Application of bacterial strain for improving 5-hydroxytryptophan secreted by intestinal pheochromophilic cells

Abstract

The invention provides a bacterial strain application, the bacterial strain is used for preparing a composition for improving the secretion of 5-hydroxytryptophan acid by intestinal pheochromophilic cells, the bacterial strain comprises a bifidobacterium animalis subsp. Lactis BB-115 bacterial strain, the BB-115 bacterial strain is deposited in the China General Microbiological Culture Collection Center, and the depositing number is CGMCC21840.

Description

Technical Field

[0001] The present invention relates to the use of strains, and particularly to the use of strains for enhancing the secretion of 5-hydroxytryptophan by enterochromaffin cells. Background Art

[0002] Serotonin affects the body's emotional stability, well-being, concentration, and sleep quality. Therefore, maintaining the concentration of serotonin in the body is crucial for physical and mental health.

[0003] Enterochromaffin cells are a subtype of enteroendocrine cells, accounting for less than 1% of the total intestinal epithelium, but responsible for more than 90% of the body's serotonin production and storage. Enterochromaffin cells can directly contact the gut microbiota and microbial metabolites on one side of the intestinal lumen and interact with the afferent and efferent nerve endings located in the lamina propria of the intestine through synaptic connections. Enterochromaffin cells are electrically excitable and can express functional voltage-gated sodium and calcium channels, so they are similar to primary sensory cells. All of the above characteristics enable enterochromaffin cells to serve as a two-way communication medium between the intestinal lumen, intestinal epithelial cells, and specific primary afferent nerve fibers. It is also considered to participate in the interaction of the microbiome-gut-brain axis in human cognition, depression, and chronic pain. + and Ca 2+ channels), so they are similar to primary sensory cells. All of the above characteristics enable enterochromaffin cells to serve as a two-way communication medium between the intestinal lumen, intestinal epithelial cells, and specific primary afferent nerve fibers. It is also considered to participate in the interaction of the microbiome-gut-brain axis in human cognition, depression, and chronic pain.

[0004] Enterochromaffin cells contain tryptophan hydroxylase 1, which can convert L-tryptophan into 5-hydroxytryptophan. 5-Hydroxytryptophan can pass through the blood-brain barrier via the systemic blood circulation system and be converted into a precursor of the neurotransmitter serotonin in the brain. Tryptophan is an essential amino acid that the human body cannot synthesize on its own and is usually obtained by ingesting foods rich in tryptophan. When the intake of tryptophan-containing foods is insufficient or the body's serotonin secretion decreases, it is easy to cause related physical and mental disorders such as anxiety, depression, mood instability, inattention, and sleep disorders.

[0005] Currently, it is recommended to consume more tryptophan-rich foods or take supplements containing 5-hydroxytryptophan to increase the concentration of serotonin in the body. However, excessive supplementation of 5-hydroxytryptophan may cause side effects such as nausea, diarrhea, and serotonin syndrome. Therefore, those skilled in the art are committed to developing ways to effectively enhance the secretion of 5-hydroxytryptophan by enterochromaffin cells without producing side effects. Summary of the Invention

[0006] Through experiments, it was found that the Bifidobacterium animalis subsp. lactis BB-115 strain has a better activity in promoting the secretion of 5-hydroxytryptophan by enterochromaffin cells than other strains, thereby increasing the precursor of 5-hydroxytryptamine in the brain. Therefore, it is expected to use the BB-115 strain in the application of enhancing the secretion of 5-hydroxytryptophan in individuals.

[0007] One object of the present invention relates to a use of a strain for preparing a composition for promoting the secretion of 5-hydroxytryptophan by enterochromaffin cells; wherein the strain includes the Bifidobacterium animalis subsp. lactis BB-115 strain, and the BB-115 strain is deposited in the China General Microbiological Culture Collection Center with the deposit number CGMCC21840.

[0008] Preferably, the BB-115 strain is an active strain or an inactivated strain.

[0009] Preferably, the composition is further used for enhancing immunity, treating insomnia, treating depression, treating hypertension, improving sleep disorders, improving anxiety, improving inattention, treating menopausal syndrome, or treating gastrointestinal diseases.

[0010] Preferably, the composition is a drug, food, or health product.

[0011] Another object of the present invention relates to a use of a strain for preparing a composition for promoting the secretion of 5-hydroxytryptophan by enterochromaffin cells; wherein the strain includes the Bifidobacterium animalis subsp. lactis BB-115 strain, the Lactobacilus rhamnosus MP108 strain, and the Bifidobacterium longum subsp. infantis BLI-02 strain; wherein the BB-115 strain is deposited in the China General Microbiological Culture Collection Center with the deposit number CGMCC21840, the MP108 strain is deposited in the China General Microbiological Culture Collection Center with the deposit number CGMCC21225, and the BLI-02 strain is deposited in the China General Microbiological Culture Collection Center with the deposit number CGMCC15212.

[0012] Preferably, the BB-115 strain, the MP108 strain, and the BLI-02 strain are each an active strain or an inactivated strain.

[0013] Preferably, the ratio of colony-forming units (CFU) among strain BB-115, strain MP108, and strain BLI-02 is (1 to 2):1:1.

[0014] Preferably, the ratio of colony-forming units among strain BB-115, strain MP108, and strain BLI-02 is 2:1:1.

[0015] Preferably, the composition is more used for enhancing immunity, treating insomnia, treating depression, treating hypertension, improving sleep disorders, improving anxiety, improving inattention, treating menopausal syndrome, or treating gastrointestinal diseases.

[0016] Preferably, the composition is a medicine, food, or health product. Description of the Drawings

[0017] Figure 1 Showing the expression level of tryptophan in rat insulinoma cells RIN-14B after different treatments; wherein, ＊ P < 0.05, ＊＊ P < 0.01, ＊＊＊ P < 0.001, ＊＊＊＊ P < 0.0001. Detailed Embodiments

[0018] To make the above and / or other objects, effects, and features of the present invention more obvious and understandable, the following preferred embodiments are specifically described in detail as follows:

[0019] The intestines of mammals inhabit a large number and variety of microorganisms, and these microorganisms are closely related to health and diseases. An unbalanced microecology is closely related to the formation of many diseases. For example, gastrointestinal pathology and intestinal microbiota dysregulation have been found in autism, neurodegenerative diseases, and schizophrenia. Studies have found that the composition of the microbial flora can regulate the serotonin level in the blood. Generally, the serotonin concentration in the blood of mice with a normal gut microbiota is higher than that of germ-free mice. Compared with rats with a normal gut microbiota, the enterochromaffin cells in the intestines of germ-free rats have a larger morphology, indicating that the composition of intestinal microorganisms may affect the development or function of enterochromaffin cells.

[0020] Lactic acid bacteria are generally recognized as safe, familiar, and widely used probiotics. Common ones include: Lactobacillus, Lactococcus, Pediococcus, Enterococcus, Streptococcus, Bifidobacterium, Bacillus, and Leuconostoc.

[0021] The research explores the related applications of lactic acid bacteria strains in enhancing the secretion of 5-hydroxytryptophan by enterochromaffin cells. Food Funct. 2019 Nov 1;10(11):7588 - 7598 tested the effects of several different strains on the secretion of 5-hydroxytryptophan by rat insulinoma cells RIN-14B and found that Bifidobacterium longum subsp. infantis CCFM687 could enhance the secretion of 5-hydroxytryptophan and reduce depressive behavior in mice. However, Bifidobacterium longum HAN4-25, HUB2-25, GX17-A9, FSHHK25M1, and FSHHK27M1 did not have the activity to promote the secretion of 5-hydroxytryptophan. Front Microbiol. 2023 May 10:14:1174800 proposed probiotics related to the prevention and treatment of depression and found that strains such as Bifidobacterium longum R0175, Bifidobacterium infantis 35624, Lactobacillus helveticus R0052, Bifidobacterium longum R0175, Bifidobacterium adolescentis NK98, Bifidobacterium breve M-16V, and M2CF22M7 had a positive impact on the treatment of depression. The anti-depressant mechanism of the strains may be the regulation of inflammatory responses, tryptophan metabolism, 5-hydroxytryptamine synthesis, and the hypothalamus-pituitary-adrenal axis by the gut-brain axis.

[0022] The above description shows that different strains of the same bacterial species cannot be reasonably expected to have the effect of increasing the secretion of 5-hydroxytryptophan. That is to say, the effect of increasing the secretion of 5-hydroxytryptophan is a strain-specific function. Generally, lactic acid bacteria are recognized as safe strains and mental diseases are modern civilization diseases. Therefore, developing the application of lactic acid bacteria in mental diseases is one of the actively pursued topics.

[0023] The first embodiment of the present invention provides a use of a strain for preparing a composition that enhances the secretion of 5-hydroxytryptophan by enterochromaffin cells. The strain includes the Bifidobacterium animalis subsp. lactis BB-115 strain, and the BB-115 strain is deposited in the China General Microbiological Culture Collection Center with the deposit number CGMCC21840.

[0024] The BB-115 strain can be an active strain or an inactivated strain and can be obtained by culturing in a suitable growth medium. Suitable growth media are well-known to those skilled in the art, such as commercially available or self-prepared. Suitable growth media can be MRS broth medium, MRS broth medium supplemented with cysteine, medium supplemented with glucose, yeast extract, and soy peptone. The culturing operation is also well-known to those skilled in the art, such as referring to New Microbiol. 2013 Apr; 36(2): 167-79. Before culturing, the BB-115 strain can be inoculated into the medium at a cell concentration of 10 6 to 10 12 CFU / mL, preferably at a cell concentration of 10 6 to 10 10 CFU / mL, more preferably at a cell concentration of 2x10 7 to 10 8 CFU / mL. During culturing, the BB-115 strain can be placed at a temperature range of 25 to 40 °C for 24 to 48 hours, preferably at 37 °C for 24 hours.

[0025] The composition can be administered to an individual in need of enhancing the secretion of 5-hydroxytryptophan by enterochromaffin cells to increase the secretion of 5-hydroxytryptophan in the individual. The individual can be a mammal, and the mammal can be a primate, cat, dog, mouse, rat, rabbit, cow, horse, goat, sheep, or pig, and the primate can be a chimpanzee, human, gorilla, bonobo, orangutan, or monkey. In addition, the BB-115 strain of 10 6 to 10 10 CFU per kilogram of body weight of the individual can be administered to the individual in the composition, preferably 10 7 to 10 9 CFU per kilogram of body weight of the individual per day, more preferably 108 The BB-115 strain of CFU. Based on the property of the BB-115 strain to enhance the secretion of 5-hydroxytryptophan by enterochromaffin cells, the composition can further enhance immunity, treat insomnia, treat depression, treat hypertension, improve sleep disorders, improve anxiety, improve inattention, treat menopausal syndrome, or treat gastrointestinal diseases.

[0026] The composition may further comprise a physiologically acceptable excipient, diluent, or carrier, and the composition can be a medicine, food, or health product.

[0027] Under the condition that the composition is a medicine or health product, the physiologically acceptable excipient, diluent, or carrier can be a pharmacologically acceptable excipient, diluent, or carrier, which can be a solvent, buffer, emulsifier, suspending agent, decomposing agent, disintegrating agent, dispersing agent, binder, stabilizer, chelating agent, gelling agent, wetting agent, lubricant, absorption delaying agent, or liposome. Based on this, the medicine or health product can be in a suitable dosage form, such as: powder, lozenge, tablet, buccal tablet, pill, capsule, solution, suspension, emulsion, syrup, elixir, thick paste, drops, gel, ointment, spray, or cream. Based on this, the medicine or health product can be administered to an individual in a suitable manner, such as: orally, sublingually, rectally, nasally, vaginally, intraperitoneally, transdermally, epidermally, intra-articularly, intra-ocularly, or on the ocular surface.

[0028] Under the condition that the composition is a food or health product, the physiologically acceptable excipient, diluent, or carrier can be an edible acceptable excipient, diluent, or carrier, which can be fluid dairy products (such as: milk or condensed milk), fermented dairy products (such as: yogurt), milk powder, ice cream, cheese, cottage cheese, soy milk, fermented soy milk, vegetable juice, fruit juice, sports drink, jelly, biscuit, energy bar, health food, animal feed, or dietary supplement.

[0029] The second embodiment of the present invention provides a use of strains, which is for preparing a composition that enhances the secretion of 5-hydroxytryptophan by enterochromaffin cells; wherein the strains include Bifidobacterium animalis subsp. lactis BB-115 strain, Lactobacillus rhamnosus MP108 strain, and Bifidobacterium longum subsp. infantis BLI-02 strain; wherein the BB-115 strain is deposited in the China General Microbiological Culture Collection Center with the deposit number CGMCC21840, the MP108 strain is deposited in the China General Microbiological Culture Collection Center with the deposit number CGMCC21225, and the BLI-02 strain is deposited in the China General Microbiological Culture Collection Center with the deposit number CGMCC15212. The colony-forming unit ratio among the BB-115 strain, MP108 strain, and BLI-02 strain can be (1 to 2):1:1, preferably 2:1:1.

[0030] The BB-115 strain can be a live strain or an inactivated strain, the MP108 strain can be a live strain or an inactivated strain, and the BLI-02 strain can be a live strain or an inactivated strain. Each strain can be obtained by culturing in a suitable growth medium. Suitable growth media are well-known to those skilled in the art, such as those obtained commercially or prepared by oneself. Suitable growth media can be MRS broth medium, MRS broth medium supplemented with cysteine, or a medium supplemented with glucose, yeast extract, and soy peptone. The culturing operation is also well-known to those skilled in the art, such as referring to New Microbiol. 2013 Apr; 36(2): 167-79. Before culturing, each strain can be inoculated into the medium at a cell concentration of 10 6 to 10 12 CFU / mL, preferably at a cell concentration of 10 6 to 10 10 CFU / mL, and more preferably at a cell concentration of 2x10 7 to 10 8 CFU / mL. During culturing, each strain can be placed at a temperature range of 25 to 40 °C for 24 to 48 hours, preferably at 37 °C for 24 hours.

[0031] The composition can be administered to an individual in need of enhancing the secretion of 5-hydroxytryptophan by enterochromaffin cells to enhance the secretion of 5-hydroxytryptophan in the individual. The individual can be a mammal, and the mammal can be a primate, cat, dog, mouse, rat, rabbit, cow, horse, goat, sheep, or pig, and the primate can be a chimpanzee, human, gorilla, bonobo, orangutan, or monkey. In addition, the composition containing the BB-115 strain, MP108 strain, and BLI-02 strain can be administered to the individual at a dose of 10 6 to 10 10 CFU per kg of body weight per day of the individual, preferably at a dose of 10 7 to 10 9 CFU per kg of body weight per day of the individual, and more preferably at a dose of 10 8 CFU per kg of body weight per day of the individual of the BB-115 strain, MP108 strain, and BLI-02 strain. Based on the property that the combination of the BB-115 strain, MP108 strain, and BLI-02 strain can enhance the secretion of 5-hydroxytryptophan by enterochromaffin cells, the composition can further enhance immunity, treat insomnia, treat depression, treat hypertension, improve sleep disorders, improve anxiety, improve inattention, treat menopausal syndrome, or treat gastrointestinal diseases.

[0032] The composition may further comprise a physiologically acceptable excipient, diluent, or carrier, and the composition may be a pharmaceutical, food, or health product.

[0033] When the composition is a pharmaceutical or health product, the physiologically acceptable excipient, diluent, or carrier may be a pharmacologically acceptable excipient, diluent, or carrier, which may be a solvent, buffer, emulsifier, suspending agent, decomposer, disintegrant, dispersant, binder, stabilizer, chelating agent, gelling agent, wetting agent, lubricant, absorption retardant, or liposome. Based on this, the pharmaceutical or health product may be in a suitable dosage form, such as: powder, lozenge, tablet, troche, pill, capsule, solution, suspension, emulsion, syrup, elixir, thick paste, drops, gel, ointment, spray, or cream. Based on this, the pharmaceutical or health product may be administered to an individual in a suitable manner, such as: orally, sublingually, rectally, nasally, vaginally, intraperitoneally, transdermally, epidermally, intra-articularly, intraocularly, or on the ocular surface.

[0034] When the composition is a food or health product, the physiologically acceptable excipient, diluent, or carrier may be an edible acceptable excipient, diluent, or carrier, which may be fluid dairy products (such as: milk or condensed milk), fermented dairy products (such as: yogurt), milk powder, ice cream, cheese, cottage cheese, soy milk, fermented soy milk, vegetable juice, fruit juice, sports drink, jelly, biscuit, energy bar, health food, animal feed, or dietary supplement.

[0035] The present invention will be illustrated by the following examples.

[0036] <Example 1: Cell Obtaining and Cultivation>

[0037] Proc Natl Acad Sci U S A. 2009 Mar 3; 106(9):3408-13 proposed that rat insulinoma cell RIN-14B has similar functions to enterochromaffin cells, so rat insulinoma cell RIN-14B can be used as a model for studying the functions of enterochromaffin cells.

[0038] Rat insulinoma cell RIN-14B was purchased from the American Type Culture Collection (ATCC). Rat insulinoma cell RIN-14B was inoculated into RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin, and placed in an incubator at 37 °C and 5% CO 2 for cultivation. Thereafter, fresh medium was replaced every 2 to 3 days, and subculture was carried out when the cell density reached about 80 to 90% confluence.

[0039] <Example 2: Obtaining Strains>

[0040] The strains for efficacy evaluation were deposited in the China General Microbiological Culture Collection Center in accordance with the provisions of the Budapest Treaty at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing. The detailed deposit information is listed in Table 1.

[0041] Table 1. Strain Deposit Information

[0042]

[0043] The following strains were used as control strains: Bifidobacterium animalis subsp. lactis HN019 strain (purchased from Fonterra Co-operative Group Limited), Lactobacillus rhamnosus LGG strain (purchased from Chr. Hansen A / S), and Bifidobacterium longum subsp. infantis M-63 strain (purchased from Morinaga Milk Industry Co., Ltd.).

[0044] <Example 3: Strain Cultivation>

[0045] The strains were inoculated into MRS broth medium and placed in an incubator at 37 °C and 5% CO 2 to culture and activate the strains for 24 hours. Then, the activated strains were inoculated into MRS broth medium at a concentration of 2 v / v% and placed in an incubator at 37 °C and 5% CO 2 to culture for 24 hours. After that, the obtained culture was centrifuged at 3,000 rpm for 10 minutes at 4 °C to precipitate the cells. After removing the supernatant, the precipitate was resuspended in RPMI medium supplemented with 10% fetal bovine serum, 2 μM fluoxetine, and 1% penicillin-streptomycin to obtain a bacterial suspension with an appropriate strain concentration.

[0046] <Example 4: Statistical Analysis>

[0047] The obtained experimental data were expressed as "mean ± standard deviation". The differences between the experimental data were evaluated by Student's t-test.

[0048] <Example 5: Analysis of 5-Hydroxytryptophan Secretion>

[0049] After subculture, rat insulinoma cells RIN-14B were divided into three single-strain experimental groups, three single-strain comparison groups, four multi-strain experimental groups, and three multi-strain comparison groups. In addition, the total number of bacteria in the subsequent treatments of each group was the same.

[0050] The three single-bacteria experimental groups were respectively named "BB-115 group", "MP108 group", and "BLI-02 group"; in the "BB-115 group", only the Bifidobacterium animalis subsp. lactis BB-115 strain was used to treat cells subsequently, in the "MP108 group", only the Lactobacillus rhamnosus MP108 strain was used to treat cells subsequently, and in the "BLI-02 group", only the Bifidobacterium longum subsp. infantis BLI-02 strain was used to treat cells subsequently.

[0051] The three single-bacteria comparison groups were respectively named "HN019 group", "LGG group", and "M-63 group"; in the "HN019 group", only the Bifidobacterium animalis subsp. lactis HN019 strain was used to treat cells subsequently, in the "LGG group", only the Lactobacillus rhamnosus LGG strain was used to treat cells subsequently, and in the "M-63 group", only the Bifidobacterium longum subsp. infantis M-63 strain was used to treat cells subsequently.

[0052] The four multi-bacteria experimental groups were respectively named "BB-115:MP108:BLI-02 = 1:1:1 group", "BB-115:MP108:BLI-02 = 2:1:1 group", "BB-115:MP108 = 1:1 group", and "BB-115:BLI-02 = 1:1 group"; in the "BB-115:MP108:BLI-02 = 1:1:1 group", the Bifidobacterium animalis subsp. lactis BB-115 strain, the Lactobacillus rhamnosus MP108 strain, and the Bifidobacterium longum subsp. infantis BLI-02 strain were used to treat cells at a colony-forming unit ratio of 1:1:1 subsequently, in the "BB-115:MP108:BLI-02 = 2:1:1 group", the Bifidobacterium animalis subsp. lactis BB-115 strain, the Lactobacillus rhamnosus MP108 strain, and the Bifidobacterium longum subsp. infantis BLI-02 strain were used to treat cells at a colony-forming unit ratio of 2:1:1 subsequently, in the "BB-115:MP108 = 1:1 group", the Bifidobacterium animalis subsp. lactis BB-115 strain and the Lactobacillus rhamnosus MP108 strain were used to treat cells at a colony-forming unit ratio of 1:1 subsequently, and in the "BB-115:BLI-02 = 1:1 group", the Bifidobacterium animalis subsp. lactis BB-115 strain and the Bifidobacterium longum subsp. infantis BLI-02 strain were used to treat cells at a colony-forming unit ratio of 1:1 subsequently.

[0053] The three combined bacteria comparison groups were respectively named "HN019:MP108 = 1:1 group", "HN019:BLI-02 = 1:1 group", and "HN019:MP108:BLI-02 = 1:1:1 group"; in the "HN019:MP108 = 1:1 group", the subsequent Lactobacillus paracasei subsp. paracasei HN019 strain and Lactobacillus rhamnosus MP108 strain were used to treat cells at a colony-forming unit ratio of 1:1. In the "HN019:BLI-02 = 1:1 group", the subsequent Lactobacillus paracasei subsp. paracasei HN019 strain and Bifidobacterium longum subsp. infantis BLI-02 strain were used to treat cells at a colony-forming unit ratio of 1:1. In the "HN019:MP108:BLI-02 = 1:1:1 group", the subsequent Lactobacillus paracasei subsp. paracasei HN019 strain, Lactobacillus rhamnosus MP108 strain, and Bifidobacterium longum subsp. infantis BLI-02 strain were used to treat cells at a colony-forming unit ratio of 1:1:1.

[0054] Cells were inoculated into 0.5 mL of RPMI medium supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin at a cell number of 2x10 5 and placed in an incubator at 37 °C and 5% CO 2 for 72 hours for culture. After removing the medium, the cells were washed twice with phosphate buffered saline (PBS). Then, the corresponding strain samples were added to the cells at a total bacterial count of 10 8 CFU, and then placed in an incubator at 37 °C and 5% CO 2 for 1 hour for culture. When adding the strain samples, RPMI medium was added to the cells as a control group.

[0055] The cell culture supernatants of each group were analyzed using an enzyme-linked immunosorbent assay kit. Specifically, the total amount of 5-hydroxytryptophan in the cell culture supernatants of each group was measured according to the operation instructions provided by the kit manufacturer. Based on the total amount of 5-hydroxytryptophan in the control group, the relative amount of 5-hydroxytryptophan in each group was calculated; that is,

[0056]

[0057] As Figure 1As shown in Table 2, both the BB-115 strain and the HN019 strain belong to the subspecies Bifidobacterium animalis subsp. lactis, but the BB-115 strain has a better activity in promoting the secretion of 5-hydroxytryptophan by cells than the HN019 strain; although both the MP108 strain and the LGG strain belong to the species Lactobacillus rhamnosus, the MP108 strain has a better activity in promoting the secretion of 5-hydroxytryptophan by cells than the LGG strain; although both the BLI-02 strain and the M-63 strain belong to the subspecies Bifidobacterium longum subsp. infantis, the BLI-02 strain has a better activity in promoting the secretion of 5-hydroxytryptophan by cells than the M-63 strain. The activity of the BB-115 strain in promoting the secretion of 5-hydroxytryptophan by cells is at least twice that of other strains. The above shows that the activity of promoting the secretion of 5-hydroxytryptophan by cells is strain-specific.

[0058] As Figure 1 As shown in Table 2, the combination of the BB-115 strain, the MP108 strain and the BLI-02 strain, the combination of the BB-115 strain and the MP108 strain, and the combination of the BB-115 strain and the BLI-02 strain can bring better activity in promoting the secretion of 5-hydroxytryptophan by cells than the BB-115 strain alone. This shows that the combination of the BB-115 strain with the MP108 strain and / or the BLI-02 strain can produce a synergistic effect on the activity of promoting the secretion of 5-hydroxytryptophan by cells. In addition, the combination of the BB-115 strain, the MP108 strain and the BLI-02 strain can bring better activity in promoting the secretion of 5-hydroxytryptophan by cells than the combination of the BB-115 strain and the MP108 strain and the combination of the BB-115 strain and the BLI-02 strain.

[0059] As Figure 1 As shown in Table 2, the combination of the HN019 strain, the MP108 strain and the BLI-02 strain, the combination of the HN019 strain and the MP108 strain, and the combination of the HN019 strain and the BLI-02 strain can bring better activity in promoting the secretion of 5-hydroxytryptophan by cells than the HN019 strain alone. This shows that the combination of the HN019 strain with the MP108 strain and / or the BLI-02 strain can produce a synergistic effect on the activity of promoting the secretion of 5-hydroxytryptophan by cells. However, the activities of promoting the secretion of 5-hydroxytryptophan caused by the combination of the HN019 strain, the MP108 strain and the BLI-02 strain, the combination of the HN019 strain and the MP108 strain, and the combination of the HN019 strain and the BLI-02 strain are comparable. The above shows that the BB-115 strain, the MP108 strain and the BLI-02 strain will act together to produce a synergistic effect on the activity of promoting the secretion of 5-hydroxytryptophan by cells.

[0060] Table 2. Relative expression level of tryptophan

[0061] Group Average relative expression level of tryptophan Control group 1.0 BB-115 group 12.50 HN019 group 6.08 MP108 group 6.89 LGG group 2.71 BLI-02 group 7.03 M-63 group 5.94 BB-115:MP108:BLI-02 = 1:1:1 group 14.33 BB-115:MP108:BLI-02 = 2:1:1 group 18.36 BB-115:MP108 = 1:1 group 13.15 BB-115:BLI-02 = 1:1 group 13.08 HN019:MP108 = 1:1 group 8.89 HN019:BLI-02 = 1:1 group 8.75 HN019:MP108:BLI-02 = 1:1:1 group 9.07

[0062] In summary, the Bifidobacterium animalis subsp. lactis BB-115 strain has the characteristic of enhancing the secretion of 5-hydroxytryptophan by enterochromaffin cells, and it can effectively and synergistically enhance the activity of enterochromaffin cells to secrete 5-hydroxytryptophan with the Lactobacillus rhamnosus MP108 strain and the Bifidobacterium longum subsp. infantis BLI-02 strain, and thus it is expected to be used to solve emotional problems such as insomnia, depression, anxiety, and inattention.

[0063] However, the above are only the preferred embodiments of the present invention, but the protection scope of the present invention cannot be limited thereby; therefore, all simple equivalent changes and modifications made according to the protection scope of the present invention and the content of the specification still fall within the protection scope of the present invention.

[0064]

Deposit of Biological Material

[0065] 1. CGMCC No. 21840, deposited with China General Microbiological Culture Collection Center on February 26, 2021;

[0066] 2. CGMCC No. 21225, deposited with China General Microbiological Culture Collection Center on November 23, 2020;

[0067] 3. CGMCC No. 15212, deposited with China General Microbiological Culture Collection Center on January 15, 2018.

Claims

1. A strain use, characterized in that: The invention is used for preparing a composition for enhancing the secretion of 5-hydroxytryptophan by enterochromaffin cells, wherein the strain comprises the animal Bifidobacterium lactis subsp. BB-115 strain, and the BB-115 strain is deposited in the China General Microbiological Culture Collection Center with the deposit number CGMCC21840.

2. The use according to claim 1, characterized in that The BB-115 strain is an active strain or a deactivated strain.

3. The use according to claim 1, characterized in that The composition is further used for enhancing immunity, treating insomnia, treating depression, treating hypertension, improving sleep disorders, improving anxiety, improving lack of concentration, treating menopausal syndrome, or treating gastrointestinal diseases.

4. The use according to claim 1, characterized in that The composition is a medicine, a food, or a health product.

5. A use of a strain, characterized in that: Used to prepare a composition for enhancing the secretion of 5-hydroxytryptophan by enterochromaffin cells; wherein the strains include animal Bifidobacterium lactis subsp. BB-115 strain, rhamnosus Lactobacillus MP108 strain, and Bifidobacterium longum subsp. infantis BLI-02 strain; wherein the BB-115 strain is deposited in the China Center for the Collection of General Microorganisms with a deposit number of CGMCC21840, the MP108 strain is deposited in the China Center for the Collection of General Microorganisms with a deposit number of CGMCC21225, and the BLI-02 strain is deposited in the China Center for the Collection of General Microorganisms with a deposit number of CGMCC15212.

6. The use according to claim 5, characterized in that The BB-115 strain, the MP108 strain, and the BLI-02 strain are individually active strains or inactivated strains.

7. The use according to claim 5, characterized in that The colony forming unit ratio among the BB-115 strain, the MP108 strain, and the BLI-02 strain is (1 to 2):1:

1.

8. The use according to claim 5, characterized in that The colony forming unit ratio among the BB-115 strain, the MP108 strain, and the BLI-02 strain is 2:1:

1.

9. The use according to claim 5, characterized in that The composition is further used for enhancing immunity, treating insomnia, treating depression, treating hypertension, improving sleep disorders, improving anxiety, improving lack of concentration, treating menopausal syndrome, or treating gastrointestinal diseases.

10. The use according to claim 5, characterized in that The composition is a medicine, a food, or a health product.

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