Bifidobacterium longum subsp. infantis strain ylgb-1496 and its use in reducing cortisol levels and / or improving appetite

Abstract

The application provides bifidobacterium longum infantis subsp. YLGB-1496 strain and application thereof in reducing cortisol content and / or improving appetite.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of microbiology, in particular to Bifidobacterium longum subsp. infantis YLGB-1496 strain and its application in reducing cortisol content and / or improving appetite. BACKGROUND

[0002] Cortisol is a stress hormone secreted by the adrenal glands under stress conditions, which plays a key role in regulating the body's stress response, metabolic function, and immune system. Although cortisol helps cope with stress in the short term, long-term high cortisol levels are associated with a variety of health problems, including anxiety, depression, obesity, high blood pressure, and decreased immune function. In addition, changes in cortisol levels are closely related to fluctuations in appetite, and cortisol also plays an important role in regulating appetite. Therefore, regulating cortisol levels is crucial for maintaining physical and mental health.

[0003] In recent years, probiotics have received widespread attention due to their positive effects on gut health. However, there are currently limited probiotic products on the market for reducing cortisol.

[0004] Therefore, developing a probiotic product that can effectively reduce cortisol levels will help meet the growing demand for health. SUMMARY

[0005] The present application provides Bifidobacterium longum subsp. infantis YLGB-1496 strain and its application in reducing cortisol content and / or improving appetite, which has good effects in reducing cortisol content and / or improving appetite.

[0006] The present application provides the use of Bifidobacterium longum subsp. infantis YLGB-1496 strain in the preparation of a product for reducing cortisol content and / or improving appetite.

[0007] Optionally, the Bifidobacterium longum subsp. infantis YLGB-1496 strain is preserved in the China Center for Type Culture Collection of Wuhan University, with a preservation number of CCTCC NO: M2011122.

[0008] Optionally, the product comprises the Bifidobacterium longum subsp. infantis YLGB-1496 strain.

[0009] Optionally, in the product, the viable count of the Bifidobacterium longum subsp. infantis YLGB-1496 strain is 10 6 CFU / g ~ 10 11 CFU / g.

[0010] Optionally, the product is administered to the subject through the gastrointestinal tract.

[0011] Optionally, the subject includes infants aged 0-3 years.

[0012] Optionally, the product comprises one or more of a health food, a special medical purpose formula food.

[0013] The present application provides a composition for reducing cortisol content and / or improving appetite, the composition comprising Bifidobacterium longum subsp. infantis YLGB-1496 strain.

[0014] Optionally, the composition further comprises an excipient, wherein the excipient comprises maltodextrin.

[0015] The present application provides a preparation method of the composition as described above, comprising: mixing a raw material system comprising the Bifidobacterium longum subsp. infantis YLGB-1496 strain to obtain the composition.

[0016] The present application provides Bifidobacterium longum subsp. infantis YLGB-1496 strain and its application in reducing cortisol content and / or improving appetite, YLGB-1496 strain is preserved in China Center for Type Culture Collection of Wuhan University, the preservation number is CCTCC NO: M2011122, and test proves that YLGB-1496 strain has a good effect in reducing cortisol content and / or improving appetite, and in addition, has a good improvement effect on body weight. BRIEF DESCRIPTION OF DRAWINGS

[0017] Figure 1 It is a specific technical roadmap for clinical trials;

[0018] Figure 2 It is a column chart of the number of days of self-reported anorexia of the subjects in Example 1 in the past month;

[0019] Figure 3 It is a statistical chart of cumulative days of appetite decline of the subjects in Example 2 in 3 months (N is the sample number, and the rank average is the average of cumulative days of appetite decline);

[0020] Figure 4 It is a column chart of cortisol content in saliva of the subjects in Example 1 (Probiotic is a probiotic intervention group, and Placebo is a placebo group). DETAILED DESCRIPTION

[0021] In order for those skilled in the art to better understand the scheme of the present application, the present application is further described in detail as follows. The following specific embodiments are only used to describe the principles and characteristics of the present application, and the examples are only used to explain the present application, and do not limit the scope of the present application. Based on the embodiments of the present application, all other embodiments obtained by those skilled in the art without creative labor are within the scope of protection of the present application.

[0022] Intestinal microecology refers to the complex ecological system formed by the interaction between the microorganisms parasitic in the human intestinal tract and the human body.

[0023] The formation of intestinal microecology is a complex process. The initial microorganisms in the body of a newborn mainly come from the mother's birth canal, feces and the surrounding environment. The period from 0 to 3 years old is a critical time window for the establishment and evolution of intestinal flora, and breastfeeding is the most important factor in determining the early intestinal microecology of infants. Breastfeeding of full-term, vaginal delivery newborns is more likely to form a healthy intestinal flora, such as a suitable content and species of Bifidobacterium and Lactobacillus.

[0024] Breast milk contains prebiotics such as probiotics and human milk oligosaccharides (HMOs). In 2003, a literature first reported that breast milk contains bacteria and directly affects the intestinal flora of infants; many subsequent studies have provided solid evidence that the intestinal microorganisms of breastfed infants come from breast milk; there have been many studies on probiotics in breast milk, including the discovery, isolation and study of probiotics in breast milk, and patents have been filed for them, for example, the University of Granada in Spain has applied for a global and even Chinese patent, which has discovered three probiotic strains (two Lactobacillus and one Bifidobacterium Lactobacillus rhamnosus HERO 22A (CNCM I-4036), Lactobacillus paracasei HERO 7 (CNCM I-4034) and Bifidobacterium breve HERO 15B (CNCM I-4035)) in breast milk.

[0025] The list of bacteria that can be added to food in many countries includes Bifidobacterium, Lactobacillus, Streptococcus, etc., and specifies to the genus. The list of probiotic bacteria that can be used in health food and the list of bacteria that can be used in food specify edible probiotics to the species. The list of bacteria that can be used in infant food published in 2011 specifies each probiotic bacteria that can be used in food to the strain. Lactobacillus and Bifidobacterium have been recognized as belonging to probiotics.

[0026] Currently, there is only one strain of bacteria in the list of bacteria that can be used in infant food, which is derived from human breast milk, which is the Spanish-originated Lactobacillus muci- de CECT5716, and the remaining strains are not derived from breast milk. In addition, in the latest list of bacteria that can be used in food released in August 2022, Bifidobacterium longum subsp. infantis is included, and the European Food Safety Authority qualification (QPS) list also includes Bifidobacterium longum subsp. infantis, indicating that this species is safe for consumption, and in recent years, the importance of Bifidobacterium longum subsp. infantis (infant Bifidobacterium) to the intestinal tract of infants has been confirmed by many studies.

[0027] In view of the fact that breast milk contains microorganisms and is very important for the establishment of the early life intestinal flora, especially Bifidobacterium longum subsp. Infantis in breast milk has important significance for infant health. Therefore, Bifidobacterium longum subsp. Infantis in breast milk has become a research hotspot.

[0028] Based on this, the embodiments of the present application provide the use of the Bifidobacterium longum subsp. Infantis YLGB-1496 strain in the preparation of a product for reducing the content of cortisol and / or improving appetite.

[0029] The above-mentioned Bifidobacterium longum subsp. Infantis YLGB-1496 strain (Bifidobacterium longum subsp. Infantis GB-1496) was deposited in the China Center for Type Culture Collection of Wuhan University on April 10, 2011, and the deposit number is CCTCC NO: M2011122.

[0030] The Bifidobacterium longum subsp. Infantis (infant Bifidobacterium) YLGB-1496 strain is a strain of infant Bifidobacterium cultured and isolated from the milk of a mother who is exclusively breastfed. By comparing the gene sequencing result thereof with the published gene sequence of infant Bifidobacterium, it is determined that it is a new infant Bifidobacterium.

[0031] It is proved by experiments that the Bifidobacterium longum subsp. Infantis YLGB-1496 strain has good effects in reducing the content of cortisol and / or improving appetite, and in addition, it also has good effects in improving body weight. Therefore, the Bifidobacterium longum subsp. Infantis YLGB-1496 strain can be used for preparing a product for reducing the content of cortisol and / or improving appetite.

[0032] In some embodiments, the above-mentioned product comprises the Bifidobacterium longum subsp. Infantis YLGB-1496 strain.

[0033] Specifically, in the above-mentioned product, the viable count of the Bifidobacterium longum subsp. Infantis YLGB-1496 strain can be greater than or equal to 10 6 CFU / g.

[0034] Further, in the above-mentioned product, the viable count of the Bifidobacterium longum subsp. Infantis YLGB-1496 strain can be 10 6 CFU / g to 10 11 CFU / g.

[0035] In some embodiments, the above-mentioned Bifidobacterium longum subsp. Infantis YLGB-1496 strain is administered to the subject through the gastrointestinal tract.

[0036] It has been confirmed by many studies that the intestinal micro-ecology in early life plays a crucial role in the maturation process of the immune system, and the diversity and maturity of the intestinal flora in infancy are closely related to health. The above B. longum infantis YLGB-1496 strain can tolerate strong acidity and bile salts in the artificial gastrointestinal tract, which means that it can pass through the stomach and the upper part of the small intestine of the human body, and finally reach the lower part of the small intestine and the colon to exert activity. For infants and young children (for example, infants and young children aged 0-3 years), the acidity of gastric acid and the concentration of bile acid salt are lower than those of adults, so the above B. longum infantis YLGB-1496 strain is more likely to colonize in the early stage in the intestinal tract of infants and young children as probiotics.

[0037] In a specific embodiment, the above subject includes infants and young children aged 0-3 years (or 0-36 months), specifically, the infants and young children aged 0-3 years can include infants and young children aged 0-3 years, infants and young children aged 1-3 years.

[0038] Clinical trial data have confirmed that the above B. longum infantis YLGB-1496 strain can effectively reduce the content of cortisol in the saliva of infants and young children aged 0-3 years, improve appetite and appetite, and also has a good effect on improving body weight.

[0039] It can be understood that the embodiments of the present application do not limit the types of the above products, for example, the above products can include one or more of health foods, special medical use formula foods.

[0040] The embodiments of the present application also provide a composition for reducing the content of cortisol and / or improving appetite, which includes the above B. longum infantis YLGB-1496 strain. Based on the above B. longum infantis YLGB-1496 strain, the composition can effectively reduce the content of cortisol and / or improve appetite.

[0041] In addition, the composition can also include excipients, such as malt dextrin.

[0042] Further, in the above composition, the viable bacterial count of the B. longum infantis YLGB-1496 strain can be greater than or equal to 10 6 CFU / g.

[0043] Further, in the above composition, the viable bacterial count of the B. longum infantis YLGB-1496 strain can be greater than or equal to 10 6 CFU / g to 10 11 CFU / g.

[0044] The embodiments of the present application also provide a preparation method of the above composition, which includes: mixing a raw material system including the B. longum infantis YLGB-1496 strain to obtain the above composition.

[0045] In particular implementation, the B. longum subsp. infantis YLGB-1496 strain powder can be prepared first, and then the raw material system including the powder is uniformly mixed to obtain the composition.

[0046] The B. longum subsp. infantis YLGB-1496 strain powder needs to meet the requirements of the number of viable lactic acid bacteria ≥1.5×10 11 CFU / g, and the water activity is 0.05-0.1, for example, 0.05, 0.07, 0.1, or a range formed by any two of them.

[0047] In addition, the raw material system can further include an excipient, such as malt dextrin.

[0048] The present application will be further described below through specific examples and comparative examples. Unless otherwise specified, the reagents, materials and instruments used in the following are conventional reagents, conventional materials and conventional instruments, which are commercially available, and the reagents and materials involved can also be synthesized by conventional synthesis methods.

[0049] A clinical trial was conducted on a population of infants and young children aged 0-3 years, including targeting the B. longum subsp. infantis YLGB-1496 strain, investigating the clinical manifestations after supplementing the strain and intervention in the population of infants and young children aged 0-3 years, and studying the effects of the YLGB-1496 strain on the cortisol content in saliva and appetite of the population of infants and young children aged 0-3 years (early life population). The trial used a multi-center randomized double-blind controlled study. The specific technical route is shown in Figure 1 .

[0050] Example 1

[0051] This study has been registered on ClinicalTrials.gov (identifier NCT05794815), and the inclusion and exclusion criteria for subjects are as follows:

[0052] 1) Recruit children aged 1-3 years as subjects, and all parents of the subjects sign a written informed consent before the start of the study. The inclusion and exclusion criteria for subjects are as follows,

[0053] The inclusion criteria include: the current weight is P20-P80 of the children percentile table, the infant formula milk powder currently used does not contain probiotics, the use of prebiotics and probiotics is not allowed, and the willingness to participate in the entire experiment;

[0054] Exclusion criteria included: having any deformity, mother with metabolic and / or chronic diseases, currently or previously suffering from conditions or interventions that could interfere with the study, taking oral antibiotics in the two weeks prior to the intervention, having known allergy to probiotics or possible food allergies, participating in other clinical studies in the four weeks prior to the intervention, or unwilling to participate for any special reason.

[0055] 2) Preparation of probiotic product and placebo

[0056] The intervention used in this study was Bifidobacterium longum subsp. infantis YLGB-1496 powder, with maltodextrin as excipient, in a 2 g / pack format; the placebo was maltodextrin, 2 g / pack, with identical appearance, weight, and color to the intervention. The intervention and placebo were tested for Escherichia coli, Salmonella, Staphylococcus aureus, and mold by the Shanghai Standard Technology Service Co., Ltd., and the results met the national standards. In addition, heavy metals, additives such as lead (Pb), benzoic acid, sorbic acid, sodium saccharin, sodium cyclamate (sweetener), and lemon yellow were not detected.

[0057] The intervention (bacterial powder) was stored in a -20°C freezer; after the intervention was distributed to the subjects (study participants), it was stored in a home freezer at -18°C.

[0058] 3) Study implementation

[0059] After the subjects were enrolled, they were randomly grouped according to the inclusion and exclusion criteria. According to the computer-generated list, the eligible subjects were randomly assigned to the two groups of the study (probiotic group or placebo group) in a 1:1 ratio and identified with separate codes. The random grouping was performed by the study statistician, who had no contact with the participants. Before the completion of the study, no member of the study team could obtain the allocation sequence.

[0060] The researchers distributed the intervention to all enrolled infants and young children according to the grouping information.

[0061] Probiotic intervention group (intervention group or control group): 1 pack (bag) per day (containing 1.5 x 10 10 CFU / pack of YLGB-1496 live bacteria), starting from the day the probiotic product was used, and continuing for 3 months.

[0062] Placebo control group (placebo group): 1 pack (maltodextrin, not containing Bifidobacterium infantis YLGB-1496 strain) per day, starting from the day the placebo product was used, and continuing for 3 months.

[0063] At 3 follow-up points (0w, 6w, 12w), both groups of infants and primary caregivers were visited to collect information such as dietary and health records, and saliva samples. The content of cortisol in saliva, appetite and weight improvement of the subjects at 3 follow-up points were recorded and tested.

[0064] 4) Data entry and statistical analysis

[0065] The entry of research data used Epidata software double entry, and then the database was compared twice, and if problems were found during the period, the inspector was notified in time, and the researcher was required to answer. The question table was used to record various questions and answer processing results during the entry process, and the question table was kept for reference.

[0066] The study data was analyzed using SPSS software version 20.0. Considering the skewness distribution and non-parametric nature of the data, Mann-Whitney U test was used to compare the differences between groups, and chi-square test was used to compare the conventional data. Spearman rank correlation was used in non-parametric correlation analysis, and the correlation coefficient was rho (r). Negative binomial regression was also used to evaluate the effect of probiotic intervention on the content of cortisol, appetite and weight improvement. All tests were two-sided, *P<0.05 was considered statistically significant, unless otherwise stated, the data was expressed as mean ± standard error.

[0067] Example 2

[0068] 1) Recruit healthy infants aged 0-3 years as subjects, all parents of the subjects are informed and sign the informed consent form; this study has been registered in ISRCTN registry (registration number ISRCTN12824613); the inclusion and exclusion criteria for the subjects are as follows,

[0069] The inclusion criteria include:

[0070] Infants aged 0-3 years born at 37-42 weeks of gestation, birth weight greater than or equal to 2500 and less than 4000 g, no gender restriction;

[0071] The mother of the infant has no diagnosed metabolic diseases such as diabetes, and no infectious diseases such as hepatitis B and HIV;

[0072] The parents or primary caregivers of the infant agree to collect the infant's fecal samples during the study period;

[0073] No clinically diagnosed allergic diseases (including but not limited to eczema, asthma, allergic proctocolitis, allergic rhinitis, hay fever, etc.) at the time of enrollment;

[0074] The family or primary caregiver agrees not to add additional probiotic products (including formula with probiotics) to the infant during the intervention period;

[0075] The guardian of the enrolled subject agrees to participate in this interventional study and signs the written informed consent form, and is able to understand and fill out the forms such as infant diary as required.

[0076] Exclusion criteria include:

[0077] Infants included due to missed diagnosis;

[0078] Found to violate the inclusion criteria during the trial period;

[0079] Infants with no test records available for evaluation;

[0080] During the trial period, self-administration of other probiotics not related to the study, which cannot be evaluated for the effect of the probiotics used in the study;

[0081] Poor compliance, taking less than 80% of the full dose of probiotics;

[0082] The subject develops AEs, laboratory abnormalities or other medical conditions related to Bifidobacterium infantis YLGB-1496, and continued participation in the study would not be in the best interest of the subject;

[0083] Withdrawal of informed consent during the intervention period;

[0084] Subjects who, with informed consent, are screened and eligible for randomization, do not complete the treatment period, observation period and data collection required by the program for any reason, are treated as dropouts;

[0085] Infants who require the use of antibiotics or immunosuppressive drugs due to illness during the intervention period;

[0086] Infants who develop an allergy to the probiotic strain and / or maltodextrin ingredient used in the study during the intervention period.

[0087] 2) The preparation of probiotic products and placebo, the implementation of the study, data entry and statistical analysis are carried out according to Example 1, and the number of days of decreased appetite and body weight improvement within 3 months of the above infants are statistically analyzed.

[0088] Test results

[0089] Table 1 Number of days of self-reported anorexia in subjects in Example 1 in the past month (days / week)

[0090]

[0091] Combined with Table 1, Figure 2 , Figure 3It can be seen that the number of anorexia days of the probiotic intervention group (intervention group) is significantly reduced, which can prove that B. longum infantis YLGB-1496 can effectively improve the appetite of the subjects.

[0092] Table 2: Cortisol content in saliva of subjects in Example 1 (unit: ng / μg)

[0093]

[0094] In combination with Table 2, Figure 4 It can be seen that the saliva cortisol content of the probiotic intervention group (intervention group) is significantly reduced, which can prove that B. longum infantis YLGB-1496 can effectively reduce the saliva cortisol content of the subjects.

[0095] Table 3: Body weight of subjects in Example 1 (unit: kg)

[0096]

[0097] Table 4: Body weight of subjects in Example 2 (unit: kg)

[0098]

[0099] In combination with Table 3 and Table 4, it can be seen that the weight gain of the probiotic intervention group (intervention group) is obvious, which can prove that B. longum infantis YLGB-1496 can effectively improve the body weight of the subjects.

[0100] Finally, it should be noted that: the above examples are only used to illustrate the technical solutions of the present application, but not to limit them; although the present application has been described in detail with reference to the foregoing examples, those skilled in the art should understand that: it can still modify the technical solutions recorded in the foregoing examples, or make equivalent replacement for part or all of the technical features; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the scope of the technical solutions of the embodiments of the present application.

Claims

1. Use of Bifidobacterium longum subsp. infantis YLGB-1496 strain for the preparation of a product for reducing the cortisol level and for improving the appetite in healthy infants aged 0-3 years, said product being a health food or a food for special medical purposes; wherein, The B. longum subsp. infantis YLGB-1496 strain is preserved in the China Center Type Culture Collection of Wuhan University, and the preservation number is CCTCC NO: M2011122. The B. longum subsp. infantis YLGB-1496 strain is preserved in the China Center Type Culture Collection of Wuhan University, and the preservation number is CCTCC NO: M2011122.

2. Use according to claim 1, characterized in that, In the product, the viable cell count of the Bifidobacterium longum subsp. infantis YLGB-1496 strain is 10 6 CFU / g ~ 10 11 CFU / g.

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