Application of active folic acid substances to regulation of intestinal flora of mother and child

Abstract

The invention belongs to the field of food, and particularly relates to application of active folic acid substances to regulation of intestinal flora of a mother and a child. The invention provides an application of active folic acid substances in preparation of food which is taken by a mother and contributes to regulation of intestinal flora of the mother and offspring of the mother; wherein the active folic acid substances are selected from 6S-5-methyltetrahydrofolic acid and salts thereof, the mother is in a gestation period and / or a lactation period, and the offspring is in a fetal period, an infant period and / or a children period. The invention accidentally finds that the active folic acid substances are taken by the mother in the gestation period, so that the intestinal flora of the mother and the offspring can be obviously improved, and the active folic acid substances can promote the increase of beneficial bacteria in the intestinal tracts of the mother in the gestation period and the lactation period and the offspring of the mother and reduce the relative content of conditioned pathogens; furthermore, by adjusting the intestinal flora, the health of nervous systems of the mother and the offspring of the mother can be maintained through the intestinal brain axis.

Description

Technical Field

[0001] The invention belongs to the field of food, and in particular relates to the use of active folic acid substances in helping to regulate the intestinal flora of mother and child. Background Art

[0002] The intestinal flora is considered to be one of the key factors in regulating human health, which directly affects the body's metabolic system, immune system, and nervous system. It is known that the intestinal flora is affected by many factors, including diet, age, lifestyle, medication, antibiotic use, and disease status. Different populations and individuals have different intestinal flora. Studies have shown that the mother's intestinal flora can be transported to the placenta through the immune lymphatic system, and can be transmitted to breast milk through the immune mammary system to further affect the development of the newborn's intestinal flora. At the same time, studies have also shown that regardless of the mode of delivery and whether breastfeeding is performed, mothers can affect the baby's intestinal flora through various channels, such as contact with the baby.

[0003] Reference document 1 (CN117882771A) discloses a nutritional composition, food and use containing bovine colostrum and bifidobacterium. Studies have shown that by supplementing bovine colostrum and / or Bifidobacterium longum BB536 during pregnancy and / or lactation, the immunity and intestinal flora (such as lactobacilli and / or bifidobacteria) of offspring fed by older pregnant women can be improved through mother-to-child transmission.

[0004] Folic acid is a B vitamin that the human body cannot synthesize on its own and must be obtained through additional intake. It is known that folic acid supplementation for mothers during pregnancy can effectively prevent the occurrence of fetal neural tube defects and reduce the incidence of placental malformations, intrauterine growth retardation, premature infants and low birth weight infants. In the process of folic acid being utilized in the body, methylenetetrahydrofolate reductase plays an important role, and the gene mutation that determines the synthesis of this enzyme can lead to the occurrence of corresponding diseases. 6S-5-Methyltetrahydrofolate, also known as active folic acid, is the most active form of folic acid formed after metabolism after entering the human body. It can be directly absorbed and utilized by the human body and is not affected by the tetrahydrofolate reductase genotype in the body. It is a better form of folic acid supplementation.

[0005] Reference 2 (CN114340635B) discloses the use of 6S-5-methyltetrahydrofolate (active folic acid) for preparing medicines or health foods for preventing congenital heart disease defects in newborns for women during the peri-pregnancy and / or pregnancy period. Studies have shown that when mothers take folic acid during pregnancy, the unmetabolized folic acid will be teratogenic to the embryo, and its long-term or excessive supplementation can induce the occurrence of congenital heart disease, while 6S-5-methyltetrahydrofolate (active folic acid) is not teratogenic and can prevent the occurrence of congenital heart disease when taken during pregnancy.

[0006] Reference document 3 (CN118252800A) discloses a method for modifying 5-methyltetrahydrofolate liposomes and its use in improving intestinal flora, wherein soybean lecithin and cholesterol are used to prepare liposomes to embed 5-methyltetrahydrofolate, and chitosan and pectin are used for step-by-step modification, and a special fermented enzymatically hydrolyzed green plum extract is used to spray-coat the modified embedded material again, and Guang Chenpi powder, Laoyaoju powder and Laojiang powder are further compounded, thereby achieving the effect of regulating intestinal microorganisms. Summary of the invention

[0007] Problem that the invention aims to solve

[0008] Existing studies believe that the initial flora in the gastrointestinal tract of newborns comes from the mother. The mother's healthy flora can pass on better flora to the offspring and affect the establishment of the offspring flora, laying a good foundation for the healthy establishment of the offspring flora. Therefore, the present invention believes that improvements should be made from the source, that is, intervention should be carried out from the pregnancy period to improve the health of the mother during pregnancy and lactation, especially to improve the health of the mother's flora, thereby regulating the offspring flora and improving the health of the offspring. This improvement method is better than the method of intervening after the fetus is born. However, existing studies have shown that even if nutrients are considered to be directly given to the body to exert specific health benefits, they may not necessarily promote the health of offspring when consumed by the mother.

[0009] Although the above-mentioned reference 3 discloses a use of 5-methyltetrahydrofolate liposomes to improve intestinal flora, such an effect is not based on 5-methyltetrahydrofolate alone, but on the combination of 5-methyltetrahydrofolate liposomes sprayed with special fermented enzymatic green plum extract and Guang Chenpi powder, Lao Yaoju powder and Lao Jiang powder. At present, no study has ever shown the effect of direct supplementation of active folic acid substances by the mother on her own and her offspring's intestinal flora.

[0010] In the process of in-depth research on the physiological functions of active folic acid substances, the present invention unexpectedly discovered that mothers who start taking active folic acid substances during pregnancy can significantly improve their intestinal flora and can also improve the intestinal flora of their offspring at the same time. Active folic acid substances can promote the increase of beneficial bacteria in the intestines of pregnant and lactating mothers and reduce the relative content of conditional pathogens. At the same time, they can increase the content of probiotics in the intestines of their offspring and inhibit the relative abundance of conditional pathogens in the intestines of their offspring.

[0011] Therefore, the object of the present invention is to provide the use of active folic acid substances for non-therapeutic purposes in simultaneously regulating the intestinal flora of pregnant and / or lactating mothers and their offspring. Furthermore, the object of the present invention is to provide the use of active folic acid substances in preparing foods that help regulate the intestinal flora of mothers and their offspring through ingestion by mothers.

[0012] Solutions for solving problems

[0013] [1] Use of active folic acid substances in the preparation of foods that help regulate the intestinal flora of the mother and her offspring through ingestion; wherein the active folic acid substances are selected from 6S-5-methyltetrahydrofolate and its salts, the mother is in pregnancy and / or lactation, and the offspring is in the fetal period, infancy, and / or childhood.

[0014] [2] The use according to [1], wherein the active folic acid substance is 6S-5-methyltetrahydrofolate, 6S-5-methyltetrahydrofolate calcium or 6S-5-methyltetrahydrofolate glucosamine salt.

[0015] [3] The use according to [1] or [2], wherein the method for helping to regulate the intestinal flora of the mother and her offspring comprises at least one of: increasing the relative abundance of Akkermansia in the intestinal tract of pregnant mothers, increasing the relative abundance of Bifidobacterium in the intestinal tract of pregnant mothers, decreasing the relative abundance of Escherichia-Shigella in the intestinal tract of pregnant mothers, and decreasing the relative abundance of Streptococcus in the intestinal tract of pregnant mothers.

[0016] [4] The use according to any one of [1] to [3], wherein the method for helping to regulate the intestinal flora of the mother and her offspring includes: increasing the relative abundance of Akkermansia in the intestines of lactating mothers, increasing the relative abundance of Clostridia UCG-014 unclassified in the intestines of lactating mothers, increasing the relative abundance of Muribaculaceae unclassified in the intestines of lactating mothers, decreasing the relative abundance of Escherichia-Shigella in the intestines of lactating mothers, and decreasing the relative abundance of Streptococcus in the intestines of lactating mothers.

[0017] [5]. The use according to any one of [1] to [4], wherein the method helps to regulate the intestinal flora of the mother and her offspring, including: increasing the relative abundance of Akkermansia in the offspring's intestine, increasing the relative abundance of Muribaculaceae unclassified in the offspring's intestine, increasing the relative abundance of Bacteroides in the offspring's intestine, decreasing the relative abundance of Escherichia-Shigella in the offspring's intestine, and decreasing the relative abundance of Streptococcus in the offspring's intestine.

[0018] [6]. The use according to any one of [1] to [5], wherein the food affects the gut-brain axis of the mother and her offspring by helping to regulate their intestinal flora, thereby helping to maintain the health of the nervous system of the mother and her offspring.

[0019] [7]. The use according to any one of [1] to [6], wherein the food is infant food, children's food, adolescent food or adult food.

[0020] [8] The use according to any one of [1] to [7], wherein the food is a beverage, candy, dairy product, baked food or dietary supplement.

[0021] [9]. The use according to any one of [1] to [8], wherein the food is in liquid form, solid-liquid mixed form, solid block form or solid powder form at room temperature.

[0022]

[10] . The use according to any one of [1] to [9], wherein the food contains any one or more of the following ingredients: plant product ingredients, animal dairy product ingredients, animal meat product ingredients, functional additives and any excipients acceptable to food.

[0023]

[11] . The use according to any one of [1] to

[10] , wherein the mass content of the active folic acid substance in the food is 0.0001% to 0.1%.

[0024] Effects of the Invention

[0025] After extensive research, the present invention proposes that supplementing a certain amount of active folic acid substances during pregnancy and / or lactation can significantly increase the abundance of beneficial bacteria in the intestines of mothers and reduce the content of harmful bacteria. At the same time, it can significantly increase the abundance of beneficial bacteria in the intestines of their offspring and reduce the content of harmful bacteria. In particular, it can regulate the abundance of some non-directly edible beneficial bacteria in the intestines of mothers and offspring, such as Akkermansia. Furthermore, regulating the intestinal flora will affect the gut-brain axis of the body. Therefore, the present invention proposes that supplementing a certain amount of active folic acid substances during pregnancy and / or lactation is also beneficial to maintaining the health of the nervous system of mothers and their offspring.

[0026] Experimental data show that supplementing mothers with active folic acid during pregnancy and lactation can significantly increase the relative abundance of Akkermansia and Bifidobacterium in the mother's intestines during pregnancy, and reduce the relative abundance of Escherichia-Shigella and Streptococcus in the mother's intestines during pregnancy; increase the relative abundance of Akkermansia, Clostridia UCG-014 unclassified, and Muribaculaceae unclassified in the mother's intestines during lactation, and reduce the relative abundance of Escherichia-Shigella and Streptococcus in the mother's intestines during lactation; at the same time, it can significantly increase the abundance of Akkermansia and Muribaculaceae in the mother's offspring's intestines. unclassified and Bacteroides, and reduce the relative abundance of Escherichia-Shigella and Streptococcus in the intestines of the mother's offspring. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 :Changes of Akkermansia in the intestines of female mice.

[0028] Figure 2 :Changes of Bifidobacterium in the intestine of female mice.

[0029] Figure 3 :Changes of Clostridia UCG-014unclassified in the intestines of female mice.

[0030] Figure 4 : Changes of Muribaculaceae unclassified in the intestine of female mice.

[0031] Figure 5 :Changes of Escherichia-Shigella in the intestines of female mice.

[0032] Figure 6 :Changes of Streptococcus in the intestines of female mice.

[0033] Figure 7 :Changes of Akkermansia in the intestine of offspring mice.

[0034] Figure 8 : Changes of Muribaculaceae unclassified in the intestine of offspring mice.

[0035] Fig. 9 :Changes of Bacteroides in the intestine of offspring mice.

[0036] Fig.10 :Changes of Escherichia-Shigella in the intestine of offspring mice.

[0037] Fig.11 :Changes of Streptococcus in the intestine of offspring mice. DETAILED DESCRIPTION

[0038] The following describes the embodiments of the present invention, but the present invention is not limited thereto. Various changes can be made within the scope of the present invention, and the embodiments and examples obtained by appropriately combining the technical means disclosed in different embodiments and examples are also included in the technical scope of the present invention.

[0039] I. Definition of Terms

[0040] In the present invention, the term "a" or "an" or "the" may mean "one", and may also mean "one or more", "at least one" and "one or more than one".

[0041] In the present invention, the numerical ranges expressed using “value A to value B”, “value A - value B”, “above value A”, “below value A”, “at least value A” and “at most value A” refer to the range including the endpoint values ​​A and B.

[0042] In the present invention, "optional" or "optionally" means that the subsequently described event or circumstance may or may not occur, and the description includes instances where the event occurs and instances where it does not occur.

[0043] In the present invention, the word "may" includes both performing a certain process and not performing a certain process.

[0044] In the present invention, the terms "comprising", "having", "including" or "containing" may mean inclusive or open-ended, and do not exclude additional, uncited elements or method steps. At the same time, "comprising", "having", "including" or "containing" may also mean closed, excluding additional, uncited elements or method steps.

[0045] In the present invention, the term "about" is used to define the numerical range and parameters of the present invention are approximate values, and the specific relevant values ​​are presented as accurately as possible. Unless otherwise expressly stated, it should be understood that all ranges, quantities, values ​​and percentages used in the present invention are modified by "about". Here, "about" usually means that the actual value is within ±5%, ±3%, ±1% or ±0.5% of a particular value or range.

[0046] In the present invention, "normal temperature" refers to an indoor ambient temperature of 23±2°C.

[0047] In the present invention, "animal milk" is used to refer to the liquid obtained from the mammary gland of a mammal in the lactation period. The term "animal milk" should be interpreted broadly and covers both raw milk (ie, liquid obtained directly from the mammary gland) and standardized milk products.

[0048] In the present invention, "gestation period" and "pregnancy period" are used interchangeably and refer to the period from fertilization to birth.

[0049] In the present invention, the "breastfeeding period" refers to the period from the start of breastfeeding to the cessation of breastfeeding after a mother gives birth.

[0050] In the present invention, "infants" refers to a human group under 36 months of age.

[0051] In the present invention, "infant" refers to a human group under 12 months of age.

[0052] In the present invention, "infants" refers to a human group of 13 to 36 months old.

[0053] In the present invention, "children" refers to a human group older than 3 years and younger than 12 years.

[0054] In the present invention, "teenagers" refers to a human group aged 12 years or older and younger than 18 years old.

[0055] In the present invention, "adult" refers to a human group of people aged 18 years or older.

[0056] In the present invention, "middle-aged and elderly" refers to a human group over the age of 45.

[0057] Unless otherwise defined, other technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0058] II. Active folic acid substances

[0059] In some embodiments, the active folate described in the present invention is selected from 6S-5-methyltetrahydrofolate and its salts.

[0060] In some specific embodiments, the active folic acid substance described in the present invention is 6S-5-methyltetrahydrofolate, 6S-5-methyltetrahydrofolate calcium or 6S-5-methyltetrahydrofolate glucosamine salt.

[0061] From the perspective of the convenience of obtaining raw materials and the universality of consumption, in some preferred embodiments, the active folic acid substance described in the present invention is 6S-5-methyltetrahydrofolate calcium.

[0062] The present invention does not particularly limit the source of the above-mentioned active folic acid substances. Typically, they can be synthesized by common chemical synthesis methods in the art, such as using folic acid as a raw material, through reduction, methylation, chiral resolution and optional salt formation steps.

[0063] II. Foods containing active folic acid (including health foods)

[0064] The food containing active folic acid substances of the present invention is suitable for people of all ages, especially for people who need to regulate intestinal flora, and especially for mothers in pregnancy and / or lactation. In some specific embodiments, the food of the present invention is infant food, children's food, adolescent food or adult food; exemplary adult food includes pregnant women food, maternal food, maternal food, middle-aged and elderly food, etc.

[0065] The food containing active folic acid substances described in the present invention can be any kind of food. In some specific embodiments, the food described in the present invention is a beverage, candy, dairy product, baked food or dietary supplement; exemplary beverages include carbonated beverages, tea beverages, coffee beverages, fruit and vegetable juice beverages and lactic acid bacteria beverages; exemplary candies include hard candies, gel candies, crisp candies, pressed candies and inflated candies; exemplary dairy products include fermented milk, cheese and milk powder; exemplary baked food includes bread, cakes and biscuits; exemplary dietary supplements include hard capsules, soft capsules, tablets, oral liquids, granules and powders.

[0066] The food containing active folic acid substances of the present invention can be in any form. In some specific embodiments, at room temperature, the food of the present invention is in liquid form, solid-liquid mixed form, solid block form or solid powder form.

[0067] In the food of the present invention, in addition to containing active folic acid substances, other arbitrary food acceptable substances may also be included, such as protein / amino acids, carbohydrates, fats, vitamins, minerals, etc., and those skilled in the art can select according to the actual food types and target populations. In some specific embodiments, the food of the present invention contains any one or more of the following ingredients: plant product ingredients, animal dairy product ingredients, animal meat product ingredients, functional additives, and any acceptable excipients in food. Exemplary plant product ingredients include various types of vegetables, fruits, grains, nuts, coffee, tea leaves and their extracts, as well as some medicinal and edible botanical Chinese medicinal materials and their extracts. Exemplary animal dairy product ingredients include fresh milk from animals, as well as whole milk powder, skim milk powder, concentrated whey protein powder, desalted whey powder, whey protein powder, hydrolyzed whey protein powder, casein powder and other reprocessed dairy products. Exemplary animal meat product ingredients include meat product ingredients of pigs, cattle, sheep, aquatic products or poultry. Exemplary functional additives include vitamin supplements, mineral supplements, nucleotide supplements, dietary fiber, functional polyunsaturated fatty acid supplements, etc. Exemplary acceptable excipients in foods include solvents, antioxidants, antibacterial agents, thickeners, diluents, cosolvents, stabilizers, emulsifiers, fillers, disintegrants, lubricants, coating materials, anti-caking agents, flavoring agents, sweeteners, edible flavors, edible colorings, and the like.

[0068] In some more specific embodiments, the food of the present invention is infant formula milk powder, baby complementary food, children's formula milk powder, children's snacks, maternal formula milk powder, pregnant woman formula milk powder, middle-aged and elderly milk powder, maternal oral liquid or pregnant woman oral liquid.

[0069] In the food of the present invention, the content of active folic acid substances meets the requirements of relevant laws and regulations. In some specific embodiments, in the food described in the present invention, the mass content of the active folic acid substance is 0.0001% to 0.1%, for example, it can be 0.0001%, 0.0002%, 0.0003%, 0.0004%, 0.0005%, 0.0006%, 0.0007%, 0.0008%, 0.0009, 0.001%, 0.002%, 0.003%, 0.004%, 0.005%, 0.006%, 0.007%, 0.008%, 0.009%, 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09% or 0.1%, etc., preferably 0.0003% to 0.01%.

[0070] III. Use of the drug to help regulate the intestinal flora of the mother and her offspring through maternal ingestion

[0071] The present invention finds that the intake of a certain amount of active folic acid substances by the mother during pregnancy and / or lactation (consumption) can help regulate the intestinal flora of the mother and her offspring, which is specifically manifested in that the relative abundance of beneficial bacteria in the intestines of the mother and her offspring can be increased and the relative abundance of conditional pathogens can be reduced. At the same time, the present invention finds that the effect of active folic acid substances ingested by the mother to regulate the intestinal flora of her offspring is not only for the fetus in the mother's body and infants who consume breast milk, but also for infants who do not consume breast milk but have full contact with the mother. At the same time, this regulatory effect can also continue to the childhood of the offspring, and even have a lasting impact on the offspring.

[0072] Based on this, the present invention provides the non-therapeutic use of active folic acid substances and foods containing active folic acid substances, which help regulate the intestinal flora of mothers and their offspring through ingestion by mothers. At the same time, the present invention provides the use of active folic acid substances in the preparation of foods that help regulate the intestinal flora of mothers and their offspring through ingestion by mothers.

[0073] In some embodiments, the mother is in pregnancy and / or lactation; preferably, the mother is in pregnancy and lactation. In some embodiments, the offspring is in the fetal period, infant period, and / or childhood; preferably, the offspring is in the fetal period and / or infant period and optional childhood; more preferably, the offspring is in the infant period and optional childhood; even more preferably, the offspring is in the infant period. In some embodiments, the offspring in the infant period can be fed with breast milk, preferably completely fed with breast milk or mainly fed with breast milk. In the present invention, "completely fed with breast milk" means that the vast majority (at least 90%, preferably at least 95%, more preferably at least 98%) of the nutrients and / or energy consumed by the offspring comes from breast milk. In the present invention, the use of "mainly fed with breast milk" means that the nutrients and / or energy consumed by the offspring mainly (at least 50%, preferably at least 65%, more preferably at least 75%) come from breast milk.

[0074] In some embodiments, the "mother" and "offspring" described in the present invention include mammals, including but not limited to humans, monkeys, gorillas, cows, sheep, cats, dogs, horses, rabbits, mice, rats or guinea pigs, etc.

[0075] The present invention finds that although the active folic acid substances are all ingested by the mother, the intestinal regulatory effects of the active folic acid substances on the mothers and offspring at different stages are not exactly the same. In some embodiments, the mother's supplementary intake of active folic acid substances during pregnancy and / or lactation can regulate different flora in the intestines of pregnant mothers, lactating mothers and their offspring respectively.

[0076] In some embodiments, the method of helping to regulate the intestinal flora of the mother and her offspring includes: increasing the relative abundance of beneficial bacteria in the intestine of the pregnant mother, decreasing the relative abundance of conditional pathogens in the intestine of the pregnant mother, increasing the relative abundance of beneficial bacteria in the intestine of the lactating mother, decreasing the relative abundance of conditional pathogens in the intestine of the lactating mother, increasing the relative abundance of beneficial bacteria in the intestine of the offspring, and decreasing the relative abundance of conditional pathogens in the intestine of the offspring.

[0077] In some embodiments, the method of helping to regulate the intestinal flora of the mother and her offspring includes: increasing the relative abundance of beneficial bacteria in the intestine of the pregnant mother, decreasing the relative abundance of conditional pathogens in the intestine of the pregnant mother, increasing the relative abundance of beneficial bacteria in the intestine of the lactating mother, decreasing the relative abundance of conditional pathogens in the intestine of the lactating mother, increasing the relative abundance of beneficial bacteria in the intestine of the offspring, and decreasing the relative abundance of conditional pathogens in the intestine of the offspring.

[0078] In some embodiments, the method helps to regulate the intestinal flora of the mother and her offspring, including: increasing the relative abundance of Akkermansia in the intestinal tract of the pregnant mother, increasing the relative abundance of Bifidobacterium in the intestinal tract of the pregnant mother, reducing the relative abundance of Escherichia-Shigella in the intestinal tract of the pregnant mother, and reducing at least one of the relative abundance of Streptococcus in the intestinal tract of the pregnant mother.

[0079] In some embodiments, the helping to regulate the intestinal flora of the mother and her offspring includes: increasing the relative abundance of Akkermansia in the intestinal tract of pregnant mothers, increasing the relative abundance of Bifidobacterium in the intestinal tract of pregnant mothers, reducing the relative abundance of Escherichia-Shigella in the intestinal tract of pregnant mothers, and reducing the relative abundance of Streptococcus in the intestinal tract of pregnant mothers.

[0080] In some embodiments, the method helps to regulate the intestinal flora of the mother and her offspring, including: increasing the relative abundance of Akkermansia in the intestine of lactating mothers, increasing the relative abundance of Clostridia UCG-014unclassified in the intestine of lactating mothers, increasing the relative abundance of Muribaculaceae unclassified in the intestine of lactating mothers, reducing the relative abundance of Escherichia-Shigella in the intestine of lactating mothers, and reducing at least one of the relative abundance of Streptococcus in the intestine of lactating mothers.

[0081] In some embodiments, the method helps to regulate the intestinal flora of the mother and her offspring, including: increasing the relative abundance of Akkermansia in the intestine of lactating mothers, increasing the relative abundance of Clostridia UCG-014unclassified in the intestine of lactating mothers, increasing the relative abundance of Muribaculaceae unclassified in the intestine of lactating mothers, reducing the relative abundance of Escherichia-Shigella in the intestine of lactating mothers, and reducing the relative abundance of Streptococcus in the intestine of lactating mothers.

[0082] In some embodiments, the method helps to regulate the intestinal flora of the mother and her offspring, including: increasing the relative abundance of Akkermansia in the offspring's intestine, increasing the relative abundance of Muribaculaceaeunclassified in the offspring's intestine, increasing the relative abundance of Bacteroides in the offspring's intestine, reducing the relative abundance of Escherichia-Shigella in the offspring's intestine, and reducing at least one of the relative abundance of Streptococcus in the offspring's intestine.

[0083] In some embodiments, the helping to regulate the intestinal flora of the mother and her offspring includes: increasing the relative abundance of Akkermansia in the offspring's intestine, increasing the relative abundance of Muribaculaceae unclassified in the offspring's intestine, increasing the relative abundance of Bacteroides in the offspring's intestine, reducing the relative abundance of Escherichia-Shigella in the offspring's intestine, and reducing the relative abundance of Streptococcus in the offspring's intestine.

[0084] Akkermansia microorganisms are Gram-negative anaerobic bacteria in the intestine. They mainly colonize in the mucus layer in the gastrointestinal tract and protect the mucus layer and connective tissue (Derrien M, Vaughan EE, Plugge CM, de Vos WM. Akkermansia muciniphila gen. nov., sp. nov., a human intestinal mucin-degrading bacterium. Int J Syst Evol Microbiol. 2004 Sep; 54 (Pt5): 1469-1476.); as a new generation of probiotics, they are a biomarker of health status (Zhang T, Li Q, Cheng L, Buch H, Zhang F. Akkermansia muciniphila is a promising probiotic. Microb Biotechnol. 2019 Nov; 12 (6): 1109-1125.). Studies have shown that Akkermansia microorganisms play an important role as probiotics in metabolic diseases (such as diabetes, obesity, etc.), intestinal diseases (such as inflammatory bowel disease, ulcerative colitis, etc.) and neuropsychiatric systems (Huang Qiaoshen, Zhang Yongkang, Wang Shuiping, et al. Research progress on the potential and mechanism of action of Akkermansia muciniphila in treating diseases [J]. Journal of Microbiology, 2023, 63(09): 3360-3373.). With the continuous deepening of research on the gut-brain axis, studies have found that Akkermansia plays an important role in a variety of neuropsychiatric diseases (such as depression and anxiety, autism spectrum disorders (ASDs), Alzheimer's disease and cognitive impairment, etc.).

[0085] Bifidobacterium microorganisms are Gram-negative anaerobic bacteria in the intestine. Studies have shown that Bifidobacterium microorganisms have a variety of probiotic functions, such as improving intestinal diseases caused by immune system disorders; inhibiting pathogens from invading the intestine; improving the activity of host antioxidant enzymes to alleviate oxidative damage to the body; improving diseases caused by intestinal flora imbalance, such as constipation and diarrhea; secreting β-galactosidase to alleviate lactose intolerance, etc. Studies have found that Bifidobacterium microorganisms have effects on memory and learning behavior, long-term potentiation (LTP) and some biochemical parameters in β-Amyloid-Induced Rat's Model of Alzheimer's Disease. Prev Nutr Food Sci. 2019 Sep; 24 (3): 265-273.).

[0086] Clostridia UCG-014unclassified is an unnamed genus under the class Clostridia. It is a group of Gram-positive anaerobic or microaerophilic bacteria, and a large number of butyrate-producing bacteria belong to this genus. Studies have found that compared with healthy people, the relative abundance of Clostridia UCG-014unclassified in the intestines of patients with thyroiditis is reduced. Studies have found that the relative abundance of Clostridia UCG-014unclassified in the intestines during pregnancy is negatively correlated with dyslipidemia during pregnancy.

[0087] Muribaculaceae unclassified is an unnamed genus under the family Muribaculaceae. Muribaculaceae is a bacterial family in the order Bacteroidetes. Muribaculaceae produces short-chain fatty acids through endogenous (mucin polysaccharides) and exogenous polysaccharides (dietary fiber). In recent years, Muribaculaceae has begun to be paid attention to and studied as a potential probiotic family. Studies have found that this family has a cross-feeding relationship with probiotics such as Bifidobacterium and Lactobacillus. Muribaculaceae can effectively decompose dietary fiber into monosaccharides and then serve as a food source for Bifidobacterium and Lactobacillus to promote the proliferation of Lactobacillus and Bifidobacterium (Zhu Y, Chen B, Zhang X, Akbar MT, Wu T, Zhang Y, ZhiL, Shen Q. Exploration of the Muribaculaceae Family in the Gut Microbiota: Diversity, Metabolism, and Function. Nutrients. 2024Aug12; 16(16): 2660.).

[0088] Bacteroides microorganisms are Gram-negative anaerobic bacteria in the intestine, mainly found in the gastrointestinal tract of humans and animals. They can participate in many important metabolic activities in the human colon, including carbohydrate fermentation, utilization of nitrogenous substances, and biotransformation of bile acids and other steroids. Studies have shown that the number of Bacteroides in the feces of children with autism is different from that of healthy controls (Liu J, Gao Z, Liu C, Liu T, Gao J, Cai Y, Fan X. Alteration of Gut Microbiota: New Strategy for Treating Autism Spectrum Disorder. Front Cell Dev Biol. 2022 Mar 3; 10: 792490.).

[0089] Escherichia-Shigella is a genus of the Enterobacteriaceae family. It is one of the common pathogenic bacteria in the intestine. It is a Gram-negative facultative anaerobic bacterium. Its growth does not require high nutrition. It can produce enterotoxins with high pathogenicity. Its large-scale reproduction can cause intestinal flora disorders and cause bacillary dysentery. Studies have shown that Escherichia-Shigella is more abundant in patients with generalized anxiety disorder and is positively correlated with the severity of anxiety (Chen YH, Bai J, Wu D, Yu SF, Qiang XL, Bai H, Wang HN, Peng ZW. Association between fecal microbiota and generalized anxiety disorder: Severity and early treatment response. J Affect Disord. 2019 Dec 1; 259: 56-66.).

[0090] Streptococcus is another common Gram-positive cocci in pyogenic cocci, which is widely distributed in the nasopharynx and gastrointestinal tract of the human body and in nature. It contains normal flora and conditional pathogens. Pathogenic streptococci can cause suppurative inflammation and hypersensitivity diseases in humans.

[0091] Based on the experimental results of the present invention and existing literature reports, the present invention believes that by allowing mothers to supplement the intake of active folic acid substances during pregnancy and / or lactation, it is not only beneficial to the health of the intestinal flora of the mother and her offspring, but also will affect the gut-brain axis of the mother and her offspring, and thus have a certain beneficial effect on the nervous system health of the mother and her offspring. The invention's help in maintaining the nervous system health of the mother and her offspring is not intended to prevent and treat diseases.

[0092] Through a large number of studies, the present invention has found that supplementary intake of active folic acid substances by mothers during pregnancy and / or lactation can simultaneously regulate some of the same flora in the intestines of pregnant and / or lactating mothers and their offspring.

[0093] In some embodiments, the method helps to regulate the intestinal flora of the mother and her offspring, including: simultaneously increasing the relative abundance of Akkermansia in the intestines of pregnant and lactating mothers and their offspring, simultaneously reducing the relative abundance of Escherichia-Shigella in the intestines of pregnant and lactating mothers and their offspring, and simultaneously reducing the relative abundance of Streptococcus in the intestines of pregnant and lactating mothers and their offspring.

[0094] In some embodiments, the method helps to regulate the intestinal flora of the mother and her offspring, including: simultaneously increasing the relative abundance of Muribaculaceae unclassified in the intestinal tract of the mother and her offspring during lactation.

[0095] Example

[0096] The embodiments of the present invention will be described in detail below in conjunction with examples, but those skilled in the art will appreciate that the following examples are only used to illustrate the present invention and should not be considered to limit the scope of the present invention. If no specific conditions are specified in the examples, they are carried out under conventional conditions or conditions recommended by the manufacturer. Unless otherwise specified, the materials or instruments used are all conventional products that can be obtained commercially.

[0097] 1. Test materials

[0098] 1.1. Experimental animals and groups

[0099] 40 8-week-old sexually mature SPF female rats and 40 SPF male rats were selected; purchased from Hangzhou Medical College, animal certificate number 20240318Aazz0100000836, production license number: SCXK (Zhejiang) 2024-002. Feeding conditions: Animals were raised in a barrier environment, with an indoor temperature of 25±1℃, humidity of 45±5%, free access to drinking water, and 12h light and dark alternation every day. This experiment passed the animal experiment ethics review of Southeast University, ethics number: 20240316003.

[0100] After 3 days of adaptive feeding, female and male rats were placed in the same cage at a ratio of 1:1. The intervention was started when the mother rat showed vaginal plugs, and the mother rats were continuously gavaged until the offspring rats were weaned. The intervention group was divided into groups and given the corresponding dose of sample aqueous solution, and the control group was given the corresponding dose of normal saline, gavaged once a day for 6 consecutive weeks. All groups were given maintenance feed, and the low and high doses of active folic acid in the intervention group were 5.3 and 10.1 (μg / d / rat). The specific doses of each group are shown in Table 1 below. The feces of the mother rats before gavage, after delivery, and after weaning, and the feces of the offspring rats one week and three weeks after birth (after weaning) were collected for intestinal flora sequencing.

[0101] Table 1 Oral doses for experimental animals

[0102] Group Dosage Group Active folic acid μg / d / each Blank control Basic feed + saline Example 1 Basic feed + low dose of active folic acid 5.3 Example 2 Basic feed + high dose of active folic acid 10.1

[0103] 1.2. Instruments, consumables and reagents

[0104] The consumables, reagents and experimental instruments used in the experiment are shown in Tables 2 and 3.

[0105] Table 2 Main experimental consumables and reagents

[0106]

[0107] Table 3 Main experimental instruments

[0108] Instrument Name company Electronic balance PTX-FA110S Huazhi (Fujian) Electronic Technology Co., Ltd. Desktop high speed refrigerated centrifuge Xiangyi Agilent 2100 Bioanalyzer Agilent Qubit Fluorometer Invitrogen NovaSeq 6000 Illumina

[0109] 2. Detection Method

[0110] 2.1. Feces collection of mother mice and offspring

[0111] (1) Feces collection of maternal mice: Fresh feces of each maternal mouse were collected in a clean environment before oral gavage, after delivery, and after weaning. The feces were numbered and stored at -80°C.

[0112] (2) Collection of feces of offspring: Fresh feces of each litter were collected at 1 week and 3 weeks after birth (after weaning), numbered, and stored in a frozen state at -80°C.

[0113] 2.2. Detection of fecal flora of mother mice and offspring

[0114] High-throughput sequencing of microorganisms in fecal samples of mother mice and offspring using 16S rDNA:

[0115] (1) Extraction of total DNA of the microbiome: A DNA extraction kit was used to extract the total DNA of the microbiome in the feces, and the quality of DNA extraction was tested by agarose gel electrophoresis. The DNA was quantified using an ultraviolet spectrophotometer.

[0116] (2) PCR amplification: The V3-V4 region of bacterial 16s rDNA was selected for gene amplification and sequencing. The primer sequences are shown in Table 4 below:

[0117] Table 4 Primer sequences

[0118]

[0119] The PCR reaction system is shown in Table 5 below:

[0120] Table 5 Reaction system

[0121] PCR reaction components PCR reaction volume Phusion Hot Start Flex 2X Master Mix 12.5μL Forward primer 2.5μL Reverse primer 2.5μL Template DNA 50ng <![CDATA[Add ddH2O to]]> 25μL

[0122] The PCR reaction conditions are shown in Table 6 below:

[0123] Table 6 Reaction conditions

[0124]

[0125] (3) PCR products were purified using AMPure XT beads (Beckman Coulter Genomice, MA, USA) and quantified using Qubit (Invitrogen, USA).

[0126] (4) The purified PCR products were evaluated using the Agilent 2100 bioanalyzer (Agilent, USA) and the Illumina (Kapa Biosciences, Wobum, MA, USA) library quantification kit. The qualified library concentration was above 2 nM. The qualified libraries for sequencing were gradient diluted, mixed according to the required sequencing amount, and denatured into single strands by NaOH for sequencing. The NovaSeq 6000 sequencer was used for 2×250 bp double-end sequencing, and the corresponding reagent was the NovaSeq6000SP Reagent Kit (500 cycles).

[0127] (5) For the paired-end data obtained by sequencing, the sample data is first split according to the Barcode information, and the adapter and Barcode sequence are removed, and then the sequence is spliced ​​to obtain the optimized sequence. After removing the chimera sequence, OUT cluster analysis is performed, and taxonomic analysis is performed on the representative OUT sequence. Based on the OUT cluster analysis results, multiple diversity index analyses and sequence depth detection are performed on OUT; based on the taxonomic information, statistical analysis of colony structure is performed at each classification level.

[0128] 3. Test results

[0129] 3.1. Results of dominant flora in the intestines of maternal mice

[0130] 3.1.1. Changes of Akkermansia in the intestines of female mice

[0131] Changes of Akkermansia in the intestines of female mice Figure 1 As shown, compared with the control group, different doses of active folic acid substances can increase the relative abundance of Akkermansia in the intestines of pregnant and lactating mice, and the effect of high-dose active folic acid substances is significantly higher than that of low-dose active folic acid substances.

[0132] 3.1.2. Changes in Bifidobacterium in the intestines of female mice

[0133] Changes of Bifidobacterium in the intestines of female mice Figure 2 As shown, compared with the control group, different doses of active folic acid substances can increase the relative abundance of Bifidobacterium in the intestines of pregnant rats; the relative abundance of Bifidobacterium in the intestines of lactating rats in the intervention groups of different doses of active folic acid substances showed a downward trend and the relative abundance was lower than that in the control group.

[0134] 3.1.3. Changes of Clostridia UCG-014unclassified in the intestines of female mice

[0135] Changes of Clostridia UCG-014unclassified in the intestines of female mice Figure 3 As shown, compared with the control group, different doses of active folic acid substances had no significant effect on the relative abundance of Clostridia UCG-014unclassified in the intestines of pregnant mice; different doses of active folic acid substances could significantly increase the relative abundance of Clostridia UCG-014unclassified in the intestines of lactating mice, and there was no significant difference in the increasing effect among different dose groups.

[0136] 3.1.4. Changes of Muribaculaceae unclassified in the intestine of female mice

[0137] Changes of Muribaculaceae unclassified in the intestine of female mice Figure 4As shown, compared with the control group, different doses of active folic acid substances can increase the relative abundance of Muribaculaceae unclassified in the intestines of pregnant mice, but the degree of increase is not significantly different from that of the control group; during the lactation period, the relative abundance of Muribaculaceae unclassified in the intestines of maternal mice in the control group showed a downward trend, while the relative abundance of Muribaculaceae unclassified in the intestines of maternal mice in the low-dose active folic acid intervention group was not significantly different from the relative abundance after delivery, and the relative abundance of Muribaculaceae unclassified in the intestines of maternal mice in the high-dose active folic acid intervention group was higher than its relative abundance after delivery, and higher than the control group and the low-dose active folic acid intervention group; The results showed that active folic acid intervention can help increase the relative abundance of Muribaculaceae unclassified in the intestines of lactating maternal mice.

[0138] 3.1.5. Changes in Escherichia-Shigella in the intestines of female mice

[0139] Changes in Escherichia-Shigella in the intestines of female mice Figure 5 As shown in the data, the relative abundance of Escherichia-Shigella in the intestines of the mothers in the control group increased with pregnancy and delivery, while the relative abundance of Escherichia-Shigella in the intestines of the mothers in the different doses of active folic acid intervention groups decreased to varying degrees after pregnancy and delivery; compared with the control group, the relative abundance of Escherichia-Shigella in the intestines of the lactating mothers in the different doses of active folic acid intervention groups was lower than that in the control group, and there was no significant difference in the degree of reduction among the different doses of active folic acid intervention groups; the results showed that supplementation of active folic acid during pregnancy can reduce the relative abundance of Escherichia-Shigella in the intestines of pregnant and lactating mothers.

[0140] 3.1.6. Changes of Streptococcus in the intestines of female mice

[0141] Changes of Streptococcus in the intestines of female mice Figure 6As shown, compared with the control group, the relative abundance of Streptococcus in the intestines of pregnant rats in the different doses of active folic acid intervention groups was reduced to varying degrees, and the high dose of active folic acid intervention group was lower than that in the low dose of active folic acid intervention group; the relative abundance of Streptococcus in the intestines of lactating rats in the different doses of active folic acid intervention groups was lower than that in the control group, and there was no significant difference between the different doses of active folic acid intervention groups; the results showed that active folic acid can inhibit the proliferation of Streptococcus in the intestines of pregnant rats and reduce its relative abundance in the intestines during lactation.

[0142] 3.2. Results of dominant bacterial flora in the intestines of offspring mice

[0143] 3.2.1. Changes of Akkermansia in the intestines of offspring mice

[0144] Changes of Akkermansia in the intestines of offspring mice Figure 7 As shown, compared with the control group, there was no significant difference in the relative abundance of Akkermansia in the intestine of the offspring in the different doses of active folic acid intervention groups 1 week after birth. With the continuation of breastfeeding, the relative abundance of Akkermansia in the intestine of the offspring in the different doses of active folic acid intervention groups 3 weeks after birth was significantly higher than that in the control group, and the high-dose intervention group was higher than the low-dose intervention group.

[0145] 3.2.2. Changes of Muribaculaceae unclassified in the intestine of offspring mice

[0146] Changes of Muribaculaceae unclassified in the intestine of offspring mice Figure 8 As shown, compared with the control group, there was no significant difference in the relative abundance of Muribaculaceae unclassified in the intestine of the offspring in the different doses of active folic acid intervention groups 1 week after birth. With the continuation of breastfeeding, the relative abundance of Muribaculaceae unclassified in the intestine of the offspring in the different doses of active folic acid intervention groups 3 weeks after birth was significantly higher than that in the control group, and there was no significant difference in the relative abundance of Muribaculaceae unclassified between the different doses of active folic acid intervention groups.

[0147] 3.2.3. Changes in Bacteroides in the intestines of offspring mice

[0148] Changes of Bacteroides in the intestine of offspring mice Fig. 9 As shown, compared with the control group, there was no significant difference in the relative abundance of Bacteroides in the intestine of the offspring in the intervention groups with different doses of active folic acid substances one week after birth. With the continuation of breastfeeding, the relative abundance of Bacteroides in the intestine of the offspring in the high-dose active folic acid intervention group was significantly higher than that in the control group three weeks after birth. The results showed that giving high doses of active folic acid substances to mothers during pregnancy and lactation can increase the relative abundance of Bacteroides in the intestine of the offspring.

[0149] 3.2.4. Changes of Escherichia-Shigella in the intestines of offspring mice

[0150] Changes of Escherichia-Shigella in the intestines of offspring mice Fig.10 As shown in the data, Eschrichia-Shigella were the dominant genera in the intestines of the offspring in the different doses of active folic acid intervention groups one week after birth. With the continuation of breastfeeding, the relative abundance of Escherichia-Shigella in the intestines of the offspring decreased significantly three weeks after birth, and the relative abundance of Escherichia-Shigella in the intestines of the offspring in the high dose of active folic acid intervention group was significantly lower than that in the control group.

[0151] 3.2.5. Changes of Streptococcus in the Intestines of Offspring Mice

[0152] Changes of Streptococcus in the intestines of offspring mice Fig.11 As shown in the data, the relative abundance of Streptococcus in the intestines of the offspring in the intervention groups with different doses of active folic acid substances was higher than that in the control group one week after birth. With the continuation of breastfeeding, the relative abundance of Streptococcus in the intestines of the offspring was significantly reduced three weeks after birth, and the reduction in the intervention groups with different doses of active folic acid substances was higher than that in the control group.

Claims

1. Use of active folic acid substances in the preparation of foods that are ingested by mothers and help regulate the intestinal flora of mothers and their offspring; wherein: The active folic acid substance is selected from 6S-5-methyltetrahydrofolate and its salts, the mother is in pregnancy and / or lactation, and the offspring is in fetal period, infant period, and / or childhood period.

2. The use according to claim 1, characterized in that The active folic acid substance is 6S-5-methyltetrahydrofolate, 6S-5-methyltetrahydrofolate calcium or 6S-5-methyltetrahydrofolate glucosamine salt.

3. The use according to claim 1 or 2, characterized in that The method of helping to regulate the intestinal flora of the mother and her offspring includes: increasing the relative abundance of Akkermansia in the intestinal tract of the pregnant mother, increasing the relative abundance of Bifidobacterium in the intestinal tract of the pregnant mother, reducing the relative abundance of Escherichia-Shigella in the intestinal tract of the pregnant mother, and reducing the relative abundance of Streptococcus in the intestinal tract of the pregnant mother.

4. The use according to any one of claims 1 to 3, characterized in that The intestinal flora that helps regulate the mother and her offspring includes: increasing the relative abundance of Akkermansia in the intestines of lactating mothers, increasing the relative abundance of Clostridia UCG-014unclassified in the intestines of lactating mothers, increasing the relative abundance of Muribaculaceae unclassified in the intestines of lactating mothers, reducing the relative abundance of Escherichia-Shigella in the intestines of lactating mothers, and reducing at least one of the relative abundance of Streptococcus in the intestines of lactating mothers.

5. The use according to any one of claims 1 to 4, characterized in that: The method of helping to regulate the intestinal flora of the mother and her offspring includes: increasing the relative abundance of Akkermansia in the offspring's intestine, increasing the relative abundance of Muribaculaceae unclassified in the offspring's intestine, increasing the relative abundance of Bacteroides in the offspring's intestine, reducing the relative abundance of Escherichia-Shigella in the offspring's intestine, and reducing at least one of the relative abundance of Streptococcus in the offspring's intestine.

6. The use according to any one of claims 1 to 5, characterized in that: The food helps to regulate the intestinal flora of the mother and her offspring, thereby affecting the gut-brain axis of the mother and her offspring and thus helping to maintain the health of the nervous system of the mother and her offspring.

7. The use according to any one of claims 1 to 6, characterized in that The food is infant food, children's food, adolescent food or adult food.

8. The use according to any one of claims 1 to 7, characterized in that The food is a beverage, candy, dairy product, bakery product or dietary supplement.

9. The use according to any one of claims 1 to 8, characterized in that At room temperature, the food is in liquid form, solid-liquid mixed form, solid block form or solid powder form.

10. The use according to any one of claims 1 to 9, characterized in that The food contains any one or more of the following ingredients: plant product ingredients, animal dairy product ingredients, animal meat product ingredients, functional added ingredients and any auxiliary materials acceptable in food.

11. The use according to any one of claims 1 to 10, characterized in that The mass content of the active folic acid substance in the food is 0.0001% to 0.1%.

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