Nutritional composition and application thereof for improving intestinal microecology

Abstract

The invention belongs to the field of food, and particularly relates to a nutritional composition and application thereof in improving intestinal microecology. The invention provides application of a nutritional composition in preparation of food for improving offspring intestinal microecological imbalance caused by maternal immune activation. The nutritional composition comprises essential active ingredients shown as the following (i) and (ii): (i) a neutral fucosylated breast milk oligosaccharide, (ii) a neutral non-fucosylated breast milk oligosaccharide; wherein the neutral fucosylated breast milk oligosaccharide at least comprises 2 '-fucosylated lactose, and the neutral non-fucosylated breast milk oligosaccharide at least comprises lactose-N-neotetraose; moreover, in the nutritional composition, the mass ratio of the lactose-N-neotetraose to the 2 '-fucosyllactose is 1: (1-9), and the 2'-fucosyllactose and the lactose-N-neotetraose can achieve a synergistic effect to improve the intestinal microecology of the offspring and provide benefits for the development of the nervous system of the offspring.

Description

Technical Field

[0001] This invention belongs to the food field, specifically relating to nutritional compositions and their use in improving intestinal microecology. Background Technology

[0002] During pregnancy, the mother's immune system undergoes dramatic changes to maintain immune balance with the fetus, prevent rejection reactions, and preserve innate and adaptive immune mechanisms to fight infection. Exposure to various environmental factors during pregnancy can lead to an imbalance in inflammatory cytokine levels, resulting in maternal immune activation (MIA). The effects of MIA on offspring are heterogeneous. In recent years, the role of maternal immune activation induced by infection during pregnancy in offspring neurodevelopment has become a research hotspot, and some progress has been made. Studies have found that the immature fetal brain is particularly vulnerable to MIA damage. MIA can affect offspring neurodevelopment by altering genomic susceptibility, influencing epigenetic modifications, microglia activation, and intracranial inflammation.

[0003] In addition, some studies have found that gut microbiota can alter the body's metabolic homeostasis, and an imbalance in gut microbiota homeostasis can affect neurodevelopment by regulating gastrointestinal physiology and immune function through the gut-brain axis.

[0004] Human milk oligosaccharides (HMOs) are the third most abundant solid component in breast milk, after lactose and fat, and their content is even higher than that of proteins. HMOs are a class of complex sugars with various structural types. Based on their molecular structure, HMOs can be classified into neutral fucosylated HMOs, neutral non-fucosylated HMOs, and acidic sialylated HMOs.

[0005] Studies have found that human milk oligosaccharides (HMOs) have a regulatory effect on gut health in subjects. For example, Reference 1 discloses that human milk oligosaccharides (including 2'-FL, 3-FL, DFL, LNT, LNnT, 3'-SL, 6'-SL, LNFP-I, or mixtures thereof) can be used to increase the abundance of Akkermansia bacteria in the human gastrointestinal tract, thereby helping to alleviate brain-gut symptoms such as stress, anxiety, and depression. Reference 2 discloses a nutritional composition containing at least two types of human milk oligosaccharides, which can reduce the concentration of harmful proteolytic metabolites such as branched-chain short-chain fatty acids in the digestive tract of infants or young children, thereby helping to resist intestinal inflammation and intestinal infections. However, studies like References 1 and 2 are still not considered sufficient, as their intervention subjects are mainly healthy humans, and Reference 1 also emphasizes that its intervention subjects are non-infants. Whether human milk oligosaccharides have an effect on gut health under specific conditions (such as the gut health of offspring born to females who have experienced or are currently in a state of maternal immune activation) has not been studied in the prior art.

[0006] Citation:

[0007] Citation 1: CN111683665A;

[0008] Citation 2: CN110650635A. SUMMARY

[0009] Problem to be solved by the invention

[0010] Studies have shown that maternal immune activation affects the neural development of offspring, and intestinal flora homeostasis imbalance can affect neural development through the gut-brain axis by regulating gastrointestinal physiology, immune function, etc.

[0011] In view of this, the present application has carried out a large amount of research, and found that maternal immune activation can lead to intestinal microecological imbalance in offspring, including changes in intestinal flora and its metabolites, and changes in intestinal structure and function. Based on this, the present application believes that how to reduce, improve or inhibit the adverse effects of maternal immune activation on the intestinal health of offspring, so as to provide help for the neural development of offspring through the gut-brain axis, has important research significance.

[0012] Although the prior art such as the above-mentioned citation 1, 2, etc. studies the effect of breast milk oligosaccharides on intestinal health, similar studies do not consider the triggering factors of intestinal abnormal conditions, and the development period damage (i.e. congenital, systemic and diffuse damage, rather than acquired, acquired, and local damage) is rarely involved in the prior art.

[0013] During the research of the present application, it is accidentally found that the combination of 2'-fucosyllactose and lacto-N-neotetraose can improve the intestinal microecological imbalance of offspring caused by maternal immune activation, including improving the changes of intestinal microorganisms and their metabolites, and the damage of intestinal structure and function. Based on the concept of gut-brain axis, this improvement will be beneficial to the healthy development of the nervous system of offspring, and the present application accidentally finds that lacto-N-neotetraose can significantly amplify the improvement effect of 2'-fucosyllactose on the intestinal microecological imbalance of 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.

[0014] Solution to the problem

[0015] In order to solve the above technical problems, the present application provides the following technical solutions:

[0016] [1]. Use of a nutritional composition in the manufacture of a food for improving intestinal microflora imbalance in offspring caused by maternal immune activation; wherein the nutritional composition comprises essential active ingredients represented by (i) and (ii) below:

[0017] (i) neutral fucosylated human milk oligosaccharides,

[0018] (ii) neutral non-fucosylated human milk oligosaccharides;

[0019] wherein,

[0020] the neutral fucosylated human milk oligosaccharides comprise at least 2'-fucosyllactose,

[0021] the neutral non-fucosylated human milk oligosaccharides comprise at least lacto-N-neotetraose; and

[0022] and, in the nutritional composition, the mass ratio of the lacto-N-neotetraose to the 2'-fucosyllactose is 1 : (1-9).

[0023] [2]. The use according to [1], wherein the improvement of intestinal microflora imbalance in offspring caused by maternal immune activation comprises (a1) below:

[0024] (a1) improvement of reduced abundance of beneficial genera in the intestine of offspring caused by maternal immune activation, the beneficial genera comprising any one or more of Lactobacillus, Limosilactobacillus, and Butyricicoccus.

[0025] [3]. The use according to [1] or [2], wherein the improvement of intestinal microflora imbalance in offspring caused by maternal immune activation comprises (a2) below:

[0026] (a2) improvement of reduced content of short-chain fatty acids in the intestine of offspring caused by maternal immune activation, the short-chain fatty acids comprising any one or more of acetic acid, propionic acid, and butyric acid.

[0027] [4]. The use according to any one of [1] to [3], wherein the improvement of intestinal microflora imbalance in offspring caused by maternal immune activation comprises (a3) below:

[0028] (a3) improvement of structural and / or functional impairment of the intestine of offspring caused by maternal immune activation;

[0029] the improvement of structural and / or functional impairment of the intestine of offspring caused by maternal immune activation comprises any one or more of (b1) to (b3) below:

[0030] (b1) improving the reduced thickness of the intestinal muscularis layer of the offspring caused by maternal immune activation;

[0031] (b2) improving the reduced crypt depth of the intestine of the offspring caused by maternal immune activation;

[0032] (b3) improving the reduced amount of salivary mucin secretion from goblet cells of the intestine of the offspring caused by maternal immune activation.

[0033] [5]. The use according to any one of [1] to [4], wherein the neutral fucosylated human milk oligosaccharide further comprises any one or more of 3-fucosyllactose and difucosyllactose.

[0034] [6]. The use according to any one of [1] to [4], wherein the neutral non-fucosylated human milk oligosaccharide further comprises lacto-N-tetraose.

[0035] [7]. The use according to any one of [1] to [6], wherein the food product is an infant formula or a baby food supplement.

[0036] [8]. The use according to any one of [1] to [7], wherein the food product is a confectionery, a beverage, a dairy product, a bakery product or a dietary supplement.

[0037] [9]. The use according to any one of [1] to [8], wherein the food product comprises any one or more of a plant product ingredient, an animal dairy product ingredient, an animal meat product ingredient, a functional additive ingredient and any acceptable excipient.

[0038]

[10] . The use according to any one of [1] to [9], wherein the food product comprises 2’-fucosyllactose in an amount of 0.4 g / 100 g to 2 g / 100 g and lacto-N-neotetraose in an amount of 0.1 g / 100 g to 0.5 g / 100 g.

[0039]

[11] . Use of a nutritional composition for the manufacture of a food product having any one or more of the following (d1) to (d5) improving effects:

[0040] (d1) improving the reduced abundance of beneficial genera in the intestine of the offspring caused by maternal immune activation, the beneficial genera comprising any one or more of Lactobacillus, Limosilactobacillus and Butyricicoccus;

[0041] (d2) improving the reduced content of short chain fatty acids in the intestine of the offspring caused by maternal immune activation, the short chain fatty acids comprising any one or more of acetic acid, propionic acid and butyric acid;

[0042] (d3) improving the decrease in the thickness of the intestinal muscularis layer of the offspring caused by maternal immune activation;

[0043] (d4) improving the decrease in the depth of the intestinal crypt of the offspring caused by maternal immune activation;

[0044] (d5) improving the decrease in the amount of salivary mucin secretion from the intestinal goblet cells of the offspring caused by maternal immune activation;

[0045] wherein the nutritional composition comprises essential active ingredients shown in (i) and (ii) below:

[0046] (i) neutral fucosylated human milk oligosaccharides, at least comprising 2'-fucosyllactose;

[0047] (ii) neutral non-fucosylated human milk oligosaccharides, at least comprising lacto-N-neotetraose;

[0048] and, in the nutritional composition, the mass ratio of the lacto-N-neotetraose to the 2'-fucosyllactose is 1 : (1-9).

[0049]

[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.

[0050]

[13] . The use according to

[11] or

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

[0051]

[14] . The use according to any one of

[11] to

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

[0052]

[15] . The use according to any one of

[11] to

[14] , wherein the food contains any one or more of the following ingredients: a plant product ingredient, an animal dairy product ingredient, an animal meat product ingredient, a functional additive ingredient, and any acceptable excipient.

[0053]

[16] . The use according to any one of

[11] to

[15] , wherein, in the food, 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.

[0054] Effects of the invention

[0055] The present application proposes that the combination of 2'-fucosyllactose and lacto-N-neotetraose can improve the intestinal microecological imbalance of offspring caused by maternal immune activation, including improving the microbial population and its metabolism in the intestine of offspring, especially promoting the abundance of beneficial bacteria Lactobacillus, Limosilactobacillus and Butyricicoccus and their subordinate species in the intestine of offspring, and promoting the content of short-chain fatty acids (including acetic acid, propionic acid and butyric acid) as the main metabolic products of intestinal microbial fermentation. At the same time, the improvement of the intestinal microecological imbalance of offspring caused by maternal immune activation by the combination of 2'-fucosyllactose and lacto-N-neotetraose also includes improving the structural and functional damage of the intestine of offspring, that is, the combination of 2'-fucosyllactose and lacto-N-neotetraose can comprehensively improve the intestinal flora and the intestinal structure environment supporting the survival of the flora. The function of intestinal flora is highly dependent on the intestinal environment in which it survives, and the integrity of intestinal barrier function is regulated by flora. This comprehensive improvement of intestinal microecology can provide benefits for the development of the nervous system of offspring through the gut-brain axis. Alternatively, it can be considered that the combination of 2'-fucosyllactose and lacto-N-neotetraose can improve the intestinal microecological imbalance accompanied by abnormal neural development of offspring born by females who have experienced or are experiencing maternal immune activation.

[0056] More importantly, the improvement proposed by the present application is aimed at the intestinal microecological imbalance of offspring caused by maternal immune activation, that is, the improvement proposed by the present application is aimed at the improvement of developmental period damage that occurs earlier. This improvement provides help for the reversal of congenital developmental deficiency or abnormality.

[0057] Furthermore, 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 intestinal microecological imbalance of offspring caused by maternal immune activation.

[0058] At the same time, the nutritional composition containing the combination of 2'-fucosyllactose and lacto-N-neotetraose described in the present application can be used independently as a nutritional fortifier, or can be added to various foods and ingested through eating, and plays a role in the early life, that is, the critical period of neural development of infants and young children. DETAILED DESCRIPTION

[0059] The embodiments of the present application are 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.

[0060] In the present application, the terms "comprising", "having", "including", or "containing" can mean "including" or "open" and do not exclude additional, unrecited elements or method steps. At the same time, "comprising", "having", "including", or "containing" can also mean "closed" and exclude additional, unrecited elements or method steps.

[0061] In the present application, the meaning of "may" includes both the meaning of performing a certain process and the meaning of not performing a certain process.

[0062] In the present application, "optional" or "optionally" means that the use or non-use of certain substances, components, execution steps, applied conditions, and the like factors.

[0063] 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, B.

[0064] In the present application, the term "about" is used to define the numerical ranges and parameters of the present application as approximate values, and the relevant values in the specific examples have been presented as accurately as possible. Unless otherwise explicitly stated, it should be understood that all ranges, numbers, values, and percentages used in the present application are modified by "about". Here, "about" generally means that the actual value is within ±5%, ±3%, ±1%, or ±0.5% of a certain value or range. And the numerical values appearing in the present application, the numerical range, should be understood to include the systematic errors that are unavoidable in industrial production.

[0065] In the present application, "some specific / preferred embodiments", "other specific / preferred embodiments", "embodiments", and the like mean that the specific elements (for example, features, structures, properties, and / or characteristics) described in relation to the embodiments are included in at least one embodiment described herein, and can be present in other embodiments or can not be present in other embodiments. In addition, it should be understood that the elements can be combined in various embodiments in any suitable manner.

[0066] In the present application, the unit names used are international standard unit names, and if not specifically stated, "%" used means weight or mass percentage content.

[0067] 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 in such a trace amount as to have no function.

[0068] In the present application, "infants" means a group of humans under 3 years of age, which includes infants from 0 to 6 months of age, older infants from 6 to 12 months of age, and toddlers from 12 to 36 months of age.

[0069] 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.

[0070] I. Nutritional composition

[0071] The present application provides a nutritional composition comprising essential active ingredients as shown in (i) and (ii) below: (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 lacto-N-neotetraose to 2'-fucosyllactose is 1:(1-9).

[0072] 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-galactose via an α(1→2) bond. Its molecular formula is C 18 H 32 O 15 .

[0073] Lacto-N-neotetraose (LNnT) is a linear tetrasaccharide consisting of D-galactose, N-acetylglucosamine, D-galactose and D-glucose, with the molecular arrangement being D-galactose linked to N-acetylglucosamine (GlcNAc) via a β-(1→4) bond, then to D-galactose via a β-(1→3) bond, and to D-glucose via a β-(1→4) bond. Its molecular formula is C 26 H 45 NO 21 .

[0074] The present application does not make special limitations on the source of 2'-FL and LNnT, and typically, they can be obtained by means of ordinary chemical synthesis methods, microbial fermentation, etc. in the art.

[0075] In some embodiments, in the nutritional composition of the present application, the (i) neutral fucosylated human milk oligosaccharides and (ii) neutral non-fucosylated human milk oligosaccharides are the main effective components (active components), i.e. the nutritional composition of the present application mainly relies on the (i) neutral fucosylated human milk oligosaccharides and (ii) neutral non-fucosylated human milk oligosaccharides contained therein to exert a specific physiological activity function, such as improving the intestinal microecological imbalance of offspring caused by maternal immune activation. In other words, in some embodiments, the active components (components for exerting a specific physiological activity function, i.e. components for exerting the effect of improving the intestinal microecological imbalance of 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 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:(1-9).

[0076] In some specific embodiments, in the nutritional composition of the present application, the 2'-fucosyllactose in the (i) neutral fucosylated human milk oligosaccharides and the lacto-N-neotetraose in the (ii) neutral non-fucosylated human milk oligosaccharides are the main effective components (active components), i.e. the nutritional composition of the present application mainly relies on the 2'-fucosyllactose and lacto-N-neotetraose contained therein to exert a specific physiological activity function, such as improving the intestinal microecological imbalance of offspring caused by maternal immune activation. In other words, in some embodiments, the active components (components for exerting a specific physiological activity function, i.e. components for exerting the effect of improving the intestinal microecological imbalance of offspring caused by maternal immune activation) of the nutritional composition consist of 2'-fucosyllactose and lacto-N-neotetraose, and in the nutritional composition, the mass ratio of the lacto-N-neotetraose to the 2'-fucosyllactose is 1:(1-9).

[0077] The present application finds that, compared with neutral fucosylated human milk oligosaccharides such as 2'-fucosyllactose and neutral non-fucosylated human milk oligosaccharides such as lacto-N-neotetraose alone, the nutritional composition provided by the present application can more effectively improve the intestinal microecological imbalance of offspring caused by maternal immune activation, i.e. there is a synergistic effect between the two.

[0078] In some embodiments, the nutritional composition comprises an active ingredient (an ingredient for exerting a specific physiological activity, i.e., an ingredient for exerting an effect of ameliorating the intestinal microflora imbalance of the offspring caused by maternal immune activation) and a non-active ingredient (a substance that does not exert an effect of ameliorating the intestinal microflora imbalance of the offspring caused by maternal immune activation). Illustratively, the non-active ingredient can be other nutrients, any food-acceptable adjuvant, and / or a substance that is concomitantly produced in the production or acquisition of the active ingredient and cannot be effectively separated from the active ingredient or does not need to be separated. In some embodiments, the nutritional composition consists of the active ingredient and the non-active ingredient.

[0079] In some embodiments, the neutral fucosylated human milk oligosaccharide comprised in the nutritional composition further comprises any one or more of 3-fucosyllactose and difucosyllactose.

[0080] In other embodiments, the nutritional composition is substantially free of neutral fucosylated human milk oligosaccharides other than 2'-fucosyllactose.

[0081] In some embodiments, the neutral non-fucosylated human milk oligosaccharide comprised in the nutritional composition further comprises lacto-N-tetraose.

[0082] In other embodiments, the nutritional composition is substantially free of neutral non-fucosylated human milk oligosaccharides other than lacto-N-neotetraose.

[0083] In some embodiments, the nutritional composition comprises the necessary active ingredients as shown in (i) and (ii) below: (i) a neutral fucosylated human milk oligosaccharide, (ii) a neutral non-fucosylated human milk oligosaccharide; 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 : (1-9).

[0084] In some embodiments, the nutritional composition consists of 2'-fucosyllactose and lacto-N-neotetraose; and, in the nutritional composition, the mass ratio of the lacto-N-neotetraose to the 2'-fucosyllactose is 1 : (1-9).

[0085] 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, 1:8.0, 1:8.1, 1:8.2, 1:8.3, 1:8.4, 1:8.5, 1:8.6, 1:8.7, 1:8.8, 1:8.9, or 1:9.0, 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).

[0086] The present application does not make special limitation to the form of the nutritional composition, which can typically be liquid or 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 powder.

[0087] II. Use of the nutritional composition

[0088] The present application proposes that the combination of 2'-fucosyllactose and lacto-N-neotetraose in a certain ratio can improve the intestinal microecological imbalance of offspring caused by maternal immune activation, which is beneficial to the development of offspring's neural health through the gut-brain axis, and the two substances have a synergistic effect.

[0089] The "maternal immune activation" described in the present application mainly refers to the abnormal activation state of the immune system of a female during pregnancy, which can include immune homeostasis disorders such as inflammatory factor level disorders. Factors that can cause the immune system of a female to be abnormally activated during pregnancy include, but are not limited to, infection by bacteria, viruses, and the like.

[0090] The "intestinal microecological imbalance" described in the present application mainly refers to an imbalance state caused by changes in the composition of intestinal flora, changes in bacterial metabolic activity, or changes in the distribution of flora, as well as abnormal changes in normal intestinal structure (such as the thickness of the intestinal muscle layer, crypt structure, and the like) and physiological function (such as barrier function, and the like).

[0091] The improvement of the intestinal microecological imbalance of the offspring caused by maternal immune activation described in the present application is not for the purpose of treating or preventing diseases, and at the same time, the intestinal microecological imbalance described in the present application has not reached a level that can be recognized as a disease.

[0092] Based on this, the present application provides the use of the above-mentioned nutritional composition in the preparation of a food for improving the intestinal microecological imbalance of the offspring caused by maternal immune activation.

[0093] In some embodiments, the improvement of the intestinal microecological imbalance of the offspring caused by maternal immune activation includes the following (a1):

[0094] (a1) improving the decrease in the abundance of beneficial genera in the intestine of the offspring caused by maternal immune activation, the beneficial genera including any one or more of Lactobacillus, Limosilactobacillus, and Butyricicoccus.

[0095] In some specific embodiments, the improvement of the intestinal microecological imbalance of the offspring caused by maternal immune activation includes improving the decrease in the abundance of the beneficial genus Lactobacillus in the intestine of the offspring caused by maternal immune activation.

[0096] In some specific embodiments, the improvement of the intestinal microecological imbalance of the offspring caused by maternal immune activation includes improving the decrease in the abundance of the beneficial genus Limosilactobacillus in the intestine of the offspring caused by maternal immune activation.

[0097] In some specific embodiments, the improvement of the intestinal microecological imbalance of the offspring caused by maternal immune activation includes improving the decrease in the abundance of the beneficial genus Butyricicoccus in the intestine of the offspring caused by maternal immune activation.

[0098] In some specific embodiments, the improving the offspring intestinal dysbiosis caused by maternal immune activation comprises improving the abundance of the genus Lactobacillus in the offspring intestine caused by maternal immune activation.

[0099] In some specific embodiments, the improving the offspring intestinal dysbiosis caused by maternal immune activation comprises improving the abundance of the genus Lactobacillus and Butyricicoccus in the offspring intestine caused by maternal immune activation.

[0100] In some specific embodiments, the improving the offspring intestinal dysbiosis caused by maternal immune activation comprises improving the abundance of the genus Lactobacillus and Butyricicoccus in the offspring intestine caused by maternal immune activation.

[0101] In some specific embodiments, the improving the offspring intestinal dysbiosis caused by maternal immune activation comprises improving the abundance of the genus Lactobacillus, Limosilactobacillus and Butyricicoccus in the offspring intestine caused by maternal immune activation.

[0102] In some more specific embodiments, the improving the abundance of the genus Lactobacillus in the offspring intestine caused by maternal immune activation comprises increasing the abundance of Lactobacillus johnsonii.

[0103] In some more specific embodiments, the improving the abundance of the genus Lactobacillus in the offspring intestine caused by maternal immune activation comprises increasing the abundance of Lactobacillus johnsonii.

[0104] Studies have shown that Lactobacillus johnsonii strains can significantly improve the cognitive dysfunction of mice exposed to high altitude by regulating the intestinal flora and antioxidant capacity (Zhao Z, Zhang X, Sun N, Duan L, Xin J, Li H, Ni X, Wang H, Ma H, Bai Y. Lactobacillus johnsonii HL79 modulates the microbiota-gut-brain axis to protect cognitive function in mice chronically exposed to high altitude. Front Microbiol. 2025 Mar 7; 16: 1561400.).

[0105] Studies have shown that treatment with Lactobacillus reuteri can correct social impairment by transmitting signals from vagal neurons to the microbiota-gut-brain axis (Agirman G, Yu KB, Hsiao EY. Signaling inflammation across the gut-brain axis. Science. 2021 Nov 26; 374(6571): 1087-1092.).

[0106] Studies have shown that Butyricicoccus, a butyric acid-producing bacteria, is reduced in the feces of Alzheimer's patients, and increasing the abundance of Butyricicoccus may help prevent Alzheimer's disease (Ling Z, Zhu M, Yan X, Cheng Y, Shao L, Liu X, Jiang R, Wu S. Structural and Functional Dysbiosis of Fecal Microbiota in Chinese Patients With Alzheimer's Disease. Front Cell Dev Biol. 2021 Feb 4; 8: 634069.).

[0107] In some embodiments, the improvement of the intestinal microecological imbalance of the offspring caused by maternal immune activation comprises (a2) as follows:

[0108] (a2) improving the decrease in the content of short-chain fatty acids in the intestine of the offspring caused by maternal immune activation, the short-chain fatty acids including any one or more of acetic acid, propionic acid, and butyric acid.

[0109] In some embodiments, the improving the offspring intestinal microecological imbalance caused by maternal immune activation comprises improving the decrease in acetate content in the offspring intestine caused by maternal immune activation.

[0110] In some embodiments, the improving the offspring intestinal microecological imbalance caused by maternal immune activation comprises improving the decrease in propionate content in the offspring intestine caused by maternal immune activation.

[0111] In some embodiments, the improving the offspring intestinal microecological imbalance caused by maternal immune activation comprises improving the decrease in butyrate content in the offspring intestine caused by maternal immune activation.

[0112] In some embodiments, the improving the offspring intestinal microecological imbalance caused by maternal immune activation comprises improving the decrease in acetate and propionate content in the offspring intestine caused by maternal immune activation.

[0113] In some embodiments, the improving the offspring intestinal microecological imbalance caused by maternal immune activation comprises improving the decrease in acetate and butyrate content in the offspring intestine caused by maternal immune activation.

[0114] In some embodiments, the improving the offspring intestinal microecological imbalance caused by maternal immune activation comprises improving the decrease in propionate and butyrate content in the offspring intestine caused by maternal immune activation.

[0115] In some embodiments, the improving the offspring intestinal microecological imbalance caused by maternal immune activation comprises improving the decrease in acetate, propionate and butyrate content in the offspring intestine caused by maternal immune activation.

[0116] It has been shown that short-chain fatty acids in the intestine can regulate microglia-mediated inflammatory response through the gut-brain axis, improve neurodegenerative diseases such as Alzheimer's disease, autism, Parkinson's disease, and short-chain fatty acids can also participate in the regulation of neurotransmitter synthesis, thereby affecting the mood of the brain.

[0117] In some embodiments, the improving the offspring intestinal microecological imbalance caused by maternal immune activation comprises any one or more of the following (a3):

[0118] (a3) improving the damage to the structure and / or function of the offspring intestine caused by maternal immune activation;

[0119] The improving the damage to the structure and / or function of the offspring intestine caused by maternal immune activation comprises any one or more of the following (b1)-(b3):

[0120] (b1) improving the decrease in the muscle layer thickness of the offspring intestine caused by maternal immune activation;

[0121] (b2) ameliorating the decrease in intestinal crypt depth in the offspring caused by maternal immune activation;

[0122] (b3) ameliorating the decrease in the amount of salivary mucin secretion by intestinal goblet cells in the offspring caused by maternal immune activation.

[0123] In some specific embodiments, the ameliorating the intestinal dysbiosis in the offspring caused by maternal immune activation comprises ameliorating the damage to the structure and function of the intestine in the offspring caused by maternal immune activation.

[0124] In some specific embodiments, the ameliorating the intestinal dysbiosis in the offspring caused by maternal immune activation comprises ameliorating the decrease in the muscularis thickness of the intestine in the offspring caused by maternal immune activation.

[0125] In some specific embodiments, the ameliorating the intestinal dysbiosis in the offspring caused by maternal immune activation comprises ameliorating the decrease in intestinal crypt depth in the offspring caused by maternal immune activation.

[0126] In some specific embodiments, the ameliorating the intestinal dysbiosis in the offspring caused by maternal immune activation comprises ameliorating the decrease in the amount of salivary mucin secretion by intestinal goblet cells in the offspring caused by maternal immune activation.

[0127] In some specific embodiments, the ameliorating the intestinal dysbiosis in the offspring caused by maternal immune activation comprises ameliorating the decrease in the muscularis thickness of the intestine and the decrease in intestinal crypt depth in the offspring caused by maternal immune activation.

[0128] In some specific embodiments, the ameliorating the intestinal dysbiosis in the offspring caused by maternal immune activation comprises ameliorating the decrease in the muscularis thickness of the intestine and the decrease in the amount of salivary mucin secretion by intestinal goblet cells in the offspring caused by maternal immune activation.

[0129] In some specific embodiments, the ameliorating the intestinal dysbiosis in the offspring caused by maternal immune activation comprises ameliorating the decrease in intestinal crypt depth and the decrease in the amount of salivary mucin secretion by intestinal goblet cells in the offspring caused by maternal immune activation.

[0130] In some specific embodiments, the ameliorating the intestinal dysbiosis in the offspring caused by maternal immune activation comprises ameliorating the decrease in the muscularis thickness of the intestine, the decrease in intestinal crypt depth, and the decrease in the amount of salivary mucin secretion by intestinal goblet cells in the offspring caused by maternal immune activation.

[0131] Studies have shown that normal intestinal structure and barrier can interact with the brain through pathways such as neural transmission, mechanical signals, and inflammation, and abnormal changes in intestinal structure and barrier play an important role in neurodegenerative diseases and mental disorders.

[0132] In some specific embodiments, the improvement of the intestinal microflora imbalance of the offspring caused by maternal immune activation comprises (a1)-(a3) described above; in these three aspects, lacto-N-neotetraose and 2'-fucosyllactose can significantly amplify the improvement of each other.

[0133] As can be seen, numerous studies show that intestinal microorganisms and their metabolites, as well as intestinal structure and barrier function, can affect the central nervous system through endocrine, immune pathways and other ways, and are directly related to the structural development of the nervous system, and affect brain function. Experimental data show that 2'-FL and LNnT have an improvement effect on the imbalance of the intestinal microflora of the offspring caused by maternal immune activation. Based on this, the present application believes that by making the offspring produced by a female who has experienced or is in a state of maternal immune activation consume the above-mentioned nutritional composition, not only is beneficial to the stable and healthy intestinal microflora, but also can achieve the effect of assisting in neural development through the gut-brain axis regulation. Or, it can be considered that by making the offspring produced by a female who has experienced or is in a state of maternal immune activation consume the above-mentioned nutritional composition, the intestinal microflora imbalance accompanied by abnormal neural development in the offspring can be improved.

[0134] The food described in the present application exerts the above-mentioned effects after being consumed by the offspring (offspring produced by a female who has experienced or is in a state of maternal immune activation).

[0135] For specific foods containing or prepared using the above-mentioned nutritional composition, the present application does not make special limitations.

[0136] In some embodiments, the form of the food described in the present application is liquid or solid at room temperature.

[0137] In some embodiments, the food described in the present application is an infant formula or an infant complementary food; for example, it can be an infant formula milk powder, a children's snack, etc.

[0138] In some embodiments, the food described in the present application is a confectionery, such as a hard candy, a gel candy, a crisp candy, a pressed candy, and an aerated candy, etc.

[0139] In some embodiments, the food described in the present application is a beverage, such as a carbonated beverage, a tea beverage, a coffee beverage, a fruit and vegetable juice beverage, and a lactic acid bacteria beverage, etc.

[0140] In some embodiments, the food described in the present application is a dairy product, such as milk powder, cheese, yogurt, liquid milk, etc.

[0141] In some embodiments, the food described in the present application is a baked food, such as bread, cake, and biscuit, etc.

[0142] 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 solution, a pill, a granule, a powder, and the like

[0143] 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, and the like.

[0144] 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:(1-9); 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).

[0145] The present application does not particularly limit the specific absolute content of the lacto-N-neotetraose and the 2'-fucosyllactose in the food product, which meets the requirements of the local food-related laws and regulations.

[0146] In some embodiments, in the food product, the content of the 2'-fucosyllactose is 0.4g / 100g-2g / 100g, and the content of the lacto-N-neotetraose is 0.1g / 100g-0.5g / 100g.

[0147] In addition to the above-described components in the nutritional composition, the food product can further comprise other ingredients, such as common food ingredients, e.g., proteins / amino acids, carbohydrates, fats, vitamins, minerals, and the like.

[0148] In addition, according to the type of food product and the final needs of the target object, in some embodiments, the food product further comprises 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.

[0149] For the plant product ingredient, 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.

[0150] For the animal milk product ingredient, examples include fresh milk derived from cows, sheep, and the like, and reprocessed milk products such as whole milk powder, skim milk powder, concentrated whey protein powder, desalted whey powder, whey protein powder, hydrolyzed whey protein powder, and casein powder.

[0151] For the animal meat product ingredient, examples include meat product ingredients derived from pigs, cows, sheep, aquatic animals, or birds.

[0152] For the functional additive ingredient, examples include vitamin supplements, mineral supplements, nucleotide supplements, dietary fibers, functional polyunsaturated fatty acid supplements, and the like.

[0153] For any acceptable adjuvant, examples include solvents, antioxidants, antibacterial agents, thickening agents, diluents, co-solvents, stabilizers, emulsifiers, fillers, disintegrants, lubricants, coating materials, anti-caking agents, flavoring agents, sweetening agents, food essences, food colorants, and the like.

[0154] Examples

[0155] Hereinafter, embodiments of the present application will be described in detail with reference to Examples, but those skilled in the art will appreciate that the following Examples are for illustration only and should not be taken as limiting the scope of the present application. In the Examples, unless otherwise specified, the procedures were carried out under conventional conditions or under conditions recommended by the manufacturer. The materials or instruments used were commercially available conventional products unless otherwise specified.

[0156] Experimental Example 1: Effects of nutrients on growth indicators of MIA offspring mice

[0157] 1. Experimental Methods

[0158] 1.1 Construction of maternal immune homeostasis disorder model

[0159] Select 150 female mice and 30 male mice, the female and male mice are mainly used for offspring breeding. When caging, select small 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 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 a PBS solution, and the concentration is 5 mg / mL) into the abdominal cavity for constructing a maternal immune homeostasis disorder model, that is, a maternal immune activation model.

[0160] 1.2 Offspring intervention

[0161] During the experiment, the offspring male mice were used as experimental objects. After the maternal female mice delivered for 3 weeks, the offspring male mice were weaned and fed separately. When the offspring male mice were 4 weeks old, they were respectively given PBS solution or different proportions of HMOs solution by gavage, and the gavage period was 6 weeks.

[0162] 1.3 Intervention grouping of offspring animals

[0163] A total of 11 groups were set in the experiment, each group having 12 mice, which were control group, 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 was fed in a standard cage and placed in an environment with 12 h light / dark cycle, humidity 50±15%, temperature 22±2℃, and the mice were free to eat and drink water.

[0164] 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 intestinal microecology was investigated. The experimental design is shown in Table 1. According to the literature 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 a comparative example, and LNnT and 2'-FL are combined in different dose combinations to form four different proportion combinations of LNnT:2'-FL as 1:1, 1:2, 1:3.4 and 1:6 as four embodiments.

[0165] Table 1 Experimental intervention scheme design

[0166]

[0167] 1.4 Basic growth index monitoring

[0168] During the 6-week intervention, the body weight, food intake and water intake of the mice were recorded once a week.

[0169] 2. Experimental results

[0170] The body weight change data of the mice in the control group, model group and 9 intervention groups during the 6-week intervention are shown in Table 2. As can be seen from the table, the body weights of the mice in each group on day 0 of the intervention were not significantly different, and the average body weight of the mice in the model group was the lowest, 11.85 g, but after significant difference analysis, it was found that the average body weight of the mice in the control group, model group, 5 comparative groups and 4 example groups was not significantly different (p>0.05). With the extension of the intervention time, the average body weight of the 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, respectively. Finally, it was found that the average body weight of the mice in the 11 groups at each time point was not significantly different (p>0.05); at the same time, the weight gain of the 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 the 11 groups, including the model group and the mice after intervention with different HMOs monomers or combinations, did not affect the normal increase of the body weight of the mice, and there was no significant difference between the groups (p>0.05).

[0171] At the same time, the food intake and water intake of the mice in the 11 groups 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 the mice in the 11 groups during the entire intervention period was about 3 g / day, and after significant difference analysis, it was found that the average daily food intake of the mice in Comparative Example 1, Comparative Example 4 and Comparative Example 5 was significantly higher than that of the control group (p<0.05), and the food intake of the mice in the other groups was not significantly different from that of the control group (p>0.05). At the same time, the water intake of the mice in each group was recorded and compared, and it was found from Table 3 that the water intake of the mice in the 11 groups was not significantly different (p>0.05).

[0172] Table 2 Body weight change of mice in each group during the 6-week intervention (g)

[0173]

[0174] Table 3 Comparison of average daily food intake and water intake of mice in each group during the intervention period

[0175]

[0176] Experimental Example 2: Effect of nutrients on intestinal microorganisms in the intestinal microecosystem of MIA offspring

[0177] 1. Experimental method

[0178] 1.1 Construction of maternal immune homeostasis disorder model

[0179] The same as Example 1.

[0180] 1.2 Offspring intervention

[0181] As in Example 1.

[0182] 1.3 Offspring animal intervention grouping

[0183] As in Example 1.

[0184] 1.4 Intestinal flora detection

[0185] Intestinal flora detection was performed using 16S rRNA amplicon sequencing. First, DNA extraction kits were used to extract the intestinal content DNA of mice in each group, and the extracted genomic DNA was detected by 1% agarose gel electrophoresis to determine the concentration and purity; the 3rd to 4th regions (V3-V4 region) in the 16S rDNA hypervariable region were used as the target sequencing fragments, and the corresponding primers were designed to amplify the target fragments by PCR. The PCR products were quantified using a QuantiFluor-ST fluorometer, and then mixed in the appropriate proportion according to the sequencing requirements of each sample. After the quantitative and uniformization of the PCR amplification products, the samples were sequenced using the Illumina Miseq platform, and the sequencing analysis was outsourced to the platform of Guangzhou Kideko Biological Technology Co., Ltd. The raw data obtained by sequencing was analyzed using R language tools and related analysis software for bioinformatics analysis and data statistical analysis.

[0186] 2. Experimental results

[0187] First, the intestinal microorganisms of the offspring mice after maternal immune activation were analyzed. The results showed that the a-diversity of the intestinal flora of the model group mice was significantly different from that of the control group, such as the Shannon index and the Simpson index. On the β-diversity, principal coordinate analysis (PCoA), non-metric multidimensional scaling analysis (NMDS), and partial least squares discriminant analysis (PLS-DA) found that the intestinal flora of the model group and the control group mice showed significant independence. It can be seen that the maternal immune activation has a significant effect on the intestinal flora of the offspring. Further analysis found that the intestinal flora of the mice after maternal immune activation showed significant differences in the genus and species levels.

[0188] Tables 4 and 5 show the abundance changes of three representative different flora at the genus level in the intestinal flora of mice in each group. As can be seen from the tables, the three genera were not detected or had very low detection in the offspring after maternal immune activation (model group).

[0189] Firstly, the results of Table 4 and Table 5 show that the abundance of g_Lactobacillus in the intestines of the offspring (model group) without intervention after maternal immune activation is reduced to 1 / 10 of the control group. This bacterium belongs to lactic acid bacteria and has multidimensional regulation on human health: 1) promoting intestinal health by inhibiting pathogens, enhancing intestinal barrier function, and relieving diarrhea; 2) regulating the immune system by activating innate immunity and regulating adaptive immunity; 3) having metabolic regulation and chronic disease prevention and control functions such as improving obesity and insulin resistance, and reducing cholesterol; 4) improving and relieving anxiety and depression through the gut-brain axis, and having potential effects on delaying the occurrence and development of neurodegenerative diseases. Such an important microorganism is significantly reduced in the intestines of the offspring produced after maternal immune activation, and the abundance of g_Lactobacillus is improved after intervention with 2’-FL and LNnT in single or complex compositions. After difference significance analysis, it is found that the abundance of g_Lactobacillus in the intestines of the offspring mice in Comparative Examples 1-5 and Examples 1-4 is significantly higher than that in the model group (p<0.01 for Comparative Example 1, and p<0.0001 for the rest). Further comparison shows that the abundance of g_Lactobacillus after intervention with LNnT and 2’-FL in 1:1, 1:2, 1:3.4 and 1:6 complex is significantly higher than that in the five comparative examples (p<0.0001, p<0.001, p<0.01 or p<0.05), and significantly higher than that in the control group (p<0.0001). Therefore, 2’-FL alone can improve the abundance of g_Lactobacillus in the intestines of the offspring of maternal immune activation mice, and the addition of LNnT can significantly amplify the increment effect of 2’-FL on g_Lactobacillus. Similarly, 2’-FL can also significantly amplify the increment effect of LNnT on g_Lactobacillus. In terms of improving the abundance of g_Lactobacillus, 2’-FL and LNnT have a synergistic effect. At the same time, the abundance of Example 2 is the highest among the four examples, which can reach 12.46, and is significantly higher than that of Example 1 (p<0.0001), Example 3 (p<0.0001) and Example 4 (p<0.01).

[0190] Secondly, the results of Table 4 and Table 5 show that g_Limosilactobacillus is also significantly reduced in the intestinal tract of model mice (the average abundance of the control group is 0.93, while that of the model group is reduced to about 0.041 on average). This bacterium also belongs to the Lactobacillaceae family, and is independently classified into a genus due to its unique genetic and metabolic characteristics. It can ferment carbohydrates to generate D-lactic acid, thereby inhibiting pathogenic bacteria, and can also produce acetic acid, propionic acid and other substances to support the energy metabolism of intestinal epithelial cells and strengthen the barrier function; it can also regulate Th1 / Th2 balance, inhibit the NF-κB pathway, enhance mucosal immunity, and perform immunoregulation and anti-inflammation. The results of Table 4 and Table 5 show that the abundance of g_Limosilactobacillus is improved after the intervention of 2’-FL and LNnT in monomer or complex composition. After significant difference analysis, it is found that the abundance of g_Limosilactobacillus in the intestinal tract of the offspring mice in Comparative Examples 1-5 (p<0.01 or p<0.05) and Examples 1-4 (p<0.0001) is significantly higher than that in the model group. Further comparison shows that the abundance of g_Limosilactobacillus after the intervention of LNnT and 2’-FL in 1:1, 1:2, 1:3.4 and 1:6 complex is significantly higher than that in the five comparative examples (p<0.0001, p<0.001, p<0.01 or p<0.05). It can be seen that 2’-FL monomer can improve the abundance of g_Limosilactobacillus in the intestinal tract of the offspring of the immune-activated mother mice, and the addition of LNnT can significantly amplify the increment effect of 2’-FL on g_Limosilactobacillus. Similarly, 2’-FL can also significantly amplify the increment effect of LNnT on g_Limosilactobacillus. In terms of improving the abundance of g_Limosilactobacillus, 2’-FL and LNnT have a synergistic effect. At the same time, the abundance of Example 2 in the four examples is the highest, which can reach 2.99, and is significantly higher than that of Example 1 (p<0.0001), Example 3 (p<0.0001) and Example 4 (p<0.01).

[0191] Meanwhile, Tables 4 and 5 show the differences in *g. butyricicoccus* (a genus of bacteria in the phylum Firmicutes and class Clostridia) between groups. This bacterium has attracted much attention due to its ability to produce butyric acid. Butyric acid is one of the important short-chain fatty acids in the gut, playing a crucial role in maintaining intestinal homeostasis, energy metabolism, and immune regulation. Studies have also found that the abundance of this bacterium is significantly reduced in the intestines of patients with inflammatory bowel disease, and supplementation with *Butyricicoccus pullicaecorum* can alleviate colitis symptoms and repair mucosal damage. Furthermore, it plays an important role in the prevention and control of metabolic diseases; for example, *g. butyricicoccus* can protect the liver by reducing hepatic lipid deposition and oxidative stress. Tables 4 and 5 show that the abundance of this bacterium in the intestines of the offspring in the model group was almost zero. However, after intervention with 2'-FL and LNnT, two human milk oligosaccharides, either as monomers or in combination, the abundance of *g_Butyricicoccus* was increased. After significant difference analysis, it was found that the abundance of *g_Butyricicoccus* in the intestines of offspring mice in Comparative Examples 1-5 and Examples 1-4 was significantly higher than that in the model group (except for Comparative Example 1, p < 0.05, all others p < 0.0001). Further comparison revealed that after intervention with LNnT and 2'-FL in ratios of 1:1, 1:2, 1:3.4, and 1:6, the abundance of *g_Butyricicoccus* reached approximately 0.1, significantly higher than the five comparative examples (p < 0.0001, p < 0.001, p < 0.01, or p < 0.05). Therefore, 2'-FL monomer can increase the abundance of g_Butyricicoccus in the intestines of offspring mice with maternally activated immune systems. The addition of LNnT significantly amplifies the increasing effect of 2'-FL on g_Butyricicoccus, and similarly, 2'-FL can significantly amplify the increasing effect of LNnT on g_Butyricicoccus. There is a synergistic effect between 2'-FL and LNnT in increasing g_Butyricicoccus abundance. Furthermore, in the four examples, Example 2 showed the highest abundance, reaching 0.16, which was significantly higher than Example 1 (p<0.0001), Example 3 (p<0.0001), and Example 4 (p<0.0001).

[0192] Table 4. Comparison of genus-level abundance of differentially expressed bacterial communities in the gut of offspring mice from different groups.

[0193]

[0194] Table 5. p-values ​​after statistical analysis of differences in gut microbiota among different groups of mice.

[0195]

[0196] Further, at the species level, it was found through experiments that the abundance of two species s_Lactobacillus_johnsonii and s_Limosilactobacillus_reuteri differed among the mice in each group (results shown in Table 6), and these two species belong to g_Lactobacillus and g_Limosilactobacillus, respectively. As mentioned above, the abundance of these two genera in the intestinal tract of mice after LNnT and 2’-FL complex intervention was significantly increased.

[0197] s_Lactobacillus_johnsonii exists in breast milk and the intestinal tract of infants, and can help infants establish immune tolerance and reduce the risk of allergies and infections through vertical transmission from mother to infant. The results shown in Tables 6 and 7 show that the abundance of s_Lactobacillus_johnsonii in the offspring of the maternal immune-activated group was significantly reduced to 0.0074, but the abundance of this bacterium in the intestinal tract was increased after intervention with 2’-FL and LNnT in monomer or complex compositions. After difference significance analysis, it was found that the abundance of s_Lactobacillus_johnsonii in the intestinal tract of the offspring mice in Comparative Examples 1-5 and Examples 1-4 was significantly higher than that in the model group (p<0.0001). Further comparison found that after LNnT and 2’-FL were complexed at a ratio of 1:1, 1:2, 1:3.4 and 1:6, the abundance of s_Lactobacillus_johnsonii was all above 1, and was significantly higher than that in the five comparative examples (p<0.0001 or p<0.001). Therefore, 2’-FL monomer can increase the abundance of s_Lactobacillus_johnsonii in the intestinal tract of the offspring of the maternal immune-activated mice, and the addition of LNnT can significantly amplify the increment effect of 2’-FL on s_Lactobacillus_johnsonii, and 2’-FL can also significantly amplify the increment effect of LNnT on s_Lactobacillus_johnsonii. In terms of increasing the abundance of s_Lactobacillus_johnsonii, 2’-FL and LNnT have a synergistic effect. At the same time, the abundance of Example 2 among the four examples was the highest, reaching 2.10, and was significantly higher than that of Example 1 (p<0.0001), Example 3 (p<0.0001) and Example 4 (p<0.0001).

[0198] s_Limosilactobacillus_reuteri is also present in breast milk and can help infants establish gut microbiota, reduce the risk of necrotizing enterocolitis (NEC) and infection; it can also regulate brain development by promoting neurotransmitter synthesis, and studies have shown that supplementation with this bacterium can improve social impairment in mice. As can be seen from Tables 6 and 7, the abundance of s_Limosilactobacillus_reuteri in the offspring gut was significantly reduced to 0.031 after maternal immune activation, but the abundance of this bacterium in the gut was improved after intervention with 2’-FL and LNnT breast milk oligosaccharides in monomer or complex composition. After difference significance analysis, it was found that the abundance of s_Limosilactobacillus_reuteri in the gut of offspring mice in Comparative Examples 1-5 and Examples 1-4 was significantly higher than that in the model group (p<0.0001). Further comparison found that after LNnT and 2’-FL were complexed at a ratio of 1:1, 1:2, 1:3.4 and 1:6, the abundance of s_Limosilactobacillus_reuteri was all above 1, and was significantly higher than that in the five comparative examples (p<0.0001), and had a synergistic effect. As can be seen, 2’-FL monomer can increase the abundance of s_Limosilactobacillus_reuteri in the gut of offspring of maternal immune activation mice, and after the addition of LNnT, the increment effect of 2’-FL on s_Limosilactobacillus_reuteri can be significantly amplified, and similarly, 2’-FL can also significantly amplify the increment effect of LNnT on s_Limosilactobacillus_reuteri. In terms of improving the abundance of s_Limosilactobacillus_reuteri, 2’-FL and LNnT have a synergistic effect. At the same time, the abundance of Example 2 in the four examples is the highest, which can reach 2.07, and is significantly higher than that of Example 1 (p<0.0001), Example 3 (p<0.0001) and Example 4 (p<0.001).

[0199] From the above results, it can be seen that after LNnT and 2’-FL are complexed at a ratio of 1:1, 1:2, 1:3.4 and 1:6, the recovery of beneficial flora in the gut of offspring produced after maternal immune activation can be significantly promoted.

[0200] Table 6 Comparison of the abundance of different bacterial groups at the species level in the gut of offspring mice in each group

[0201]

[0202] Table 7 Difference significance analysis of p value after correction of bacterial groups at the species level in the gut of offspring mice

[0203]

[0204] Experimental Example 3: Effect of Nutrients on Intestinal Microbial Metabolites of Offspring of MIA Mice

[0205] 1. Experimental Methods

[0206] 1.1. Establishment of Maternal Immune Homeostasis Disorder Model

[0207] The same as Example 1.

[0208] 1.2. Intervention of Offspring

[0209] The same as Example 1.

[0210] 1.3. Grouping of Intervention of Offspring Animals

[0211] The same as Example 1.

[0212] 1.4. Sample Collection and Index Detection

[0213] 1.4.1. Collection of Fecal Samples

[0214] One day before the mice were sacrificed, the mice were placed in a high-pressure sterilized beaker individually and waited for them to defecate freely. After the mice defecated, the feces were clamped with a high-pressure sterilized forceps and transferred to a sterile EP tube, and immediately transferred to a -80°C refrigerator for detection.

[0215] 1.4.2. Determination of Short-Chain Fatty Acids in Intestines

[0216] Non-targeted metabolomics was used to determine short-chain fatty acids in mouse feces. Fecal non-targeted metabolomics testing was mainly tested by liquid chromatography-mass spectrometry and gas chromatography-mass spectrometry. The mouse fecal sample was frozen with liquid nitrogen, then ground into powder, and then extracted with an extractant. The pretreated sample was injected into the mass spectrometer instrument for positive and negative ion scanning twice. The original data file was obtained, including total ion flow chart (TIC), metabolite identification results, etc. The original data file was subjected to quality control and pretreatment, including peak alignment, denoising, normalization, etc. Appropriate statistical methods (such as PCA, PLS-DA, etc.) were used to compare and analyze the metabolite data of the experimental group and the control group, and to find the differential metabolites.

[0217] 2. Experimental Results

[0218] It can be found from the research results of Experimental Example 2 that the intestinal flora of the offspring produced after the maternal immune activation is disturbed, and in particular the abundance of beneficial flora is significantly reduced, which has a great impact on the overall intestinal microenvironment. Further, the present application analyzes the influence of breast milk oligosaccharide intervention on the production of short-chain fatty acids in the intestine. The experimental results are shown in Tables 8 to 11.

[0219] Short-chain fatty acids in the intestine are the main metabolic products after intestinal microbial fermentation, and have a variety of beneficial effects on human health. Among them, acetic acid can be absorbed by intestinal epithelial cells and enter the blood circulation through the portal vein to provide energy for peripheral tissues such as the liver and muscle. It can play an immunoregulatory role by activating G protein-coupled receptors to inhibit the release of inflammatory factors, and can also regulate appetite signals through the hypothalamus to regulate appetite. Propionic acid can inhibit the growth of pathogenic bacteria such as Salmonella, thereby protecting intestinal health, and can play an anti-inflammatory role by activating GPR41 / GPR43 to inhibit intestinal inflammation, and can also participate in the regulation of sugar metabolism, cholesterol metabolism, etc. Butyric acid is of great significance to maintaining the integrity of the intestinal barrier, and at the same time it can relieve autoimmune diseases by regulating T cell (Treg) differentiation, and can also play an important role in immune regulation by inhibiting the NF-κB pathway to reduce the production of anti-inflammatory factors.

[0220] Table 8 shows that the amount of three short-chain fatty acids, acetic acid, propionic acid and butyric acid, in the offspring intestinal tract after maternal immune activation is significantly reduced (acetic acid p<0.0001, propionic acid p<0.0001, butyric acid p<0.01). But after the intervention of LNnT and 2'-FL in monomer (comparative examples 1-5) or complex composition (examples 1-4), the three short-chain fatty acids in the intestinal tract of offspring mice are significantly increased (acetic acid p<0.0001, p<0.001 or p<0.05, propionic acid and butyric acid p<0.0001). Further analysis found that after the intervention of LNnT and 2'-FL in four ratios of 1:1, 1:2, 1:3.4 and 1:6 (examples 1-4), the production amount of short-chain fatty acids in the intestinal tract of mice is significantly higher than that of two monomers (comparative examples 1-5), and the significant difference is shown in Tables 9 to 11. As can be seen from this, 2'-FL monomer can increase the content of three short-chain fatty acids, acetic acid, propionic acid and butyric acid, in the intestinal tract of offspring of maternal immune activation mice, and after the addition of LNnT, the increment effect of 2'-FL on acetic acid, propionic acid and butyric acid can be significantly amplified, and 2'-FL can also significantly amplify the increment effect of LNnT on acetic acid, propionic acid and butyric acid. In terms of increasing the content of acetic acid, propionic acid and butyric acid, there is a synergistic effect between 2'-FL and LNnT. Further comparison of the production amount of short-chain fatty acids in the four examples found that the content of acetic acid is the highest in example 2, the content of propionic acid is the highest in example 3, and the content of butyric acid is the highest in example 2. After significant difference analysis, it was found that there was no significant difference in acetic acid and butyric acid among the four groups (p>0.05), and the propionic acid in example 2, example 3 and example 4 was significantly higher than that in example 1 (p<0.0001, p<0.0001, p<0.01).

[0221] From these results, we can see that after LNnT and 2'-FL are compounded in four ratios of 1:1, 1:2, 1:3.4 and 1:6, the production of three short-chain fatty acids in the intestinal tract of offspring can be significantly promoted.

[0222] Table 8 Production amount of short-chain fatty acids in the intestinal tract of mice in each group (μg / g)

[0223]

[0224] Table 9 Difference analysis of acetic acid production in the intestinal tract of mice in examples and comparative examples

[0225]

[0226] Table 10 Difference analysis of propionic acid production in the intestinal tract of mice in examples and comparative examples

[0227]

[0228] Table 11 Difference analysis of butyric acid production in the intestines of mice in the examples and comparative examples

[0229]

[0230] Example 4: Effect of nutrients on the intestinal tissue state and barrier function of the intestinal microecology of the offspring of MIA mice

[0231] 1. Experimental method

[0232] 1.1 Construction of maternal immune homeostasis disorder model

[0233] The same as Example 1.

[0234] 1.2 Offspring intervention

[0235] The same as Example 1.

[0236] 1.3 Grouping of intervention of offspring animals

[0237] The same as Example 1.

[0238] 1.4 Collection of tissue samples and detection of indexes

[0239] 1.4.1 Collection of tissue samples

[0240] The mice were fasted for 12 h before sacrifice, and the mice were anesthetized by intraperitoneal injection of a 1.25% tribromoethanol solution at a dose of 0.2 mL / 10 g bw, and the mice were sacrificed by cervical dislocation. Then the mouse intestinal tissue samples were collected on ice, wrapped with tin paper, quickly frozen in liquid nitrogen, and moved to a -80°C refrigerator. The tissue for pathological sectioning was placed in a 4% (v / v) paraformaldehyde solution for subsequent experiments.

[0241] 1.4.2 Quantitative analysis of intestinal morphology

[0242] Hematoxylin-eosin staining was used to analyze the thickness of the intestinal muscle layer and the depth of the crypt. The specific experimental steps are as follows:

[0243] (1) Preparation of paraffin sections

[0244] Immediately after the mice were sacrificed, the intestinal tissue was taken, cut in half, and placed in 4% (v / v) paraformaldehyde / PBS fixing solution for 24 h. After dehydration, it was embedded in paraffin, and the wax block was cut into 5 μm thin sections using a microtome. The sections were placed on the water surface at 42°C to flatten, and the APES-coated glass slides were used to collect the sections. The section holder was inserted into a 37°C incubator for drying.

[0245] (2) Hematoxylin-eosin (H&E) staining

[0246] The tissue section is dewaxed and rehydrated in xylene I for 10 min, xylene II for 10 min, 100% ethanol I for 5 min, 100% ethanol II for 5 min, 90% ethanol for 5 min, 80% ethanol for 5 min, 70% ethanol for 5 min, and 60% ethanol for 5 min. Then, the section is washed with PBS for 3 times, each for 5 min. Then, the section is stained with hematoxylin for 5 min, differentiated with 1% hydrochloric acid alcohol, and then washed with tap water for 15 min. Then, the section is counterstained with 1% diluted ammonia water, and then stained with cytoplasmic eosin for 3 min. Then, the section is dehydrated with ethanol, transparentized with xylene, and then mounted with neutral resin, and bubbles are prevented. After air-drying, the section is observed and photographed under an optical microscope.

[0247] 1.4.3 Quantitative analysis of intestinal goblet cell secreted mucin

[0248] Alcian blue staining is commonly used to observe the mucin secreted by goblet cells. The intestinal tissue section is dewaxed in xylene for 5 min, rehydrated in 100%, 90%, 80% gradient alcohol for 5 min, and distilled water for 5 min. The section is soaked in acidification solution for 3 min, and then the liquid on the section is gently shaken off. The section is stained in staining solution for 30 min, and then washed with running water for 5 min. The section is re-stained in nuclear fast red staining solution for 10 min, and then washed with running water for 3 min. The section is dehydrated in 80%, 90%, 100% gradient ethanol for 5 min, transparentized in xylene for 5 min, mounted with neutral resin, and then air-dried. The colon tissue section is observed under an optical microscope. The alcian blue staining is used for quantitative analysis of goblet cell secreted mucin by using Image J.

[0249] 2. Experimental results

[0250] The results of the experimental examples 2 and 3 show that there are significant differences between the model group and the normal control group in terms of intestinal flora and short-chain fatty acids. Since intestinal microorganisms and short-chain fatty acids can act on intestinal epithelial cells, and then affect the overall barrier function of the intestine. Therefore, the present application further investigates the state of the intestinal tissue and the barrier function, and the experimental results are shown in Tables 12 and 13.

[0251] As can be seen from Table 12 and Table 13, the thickness of the muscle layer and the depth of the crypt of the offspring after maternal immune activation are significantly reduced (p<0.0001). This shows that not only does maternal immune activation have a serious impact on the neural development of the offspring, but the intestinal tissue state is also damaged to a certain extent. However, after the intervention of LNnT and 2'-FL, the intestinal tissue state is restored. As can be seen from Table 12 and Table 13, the thickness of the muscle layer in Comparative Examples 1-5 is higher than that of the model group, but the difference significance analysis shows that the single-dose intervention of LNnT (Comparative Example 1) and the low-to-medium-dose intervention of 2'-FL (Comparative Examples 2-4) have no significant difference with the model group, and the high-dose intervention of 2'-FL (Comparative Example 5) is significantly higher than the model group; and after the intervention of LNnT and 2'-FL at a ratio of 1:1, 1:2, 1:3.4 and 1:6, the thickness of the muscle layer of the offspring mice can be further improved, and the four examples are significantly higher than the five comparative examples, and there is a synergistic effect between 2'-FL and LNnT, especially Example 2, the thickness of the muscle layer can reach about 96.98 μm, reaching the level of the control group. The crypt in the intestine can not only play a barrier function, but also is a site for absorbing nutrients. The results of Table 12 and Table 13 show that the depth of the crypt of the offspring after maternal immune activation is significantly reduced (p<0.0001) to 52.26 μm, but when LNnT and 2'-FL are intervened as single bodies, the depth of the crypt is improved to about 61 μm-67 μm, and after the intervention of LNnT and 2'-FL at a ratio of 1:1, 1:2, 1:3.4 and 1:6, the depth of the crypt of the offspring mice is further improved, and the difference significance analysis shows that the four examples are significantly higher than the five comparative examples, and there is a synergistic effect between them. Among the four examples, especially Example 2, the depth of the crypt can reach 81.28 μm. This shows that from the perspective of tissue state and barrier function, the composition formed by the combination of the two breast milk oligosaccharides can significantly improve the intestinal tissue state.

[0252] Further, the amount of mucin secreted by goblet cells in the intestinal tract of each group of pups was investigated by using Alcian Blue staining. Goblet cells and the mucin secreted by them can be considered as the first line of defense of the gastrointestinal tract barrier function, and can interact with the intestinal immune system, playing an important role in the establishment of the human immune system. The amount of mucin secreted by goblet cells in the intestinal tissue of each group of pups was quantitatively analyzed, and the amount of salivary mucin per unit tissue was used as an indicator. As can be seen from Tables 12 and 13, maternal immune activation significantly reduced the amount of salivary mucin in the intestinal tract of the offspring. However, after intervention with the nutritional composition of LNnT and 2'-FL in monomer or different proportions, this index was improved and was significantly higher than that of the model group (p<0.0001, p<0.0001 or p<0.01). Further comparison found that after intervention with LNnT and 2'-FL in 1:1, 1:2, 1:3.4 and 1:6 proportions (Examples 1-4), the amount of salivary mucin in the intestinal tract of the offspring could reach as high as 16.01 (Example 2), and was significantly higher than that of LNnT and 2'-FL monomers (Comparative Examples 1-5) (p<0.0001, p<0.0001 or p<0.01). It can be seen that 2'-FL monomer can increase the content of salivary mucin in the intestinal tract of the offspring of maternal immune activation mice, and after the addition of LNnT, the increment effect of 2'-FL on salivary mucin can be significantly amplified, and similarly, 2'-FL can also significantly amplify the increment effect of LNnT on salivary mucin. In terms of improving the content of salivary mucin, 2'-FL and LNnT have a synergistic effect.

[0253] It can be seen that after LNnT and 2'-FL are compounded in 1:1, 1:2, 1:3.4 and 1:6 proportions, the intestinal tissue state and barrier function of the offspring produced after maternal immune activation can be significantly improved.

[0254] Table 12 Comparison of intestinal tissue state and barrier function of each group of mice

[0255]

[0256] Table 13 Significant difference analysis of intestinal tissue state and barrier function of mice

[0257]

Claims

1. The use of a nutritional composition in the preparation of a food that improves the intestinal microecological imbalance 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:(1-9).

2. The use according to claim 1, characterized in that, The improvement of offspring gut microbiota imbalance caused by maternal immune activation includes the following (a1): (a1) Improves the decrease in the abundance of beneficial bacteria in the offspring gut caused by maternal immune activation, said beneficial bacteria including any one or more of Lactobacillus, Limosilactobacillus and Butyricicoccus.

3. The use according to claim 1 or 2, characterized in that, The improvement of offspring gut microbiota imbalance caused by maternal immune activation includes the following (a2): (a2) Improves the reduction of short-chain fatty acid content in the offspring gut caused by maternal immune activation, said short-chain fatty acids including any one or more of acetic acid, propionic acid and butyric acid.

4. The use according to any one of claims 1-3, characterized in that, The improvement of offspring gut microbiota imbalance caused by maternal immune activation includes the following (a3): (a3) Improves intestinal structural and / or functional damage in offspring caused by maternal immune activation; The improvement of offspring intestinal structural and / or functional impairment caused by maternal immune activation includes any one or more of the following (b1)-(b3): (b1) Improves the reduction in intestinal muscle layer thickness in offspring caused by maternal immune activation; (b2) Improves the reduction in intestinal crypt depth in offspring caused by maternal immune activation; (b3) Improves the reduction in salivary mucin secretion by offspring intestinal goblet cells caused by maternal immune activation.

5. The use according to any one of claims 1-4, characterized in that, The neutral fucosylated human milk oligosaccharide further comprises any one or more of 3-fucosylated lactose and difucosylated lactose.

6. The use according to any one of claims 1-4, characterized in that, The neutral non-fucosylated human milk oligosaccharide also contains lactose-N-tetrasaccharide.

7. The use according to any one of claims 1-6, characterized in that, The food mentioned is infant formula or infant supplementary food.

8. The use according to any one of claims 1-7, characterized in that, The food products mentioned are candies, beverages, dairy products, baked goods, or dietary supplements.

9. The use according to any one of claims 1-8, 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.

10. The use according to any one of claims 1-9, 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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