates, and indigestible oligosaccharides.

The nutritional composition of the invention is generally an enteral composition, i. E. intended for oral administration. It is accordingly administered in liquid form. For example, the composition may include water in which additional components are dissolved or suspended. Consequently, the composition is suitably liquid or solid, usually a powder or tablet, which is reconstituted with liquid, suitably water or other edible aqueous liquids, to obtain a liquid composition, or in the form of a liquid concentrate which is diluted with water. Preferably, the liquid composition has a viscosity below 100 mPa.s, suitably below 60 mPa.s, suitably below 35 mPa.s, more precisely below 6 mPa.s, as measured on a Brookfield viscometer at 20 ° C at a shear rate of 100 s ^-1 . In order to meet caloric requirements for an infant, the composition preferably comprises 45 to 200 kcal / 100 ml liquid, more preferably 60 to 90 kcal / 100 ml liquid, even more preferably 60 to 75 kcal / 100 ml liquid. This caloric content provides an optimal balance between water consumption and calories. The osmolarity of the present composition is preferably between 150 and 420 mOsmol / L, more preferably between 260 and 360 mOsmol / L.

Lipid

The nutritional composition comprises a lipid component, suitably a lipid component suitable for feeding infants, as is known in the art. The lipid component of the present composition suitably provides 2.9 to 6.0 g, preferably 4 to 6 g, per 100 kcal of the composition. When in liquid form, the nutritional composition preferably comprises 2.1 to 6.5 g lipid per 100 ml, more preferably 3.0 to 4.0 g per 100 ml. The lipid provides preferably from 30 to 60% of the total calories of the composition. More preferably, the present composition comprises a lipid providing 35 to 55% of the total calories, even more preferably, the present composition comprises a lipid providing 40 to 50% of the total calories. When in liquid form, for example a ready-to-feed liquid, the composition preferably comprises 2.1 to 6.5 g lipids per 100 ml, more preferably 3.0 to 4.0 g per 100 ml. In one embodiment, the nutritional composition comprises at least 15 wt% liquid based on the dry weight of the composition. Based on dry weight, the nutritional composition preferably comprises 10 to 50 wt%, more preferably 12.5 to 40 wt% lipid, even more preferably 15 to 35 wt% lipid, even more preferably 19 to 30 wt% lipid.

The lipid component contained in the nutritional composition includes linoleic acid (LA) and alpha-linolenic acid (ALA). Here, "linoleic acid" or "LA" refers to linoleic acid and / or acyl chain (18: 2 n6). Here, "alpha-linolenic acid" or "ALA" refers to the alpha-linolenic acid and / or acyl chain (18: 3 n3). LA is preferably present in sufficient amounts to promote healthy growth and development. Therefore, the composition preferably comprises less than 20 wt% LA based on all fatty acids, preferably 5 to 15 wt%. Preferably, the composition comprises more than 5 wt% LA based on fatty acids, preferably at least 10 wt% based on fatty acids. Preferably ALA is present in sufficient quantity to promote healthy growth and development. Therefore, the nutritional composition preferably comprises at least 0.5 wt% ALA based on all fatty acids, preferably comprises at least 1.0 wt% ALA based on all fatty acids. Preferably, the composition comprises at least 1.4 wt% ALA based on all fatty acids, more preferably at least 1.5 wt%. Preferably, the nutritional composition comprises less than 10 wt% ALA, more preferably less than 5.0 wt%, based on all fatty acids. In one embodiment, the nutritional composition comprises at least 10 wt% LA and at least 1 wt% ALA, preferably 10 to 20 wt% LA and 1 to 5 wt% ALA.

Surprisingly, it has been found that a reduced LA / ALA weight ratio, when comparing conventional nutritional compositions, in particular conventional milk formulas, has a beneficial effect on the cognitive functions of a patient when exposed to stress in early life and prevents cognitive decline induced by stress in early life. The LA / ALA weight ratio of the nutritional composition according to the present invention is in the range of 0.1-12, preferably in the range of 0.5-11, more preferably in the range of 0.8-10, more preferably in the range of 1-9, more preferably in the range 1-8.

Lipid components such as LA and ALA, as well as optionally present additional components such as (LC-) PUFA, can be provided as free fatty acids, triglycerides, diglycerides, monoglycerides, and phospholipids, or the form of a mixture of one or more of the above. Preferably, the nutritional composition comprises triglycerides. Preferably, the composition comprises at least 70 wt%, more preferably at least 80 wt%, more preferably at least 85 wt% triglycerides, even more preferably at least 90 wt% triglycerides, based on total weight. Lipids can additionally include one or more of free fatty acids, monoglycerides, and diglycerides.

It was also surprisingly found that a reduced LA / ALA weight ratio when compared with conventional nutritional compositions, in particular conventional milk formulas, favorably affects the survival of the patient's hippocampal cells when exposed to stress in the early period of life and prevents the decrease in neurogenesis induced by stress in the early period of life. ... The LA / ALA weight ratio of the nutritional composition of the present invention is in the range from 0.1-12, preferably in the range 0.2-11, preferably in the range 0.4-10, preferably in the range 0.5-10, more preferably in the range of 0.6-10, more preferably in the range of 0.8-10, more preferably in the range of 1-8.

In a preferred embodiment, the nutritional composition comprises triglycerides derived from vegetable fat. In a further preferred embodiment of the invention, the nutritional composition preferably further comprises phospholipids. Preferably, the nutritional composition comprises phospholipids derived from the milk of non-human mammals. The presence of plant lipids predominantly provides an optimal fatty acid profile, high in polyunsaturated fatty acids and more reminiscent of human milk fat. Using lipids from ruminant milk, in particular only cow's milk, or other domestic ruminants, does not provide an optimal fatty acid profile. This less optimal fatty acid profile, such as a high amount of saturated fatty acids, is known to be unfavorable. Preferably, the present composition comprises at least one, preferably at least two lipid sources selected from the group consisting of flaxseed oil (flaxseed oil), rapeseed oil (such as rapeseed oil, low erucic rapeseed oil and canola oil) , Dalmatian sage oil, perilla oil, purslane oil, lingonberry oil, sea buckthorn oil, hemp oil, sunflower oil, high oleic sunflower oil, safflower oil, high oleic safflower oil, olive oil, black currant seed oil, echium oil, palm oil, coconut oil oil and palm kernel oil. Preferably, the present composition comprises at least one, preferably at least two, lipid sources selected from the group consisting of linseed oil, canola oil, coconut oil, sunflower oil and high oleic sunflower oil. Preferably, the composition comprises 30 to 99.5 wt% plant lipids based on total lipids, more preferably 35 to 99 wt%, even more preferably 40 to 95 wt% based on total lipids.

Preferably, the nutritional composition comprises fat or lipids from mammalian milk, more preferably ruminant milk, even more preferably cow's milk, goat's milk, sheep's milk, buffalo's milk, yak's milk, reindeer's milk and camel's milk, most preferably cow's milk. Preferably, the mammalian milk is not human milk. Hence, in one embodiment, the nutritional composition comprises milk fat from non-human mammals. Preferably, the fatty component of mammalian milk comprises at least 70 wt% triglycerides, more preferably at least 90 wt%, more preferably at least 97 wt%.

Preferably, the mammalian milk fat is derived from the group consisting of butter, milk fat, milk oil and anhydrous milk fat, more preferably anhydrous milk fat and milk oil. Such milk fat lipid sources are high in triglycerides. Moreover, such lipid sources are in the form of a fat continuous phase or in the form of a water-in-oil emulsion. The use of such milk fat sources in the preparation of the nutritional composition according to the invention allows the formation of lipid globules, where each globule comprises a mixture of vegetable fat and milk fat.

Preferably, the nutritional composition comprises 5 to 70 wt% milk lipids from non-human mammals based on total lipids. In one embodiment, the composition comprises 10 to 65 wt%, even more preferably 15 to 60 wt%, even more preferably 25 to 55 wt% of non-human mammalian milk lipids, based on total lipids. Preferably, such milk lipids are selected from the group consisting of butter, milk fat, ghee and anhydrous milk fat. Preferably, the ratio of vegetable fat to milk fat ranges from 3/7 to 20/1.

The composition can also include non-vegetable lipids and non-dairy fat such as non-milk fat such as fish oil and egg lipids and microbial, algal, fungal and unicellular oils. Preferably, the non-vegetable non-dairy fat is present in an amount of no more than 10 wt% based on the total lipids, more preferably no more than 5 wt%. Preferably, the lipid in the nutritional composition of the invention comprises a fat source comprising long chain polyunsaturated fatty acids (LC-PUFA) selected from the group consisting of fish oil, sea kerosene, algae oil, microbial oil, single cell oil and egg lipids in an amount from 0, 25 to 10 wt% based on total lipids, preferably in an amount of 0.5 to 10 wt%.

In a certain preferred embodiment, the lipid is in the form of lipid beads. When in liquid form, lipid beads are emulsified in the aqueous phase. Alternatively, the lipid beads are present in the powder and the powder can be reconstituted with water or other food grade aqueous phase. Typically and preferably lipid beads include a core and a surface. The core preferably comprises vegetable fat and milk fat, and preferably comprises at least 80 wt%, more preferably at least 90 wt% triglycerides, and more preferably consists essentially of triglycerides. Not all of the triglyceride lipids that are present in the composition must necessarily be contained in the core of the lipid globules, but preferably a large portion, preferably more than 50 wt%, more preferably more than 70 wt%, even more preferably more than 85 wt%, even more preferably more than 95 wt%, most preferably more than 98 wt% of the triglyceride lipids that are present in the composition are contained in the core of the lipid beads.

According to a preferred embodiment of the invention, the lipid beads have a distribution of diameters based on volume of more than 1.0 µm, preferably more than 2.0 µm, preferably more than 2.5 µm, more preferably more than 3.0 µm. The lipid beads preferably have a diameter distribution based on volume of less than 6.0 µm, preferably less than 5.5 µm, more preferably less than 5.0 µm. Preferably, the lipid beads have a diameter distribution based on volume of 2 to 6 μm, preferably 2.0 to 6.0 μm, more preferably 2.5 to 6.0 μm, more preferably 3.0 to 6.0 μm, even more preferably 3.0 to 5.5 µm, even more preferably 3.0 to 5.0 µm. The diameter distribution refers to the diameter that is most often present based on the volume of all lipids, or the peak value plotted as the diameter on the X-axis and the volume on the Y-axis (%). A suitable method for determining the volume of lipid beads and their size distribution is using a Mastersizer particle size analyzer (Malvern Instruments, Malvern, UK), for example, as described in Michalski et al, 2001, Lait 81: 787-796. The specific surface area of lipid beads is the surface area per lipid mass and decreases with increasing bead size. The specific surface area of the lipid beads can therefore be calculated from the particle size distribution of the lipid beads and the concentration and density of the lipid. The lipid beads contained in the nutritional composition preferably have a specific surface area of 0.5 to 15 m ² / g lipid, preferably 1.0 to 10.0 m ² / g, more preferably 1.5 to 8.0 m ² / g, even more preferably from 2.0 to 7.0 m ² / g of lipid.

The nutritional composition preferably includes phospholipids derived from (or "derived from") milk from non-human mammals. Phospholipids derived from the milk of non-human mammals include glycerophospholipids and sphingomyelin. The phospholipids are preferably coated on the surface of the lipid globule. By 'shell' it is meant that the outer surface layer of the lipid bead includes phospholipids, while such phospholipids are substantially absent from the core of the lipid bead. Not all lipids that are present in the nutritional composition need to be contained in the shell, but preferably most of them. Preferably more than 30 wt%, preferably more than 50 wt%, more preferably more than 70 wt%, even more preferably more than 85 wt%, most preferably more than 95 wt% of the phospholipids that are present in the composition are contained in the shell of the lipid beads ...

In one embodiment, the nutritional composition comprises at least 0.5 wt% phospholipids based on total lipids. Preferably 0.5 to 20 wt% phospholipids based on total lipids, more preferably 0.5 to 10 wt%, more preferably 1 to 10 wt%, even more preferably 1.0 to 5 wt% even more preferably 1.0 to 2.0 wt% phospholipids based on total lipids. Preferably, at least 80 wt% phospholipids are derived from milk of non-human mammals, more preferably at least 90 wt%, even more preferably at least 95 wt% or 99 wt% or preferably all phospholipids are derived from milk of non-human mammals.

Phospholipids preferably include sphingomyelin. Sphingomyelins have a phosphorylcholine or phosphorylethanolamine molecule esterified with a 1-hydroxy group of ceramide. Preferably, the nutritional composition comprises 0.05 to 10 wt% sphingomyelin based on total lipids, more preferably 0.1 to 5 wt%, even more preferably 0.2 to 2 wt%. Preferably, the nutritional composition comprises at least 15 wt%, more preferably at least 20 wt% sphingomyelin based on the total amount of phospholipids. Preferably, the amount of sphingomyelin is less than 50 wt% based on the total amount of phospholipids.

Phospholipids preferably include glycerophospholipids. Glycerophospholipids are a class of lipids formed from fatty acids esterified at hydroxyl groups on carbon-1 and carbon-2 of the backbone of the glycerol moiety and a negatively charged phosphate group attached to carbon-3 of glycerol via an ester bond and optionally a choline group (in the case of phosphatidylcholine, PC), a serine group (in the case of phosphatidylserine, PS), an ethanolamine group (in the case of phosphatidylethanolamine, PE), an inositol group (in the case of phosphatidylinositol, PI), or a glycerol group (in the case of phosphatidylglycerol, PG) attached to the phosphate group. Preferably, the nutritional composition contains PC, PS, PI and / or PE, more preferably at least PS. Preferably, the nutritional composition comprises at least 1 wt%, preferably at least 2 wt% phosphatidylserine, based on the total amount of phospholipids. Preferably, the amount of phosphatidylserine is less than 10 wt% based on the total amount of phospholipids.

Phospholipids derived from milk of non-human mammals, cream lipids, serum lipids, oil plasma lipids, beta serum lipids, serum lipids, cheese lipids and / or buttermilk lipids. Preferably, the phospholipids are derived from milk cream. Phospholipids are preferably obtained from milk of cows, mares, sheep, goats, buffaloes, horses, camels, preferably from cow's milk. It is most preferable to use a lipid extract isolated from cow's milk. A suitable source of phospholipids derived from the milk of non-human mammals is a fraction that can be isolated from milk, called the milk fat globule membrane (MFGM). Therefore, in one embodiment, the phospholipids that are contained in the nutritional composition of the invention are provided as MFGMs.

Phospholipids are usually located on the surface of lipid globules, i.e. included in the shell or outer layer. In one embodiment of the invention, the lipid beads comprise a monolayer comprising phospholipids derived from milk fat. A suitable way to determine whether polar lipids are located on the surface of lipid beads is by laser scanning microscopy or trans-electron microscopy. The concomitant use of polar lipids in certain phospholipids derived from pet milk and triglycerides derived from plant lipids, therefore, allows the production of coated lipid beads with a membrane more similar to human milk, while at the same time providing an optimal fatty acid profile. Methods for the preparation of lipid beads with such a diameter and / or phospholipid shells are described in WO 2010/0027258 and WO 2010/0027259.

In one embodiment, the nutritional composition comprises at least 0.3 wt% butyric acid and / or acyl chain (BA), based on the weight of all fatty acids. In one embodiment, the nutritional composition comprises 0.3 to 4.0 wt%, preferably 0.3 to 3 wt%, BA based on all fatty acids. The presence of relatively high amounts of BA is characteristic of triglycerides derived from ruminant milk such as cow's milk. It is not found in vegetable fats such as coconut oil, nor is it found in polar lipids derived from milk fat. Thus, an alternative way of describing the presence of milk fat triglycerides in a composition is to determine a fatty acid profile having a BA content of 0.3 to 4.0 wt%, based on all fatty acids. Based on the total weight of the fatty acids, the composition preferably comprises at least 0.3 wt% BA, preferably at least 0.5 wt%, more preferably at least 0.6 wt%, more preferably at least 0.8 wt% ... Preferably, the composition has a wt% BA below 4 wt% based on the weight of all fatty acids, more preferably below 3 wt%, more preferably below 2.5 wt%.

Preferably, the nutritional composition comprises at least 5 wt% medium chain fatty acids and / or acyl chains (MCFA) based on all fatty acids, more preferably at least 7 wt%. In the context of the present invention, MCFA refers to fatty acids and / or acyl chains with a chain length of 8 to 12 carbon atoms. The composition advantageously comprises less than 15 wt% MCFA based on all fatty acids, more preferably less than 10 wt%.

Preferably, the nutritional composition comprises 10 to 25 wt% polyunsaturated fatty acids and / or acyl chains (PUFA) based on total fatty acids. Amounts above 25 wt% are greater than those present in human milk and cause technological problems such as stability in the nutritional composition.

Preferably, the nutritional composition comprises long chain polyunsaturated fatty acids and / or acyl chains (LC-PUFA), more preferably n-3 LC-PUFA. In the context of the present invention, PUFAs comprise at least 20 carbon atoms in the fatty acyl chain and have 2 or more unsaturated bonds. More preferably, the present composition comprises eicosapentaenoic acid and / or acyl chain (20: 5 n3, EPA), docosapentaenoic acid and / or acyl chain (22: 5 n3, DPA) and / or docosahexaenoic acid and / or acyl chain (22: 6, n3, DHA), even more preferably DHA. Since a low concentration of DHA, DPA and / or EPA is already effective and normal growth and development is important, the n-3 LC-PUFA content in the nutritional composition, more preferably DHA, preferably does not exceed 5 wt% of the total fatty acid content. Preferably, the nutritional composition comprises at least 0.15 wt%, preferably at least 0.35 wt%, more preferably at least 0.75 wt% n-3 LC-PUFA, more preferably DHA, total fatty acids. The nutritional composition preferably comprises at least 0.25 wt% LC-PUFA based on total fatty acids. Preferably, the lipid in the nutritional composition comprises a fat source comprising 0.25 wt% to 5 wt% LC-PUFA based on total fatty acids, of which at least 0.15 wt% n-3 LC-PUFA based on total fatty acids are selected from the group consisting of DHA, EPA, and DPA, more preferably DHA.

Since the n-6 fatty acid group, especially the arachidonic acid and / or acyl chain (20: 4 n6, ARA) and LA as its precursor, counteracts the n-3 fatty acid group, especially DHA and EPA and ALA as their precursor, the nutritional composition preferably comprises relatively low amounts of ARA. The n-6 LC-PUFA content is more preferably ARA, preferably does not exceed 5 wt%, more preferably does not exceed 2.0 wt%, more preferably does not exceed 0.75 wt%, even more preferably does not exceed 0.5 wt%, by based on total fatty acids. Since ARA is important in infants for optimal membrane function, especially neurological tissue membranes, the amount of n-6 LC-PUFAs, preferably ARA, is preferably 0.02 wt%, more preferably at least 0.05 wt%, more preferably at least 0.1 wt% based on total fatty acids, more preferably at least 0.2 wt%. The presence of preferably low amounts of ARA is particularly beneficial in nutrition for infants up to 6 months of age, since formula is the only source of nutrition for such infants. Preferably, the mass ratio of n-6 LC-PUFA / n-3 LC-PUFA, more preferably the mass ratio of ARA / DHA is less than 3, more preferably 2 or less, even more preferably 1 or less.

Protein

The nutritional composition preferably includes a protein component. The protein component preferably provides 5 to 15% of the total calories. Preferably, the composition comprises a protein that provides 6 to 12% of the total calories. More preferably, the protein is present in the composition at less than 9%, based on calories. Human milk contains less protein, based on total calories, than cow's milk. The protein concentration in the nutritional composition is determined by the sum of protein and free amino acids. Based on dry weight, the composition preferably comprises less than 12 wt% protein, more preferably 9.6 to 12 wt%, even more preferably 10 to 11 wt%. Based on a liquid ready-to-drink product, the composition preferably comprises less than 1.5 g protein per 100 ml, more preferably 1.2 to 1.5 g, even more preferably 1.25 to 1.35 g.

The source of the protein component must be selected in such a way that the minimum requirements for essential amino acids are met and satisfactory growth is ensured. Therefore, protein sources based on cow's milk proteins such as whey, casein and mixtures thereof, and proteins based on soybeans, potatoes or peas are preferred. Preferably, the nutritional composition of the invention comprises casein and whey protein. In the case where whey proteins are used, the protein source is preferably based on acid whey or sweet whey, whey protein isolate, or mixtures thereof, and may include α-lactalbumin and β-lactoglobulin. More preferably, the protein source is based on acid whey or sweet whey from which casein glyco-macropeptide (CGMP) has been removed. Preferably, the composition comprises casein, preferably it comprises at least 3 wt% casein based on dry weight. Preferably, the casein is intact and / or not hydrolyzed. For the present invention, the protein includes peptides and free amino acids.

Digestible carbohydrates

Preferably, the nutritional composition comprises a digestible carbohydrate component. The digestible carbohydrate component preferably provides 30 to 80% of the total calories of the compositions. Preferably, the digestible carbohydrate component provides 40 to 60% of the total calories. When in liquid form, for example, a ready-to-drink liquid, the composition preferably comprises 3.0 to 30 g digestible carbohydrates per 100 ml, more preferably 6.0 to 20, even more preferably 7.0 to 10, 0 g per 100 ml. Based on dry weight, the present composition preferably comprises 20 to 80 wt%, more preferably 40 to 65 wt% digestible carbohydrate.

Preferred sources of digestible carbohydrates are lactose, glucose, sucrose, fructose, galactose, maltose, starch, and maltodextrin. Lactose is the main digestible carbohydrate in human milk. Lactose is predominantly low on the glycemic index. The present composition preferably comprises lactose. The present composition preferably includes a digestible carbohydrate component, where at least 35 wt%, more preferably at least 50 wt%, more preferably at least 75 wt%, even more preferably at least 90 wt%, most preferably at least 95 wt% digestible carbohydrate is lactose. Based on dry weight, the present composition preferably comprises at least 25 wt% lactose, preferably at least 40 wt%.

Additional components

In one embodiment, the nutritional composition preferably comprises indigestible oligosaccharides. Preferably, the present composition comprises indigestible oligosaccharides having a degree of polymerization (DP) of 2 to 250, more preferably 3 to 60. Preferred indigestible oligosaccharides are selected from fructooligosaccharides, galactooligosaccharides and / or oligosaccharides of trans galacturonic acid, more preferably galactooligosaccharides, most preferably galactooligosaccharides, most preferably galactooligosaccharides. In a preferred embodiment, the composition comprises a mixture of transgalacto-oligosaccharides and fructo-oligosaccharides. Suitable indigestible oligosaccharides are, for example, Vivinal GOS (FrieslandCampina DOMO), Raftilin HP or Raftilose (Orafti).

Preferably, the composition comprises 80 mg to 2 g of indigestible oligosaccharides per 100 ml, more preferably 150 mg to 1.50 g, even more preferably 300 mg to 1 g per 100 ml. Based on dry weight, the composition preferably comprises 0.25 wt% to 20 wt%, more preferably 0.5 wt% to 10 wt%, even more preferably 1.5 wt% to 7.5 wt%.

The composition preferably includes other ingredients such as vitamins, minerals in accordance with international guidelines for infant nutrition, in particular infant formula.

Application

The inventors have surprisingly found that the nutritional composition of the invention is effective in the prevention of stress-induced cognitive decline in human early life. The method of the invention therefore relates to the administration of the nutritional composition of the invention during the early life of a person. When the composition is in liquid form, the preferred volume administered per day is in the range from about 80 to 2500 ml, more preferably from about 450 to 1000 ml per day.

The inventors have surprisingly found that the nutritional composition of the invention is effective in preventing the reduction of stress-induced neurogenesis in early life in humans. The method of the invention therefore relates to the administration of the nutritional composition of the invention during the early life of a person. When the composition is in liquid form, the preferred volume administered on a daily basis ranges from about 80 to 2500 ml, more preferably from about 450 to 1000 ml per day.

In the context of the present invention, "prevention" can also be referred to as "reduction", "reduction of the risk of development", "prevention" or "(prophylactic) treatment". The current use of “prevention of stress-induced cognitive decline in early life” can also be termed as “stimulation of cognitive function after exposure to stress in early life” or “improvement of cognitive function after exposure to stress in early life”. The present use of prevention of stress-induced reduction in early life ʺ can also be termed as protection against stress-induced reduction in early life, снижения or recovery of neurogenesis after exposure to stress in early life or stimulation of stress neurogenesis after exposure to early life period of lifeʺ or improvement of neurogenesis after exposure to stress in the early period of lifeʺ. In addition, the current use of "adult neurogenesis" in humans may also be referred to as "adult neurogenesis".

In the context of the present invention, the "early life" usually extends up to and including adolescence, for example, up to the age of 18 years, preferably up to the age of 12 years, more preferably up to the age of 5 years, even more preferably up to the age of 36 months, most preferably up to the age of 12 months ... Early life also includes the human prenatal stage, preferably the fetal stage, more preferably the third trimester fetal stage. In one embodiment of the invention, early life begins at birth.

The person is preferably selected from the group of unborn fetuses, infants, toddlers, pre-adolescent children, and adolescents. In a more preferred embodiment of the invention, the human is selected from unborn fetuses, infants and toddlers, more preferably from infants and toddlers, i.e., 0 to 36 months old, most preferably the human is an infant, i.e. having an age of 0-12 months. The unborn fetus is preferably in the third trimester of pregnancy. Administration to unborn fetuses is via a pregnant mother, and the nutritional composition is preferably in the form of a nutritional supplement for a pregnant woman. Since the lipid composition in a mother's diet is reflected in her breast milk, the nutritional composition can also be in the form of a nutritional supplement for a nursing woman. People under 36 months of age, or even under 12 months of age, are particularly preferred because, generally speaking, the plasticity of the brain is greatest at this early stage of life, and the administration of the nutritional composition therefore has the greatest impact at this age.

In a preferred embodiment of the invention, the person is at risk or even at high risk of developing stress early in life or experiencing stress early in life. Examples of such patients at (high) risk are patients, in particular children, with a lower socioeconomic status (for example, children from families with a level of income below average or low educational level, for example, no higher than high school), children whose parents are divorced or undergoing through divorce, children separated from one or both parents, children with both parents working, children from disadvantaged families, children who are abused, children whose parent (s) suffer from stress, depression, children with depression, sick children, undernourished children and abused children. Here, “children” include infants and toddlers, preferably an infant.

Certain patients who experience physical stress and / or mental stress are preterm and small for gestational age (SGA) infants. Premature babies refer to babies born before the end of the standard gestation period, i.e. before 37 weeks of the mother's pregnancy, i.e. up to 37 weeks from the start of the mother's last menstrual period. Premature babies are also referred to as deeply premature babies. SGA infants are those whose birth weight is below the 10th percentile for gestational age. There can be several reasons for the SGA; for example, full-term or premature infants may be born with SGA because they have undergone intrauterine growth restriction (IUGR). Many premature babies are also small for gestational age. Premature and / or SGA infants include low birth weight infants (LBW infants), very low birth weight infants (VLBW infants), and extremely low birth weight infants (ELBW infants). LBW infants are infants with a birth weight below 2500 g; this group includes term infants born to SGA. VLBW and ELBW infants are almost always prematurely born and are defined as infants with a birth weight between 1500 g or 1000 g, respectively. Therefore, in one embodiment of the method or use of the present invention, the human is a premature infant, an infant born small for gestational age, or both.

Boys are likely to be more affected by stress-induced cognitive decline early in life, and therefore boys may benefit more from genuine dietary intervention. Therefore, in one embodiment, the human is a boy.

Therefore, administration of the nutritional composition of the invention usually occurs when the patient is 18 years of age or younger, preferably the person is less than 12 years old, more preferably up to 5 years old, even more preferably up to 36 months, most preferably up to 12 months. Although early life usually ends after adolescence, the beneficial effects on cognitive function and / or neurogenesis may continue beyond early life, for example, until or even beyond adulthood. The effects can last even throughout the patient's life. According to a preferred embodiment of the invention, prevention of cognitive impairment and / or reduction in neurogenesis induced by stress in early life continues into adulthood. Preferably, prevention of cognitive decline and / or decreased neurogenesis occurs when the patient reaches adulthood or when the patient reaches the age of 18 years or older, or even 25 years of age or more. Therefore, it is preferred that the nutritional composition according to the invention has an effect on cognitive functions and / or a decrease in neurogenesis later in life, preferably when the patient reaches the age of 5 years. In accordance with a preferred embodiment of the invention, the beneficial effect on cognitive function and / or neurogenesis lasts at least 3 months, preferably at least 1 year, more preferably at least 5 years after cessation of administration of the nutritional composition according to the invention. An example of such a late-in-life effect, which is a particularly preferred embodiment of the present invention, is the administration of the composition of the invention to a fetus, infant or toddler, while a prophylactic effect on cognitive decline and / or a decrease in neurogenesis induced by stress in early life is observed 5 years old or older, more preferably 12 years old or older, most preferably 18 years old or older.

Stress in early life can take any known form. Typical stressors include psychological stressors, physiological stressors, and physical stressors. Early life stress can be mental stress, physical stress, metabolic stress, or any combination of these. Exemplary stressors include broken families (for example, divorce, separation, mixing of families), separation from one or both parents (for example, arrest of a parent), maternal depression, both working parents, lack of attention from parent (s), exposure to violence, abuse (for example , physical, mental, sexual), rejection (for example, emotional, physical), the death of a loved one (in particular the death of a parent), illness (for example, mental, physical), lack of nutrition. Here, parent (s) ʺ can also refer to guardian (s) ʺ or ʺ guardian (s) ʺ. Accordingly, early life stress is selected from malnutrition, maternal stress, depression, and violence.

Cognitive impairment induced by early life stress is a specific type of cognitive impairment that is associated with many complex brain structures, such as the cerebral cortex, frontal cortex, hypothalamus, hippocampus, and paranasal cortex. Such structures in the brain play important roles in both cognitive function and the regulation of stress responses. The hippocampus, for example, contains many receptors for stress hormones and is highly plastic. Since such structures develop rapidly in the last trimester of pregnancy and postnatally until about 16 years of age or after, they are particularly sensitive to stress in early life.

In a preferred embodiment, decreased cognitive function preferably includes decreased hippocampus-dependent cognitive function, or decreased cognitive function preferably is decreased hippocampus-dependent cognitive function. Particularly preferred embodiments of cognitive function in the context of the present invention are (a) new object recognition; (b) preference for the new; (c) attention to (changes in) the environment; (d) nonspatial (subject) memory; (e) spatial memory; and (f) spatial long-term memory, and also includes new object recognition and / or new object location, or hippocampus-dependent memory (especially long-term spatial memory).

In a preferred embodiment, the prevention of a decrease in neurogenesis occurs by preventing a decrease in the survival of hippocampal cells. Alternatively, the invention also relates to such prevention of a decrease in hippocampal cell survival induced by early life stress by administering the nutritional composition of the invention during the early life of a person. The present application of “preventing the decline in hippocampal cell survival induced by early life stress” can also be termed as “stimulating hippocampal cell survival after exposure to early life stress” or “improving the survival of hippocampal cells after exposure to early life stress”.

EXAMPLES

Example 1:

Female C57 / BL6 mice with offspring were subjected to simulated early life stress of limiting nest / bedding material as described by Naninck et al., Hippocampus 2015, 25: 309-328. Stress simulation was started 2 days after the birth of the offspring (PN2), and continued until PN9. The offspring were weaned at PN21.

Starting with PN2, the female nursing the offspring was given experimental diets up to PN42 (male offspring were kept on the appropriate diet after weaning). With PN42, all male offspring were fed the same AIN-93M food until the end of the experiment.

The experimental diets were semi-synthetic diets with macronutrient and micronutrient compositions according to the American Institute of Nutrition formulation of AIN93-G purified diets for laboratory rodents (Reeves 1993), which differed only in fatty acid composition in terms of linoleic (LA) and alpha-linolenic acid (ALA), see table 1.

As adults, male offspring were tested for cognitive function using the New Object Recognition Test, the Object Positioning Test, and the Morris Water Maze Test. Such tests are widely used in the field of behavioral neuroscience.

Table 1: Composition of fatty acids in experimental diets.

Fatty acids (% in diet) Diet A Diet B C6: 0 0.01 0.01 C8: 0 0.13 0.13 C10: 0 0.13 0.13 C12: 0 1.07 1.08 C14: 0 0.48 0.48 C16: 0 0.67 0.64 C18: 0 0.42 0.44 C20: 0 0.07 0.06 C18: 1 1.51 1.55 C20: 1 0.04 0.03 C18: 2n6 (LA) 2.14 1.21 C18: 3n3 (ALA) 0.14 1.07 LA / ALA ratio 15.29 1.13

Test for recognizing a new object and the location of a new object

The mice were subjected to the New Object Recognition Test (ORT) on day 120. This test is a behavioral technique for studying nonspatial object memory preference based on the innate preference of mice for novelty. In the test, the mice had to distinguish between a new object and an object that they had studied previously (familiar). For three days, the mice were exposed for five minutes a day to the test area, which consisted of a rectangular plastic box. On the day of the study, the mice had five minutes to examine two identical objects (glass bottles 9.5 cm high), which were placed in a box 12 cm apart and 11 cm away from the wall. On the test day, 24 hours after training, one object was replaced with a new object (consisting of yellow Lego Duplo bricks) placed in the same position, and the mice were reintroduced to the area for five minutes to explore the new and familiar object. The time spent studying each object and the ratio of the time spent studying a new / familiar object were calculated. The results are shown in Table 2.

Object Position Tests (OLT): This test is a behavioral technique for studying spatial subject memory based on the innate curiosity of mice for new locations. The test was performed on PN 130 day. For the OLT, the scope and the familiarization protocol were identical to those used in the ORT; two completely new identical objects (white coffee cups) were placed 12 cm apart and 11 cm from the wall. On the day of training, the mice had five minutes to study objects and their locations. On the day of the study, 24 hours after training, one object was moved to a new position in the region and the mice were re-placed in the region for five minutes to study. The time spent on the study was calculated, recorded as before, and the ratio of the time spent on studying a new / familiar object. The results are shown in Table 2.

Table 2: Ratio of study time for new / old object

Group The ratio of new / old object in ORT OLT new / old object ratio Control diet A 2.5 ± 0.46 1.4 ± 0.19 Stress Diet A 1.26 ± 0.07 0.96 ± 0.06 Control diet B 2.38 ± 0.33 1.40 ± 0.08 Stress Diet B 2.02 ± 0.18 1.61 ± 0.22

As can be seen from Table 2, the ratio of new / old exploration time during ORT decreases as a result of prior exposure to stress in early life in animals fed Diet A, indicating that animals are unable to distinguish between an object they have already seen and a new one. which they saw for the first time. This index is not reduced by stress mimicry in animals fed Diet B. The difference between the two stress diet groups is statistically significant and indicates that Diet B can prevent stress-induced cognitive impairment, in particular new and nonspatial memory perceptions.

OLT also decreased the time to learn new / old with prior exposure to stress in early life in animals fed Diet A, indicating that the animals were unable to distinguish between an object that was in position and an object that was moved to a new position. Conversely, there was no such difference in animals fed Diet B. This shows that Diet B can prevent stress-induced cognitive impairment, in particular new preference and spatial memory.

Morris water maze

The mice were also subjected to the PN 140 Morris Water Maze Test, also known as the Morris Water Maze Navigation Test, which is a behavioral technique for assessing cognitive function, in particular spatial learning and memory. In this experiment, animals were trained to detect a hidden platform based on external cues. Briefly, the platform was immersed below the water surface in a fixed position in a round water vessel. The vessel was in a room with fixed keys on the walls, which were visible from the water surface in the vessel. For 7 consecutive days, the mice were trained twice a day (with 10 minute intervals between studies) to detect the hidden platform. During each study, the animals were placed in the water of the vessel and they had to detect the platform within 60 seconds. Between studies, the initial position of the animal varied between one of the three quadrants without a platform. On the eighth day, the platform was removed from the vessel for a one-time exploratory study in which the time spent in the target quadrant was recorded. Such research was termed as exploratory research. The results are shown in Table 3.

Table 3: Time (%) spent in the target quadrant in the exploratory study of the Morris Water Maze Test.

Group Time (%) Control diet A 31.2 ± 4.83 Stress Diet A 26.8 ± 3.67 Control diet B 34.6 ± 5.56 Stress Diet B 37.3 ± 6.16

As can be inferred from Table 3, the percentage of time spent in the target quadrant is reduced by prior exposure to stress early in life in animals fed Diet A, indicating that the animals had little memory of where the platform was located. Exposure to diet B, however, prevented stress-induced changes in time spent in the target quadrant. This is indicative of improved cognitive performance, in particular, long-term (spatial) memory.

Example 2

The survival of adult neuronal cells was assessed using 5-bromo-2'-deoxyuridine (BrdU; Sigma-Aldrich Chemie BV, Zwijndrecht, The Netherlands). 7 months after birth, mice were injected intraperitoneally with 100 mg / kg BrdU (10 mg / ml dissolved in sterile saline + 0.007M NaOH) for 2 times a day on 4 consecutive days. Four weeks after the last injections, the mice were transcardially perfused with P230.

Tissue harvesting was performed as follows: mice were anesthetized with an intraperitoneal injection of pentobarbital (Euthasol®, 120 mg / kg) and transcardially perfused with an initial 0.9% saline solution followed by 4% paraformaldehyde (PFA) in phosphate buffered saline (PB 0.1M, pH 7.4). The fully perfused brain was carefully removed, then fixed with 4% PFA in 0.1 M PB at 4 ° C for 24 h and stored in PB with 0.01% sodium azide at 4 ° C. Prior to brain slicing, perfused brains were cryoprecipitated in 30% sucrose in 0.1 M PB. Subsequently, the frozen brains were cut into coronal sections of 40 μM thick and divided into 6 parallel series of microtome and stored in antifreeze (30% ethylene glycol, 20% glycerin, 50% 0.05M PBS; company) at -20 ° C.

BrdU-labeled cells were examined using fluorescent immunohistochemistry. Brain slices (from one of 6 parallel series) were plated on glass (Superfrost Plus slides, Menzel, Braunschweig, Germany) followed by antigen unmasking induced by heating in 0.1 M citrate buffer (pH 6.0) using a microwave oven (Samsung M6235) at ~ 95 ° C for 15 min (5 min at 800 Watt, 5 min at 400 Watt, 5 min at 200 Watt). Subsequently, preset sections were treated with blocking buffer for 30 min (1% bovine serum albumin in TBS) followed by incubation with primary monoclonal rat anti-BrdU antibody (1: 200; Accurate Chemical and Scientific Corporation, OBT 0030) in incubation mixture (1 % bovine serum albumin, 0.3% Triton X-100 in 0.05M TBS) for 24 h. After this, sections were incubated with a secondary donkey anti-rat antibody, alexafluorescent 488 conjugate (1: 1000, Invitrogen) for 2 h and covered with DAPI-containing Vectashield.

To determine the volume of the granular zone of the dentate gyrus (DG), boundary contour traces were established using 20x magnification on a Zeiss Axiophot light microscope using the Stereoinvestigator software (MicroBrightField, Germany) for eight bregmas, multiplied by 6 (for parallel series correction) and 40 μm (for section thickness). To quantify BrdU (40x objective, Leica CTR5500), cell counts were performed in the DG subgranular and granular zones in eight matched bregmas and multiplied by 6 for the total number of immunoreactive cells per DG.

The results are shown in FIG. FIG. represents the survival of hippocampal cells, measured as BrdU-labeled cells in the dentate gyrus (DG), for diet A and diet B of the control group not stressed in early life (open bars on the left), and the group that was stressed in early life (black filled columns on the right). The survival of hippocampal cells after exposure to stress in early life was significantly lower for diet compared to diet B. The survival of hippocampal cells for diet B after exposure to stress in early life was the same as for both diets without exposure to stress in early life ...

From the results obtained, it was concluded that nutritional intervention with a diet having a low LA / ALA ratio has a prophylactic effect on reducing neurogenesis after stress in early life, or in other words, protects against the decrease in neurogenesis induced by stress in early life. In addition, it can be concluded that nutritional intervention with a diet having a low LA / ALA ratio restores neurogenesis in adulthood after stress in early life.

Claims (15)

1. The use of linoleic acid (LA) and alpha-linolenic acid (ALA) to obtain a composition comprising LA and ALA in a mass ratio of LA / ALA in the range of 0.1-12, for the prevention of cognitive decline induced by stress in the early period of life in a person where the person is an unborn fetus, infant, or toddler. 2. Use according to claim 1, wherein the cognitive functions include hippocampal dependent cognitive functions, in particular recognition of a new object and / or a new location of the object. 3. Use according to claim 1 or 2, wherein the cognitive functions include hippocampal-dependent memory. 4. The use of linoleic acid (LA) and alpha-linolenic acid (ALA) in the preparation of a composition comprising LA and ALA in a LA / ALA mass ratio in the range of 0.1-12 to prevent a decrease in neurogenesis induced by stress in the early period of life in a person where the person is an unborn fetus, infant, or toddler. 5. The use according to any one of the preceding claims, wherein the stress in early life is selected from mental stress, metabolic stress, and a combination thereof. 6. The use according to any of the preceding claims, wherein the human is a premature infant, an infant born small for gestational age, or both. 7. Use according to any one of the preceding claims, wherein the composition is infant formula. 8. Use according to claim 7, wherein the composition is a first level infant formula, a second level infant formula, or a third level infant formula. 9. Use according to any of the preceding claims, wherein the LA / ALA weight ratio is in the range of 0.5-8. 10. Use according to any one of the preceding claims, wherein the composition comprises a lipid component, a protein component and a digestible carbohydrate component. 11. The use according to any one of paragraphs. 1-3 or 5-10, where the prevention of stress-induced cognitive dysfunction early in life is manifested later in life. 12. Use according to claim 11, wherein the later period of life is the period when the subject reaches the age of 5 years. 13. The use according to any one of paragraphs. 4-10, where prevention of stress-induced decline in early life is manifested in later life. 14. Use according to claim 13, wherein later life means when the human patient reaches adulthood. 15. Application of linoleic acid (LA) and alpha-linolenic acid (ALA) to obtain a composition containing LA and ALA in a mass ratio of LA / ALA in the range of 0.1-12, to prevent a decrease in the survival of hippocampal cells induced by stress in the early period life (ES) in a person where the person is an unborn fetus, infant, or toddler.

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