Nutritional compositions and their use for improving immune dysregulation

Abstract

The present application belongs to the field of food, and particularly relates to a nutritional composition and its use for improving immune disorders, more particularly to the use of a nutritional composition in the preparation of a food for improving the level disorder of inflammatory factors in offspring caused by maternal immune activation; the nutritional composition comprises essential active ingredients shown as (i) and (ii): (i) neutral fucosylated human milk oligosaccharides, (ii) neutral non-fucosylated human milk oligosaccharides; wherein the neutral fucosylated human milk oligosaccharides at least comprise 2'-fucosyllactose, and the neutral non-fucosylated human milk oligosaccharides at least comprise lacto-N-neotetraose; and in the nutritional composition, the mass ratio of the lacto-N-neotetraose to the 2'-fucosyllactose is 1:(1-8). 2'-fucosyllactose and lacto-N-neotetraose can synergistically regulate offspring immunity.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of food, in particular to a nutritional composition and its use for improving immune disorders. BACKGROUND

[0002] During pregnancy, the maternal immune system undergoes dramatic changes to maintain immune balance with the fetus, prevent rejection, and maintain natural and adaptive immune mechanisms against infection. Exposure to various factors such as environment during pregnancy will lead to an imbalance in the level of inflammatory factors in the body, resulting in a state of maternal immune activation (MIA). The impact of maternal immune activation on offspring is heterogeneous.

[0003] Human milk oligosaccharides (HMOs) are the third largest solid component in breast milk, with a content second only to lactose and fat, and even higher than that of protein. HMOs are a class of complex sugars with a variety of structures. According to the molecular structure characteristics, HMOs can be divided into neutral fucosylated HMOs, neutral non-fucosylated HMOs and acidic sialylated HMOs. Studies have shown that the total HMOs content, 2'-fucosyllactose (2'-FL), 6'-sialyllactose (6'-SL) and 3'-sialyllactose (3'-SL) in the early lactation period (i.e. 1 month after birth) are positively correlated with the cognitive, language and motor development levels of later infants (i.e. 6-24 months old). Studies on infants fed with pure breast milk found that for every 1 μg / mL increase in 2'-FL content in breast milk 1 month after birth, the cognitive score of the infant at 24 months of age increased by 0.59. In addition, animal experiments have shown that the intake of HMOs composition can significantly improve the spatial memory ability of rodents and piglets and accelerate the learning speed of operation tasks. In addition, studies have shown that breast milk oligosaccharides, as prebiotics, can improve intestinal immunity in cooperation with probiotics, for example, references 1, 2, etc.

[0004] REFERENCES

[0005] Reference 1: CN112870233B;

[0006] Reference 2: CN112869168B. SUMMARY

[0007] Problem to be solved by the invention

[0008] Studies have shown that maternal immune activation has a wide range of effects on the immune system of the offspring, and how to reduce, improve or inhibit the adverse effects of maternal immune activation on the development of the offspring has important research significance. Although some studies have shown that breast milk oligosaccharides as prebiotics can improve intestinal immunity in cooperation with probiotics, such studies are not sufficient, and the existing technology has not studied whether breast milk oligosaccharides have an effect on the immune system under a specific state (for example, the immune system of the offspring produced by a female who has experienced or is in a state of maternal immune activation), especially the development of damage (that is, congenital, systemic, diffuse damage, rather than acquired, acquired, localized damage).

[0009] In view of this, the present application has conducted a large number of researches, and it is accidentally found that the combination of 2'-fucosyllactose and lacto-N-neotetraose can improve the inflammatory factor level disorder of the offspring caused by maternal immune activation, including the inflammatory factor level disorder in the offspring's intestine, serum and cerebral cortex, and the present application accidentally finds that lacto-N-neotetraose can significantly amplify the improvement effect of 2'-fucosyllactose on the inflammatory factor level disorder of the offspring caused by maternal immune activation, especially when the mass ratio of 2'-fucosyllactose and lacto-N-neotetraose is within a certain range, the two have a synergistic effect.

[0010] Solution to the problem

[0011] In order to solve the above technical problems, the present application provides the following technical solutions:

[0012] [1]. A use of a nutritional composition in the preparation of a food for improving the inflammatory factor level disorder of the offspring caused by maternal immune activation; wherein the nutritional composition comprises the necessary active ingredients as shown in (i) and (ii) below:

[0013] (i) neutral fucosylated breast milk oligosaccharides,

[0014] (ii) neutral non-fucosylated breast milk oligosaccharides;

[0015] wherein,

[0016] the neutral fucosylated breast milk oligosaccharides at least comprise 2'-fucosyllactose,

[0017] the neutral non-fucosylated breast milk oligosaccharides at least comprise lacto-N-neotetraose;

[0018] and in the nutritional composition, the mass ratio of lacto-N-neotetraose to 2'-fucosyllactose is 1:(1-8).

[0019] [2]. The use according to [1], wherein the neutral fucosylated human milk oligosaccharide further comprises any one or more of 3-fucosyllactose and difucosyllactose.

[0020] [3]. The use according to [1] or [2], wherein the neutral non-fucosylated human milk oligosaccharide further comprises lacto-N-tetraose.

[0021] [4]. The use according to any one of [1] to [3], wherein the improvement of the disturbance of the level of inflammatory factors in the offspring organism caused by maternal immune activation comprises the improvement of the disturbance of the level of inflammatory factors in the intestine, serum and / or cerebral cortex of the offspring caused by maternal immune activation.

[0022] [5]. The use according to any one of [1] to [4], wherein the improvement of the disturbance of the level of inflammatory factors in the offspring organism caused by maternal immune activation comprises any one or more of (al) to (a5):

[0023] (al) the improvement of the increase of the level of any one or more of inflammatory factors TNF-a, IL-17A and IL-6 in the intestine of the offspring caused by maternal immune activation;

[0024] (a2) the improvement of the decrease of the level of inflammatory factor IL-10 in the intestine of the offspring caused by maternal immune activation;

[0025] (a3) the improvement of the increase of the level of inflammatory factor IL-6 in the serum of the offspring caused by maternal immune activation;

[0026] (a4) the improvement of the decrease of the level of any one or more of inflammatory factors IL-22 and IL-10 in the serum of the offspring caused by maternal immune activation;

[0027] (a5) the improvement of the increase of the level of any one or more of inflammatory factors IL-lβ, TNF-a and IL-6 in the cerebral cortex of the offspring caused by maternal immune activation.

[0028] [6]. The use according to any one of [1] to [5], wherein the foodstuff exerts its effect upon ingestion by the offspring.

[0029] [7]. The use according to any one of [1] to [6], wherein the foodstuff is an infant formula or an infant complementary food.

[0030] [8]. The use according to any one of [1] to [7], wherein the foodstuff is a confectionery, a beverage, a dairy product, a bakery product or a dietary supplement.

[0031] [9] The use according to any one of [1] to [8], wherein the food contains any one or more of the following components: a plant product component, an animal milk product component, an animal meat product component, a functional additive component, and any acceptable adjuvant.

[0032]

[10] The use according to any one of [1] to [9], wherein the content of the 2'-fucosyllactose is 0.4 g / 100 g to 2 g / 100 g and the content of the lacto-N-neotetraose is 0.1 g / 100 g to 0.5 g / 100 g in the food.

[0033]

[11] Use of a nutritional composition in the manufacture of a food having any one or more of the following (bl) to (b5) improving effects:

[0034] (bl) improving an increase in the level of any one or more of inflammatory factors TNF-a, IL-17A, and IL-6 in the intestine of an offspring caused by maternal immune activation;

[0035] (b2) improving a decrease in the level of inflammatory factor IL-10 in the intestine of an offspring caused by maternal immune activation;

[0036] (b3) improving an increase in the level of inflammatory factor IL-6 in the serum of an offspring caused by maternal immune activation;

[0037] (b4) improving a decrease in the level of any one or more of inflammatory factors IL-22 and IL-10 in the serum of an offspring caused by maternal immune activation;

[0038] (b5) improving an increase in the level of any one or more of inflammatory factors IL-lβ, TNF-a, and IL-6 in the cerebral cortex of an offspring caused by maternal immune activation;

[0039] wherein the nutritional composition comprises essential active ingredients represented by (i) and (ii) below:

[0040] (i) neutral fucosylated human milk oligosaccharides, at least comprising 2'-fucosyllactose;

[0041] (ii) neutral non-fucosylated human milk oligosaccharides, at least comprising lacto-N-neotetraose;

[0042] and, in the nutritional composition, the mass ratio of the lacto-N-neotetraose to the 2'-fucosyllactose is 1 : (1-8).

[0043]

[12] . The use according to

[11] , wherein in the nutritional composition the neutral fucosylated human milk oligosaccharides further comprise any one or more of 3-fucosyllactose and difucosyllactose, and / or the neutral non-fucosylated human milk oligosaccharides further comprise lacto-N-tetraose.

[0044]

[13] . The use according to

[11] or

[12] , wherein the food product is an infant formula or a baby food supplement.

[0045]

[14] . The use according to any one of

[11] to

[13] , wherein the food product is a confectionary, a beverage, a dairy product, a bakery product or a dietary supplement.

[0046]

[15] . The use according to any one of

[11] to

[14] , wherein the food product comprises any one or more of a plant product ingredient, an animal milk product ingredient, an animal meat product ingredient, a functional additive ingredient and any acceptable adjuvant.

[0047]

[16] . The use according to any one of

[11] to

[15] , wherein in the food product the 2'-fucosyllactose is present in an amount of 0.4 g / 100 g to 2 g / 100 g and the lacto-N-neotetraose is present in an amount of 0.1 g / 100 g to 0.5 g / 100 g.

[0048] Effects of the invention

[0049] The present application proposes that the combination of 2'-fucosyllactose and lacto-N-neotetraose can improve the offspring's inflammatory factor level disorder caused by maternal immune activation, including improving the levels of pro-inflammatory factors and anti-inflammatory factors in the intestine, serum and cerebral cortex. More importantly, the improvement proposed by the present application is for the offspring's inflammatory factor level disorder caused by maternal immune activation, that is, the improvement proposed by the present application is for the improvement of the damage that occurs earlier in the development period, which helps to reverse the insufficient or abnormal development of the innate development.

[0050] And when the mass ratio of 2'-fucosyllactose and lacto-N-neotetraose is within a certain range, there is a synergistic effect between the two. The present application unexpectedly found that lacto-N-neotetraose and 2'-fucosyllactose can significantly amplify each other's improvement effect on the offspring's inflammatory factor level disorder caused by maternal immune activation.

[0051] Meanwhile, the nutritional composition containing the combination of 2'-fucosyllactose and lacto-N-neotetraose according to the present application can be used independently as a nutritional fortifier, or can be added to various foods, and plays a role in the early life, i.e., the critical period of growth and development of infants and young children, by being ingested through food. DETAILED DESCRIPTION

[0052] Embodiments of the present application will be described below, but the present application is not limited thereto. The present application is not limited to each of the configurations described below, and various modifications can be made within the scope of the present application, and embodiments obtained by appropriately combining the technical means disclosed in each of the different embodiments and examples are also included in the technical scope of the present application.

[0053] In the present application, the terms "comprising", "having", "including" or "containing" can mean inclusive or open-ended, and do not exclude additional, unrecited elements or method steps. At the same time, "comprising", "having", "including" or "containing" can also mean closed, excluding additional, unrecited elements or method steps.

[0054] In the present application, the meaning indicated by "may" includes both the meaning of performing a certain process and the meaning of not performing a certain process.

[0055] In the present application, "optional" or "optionally" means that a certain substance, component, execution step, applied condition, etc. is used or not used.

[0056] In the present application, the numerical range indicated by "numerical value A ~ numerical value B", "numerical value A - numerical value B", "numerical value A or more / less" means a range including the end point values A and B.

[0057] In the present application, the term "about" is used to define the numerical range and parameters of the present application as approximate numerical values, and the relevant numerical values in the specific embodiments have been presented as accurately as possible. Unless otherwise explicitly stated, it should be understood that all ranges, numbers, numerical values and percentages used in the present application are modified by "about". Here, "about" generally means that the actual numerical value is within ±5%, ±3%, ±1% or ±0.5% of a certain numerical value or range. And the numerical values and numerical ranges appearing in the present application should be understood to include the systematic errors that are unavoidable in industrial production.

[0058] In the present application, references to "some specific / preferred embodiments", "further specific / preferred embodiments", "embodiments", and the like, mean that the particular element (e.g., feature, structure, property, and / or characteristic) described in relation to that embodiment is included in at least one embodiment described herein, and can or can not be present in other embodiments. In addition, it should be understood that the described elements can be combined in any suitable manner in various embodiments.

[0059] In the present application, "substantially free" means that the indicated component is not detectable in the indicated composition by ordinary analytical methods, or is present at such an insignificant level as to have no function.

[0060] In the present application, the unit names used are international standard unit names, and if not specifically stated, "%" used means weight or mass percentage.

[0061] In the present application, "infants" means the human group under 3 years of age, which includes infants aged 0-6 months, older infants aged 6-12 months, and toddlers aged 12-36 months.

[0062] Unless otherwise defined, other technical and scientific terms used in the present application have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs.

[0063] I. Nutritional composition

[0064] The present application provides a nutritional composition comprising essential active ingredients shown in (i) and (ii) as follows: (i) neutral fucosylated human milk oligosaccharides, (ii) neutral non-fucosylated human milk oligosaccharides; wherein the neutral fucosylated human milk oligosaccharides comprise at least 2'-fucosyllactose, and the neutral non-fucosylated human milk oligosaccharides comprise at least lacto-N-neotetraose; and in the nutritional composition, the mass ratio of the lacto-N-neotetraose to the 2'-fucosyllactose is 1:(1-8).

[0065] 2'-fucosyllactose (2'-FL) is a neutral trisaccharide consisting of L-fucose, D-galactose and D-glucose units, with the monosaccharide L-fucose linked to the disaccharide D-lactose via an α(1→2) bond. Its molecular formula is C 18 H 32 O 15 .

[0066] Lactose-N-neotetraose (LNnT) is a linear tetrasaccharide consisting of D-galactose, N-acetylglucosamine, D-galactose and D-glucose, arranged in the order of D-galactose linked to N-acetylglucosamine (GlcNAc) by a β-(1→4) bond, then linked to D-galactose by a β-(1→3) bond, and then linked to D-glucose by a β-(1→4) bond. Its molecular formula is C 26 H 45 NO 21 .

[0067] The source of 2’-FL and LNnT is not particularly limited in the present application, and typically, it can be obtained by means of ordinary chemical synthesis method, microbial fermentation method, etc. in the art.

[0068] In some embodiments, in the nutritional composition described in the present application, the (i) neutral fucosylated human milk oligosaccharide and the (ii) neutral non-fucosylated human milk oligosaccharide are the main active ingredients (active ingredients), that is, the nutritional composition described in the present application mainly relies on the (i) neutral fucosylated human milk oligosaccharide and the (ii) neutral non-fucosylated human milk oligosaccharide contained therein to exert a specific physiological activity function, such as improving the disturbance of inflammatory factor level in the offspring caused by maternal immune activation. In other words, in some embodiments, the active ingredients (ingredients for exerting a specific physiological activity function, i.e. ingredients for exerting the effect of improving the disturbance of inflammatory factor level in the offspring caused by maternal immune activation) of the nutritional composition consist of the components shown in (i) and (ii) as follows: (i) neutral fucosylated human milk oligosaccharide, (ii) neutral non-fucosylated human milk oligosaccharide; wherein the neutral fucosylated human milk oligosaccharide at least comprises 2’-fucosyllactose, and the neutral non-fucosylated human milk oligosaccharide at least comprises lactose-N-neotetraose, and in the nutritional composition, the mass ratio of the lactose-N-neotetraose to the 2’-fucosyllactose is 1:(1-8).

[0069] In some embodiments, the 2'-fucosyllactose and the lacto-N-neotetraose in the nutritional composition of the present application are the main active ingredients (active ingredients) for the specific physiological activity of the nutritional composition, i.e. the nutritional composition relies on the 2'-fucosyllactose and the lacto-N-neotetraose to exert the specific physiological activity, such as to improve the disturbance of inflammatory factor level in the offspring caused by maternal immune activation. In other words, in some embodiments, the active ingredients (ingredients for the specific physiological activity, i.e. ingredients for the improvement of the disturbance of inflammatory factor level in the offspring caused by maternal immune activation) of the nutritional composition are composed of the 2'-fucosyllactose and the lacto-N-neotetraose, and the mass ratio of the lacto-N-neotetraose to the 2'-fucosyllactose in the nutritional composition is 1:(1-8).

[0070] The present application has found that, compared with the neutral fucosylated human milk oligosaccharide such as 2'-fucosyllactose and the neutral non-fucosylated human milk oligosaccharide such as lacto-N-neotetraose, the nutritional composition of the present application can more effectively improve the disturbance of inflammatory factor level in the offspring caused by maternal immune activation, i.e. there is a synergistic effect between the two.

[0071] In some embodiments, the nutritional composition comprises active ingredients (ingredients for the specific physiological activity, i.e. ingredients for the improvement of the disturbance of inflammatory factor level in the offspring caused by maternal immune activation) and non-active ingredients (ingredients that do not improve the disturbance of inflammatory factor level in the offspring caused by maternal immune activation). For example, the non-active ingredients can be other nutrients, any food acceptable excipients, and / or substances that are produced together with the active ingredients in the production or obtaining process of the active ingredients and cannot be effectively separated from the active ingredients or do not need to be separated. In some embodiments, the nutritional composition is composed of the active ingredients and the non-active ingredients.

[0072] In some embodiments, the neutral fucosylated human milk oligosaccharide contained in the nutritional composition further comprises any one or more of 3-fucosyllactose and difucosyllactose.

[0073] In other embodiments, the nutritional composition contains substantially no neutral fucosylated human milk oligosaccharide other than 2'-fucosyllactose.

[0074] In some embodiments, the neutral non-fucosylated human milk oligosaccharide contained in the nutritional composition further comprises lacto-N-tetraose.

[0075] In other embodiments, the nutritional composition is substantially free of neutral non-fucosylated human milk oligosaccharides other than lacto-N-neotetraose.

[0076] In some embodiments, the nutritional composition comprises essential active ingredients as indicated in (i) and (ii) below: (i) neutral fucosylated human milk oligosaccharides, (ii) neutral non-fucosylated human milk oligosaccharides; wherein the neutral fucosylated human milk oligosaccharide is 2'-fucosyllactose and the neutral non-fucosylated human milk oligosaccharide is lacto-N-neotetraose; and, in the nutritional composition, the mass ratio of the lacto-N-neotetraose to the 2'-fucosyllactose is 1 : (1-8).

[0077] In some specific embodiments, in the nutritional composition, the mass ratio of the lacto-N-neotetraose to the 2'-fucosyllactose can be 1 : 1.0, 1 : 1.1, 1 : 1.2, 1 : 1.3, 1 : 1.4, 1 : 1.5, 1 : 1.6, 1 : 1.7, 1 : 1.8, 1 : 1.9, 1 : 2.0, 1 : 2.1, 1 : 2.2, 1 : 2.3, 1 : 2.4, 1 : 2.5, 1 : 2.6, 1 : 2.7, 1 : 2.8, 1 : 2.9, 1 : 3.0, 1 : 3.1, 1 : 3.2, 1 : 3.3, 1 : 3.4, 1 : 3.5, 1 : 3.6, 1 : 3.7, 1 : 3.8, 1 : 3.9, 1 : 4.0, 1 : 4.1, 1 : 4.2, 1 : 4.3, 1 : 4.4, 1 : 4.5, 1 : 4.6, 1 : 4.7, 1 : 4.8, 1 : 4.9, 1 : 5.0, 1 : 5.1, 1 : 5.2, 1 : 5.3, 1 : 5.4, 1 : 5.5, 1 : 5.6, 1 : 5.7, 1 : 5.8, 1 : 5.9, 1 : 6.0, 1 : 6.1, 1 : 6.2, 1 : 6.3, 1 : 6.4, 1 : 6.5, 1 : 6.6, 1 : 6.7, 1 : 6.8, 1 : 6.9, 1 : 7.0, 1 : 7.1, 1 : 7.2, 1 : 7.3, 1 : 7.4, 1 : 7.5, 1 : 7.6, 1 : 7.7, 1 : 7.8, 1 : 7.9, or 1 : 8.0, etc.; preferably, the mass ratio of the lacto-N-neotetraose to the 2'-fucosyllactose is 1 :(1-6); more preferably, the mass ratio of the lacto-N-neotetraose to the 2'-fucosyllactose is 1 :(1.5-4.5); even more preferably, the mass ratio of the lacto-N-neotetraose to the 2'-fucosyllactose is 1 :(1.5-3.5); further preferably, the mass ratio of the lacto-N-neotetraose to the 2'-fucosyllactose is 1 :(1.5-3).

[0078] The present application does not make any particular limitation on the form of the nutritional composition, and typically, the nutritional composition can be in the form of a liquid or a solid, etc. From the perspective of production, transportation, storage and use convenience, the nutritional composition of the present application is preferably in the form of a powder.

[0079] II. Use of the nutritional composition

[0080] The present application proposes that the combination of 2'-fucosyllactose and lacto-N-neotetraose in a certain ratio can improve the inflammatory factor level disorder of the offspring caused by maternal immune activation, including improving the inflammatory factor level disorder in the intestine, serum and cerebral cortex of the offspring caused by maternal immune activation, and the two substances have a synergistic effect.

[0081] The "maternal immune activation" of the present application mainly refers to the abnormal activation state of the immune system of a female during pregnancy, and its manifestations can include inflammatory factor level disorder. Factors that can cause the abnormal activation of the immune system of a female during pregnancy include but are not limited to infection of bacteria, viruses, etc.

[0082] The "inflammatory factor level disorder" of the present application mainly refers to that the level of an inflammatory factor in the body (including the content of the inflammatory factor, the mRNA expression level, etc.) has changed compared with the level of the inflammatory factor in the normal or healthy state of the body, and the change includes increase and decrease.

[0083] The improvement of the inflammatory factor level disorder of the offspring caused by maternal immune activation of the present application is not for the purpose of treating or preventing diseases, and at the same time, the inflammatory factor level disorder of the present application has not reached the degree that can be recognized as a disease.

[0084] Based on this, the present application provides the use of the above-mentioned nutritional composition in the preparation of a food for improving the inflammatory factor level disorder of the offspring caused by maternal immune activation.

[0085] In some embodiments, the improvement of the inflammatory factor level disorder of the offspring caused by maternal immune activation includes improvement of the inflammatory factor level disorder in the intestine, serum and / or cerebral cortex of the offspring caused by maternal immune activation.

[0086] In some embodiments, the improvement of the inflammatory factor level disorder of the offspring caused by maternal immune activation includes improvement of the inflammatory factor level disorder in the intestine, serum and cerebral cortex of the offspring caused by maternal immune activation.

[0087] In some embodiments, the improvement of the inflammatory factor level disorder of the offspring caused by maternal immune activation includes any one or more of (a1)-(a5) as follows:

[0088] (a1) ameliorating the increase in the level of any one or more of the inflammatory factors TNF-a, IL-17A, and IL-6 in the offspring's intestine caused by maternal immune activation;

[0089] (a2) ameliorating the decrease in the level of the inflammatory factor IL-10 in the offspring's intestine caused by maternal immune activation;

[0090] (a3) ameliorating the increase in the level of the inflammatory factor IL-6 in the offspring's serum caused by maternal immune activation;

[0091] (a4) ameliorating the decrease in the level of any one or more of the inflammatory factors IL-22 and IL-10 in the offspring's serum caused by maternal immune activation;

[0092] (a5) ameliorating the increase in the level of any one or more of the inflammatory factors IL-1 β, TNF-a, and IL-6 in the offspring's cerebral cortex caused by maternal immune activation.

[0093] In some specific embodiments, the ameliorating the disturbance in the level of inflammatory factors in the offspring's body caused by maternal immune activation comprises (a1) - (a5) above; in these five aspects, lacto-N-neotetraose and 2'-fucosyllactose can significantly amplify each other's ameliorating effect.

[0094] In some specific embodiments, the ameliorating the disturbance in the level of inflammatory factors in the offspring's body caused by maternal immune activation comprises ameliorating the increase in the level of the inflammatory factors TNF-a, IL-17A, and IL-6 in the offspring's intestine and the decrease in the level of IL-10 caused by maternal immune activation.

[0095] In some specific embodiments, the ameliorating the disturbance in the level of inflammatory factors in the offspring's body caused by maternal immune activation comprises ameliorating the increase in the level of the inflammatory factor IL-6 in the offspring's intestine and the decrease in the level of IL-10 caused by maternal immune activation.

[0096] In some specific embodiments, the ameliorating the disturbance in the level of inflammatory factors in the offspring's body caused by maternal immune activation comprises ameliorating the increase in the level of the inflammatory factors IL-17A and IL-6 in the offspring's intestine and the decrease in the level of IL-10 caused by maternal immune activation.

[0097] In some specific embodiments, the ameliorating the disturbance in the level of inflammatory factors in the offspring's body caused by maternal immune activation comprises ameliorating the increase in the level of the inflammatory factors TNF-a and IL-6 in the offspring's intestine and the decrease in the level of IL-10 caused by maternal immune activation.

[0098] In some specific embodiments, the improvement in the disturbance of the level of inflammatory factors in the offspring organism caused by maternal immune activation comprises an improvement in the increase in the level of inflammatory factor IL-6 and a decrease in the level of inflammatory factors IL-22 and IL-10 in the serum of the offspring caused by maternal immune activation.

[0099] In some specific embodiments, the improvement in the disturbance of the level of inflammatory factors in the offspring organism caused by maternal immune activation comprises an improvement in the decrease in the level of inflammatory factors IL-22 and IL-10 in the serum of the offspring caused by maternal immune activation.

[0100] In some specific embodiments, the improvement in the disturbance of the level of inflammatory factors in the offspring organism caused by maternal immune activation comprises an improvement in the increase in the level of inflammatory factors IL-1β, TNF-α and IL-6 in the cerebral cortex of the offspring caused by maternal immune activation.

[0101] In some specific embodiments, the improvement in the disturbance of the level of inflammatory factors in the offspring organism caused by maternal immune activation comprises an improvement in the increase in the level of inflammatory factors IL-1β and TNF-α in the cerebral cortex of the offspring caused by maternal immune activation.

[0102] In some preferred embodiments, the improvement in the disturbance of the level of inflammatory factors in the offspring organism caused by maternal immune activation comprises an improvement in the increase in the level of inflammatory factors TNF-α, IL-17A and IL-6 and a decrease in the level of IL-10 in the intestine of the offspring, an increase in the level of inflammatory factor IL-6 and a decrease in the level of inflammatory factors IL-22 and IL-10 in the serum of the offspring and an increase in the level of inflammatory factor IL-1β in the cerebral cortex of the offspring caused by maternal immune activation.

[0103] In some preferred embodiments, the improvement in the disturbance of the level of inflammatory factors in the offspring organism caused by maternal immune activation comprises an improvement in the increase in the level of inflammatory factors TNF-α, IL-17A and IL-6 and a decrease in the level of IL-10 in the intestine of the offspring, an increase in the level of inflammatory factor IL-6 and a decrease in the level of inflammatory factors IL-22 and IL-10 in the serum of the offspring and an increase in the level of inflammatory factors IL-1β and TNF-α in the cerebral cortex of the offspring caused by maternal immune activation.

[0104] The food of the present application exerts the above-mentioned effects after being ingested by the offspring (offspring raised by a female who has experienced or is experiencing maternal immune activation).

[0105] The present application is not particularly limited to specific foods containing or preparable with the above-mentioned nutritional composition.

[0106] In some embodiments, the food product of the present application is in the form of a liquid or a solid at room temperature.

[0107] In some embodiments, the food product of the present application is an infant formula or a complementary food for infants; illustratively, it can be an infant formula milk powder, a children snack, etc.

[0108] In some embodiments, the food product of the present application is a confectionery, such as a hard candy, a gummy candy, a crisp candy, a pressed candy, an aerated candy, etc.

[0109] In some embodiments, the food product of the present application is a beverage, such as a carbonated beverage, a tea beverage, a coffee beverage, a fruit / vegetable juice beverage, a lactic acid bacteria beverage, etc.

[0110] In some embodiments, the food product of the present application is a dairy product, such as a milk powder, a cheese, a yogurt, a liquid milk, etc.

[0111] In some embodiments, the food product of the present application is a baked food, such as a bread, a cake, a biscuit, etc.

[0112] In some embodiments, the food product of the present application is a dietary supplement, such as a hard capsule, a soft capsule, a tablet, an oral liquid, a pill, a granule, a powder, etc.

[0113] In some embodiments, in the food product of the present application, the neutral fucosylated human milk oligosaccharides and the neutral non-fucosylated human milk oligosaccharides are all derived from the nutritional composition. In other embodiments, in the food product of the present application, the neutral fucosylated human milk oligosaccharides and the neutral non-fucosylated human milk oligosaccharides are derived from the nutritional composition and other food raw materials, such as animal milk, etc.

[0114] In some embodiments, in the food product of the present application, the mass ratio of the lacto-N-neotetraose to the 2’-fucosyllactose is 1:(1-8); preferably, the mass ratio of the lacto-N-neotetraose to the 2’-fucosyllactose is 1:(1-6); more preferably, the mass ratio of the lacto-N-neotetraose to the 2’-fucosyllactose is 1:(1.5-4.5); even more preferably, the mass ratio of the lacto-N-neotetraose to the 2’-fucosyllactose is 1:(1.5-3.5); further preferably, the mass ratio of the lacto-N-neotetraose to the 2’-fucosyllactose is 1:(1.5-3).

[0115] The present application does not particularly limit the absolute content of the lacto-N-neotetraose and the 2’-fucosyllactose in the food product, as long as it meets the requirements of the local food-related laws and regulations.

[0116] In some embodiments, the content of the 2'-fucosyllactose in the food is 0.4 g / 100 g to 2 g / 100 g, and the content of the lacto-N-neotetraose is 0.1 g / 100 g to 0.5 g / 100 g.

[0117] In addition to the above-described components in the nutritional composition, the food can further include other ingredients, such as common food ingredients of proteins / amino acids, carbohydrates, fats, vitamins, minerals, and the like.

[0118] Further, depending on the kind of the food and the final needs of the target subject, in some embodiments, the food further contains any one or more of the following ingredients: plant product ingredients, animal milk product ingredients, animal meat product ingredients, functional additive ingredients, and any acceptable adjuvants.

[0119] As for the plant product ingredients, examples include fruits such as figs, pomegranates, kiwis, oranges, tangerines, pineapples, strawberries, apples, bananas, grapes, pears, cherries, blueberries, blackberries, blackcurrants, cranberries, raspberries, melons, emblics, and mulberries, or extracts thereof; fruit and vegetable materials such as onions, cucumbers, tomatoes, cauliflowers, red beetroots, spinach, kohlrabi, Brussels sprouts, garlic, basil, Oregon grass, or extracts thereof; cereals such as rice (indica rice, japonica rice, waxy rice), wheat (wheat, barley, oat, rye), corn, sorghum, millet, foxtail millet, glutinous rice, buckwheat, soybeans, fava beans, peas, mung beans, adzuki beans, kidney beans, or extracts thereof; nut materials such as walnuts, pistachios, cashews, hazelnuts, almonds, apricot kernels, pine nuts, peanuts, melon seeds, chestnuts, macadamia nuts, ginkgo nuts, or extracts thereof; coffee or extracts thereof; and some medicinal and edible plant materials or extracts thereof.

[0120] As for the animal milk product ingredients, examples include fresh milk derived from cows, sheep, and the like, as well as reprocessed milk products such as full-fat milk powder, skim milk powder, concentrated whey protein powder, desalted whey powder, whey protein powder, hydrolyzed whey protein powder, casein powder, and the like.

[0121] As for the animal meat product ingredients, examples include meat product ingredients of pigs, cows, sheep, aquatic products, or poultry.

[0122] As for the functional additive ingredients, examples include vitamin supplements, mineral supplements, nucleotide supplements, dietary fibers, functional polyunsaturated fatty acid supplements, and the like.

[0123] For any acceptable adjuvant, examples can include solvents, antioxidants, antibacterial agents, thickeners, diluents, co-solvents, stabilizers, emulsifiers, fillers, disintegrants, lubricants, coating materials, anti-caking agents, flavoring agents, sweetening agents, food flavors, food colors, and the like.

[0124] Examples

[0125] The embodiments of the present application will be described in detail below with examples, but those skilled in the art will understand that the following examples are only for illustration of the present application and should not be regarded as limiting the scope of the present application. The specific conditions are not specified in the examples, which are carried out according to the conventional conditions or the conditions recommended by the manufacturer. The materials or instruments used are commercially available conventional products unless otherwise specified.

[0126] Experimental Example 1: Effect of Nutritional Substances on Growth Indicators of Offspring of MIA Mice

[0127] 1. Experimental Method

[0128] 1.1 Construction of Maternal Immune Homeostasis Disorder Model

[0129] Select 150 female mice and 30 male mice, and the female and male mice are mainly used for offspring breeding. When caging, select mice with shiny hair and healthy body, and cage them according to the ratio of 2:1 of female to male. During the next week, weigh the female mice regularly, and when the body weight of the female mice increases significantly, separate them and inject 20 mg / kg poly(I:C) (Sigma-Aldrich, item number P9582, the injection form of poly(I:C) is a solution, the solvent is PBS solution, and the concentration is 5 mg / mL) into the abdominal cavity to construct a maternal immune homeostasis disorder model, i.e., a maternal immune activation model.

[0130] 1.2 Intervention of Offspring

[0131] During the experiment, the male offspring are used as experimental objects. After the female parent mice deliver the offspring for 3 weeks, the male offspring are weaned and fed separately. When the male offspring are 4 weeks old, they are respectively given PBS solution or different proportions of HMOs solution by gavage, and the gavage period is 6 weeks.

[0132] 1.3 Grouping of Intervention of Offspring Animals

[0133] The experiment sets 11 groups in total, each group having 12 mice, which are a control group, a model group, and 9 HMOs intervention groups. Among them, the control group mice refer to the mice delivered by the mother mice injected with PBS solution during pregnancy, and the model group mice refer to the mice delivered by the mother mice injected with poly(I:C) solution during pregnancy. During the experiment, each mouse is fed in a standard cage and placed in an environment with 12 h light / dark cycle, humidity of 50±15%, and temperature of 22±2℃, and the mice are free to eat and drink water.

[0134] Two kinds of breast milk oligosaccharides 2'-FL and LNnT were selected for different single doses or different proportions of compounding, and the improvement effect of different dose combinations on immune disorders was investigated. The experimental design is shown in Table 1. According to the reference report, the feeding dose of LNnT is fixed at 250 mg / kg body weight / day, while the dose of 2'-FL is selected in three gradients of 250 mg / kg body weight / day, 500 mg / kg body weight / day, 850 mg / kg body weight / day and 1500 mg / kg body weight / day. The above two substances are five single factor intervention doses as comparative examples, and different dose combinations of LNnT and 2'-FL are used to form LNnT:2'-FL at ratios of 1:1, 1:2, 1:3.4 and 1:6 as four examples.

[0135] Table 1 Experimental intervention scheme design

[0136]

[0137] 1.4 Basic growth index monitoring

[0138] During the 6-week intervention period, the body weight, food intake and water intake of the mice were recorded once a week.

[0139] 2. Experimental results

[0140] The body weight change data of mice in the control group, model group and 9 intervention groups within 6 weeks of intervention are shown in Table 2. As can be seen from the table, the body weights of mice in each group on day 0 of intervention were not significantly different, and the average body weight of the model group was the lowest at 11.85 g, but after significant difference analysis, it was found that the average body weight of mice in the control group, model group, 5 comparative example groups and 4 example groups was not significantly different (p>0.05). With the extension of the intervention time, the average body weight of mice in each group increased, and the body weight was monitored and recorded on the 7th day, 14th day, 21st day, 28th day, 35th day and 42nd day. Finally, it was found that the average body weight of mice in 11 groups at each time point was not significantly different (p>0.05); at the same time, the weight gain of mice in each group after 42 days of intervention was about 9 g, and after significant difference analysis, it was found that the mice in 11 groups, including the model group and different HMOs single or combined intervention, did not affect the normal increase of the body weight of mice, and there was no significant difference between groups (p>0.05).

[0141] Meanwhile, the food intake and water intake of 11 groups of mice were recorded during the intervention period, and the experimental results are shown in Table 3. As can be seen from the table, the food intake of 11 groups of mice during the whole intervention period is about 3 g / day, and after difference significance analysis, it is found that the average daily food intake of mice in Comparative Example 1, Comparative Example 4 and Comparative Example 5 is significantly higher than that of the control group (p<0.05), and the food intake of mice in the other groups has no significant difference (p>0.05) with the control group. At the same time, the water intake of each group of mice was recorded and compared, and it can be seen from Table 3 that the water intake of 11 groups of mice has no significant difference (p>0.05).

[0142] Table 2 Body weight changes of mice in each group during the intervention of 6 weeks (g)

[0143]

[0144] Table 3 Comparison of average daily food intake and water intake of mice in each group during the intervention period

[0145]

[0146] Experimental Example 2: Changes in immune levels of MIA mouse offspring

[0147] 1. Experimental method

[0148] 1.1 Construction of maternal immune homeostasis disorder model

[0149] The same as Example 1.

[0150] 1.2 Intervention of offspring

[0151] The same as Example 1.

[0152] 1.3 Grouping of offspring animals for intervention

[0153] The same as Example 1.

[0154] 1.4 Collection of tissue samples and detection of indexes

[0155] 1.4.1 Collection of tissue samples

[0156] The mice were fasted for 12 h before sacrifice, and the mice were anesthetized by intraperitoneal injection of 1.25% tribromoethanol solution at a dose of 0.2 mL / 10 g bw. After eye ball blood collection, the mice were sacrificed by cervical dislocation. The mouse serum was obtained by centrifugation of the plasma at 25°C, 3000 r / min for 15 min, and stored at -80°C. Then the mouse brain and intestinal tissue samples were collected on ice, wrapped with tin paper, quickly frozen in liquid nitrogen, and moved to a -80°C refrigerator. The tissues for pathological section were placed in a 4% (v / v) paraformaldehyde solution for subsequent experiments.

[0157] 1.4.2 qRT-PCR analysis of colon cytokines

[0158] The gene transcription level of cytokines in the colon was analyzed by real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) technology, and the specific operation was as follows:

[0159] (1) Tissue RNA extraction

[0160] TRIzol was used to extract tissue mRNA, and the steps were referred to the instruction manual. A certain amount of mouse colon tissue was taken, grinding beads were added, and tissue homogenate was prepared. Then 1 mL TRIzol was added for homogenate lysis. Then according to the steps in the instruction manual, the RNA was obtained by layering (chloroform extraction), precipitation (isopropanol), and washing (75% ethanol). Then the RNA was dissolved in DEPC water and stored at -80°C.

[0161] (2) Reverse transcription (cDNA synthesis)

[0162] The extracted mRNA was reverse transcribed using PrimeScript™ RT Master Mix reverse transcription kit (TaKaRa PrimeScript RT Master Mix, Dalian). The steps were referred to the instruction manual.

[0163] (3) Index determination

[0164] According to the operation instruction of Takara Premix Ex Taq TM II RR036A kit, the PCR amplification system was configured as follows: 1 μL of upstream primer, 1 μL of downstream primer, 6 μL of ddH2O, 10 μL of Premix Ex Taq II, and 2 μL of cDNA template. The primer of each detection index is shown in Table 4.

[0165] Two-step PCR amplification standard procedure: 95°C for 30 s, cycle 1; 95°C for 3 s, 60°C for 30 s, cycle 40 times; enter the melting curve stage. Finally, the mRNA of GAPDH was used as an internal reference, and the relative expression amount of the gene was calculated according to the 2 -△△Ct value.

[0166] Table 4 Gene primer sequence

[0167]

[0168] 1.4.3 ELISA analysis of serum cytokines

[0169] The expression levels of inflammatory factors IL-22, IL-6 and IL-10 in serum were determined by enzyme linked immunosorbent assay (ELISA), and the kit was purchased from Xinle Biological Company. The specific operation steps were carried out according to the instruction manual. Data processing and analysis were carried out by ELISA Calc software. First, the standard curve was established, and then the absorbance of the sample was input to calculate the level of inflammatory factors.

[0170] 1.4.4 qRT-PCR analysis of brain tissue cytokines

[0171] (1) Tissue RNA extraction

[0172] TRIzol was used to extract tissue mRNA. The steps were referred to the instruction manual. A certain amount of mouse cerebral cortex tissue was taken, grinding beads were added, and tissue homogenate was prepared. Then 1 mL TRIzol was added for homogenate lysis. Then according to the steps in the instruction manual, the RNA was obtained by layering (chloroform extraction), precipitation (isopropanol), and washing (75% ethanol). Then the RNA was dissolved in DEPC water and stored at -80°C.

[0173] (2) Reverse transcription (cDNA synthesis)

[0174] The extracted mRNA was reverse transcribed using PrimeScript RT Master Mix reverse transcription kit (TaKaRa PrimeScript RT Master Mix, Dalian). The steps were referred to the instruction manual.

[0175] (3) Index determination

[0176] According to the operation instruction of Takara Premix Ex Taq TM II RR036A kit, the PCR amplification system was configured as follows: 1 μL of upstream primer, 1 μL of downstream primer, 6 μL of ddH2O, 10 μL of Premix Ex Taq II, and 2 μL of cDNA template. The primer of each detection index is shown in Table 5.

[0177] Two-step PCR amplification standard procedure: 95°C for 30 s, cycle 1; 95°C for 3 s, 60°C for 30 s, cycle 40 times; enter the melting curve stage. Finally, the mRNA of GAPDH was used as an internal reference, and the relative expression amount of the gene was calculated according to the 2 -△△Ct value.

[0178] Table 5 Gene primer sequence

[0179]

[0180] 2. Experimental results

[0181] On the basis of no difference in weight gain, no difference in food intake and no difference in water intake of mice throughout the intervention period, the level and changes of inflammation of the whole body of mice in each group were further analyzed.

[0182] Infection (e.g. infection with viruses, bacteria, etc.) or autoimmune diseases of the mother during pregnancy can cause an increase in inflammatory factors (such as IL-6, IL-1β, TNF-α, etc.), which can enter the fetus through the placenta. The present application gives pregnant mice to inject Poly (I: C) to promote maternal immune activation during the critical period of fetal brain development during pregnancy. The mice produced by such a mother have a disturbed immune system, and the experimental results are shown in Table 6. As can be seen from Table 6, the mRNA expression levels of pro-inflammatory factors TNF-α, IL-6 and IL-1β in the cerebral cortex of the offspring mice were significantly higher than those in the control group. This indicates that the offspring mice have severe inflammation in the brain. At the same time, it was found that the pro-inflammatory factor IL-6 in the serum of the offspring mice was significantly higher than that in the control group, the cytokine IL-22 responsible for tissue repair was significantly reduced, and the anti-inflammatory factor IL-10 was significantly reduced. Further analysis of the level of inflammatory factors in the colon found that the pro-inflammatory factors TNF-α, IL-17A and IL-6 in the model group were significantly higher than those in the control group, while the anti-inflammatory factor IL-10 was significantly lower than that in the control group. These results show that the maternal immune activation transmits inflammation disorder to the offspring, and the inflammation levels in the offspring from the intestine to the blood and then to the brain are found to be significantly changed, which will have a serious adverse effect on the development of the offspring's brain and body behavior.

[0183] Table 6 Inflammation level of offspring after maternal immune activation

[0184]

[0185] Experimental Example 3: Effect of nutrients on the intestinal immune level of MIA offspring mice

[0186] 1. Experimental method

[0187] 1.1 Construction of maternal immune homeostasis disorder model

[0188] The same as in Example 1.

[0189] 1.2 Intervention of offspring

[0190] The same as in Example 1.

[0191] 1.3 Intervention grouping of offspring animals

[0192] The same as in Example 1.

[0193] 1.4 Collection of tissue samples and detection of indicators

[0194] 1.4.1 Collection of tissue samples

[0195] The same as in Example 2.

[0196] 1.4.2 qRT-PCR analysis of colon cytokines

[0197] The same as Example 2.

[0198] 2. Experimental results

[0199] The present application also investigated the regulation of intestinal inflammation in offspring mice after consuming two HMOs (2'-FL and LNnT) monomers or combined intervention, and the results are shown in Tables 7 and 8.

[0200] (1) Tumor necrosis factor TNF-α (TNF-α) is mainly secreted by activated macrophages, T cells, NK cells, mast cells and some tissue cells (such as adipocytes). TNF-α is the core initiator of the inflammatory cascade, a strong pro-inflammatory and inducer of apoptosis, and plays a key role in autoimmune diseases, sepsis and metabolic diseases. The present application found that TNF-α in the colon of the offspring mice was significantly increased, but when 2'-FL and LNnT were used alone or in combination (Comparative Examples 1-5, Examples 1-4) to intervene, the mRNA expression of TNF-α was significantly reduced (p<0.0001); further observation found that the mRNA expression of TNF-α in Comparative Examples 1-5 was between 2.12-2.51, while the mRNA expression of TNF-α in Examples 1-4 was between 1.00-1.28, and after significant difference analysis, it was found that the mRNA expression of TNF-α in Examples 1-4 was significantly lower than that in Comparative Examples 1-5 (p<0.0001), which indicated that the two HMOs could significantly inhibit the level of TNF-α, the core factor of the inflammatory cascade, after being compounded at four ratios of 1:1, 1:2, 1:3.4 and 1:6. And the inhibition of TNF-α expression in the intestinal tract of the offspring of the maternal immune activated mice by the two HMOs after compounding had a synergistic effect.

[0201] (2) IL-17A is secreted by Th17 cells, γδ T cells, NK cells and innate lymphocytes (ILC3), its main function is to promote the occurrence of inflammatory response, and plays a major role in host defense and autoimmune diseases. The present application found that the mRNA expression of IL-17A in the offspring colon was significantly increased after maternal immune activation, but the mRNA expression of IL-17A could be significantly reduced after intervention of 2'-FL and LNnT alone or in combination (p<0.0001). After intervention of LNnT alone or 2'-FL alone at different doses, the mRNA expression could be reduced to about 3, and after the two HMOs were compounded at four ratios of 1:1, 1:2, 1:3.4 and 1:6, the mRNA expression of IL-17A was reduced to below 1.5, and after difference significance analysis, it was found that examples 1-4 were significantly lower than comparative examples 1-5 (p<0.0001). It can be known that in terms of the inhibitory effect of IL-17A expression in the offspring intestinal tract of the maternal immune activated mice, the combined use of the two HMOs has a synergistic effect.

[0202] (3) IL-6 is a core molecule of inflammatory cascade reaction, and has pro-inflammatory and anti-inflammatory effects, and is involved in acute inflammatory reaction and metabolic regulation. The present application found that the mRNA expression of IL-6 in the model group was significantly increased, indicating that IL-6 mainly played a pro-inflammatory role at this time, but the pro-inflammatory effect was inhibited after intervention of LNnT and 2'-FL, and the mRNA expression of IL-6 was significantly reduced (p<0.0001), especially when LNnT and 2'-FL were compounded at four ratios of 1:1, 1:2, 1:3.4 and 1:6 (examples 1-4), the mRNA expression of IL-6 was significantly lower than that of the two HMOs alone (comparative examples 1-5). It can be known that in terms of the inhibitory effect of IL-6 expression in the offspring intestinal tract of the maternal immune activated mice, the combined use of the two HMOs has a synergistic effect.

[0203] (4) IL-10 is mainly secreted by regulatory T cells (Treg), macrophages, B cells and dendritic cells. It is an anti-inflammatory cytokine that inhibits excessive immune response and maintains immune homeostasis. The present application found that the mRNA expression of the anti-inflammatory factor IL-10 in the intestinal tract of the offspring was significantly reduced after maternal immune activation, but was improved after LNnT and 2'-FL intervention, and was significantly higher than that of the model group (p<0.0001). Further comparison found that the mRNA expression of IL-10 was significantly higher after LNnT and 2'-FL were compounded at four ratios of 1:1, 1:2, 1:3.4 and 1:6 (Examples 1-4) than after the two HMOs were intervened alone (Comparative Examples 1-5) (p<0.0001, p<0.001 or p<0.01). This shows that intervention after the two HMOs are compounded can significantly promote the production of the anti-inflammatory factor IL-10, and there is a synergistic effect between the two. It was further found that Example 2 was higher than Examples 1, 3 and 4 in the four examples.

[0204] The above results show that maternal immune activation causes an inflammatory explosion in the intestinal tract of the offspring, but after early intervention of LNnT and 2'-FL, the inflammatory explosion can be significantly inhibited, the mRNA expression of the inflammatory cascade initiator TNF-α is inhibited, the mRNA expression of the pro-inflammatory factor IL-17A is inhibited, the mRNA expression of the core molecule IL-6 of the inflammatory cascade is inhibited, and the mRNA expression of the anti-inflammatory factor IL-10 is promoted. The effect of the two HMOs compounded is better than that of the single body.

[0205] Table 7 mRNA expression levels of intestinal inflammatory factors of offspring mice after intervention of two HMOs

[0206]

[0207] Table 8 Significance analysis of differences between groups after correction of mRNA expression levels of intestinal inflammatory factors of offspring mice in each group

[0208]

[0209] Experimental Example 4: Effect of nutrients on serum immune level of MIA offspring

[0210] 1. Experimental method

[0211] 1.1 Construction of maternal immune homeostasis disorder model

[0212] The same as Example 1.

[0213] 1.2 Intervention of offspring

[0214] The same as Example 1.

[0215] 1.3 Intervention grouping of offspring animals

[0216] The same as Example 1.

[0217] 1.4 Tissue sample collection and index detection

[0218] 1.4.1 Tissue sample collection

[0219] The same as Example 2.

[0220] 1.4.2 ELISA analysis of serum cytokines

[0221] The same as Example 2.

[0222] 2. Experimental results

[0223] The present application analyzes the production of inflammatory factors in the serum of each group of offspring mice after the intervention of two HMOs, and the results are shown in Tables 9 and 10.

[0224] (1) From Tables 9 and 10, it can be seen that after the offspring mice were intervened by LNnT and 2'-FL oligosaccharides in monomer (Comparative Examples 1-5) or after the intervention of the four ratios of 1:1, 1:2, 1:3.4 and 1:6 (Examples 1-4), the production of pro-inflammatory factor IL-6 was significantly reduced (p<0.0001 or p<0.001), and further analysis found that the IL-6 production of the four examples was significantly lower than that of the five comparative examples (p<0.0001, p<0.001 or p<0.01 or p<0.05). This shows that after LNnT and 2'-FL are mixed at a ratio of 1:1, 1:2, 1:3.4 and 1:6, the production of pro-inflammatory factor IL-6 in the offspring body caused by maternal immune activation can be significantly reduced. And in terms of reducing the level of serum IL-6, there is a synergistic effect between 2'-FL and LNnT.

[0225] (2) At the same time, the present application also analyzes the production amount of IL-22 in the serum, IL-22 is mainly secreted by Th17 cells, has tissue repair and barrier protection effects, and has pro-inflammatory and anti-inflammatory effects. The present application finds that the amount of IL-22 in the serum of the offspring is significantly reduced after maternal immune activation, but the expression amount of IL-22 is increased after the intervention of the two oligosaccharides, and after the difference significance analysis, it is found that the production amount of IL-22 after the intervention of LNnT and 2'-FL in the form of single or complex is significantly higher than that of the model group (p<0.0001, p<0.01 or p<0.05). Further analysis found that the production amount of IL-22 after the complexing of LNnT and 2'-FL in the four ratios of 1:1, 1:2, 1:3.4 and 1:6 (Examples 1-4) is significantly higher than that of the two single interventions (Comparative Examples 1-5), and after the difference significance analysis, it is found that there is a significant difference (p<0.0001 or p<0.05). And in terms of improving the level of serum IL-22, there is a synergistic effect between 2'-FL and LNnT. At the same time, the production amount of IL-22 in Example 2 is significantly higher than that in the other three examples (p<0.0001 or p<0.05).

[0226] (3) At the same time, we also analyze the production amount of anti-inflammatory factor IL-10 in the serum, from Table 9 and Table 10, it can be seen that the production amount of IL-10 in the serum of the offspring is significantly increased (p<0.0001, p<0.01 or p<0.05) after the intervention of LNnT and 2'-FL in the form of single (Comparative Examples 1-5) or complex (Examples 1-4), and the production amount of IL-10 in the serum of the offspring in Examples 1-4 is significantly higher than that in Comparative Examples 1-5 (p<0.0001, p<0.001, p<0.01 or p<0.05). And in terms of improving the level of serum IL-10, there is a synergistic effect between 2'-FL and LNnT. Also found that Example 2 is significantly higher than Example 1 (p<0.0001), Example 3 (p<0.0001) and Example 4 (p<0.001).

[0227] The above results show that maternal immune activation causes inflammation level disorder in the serum of the offspring, but after the intervention of LNnT and 2'-FL in the early stage, the inflammation outbreak can be significantly inhibited, the production of the core molecule IL-6 of the inflammation cascade reaction is inhibited, the production of the inflammation factor IL-22 with tissue repair and barrier protection function is promoted, and the production of the anti-inflammatory factor IL-10 is promoted. And the effect of the complex of the two HMOs is better than that of the single body effect.

[0228] Table 9 Production amount of inflammatory factor in serum of offspring after intervention of two HMOs (pg / mL)

[0229]

[0230] Table 10 Analysis of the significance of the difference in the amount of inflammatory factor production in the serum of each group of offspring mice

[0231]

[0232] Experimental Example 5: Effect of nutrients on the immune level of the offspring of MIA mice

[0233] 1. Experimental method

[0234] 1.1 Construction of a maternal immune homeostasis disorder model

[0235] The same as Example 1.

[0236] 1.2 Offspring intervention

[0237] The same as Example 1.

[0238] 1.3 Grouping of offspring animal intervention

[0239] The same as Example 1.

[0240] 1.4 Tissue sample collection and index detection

[0241] 1.4.1 Tissue sample collection

[0242] The same as Example 2.

[0243] 1.4.2 qRT-PCR analysis of brain tissue cytokines

[0244] The same as Example 2.

[0245] 2. Experimental results

[0246] The present application analyzed the expression of inflammatory factors in the cerebral cortex of each group of offspring mice after intervention with two HMOs, and the results are shown in Tables 11 and 12.

[0247] IL-1β is produced by activated immune cells (such as macrophages, monocytes, dendritic cells) and some non-immune cells (such as epithelial cells). It acts as a core pro-inflammatory factor, involved in innate immunity and adaptive immunity. IL-1β and TNF-α are the starting factors of early inflammation, and IL-6 subsequently amplifies inflammation. In the present invention, it was found that when maternal immune activation caused the activation of offspring microglia, the mRNA expression levels of the three inflammatory factors in the cerebral cortex of the offspring mice were significantly higher than those in the control group (p<0.0001). However, after 42 days of intervention with LNnT and 2’-FL monomers or complexes, it was found that the expression of the three inflammatory factors was inhibited to varying degrees. And after significant difference analysis, it was found that the mRNA expression levels of the three inflammatory factors of Comparative Examples 1-5 and Examples 1-4 were significantly lower than those of the model group (p<0.0001, p<0.001 or p<0.01). Further comparison of the differences between the comparative examples and the examples found that for the inflammatory factor IL-1β, Examples 1-4 were significantly lower than Comparative Examples 1-5 (p<0.0001), for the inflammatory factor TNF-α, Examples 1-4 were significantly lower than Comparative Examples 1-5 (p<0.0001 or p<0.001 or p<0.01 or p<0.05), and for the inflammatory factor IL-6, Examples 1-4 were significantly lower than Comparative Examples 1-5 (p<0.0001). This shows that the inhibitory effect of the two kinds of human milk oligosaccharides on inflammatory factors is higher than that of LNnT and 2’-FL monomers after compounding at a ratio of 1:1, 1:2, 1:3.4 and 1:6, and the two have a synergistic effect. Moreover, LNnT can significantly enhance the inhibitory effect of 2’-FL on the expression of IL-1β, TNF-α and IL-6, three inflammatory factors in the cerebral cortex of the offspring of maternal immune activation mice, and similarly, 2’-FL can also significantly promote the inhibitory effect of LNnT on the expression of IL-1β, TNF-α and IL-6, three inflammatory factors in the cerebral cortex of the offspring of maternal immune activation mice.

[0248] Further comparison of the production amounts of pro-inflammatory factors of the four examples found that Example 2 had a stronger inhibitory effect on the production of inflammatory factors IL-1β, TNF-α and IL-6 than Examples 1, 3 and 4, and after significant difference analysis, it was found that Example 2 had a more significant inhibitory effect on the production of the three inflammatory factors than Example 1 (p<0.01, p<0.01 or p<0.05), and Example 2 had a more significant inhibitory effect on the production of inflammatory factor IL-1β than Example 4 (p<0.05).

[0249] The above results show that maternal immune activation causes a large amount of pro-inflammatory factors in the offspring brain cortex, but early intervention with LNnT and 2'-FL can significantly inhibit the mRNA expression of pro-inflammatory factors IL-1β, TNF-α and IL-6, and the effect of the combination of the two HMOs is better than that of a single body, and the two have a synergistic effect.

[0250] Table 11 mRNA expression levels of inflammatory factors in the cerebral cortex of offspring mice after intervention of two HMOs

[0251]

[0252] Table 12 Analysis of significant difference in mRNA expression levels of inflammatory factors in the cerebral cortex of offspring mice in each group after correction

[0253]

[0254] It can be seen from Experimental Examples 2-5 that maternal immune activation during pregnancy causes an imbalance in inflammation in the brain, serum and intestine of the offspring, which has a serious adverse effect on the health of the offspring. The present application found that after LNnT and 2'-FL were compounded in different proportions and fed to the offspring in the early life, it was found that the direct improvement of the intestinal inflammation level could further improve the serum and brain inflammation levels through the inflammation cascade, that is, although LNnT and 2'-FL are non-digestible nutrients, the improvement of the inflammation level after eating can penetrate the blood-brain barrier, not only affecting the inflammation level of the intake site, i.e. the intestine, but also affecting the inflammation levels of the serum and the brain.

Claims

1. The use of a nutritional composition in the preparation of a food that improves the disordered levels of inflammatory factors in offspring caused by maternal immune activation; wherein, The nutritional composition contains the essential active ingredients shown in (i) and (ii) below: (i) Neutral fucoidan-based human milk oligosaccharides, (ii) Neutral non-fucosylated human milk oligosaccharides; in, The neutral fucosylated human milk oligosaccharide contains at least 2'-fucosylated lactose. The neutral non-fucosylated human milk oligosaccharide contains at least lactose-N-neotetrasaccharide; Furthermore, in the nutritional composition, the mass ratio of lactose-N-neotetrasaccharide to 2'-fucosylated lactose is 1:(6-8); The food has its effects after being ingested by the offspring; The improvement of the disordered levels of inflammatory factors in offspring caused by maternal immune activation includes improving the disordered levels of inflammatory factors in the offspring gut, serum, and cerebral cortex caused by maternal immune activation. The improvement of the disordered levels of inflammatory factors in offspring caused by maternal immune activation includes improving the increased levels of inflammatory factors TNF-α, IL-17A, and IL-6 in the offspring gut and the decreased level of IL-10, the increased level of inflammatory factor IL-6 in the offspring serum and the decreased levels of IL-22 and IL-10, and the increased level of inflammatory factor IL-1β in the offspring cerebral cortex.

2. The use according to claim 1, characterized in that, The neutral fucosylated human milk oligosaccharide further comprises any one or more of 3-fucosylated lactose and difucosylated lactose.

3. The use according to claim 1, characterized in that, The neutral non-fucosylated human milk oligosaccharide also contains lactose-N-tetrasaccharide.

4. The use according to claim 1, characterized in that, The improvement of the disordered levels of inflammatory factors in offspring caused by maternal immune activation includes improving the elevation of any one or more of the inflammatory factors TNF-α and IL-6 in the offspring cerebral cortex caused by maternal immune activation.

5. The use according to any one of claims 1-4, characterized in that, The food mentioned is infant formula or infant supplementary food.

6. The use according to any one of claims 1-4, characterized in that, The food products mentioned are candies, beverages, dairy products, baked goods, or dietary supplements.

7. The use according to any one of claims 1-4, characterized in that, The food contains any one or more of the following ingredients: plant-based ingredients, animal dairy ingredients, animal meat ingredients, functional additives, and any acceptable excipients.

8. The use according to any one of claims 1-4, characterized in that, In the food product, the content of 2'-fucosylated lactose is 0.4g / 100g-2g / 100g, and the content of lactose-N-neotetrasaccharide is 0.1g / 100g-0.5g / 100g.

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