Application of lactic acid bacteria strain composition in preparation of composition for relieving mood and promoting neuronal growth

Abstract

The invention relates to the technical field of probiotics, in particular to application of a lactic acid bacteria strain composition in preparation of a composition for relieving mood and promoting neuronal growth. The lactic acid bacteria strain composition comprises a bifidobacterium breve BBr-28 strain, a bifidobacterium animalis subsp. Lactis PS23 strain, a bifidobacterium longum subsp. Infantis BR631 strain, a bifidobacterium bifidum BD-188 strain and a plant lactobacillus PQQ-1 strain. The lactic acid bacteria strain composition has physiological activity functions of relieving mood and promoting neuronal growth, so that the lactic acid bacteria strain composition can be used for preparing food or medicine compositions for relieving mood, and the problems of low mood and depression of melancholia patients can be relieved.

Description

Technical Field

[0001] The present invention relates to the technical field of probiotics, and in particular to application of a lactic acid bacteria strain composition in preparing a composition for relieving emotions and promoting neuron growth. Background Art

[0002] According to statistics from the World Health Organization, depression will be the disease that causes the world's number one socioeconomic burden in 2030. This means that as society changes, more people may suffer from depression, and the socioeconomic burden it causes will be more serious. However, many patients with depression may be unwilling to admit that they have depression or resist treatment. Therefore, most patients with depression do not receive appropriate medical care.

[0003] Depression is a normal emotion. For example, when people encounter setbacks or unpleasant situations, they will inevitably feel depressed. However, if the low mood is more severe and lasts for a long time without recovery, or even out of control, it may develop into depression.

[0004] In the prior art, there are extensive studies on the effects of probiotics on human health. Generally speaking, strains with special effects on human health are called functional probiotics (Guidelines for the evaluation of probiotics in food; Report of joint FAO / WHO working group on drafting guidelines for the evaluation of probiotics in food; London Ontario, Canada April 30 and May 1, 2002: 1-7). The prior art also discloses various functional probiotics or, however, the special effects of functional probiotics lie in the specificity of the strain rather than the species, that is, even if the same species, different strains may still have different effects, or even opposite effects.

[0005] In view of this, it is urgent to develop a safe and long-term edible nutritional supplement with soothing effects. Lactic acid bacteria are generally considered safe, so finding a lactic acid bacteria strain composition with soothing effects is a goal that needs to be worked on. Summary of the invention

[0006] The purpose of the present application is to address the deficiencies of the prior art and to provide an application of a lactic acid bacteria strain composition in the preparation of a composition for relieving emotions and promoting neuron growth.

[0007] The above invention objectives of the present application are achieved through the following technical solutions: The present invention aims to provide an application of a lactic acid bacteria strain composition in preparing a composition for soothing emotions and promoting neuron growth. The lactic acid bacteria strain composition comprises a Bifidobacterium breve BBr-28 strain with a preservation number of CGMCC NO.32485, a Bifidobacterium animalis subsp. lactis PS23 strain with a preservation number of CGMCC NO.32691, a Bifidobacterium longum subsp. infantis BR631 strain with a preservation number of CGMCC NO.26243, a Bifidobacterium bifidum BD-188 strain with a preservation number of CGMCC NO.32484, and a Lactiplantibacillus plantarum PQQ-1 strain with a preservation number of CGMCC NO.29303.

[0008] In the above technical solution, the effective viable count of the lactic acid bacteria composition is equal to or greater than 5×10 9 CFU / g.

[0009] In the above technical scheme, the strain quantity ratio of the Bifidobacterium breve BBr-28 strain, the Bifidobacterium animalis lactis subspecies PS23 strain, the Bifidobacterium longum infantis subspecies BR631 strain, the Bifidobacterium bifidum BD-188 strain and the Lactobacillus plantarum PQQ-1 strain is (0.5-1.5):(1-2.5):(1-2.5):(1-2.5):(0.5-1.5).

[0010] In the above technical scheme, the strain amount ratio of the Bifidobacterium breve BBr-28 strain, the Bifidobacterium animalis lactis subspecies PS23 strain, the Bifidobacterium longum infantis subspecies BR631 strain, the Bifidobacterium bifidum BD-188 strain and the Lactobacillus plantarum PQQ-1 strain is 1:2:2:2:1.

[0011] In the above technical solution, the composition for soothing emotions and promoting neuron growth is a food composition or a pharmaceutical composition.

[0012] In the above technical solution, the composition includes the lactic acid bacteria strain composition and a carrier.

[0013] In the above technical solution, the carrier is a physiologically or pharmaceutically acceptable excipient or diluent.

[0014] In the above technical solution, the dosage form of the composition is tablet, capsule or powder.

[0015] Beneficial effects of this application: The invention provides an application of a lactic acid bacteria strain composition in preparing a composition for soothing emotions and promoting neuron growth. The lactic acid bacteria strain composition comprises a Bifidobacterium breve BBr-28 strain with a preservation number of CGMCC NO.32485, a Bifidobacterium animalis subsp. lactis PS23 strain with a preservation number of CGMCC NO.32691, a Bifidobacterium longum subsp. infantis BR631 strain with a preservation number of CGMCC NO.26243, a Bifidobacterium bifidum BD-188 strain with a preservation number of CGMCC NO.32484, and a Lactiplantibacillus plantarum PQQ-1 strain with a preservation number of CGMCC NO.29303. The present invention uses the composition capsule made of the powder of the above five lactic acid bacteria strains and the carrier to conduct experiments. Before taking the composition of the embodiment group and the control group and after taking it for 2 weeks, the blood of the subjects is collected to detect the changes of serotonin, dopamine, brain-derived neurotrophic factor (BDNF) and melatonin. The test results show that: 1) the increase of serotonin in the subjects of the embodiment group (the composition contains five lactic acid bacteria strains) is significantly better than that of the control group (the composition contains only a single lactic acid bacteria strain); 2) the dopamine in the blood of the subjects of the embodiment group before and after taking the composition is significantly increased, and there is a significant difference compared with before taking it, indicating that the composition of the present invention has a physiological activity effect of promoting the production of dopamine in the human body, and thus has the effect of soothing the user's emotions, especially the problem of depression caused by insufficient dopamine; 3) the melatonin in the blood of the subjects of the embodiment group before and after taking the composition is significantly reduced, and there is a significant difference compared with before taking it, indicating that the composition of the present invention has a physiological activity effect of adjusting the physiological clock, and thus has the effect of improving sleep quality and soothing emotions. It can be seen from this that the lactic acid bacteria strain composition of the present invention has the physiological activity function of soothing emotions and promoting neuronal growth. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 The test results are for the changes in serotonin levels before and after the subjects took the compositions of the example group and the control group, respectively.

[0017] Figure 2The figures are comparison results of the changes in serotonin levels before and after the subjects took the compositions of the example group and the control group, respectively.

[0018] Figure 3 The test results are for the changes in dopamine before and after the subjects took the composition of the Example group.

[0019] Figure 4 The test results are for the changes in BDNF before and after the subjects took the composition of the Example group.

[0020] Figure 5 The test results are for the changes in melatonin before and after the subjects took the composition of the Example group. DETAILED DESCRIPTION

[0021] The microbial strain materials involved in the following examples are as follows: 1. Bifidobacterium breve BBr-28 strain Storage date: 2024 / 11 / 04 Depository: China National Microbiological Culture Collection Administration General Microbiology Center Address of the depository: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, China 100101, China Deposit number: CGMCC No.32485 Classification name: Bifidobacterium breve 2. Bifidobacterium animalis subsp. lactis PS23 strain Storage date: 2024 / 11 / 18 Depository: China National Microbiological Culture Collection Administration General Microbiology Center Address of the depository: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, China 100101, China. Deposit number: CGMCC No.32691 Classification and nomenclature: Bifidobacterium animalis subsp. lactis 3. Bifidobacterium longum subspecies infantis BR631 strain Storage date: 2022 / 12 / 25 Depository: China National Microbiological Culture Collection Administration General Microbiology Center Address of the depository: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, China 100101, China. Deposit number: CGMCC No.26243 Classification name: Bifidobacterium longum subsp. infantis 4. Bifidobacterium bifidum BD-188 strain Storage date: 2024 / 11 / 04 Depository: China National Microbiological Culture Collection Administration General Microbiology Center Address of the depository: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, China 100101, China. Deposit number: CGMCC NO.32484 Classification name: Bifidobacterium bifidum 5. Lactobacillus plantarum PQQ-1 strain Storage date: 2023 / 12 / 12 Depository: China National Microbiological Culture Collection Administration General Microbiology Center Address of the depository: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, China 100101, China. Deposit number: CGMCC NO.29303 Classification name: Lactiplantibacillus plantarum The freeze-dried culture of the above strains has been deposited in the General Microbiological Center of China Microbiological Culture Collection Administration, located at No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, China. The detailed information of the deposit is shown in Table 1: Table 1. Deposit information of lactic acid bacteria strains of the present invention There are extensive studies on the effects of probiotics on human health. Generally speaking, strains that have special effects on human health are called functional probiotics (Guidelines for the evaluation of probiotics information; Report of joint FAO / WHO working group on drafting guidelines for the evaluation of probiotics in food; London Ontario, Canada April 30 and May 1, 2002: 1-7). It should be noted that the special effects of the above-mentioned functional probiotics lie in the specificity of the strain rather than the species, that is, even if they are of the same species, different strains may still have different effects, or even opposite effects.

[0022] On this basis, the present invention proposes a new use of a lactic acid bacteria strain composition in preparing a composition for soothing emotions and promoting neuron growth. The lactic acid bacteria strain composition includes Bifidobacterium breve BBr-28 strain, Bifidobacterium animalis lactis subsp. PS23 strain, Bifidobacterium longum infantis subsp. BR631 strain, Bifidobacterium bifidum BD-188 strain and Lactobacillus plantarum PQQ-1 strain.

[0023] The strain quantity ratio of Bifidobacterium breve BBr-28, Bifidobacterium animalis lactis subspecies PS23, Bifidobacterium longum infantis subspecies BR631, Bifidobacterium bifidum BD-188 and Lactobacillus plantarum PQQ-1 is (0.5-1.5):(1-2.5):(1-2.5):(1-2.5):(0.5-1.5); preferably, the strain quantity ratio of the above five strains is 1:2:2:2:1, or 1:1.5:1.5:1.5:1, or 1:1:1:1:1, or 0.5:1.5:1.5:1.5:0.5, or 1.5:2:2:2:1.5.

[0024] The above lactic acid bacteria strain composition has the physiological activity function of soothing emotions and promoting neuron growth, so the above lactic acid bacteria strain composition can be used to prepare food or pharmaceutical compositions for soothing emotions and promoting neuron growth. Among them, the effective viable count of the lactic acid bacteria strain composition is equal to or greater than 1×10 6 CFU / g, preferably, the effective viable count of the lactic acid bacteria composition is equal to or greater than 5×10 9 CFU / g.

[0025] The properties and characteristics of the lactic acid bacteria strain of the present invention are as follows: The taxonomic characteristics of the strains were confirmed based on the results of 16S rRNA sequence analysis and API bacterial identification system analysis. The morphological and general characteristics of the lactic acid bacteria strains of the present invention are listed in detail in Table 2: Table 2. Morphology and general properties of the lactic acid bacteria strains of the present invention The present application is further described in detail below in conjunction with the accompanying drawings. Example

[0026] The preparation method of the composition for soothing emotions and promoting neuron growth is as follows: Step S1, preparing a lactic acid bacteria strain composition Bifidobacterium breve BBr-28, Bifidobacterium animalis subsp. lactis PS23, Bifidobacterium longum subsp. infantis BR631, and Bifidobacterium bifidum BD-188 were isolated from breast milk; Lactobacillus plantarum PQQ-1 was isolated from snow lotus. The above five isolated lactic acid bacteria strains were stored at -80°C with 20% glycerol. When used, each was inoculated into Difco TM MRS (De Man, Rogosa and Sharpe) medium and cultured at 37°C for 20 hours. Afterwards, the five cultures were centrifuged at 4°C for 20 minutes, the supernatant was removed, and the precipitate was freeze-dried to obtain the bacterial powder of each lactic acid bacteria strain. The precipitate after centrifugation can be mixed with a protective carrier and then freeze-dried to maintain better bacterial activity.

[0027] The bacterial powders of the above-obtained Bifidobacterium breve BBr-28 strain, Bifidobacterium animalis lactis subspecies PS23 strain, Bifidobacterium longum infantis subspecies BR631 strain, Bifidobacterium bifidum BD-188 strain and Lactobacillus plantarum PQQ-1 strain were mixed in a mass ratio of 1:2:2:2:1 to obtain a lactic acid bacteria strain composition.

[0028] Step S2: preparing a composition for relieving emotions and promoting neuron growth The lactic acid bacteria strain composition of step S1 is evenly mixed with a carrier to obtain a composition for soothing emotions and promoting neuron growth. The dosage form of the composition can be a tablet, capsule or powder to facilitate oral use by the user. Wherein: the carrier can be a physiologically or pharmaceutically acceptable excipient or diluent. Specifically, the carrier comprises oligosaccharides, sugar alcohols, sodium carboxymethyl cellulose, ethyl cellulose, galacto-oligosaccharides, stearic acid, magnesium stearate, potato starch or a combination thereof. Further, the oligosaccharides include fructooligosaccharides, isomaltooligosaccharides, xylo-oligosaccharides, stachyose, mannose, arabinose or a combination thereof; the sugar alcohols include erythritol, sorbitol, isomaltulose, xylitol, mannitol or a combination thereof.

[0029] The effective viable count of the lactic acid bacteria composition is equal to or greater than 1×10 6 CFU / g, preferably, the effective viable count of the lactic acid bacteria composition is equal to or greater than 5×10 9 CFU / g.

[0030] Physiological activity experiments and analysis of the composition of this example: In the following experiments, the composition was prepared into capsule form to facilitate oral administration by users.

[0031] The 30 subjects who participated in the experiment were divided into two groups, one of which took the composition capsule of this example, and the other took the composition capsule containing only the bacterial powder of animal Bifidobacterium lactis subsp. PS23 strain. The specific experimental contents are as follows: Example group: The subjects took the composition capsule of this example twice a day, once between 2-3 pm and once 2 hours before going to bed, for a period of 2 weeks.

[0032] Control group: The composition capsules taken by the subjects only contained bacterial powder of Bifidobacterium animalis subsp. lactis PS23 strain, and the administration method was the same as that of the example group.

[0033] Before and 2 weeks after taking the composition of the Example Group and the Control Group, blood samples were collected from the subjects to detect changes in serotonin, and changes in dopamine, brain-derived neurotrophic factor (BDNF) and melatonin were detected for the subjects of the Example Group.

[0034] (I) Effect of taking the composition of this example on serotonin: Serotonin, also known as 5-hydroxytryptamine or serotonin (5-HT), is a monoamine neurotransmitter that is responsible for transmitting emotion-related signals in the brain. Serotonin is responsible for regulating emotional responses. When serotonin is deficient, people will experience depression, sadness, anxiety and other emotions. When serotonin is sufficient, people's emotions are more stable and easier to relax, and they can even feel happy.

[0035] Before taking the composition of the embodiment group and the control group and after taking it for 2 weeks, about 5 ml of blood was collected from the subjects for serotonin detection. The collected blood was centrifuged to separate the serum. The serum was processed using serotonin ELISA kits (abnova) according to the original manufacturer's operating manual, and after using HRP (horseradish peroxidase) to catalyze TMB (3,3',5,5'-Tetramethyl-benzidine) for color development, the absorption light was measured using ELISA Reader O.D450nm, and the serotonin concentration was converted with the standard curve. The test results are shown in Figure 1 and Figure 2 .

[0036] Figure 1 a is the test result of the change of serotonin before and after the subject took the composition of the Example group; Figure 1 b is the test result of the changes of serotonin before and after the subjects took the composition of the control group. Figure 1 a and Figure 1 b It can be seen that after the subjects took the composition of the example group and the control group for 2 weeks, the serotonin in the blood increased, and there was a significant difference compared with before taking the composition (p<0.001).

[0037] Figure 2 The comparison results of the changes in serotonin levels after the subjects took the composition of the example group and the control group are presented as Mean±SD. Statistics were calculated using paired t-test, and p<0.05 was considered to be significantly different. Figure 2 As shown, the change in the amount of serotonin in the subject before and after taking the composition of the example group (i.e., the difference in serotonin before and after taking) was compared with the control group. The increase in the amount of serotonin in the subject taking the composition of the example group was better than that in the control group, and there was a significant difference (p=0.019). Therefore, the increase in serotonin in the example group containing the five lactic acid bacteria strains was significantly better than that in the control group containing only a single lactic acid bacteria strain.

[0038] 1. Effect of taking the composition of this example on dopamine: Dopamine is a neurotransmitter that can be produced by the human body. The nervous system transmits messages between nerve cells through dopamine. Dopamine plays an important role in making people feel happy. It helps people strive for progress, concentrate and be interested in things. When dopamine is insufficient, people may feel tired, inattentive, lack of motivation and mood swings.

[0039] Before taking the composition of the example group and the control group and after taking it for 2 weeks, the blood of the subjects was collected for dopamine detection. The collected blood was centrifuged and then the serum was separated. The serum was processed using Dopamine ELISA kits (abnova, KA1887) according to the original manufacturer's operating manual, and the absorption light was measured using an ELISA Reader OD 450nm after color development, and the dopamine concentration was converted with a standard curve. The test results are shown in Figure 3 .

[0040] Figure 3 The test results are for the changes in dopamine before and after the subjects took the composition of the Example group. Statistics were calculated using paired t-test, and p<0.05 was considered to be significantly different. Figure 3 It can be seen that after taking the composition of the example group, the dopamine in the blood of most subjects increased significantly, and there was a significant difference compared with before taking (p = 0.0073). Therefore, the composition of this example has a physiological activity effect of promoting the production of dopamine in the human body, and thus has the effect of soothing the user's emotions, especially the problem of depression caused by insufficient dopamine.

[0041] 2. Effect of taking the composition of this example on brain-derived neurotrophic factor (BDNF): Brain-derived neurotrophic factor (BDNF) is the most abundant nerve growth factor in the brain. It can promote the growth of neurons (nerve cells) in the brain, promote the formation of synapses of brain nerve cells, and regulate the transmission of neurotransmitters and intracellular messages. Studies have shown that a lack of BDNF may cause some cognitive impairment, such as Alzheimer's disease, which is one of the most common degenerative dementias.

[0042] Before and 2 weeks after taking the composition of the Example Group, blood was collected from the subjects for BDNF testing. The collected blood was centrifuged and serum was separated. The serum was processed using BDNF ELISA kits (Aviva, OKBB00127) according to the original manufacturer's operating manual, and the absorbance was measured using an ELISA Reader OD 450nm after color development, and the BDNF concentration was converted with a standard curve. The test results are shown in Figure 4 .

[0043] Figure 4 The results are the test results of the changes in BDNF before and after the subjects took the composition of the Example group. Statistics were calculated using paired t-test, and p<0.05 was considered to be significantly different. Figure 4 It can be seen that after taking the composition of the Example group, the BDNF in the blood of most subjects increased significantly, and there was a significant difference compared to before taking the composition of the Example group (p = 0.0111). Therefore, the composition of the Example of the present invention has the physiological activity effect of promoting the production of BDNF in the human body, and thus has the effect of promoting neuronal growth, thereby reducing neurocognitive dysfunction diseases such as Alzheimer's disease.

[0044] 3. Effect of taking the composition of this example on melatonin: Depressed patients usually have sleep disorders and circadian rhythm disorders. Clinically depressed patients often experience worsening depression due to poor sleep. Melatonin has the function of adjusting the biological clock. Generally speaking, melatonin secretion is low during the day and increases rapidly at night. When the amount of melatonin secreted during the day is relatively high, it may lead to poor spirits during the day.

[0045] Before taking the composition of the example group and after taking it for 2 weeks, the blood of the subjects was collected for melatonin detection. It should be noted that the blood of the subjects was collected during the day. The collected blood was centrifuged and the serum was separated. The serum was processed using Melatonin ELISA kits (Aviva, OKEH02566) according to the original manufacturer's operating manual. After color development, the absorption light was measured using an ELISA Reader OD 450nm, and the melatonin concentration was converted with a standard curve. The test results are shown in Figure 5 .

[0046] Figure 5 The test results are for the changes in melatonin levels before and after the subjects took the composition of the Example group. Statistics were calculated using paired t-test, and p<0.05 was considered to be significantly different. Figure 5 It can be seen that after taking the composition of the example group, the melatonin in the blood of most subjects was significantly reduced, and there was a significant difference compared with before taking (p = 0.0028), indicating that better sleep quality was obtained after taking the composition of this example. Therefore, the composition of this example has a physiological activity effect of adjusting the biological clock, and thus has the effect of improving sleep quality and thus soothing emotions.

[0047] In summary, the composition of the embodiment of the present invention has physiologically active functions of soothing emotions and promoting neuron growth. It should be noted that, compared with the composition containing only a single lactic acid bacteria strain (control group), the composition of this embodiment containing 5 lactic acid bacteria strains can significantly increase serotonin and soothe emotions, which shows that the lactic acid bacteria strain composition of the present invention has unexpected effects.

[0048] The embodiments described above are only used to illustrate the technical solutions of the present application, rather than to limit them. Although the present application has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or make equivalent replacements for some of the technical features therein. Such modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present application, and should all be included in the protection scope of the present application.

Claims

1. Use of a lactic acid bacteria strain composition in the preparation of a composition for soothing emotions and promoting neuron growth, characterized in that: The lactic acid bacteria strain composition comprises Bifidobacterium breve BBr-28 strain with a preservation number of CGMCC NO. 32485, Bifidobacterium animalis lactis subspecies PS23 strain with a preservation number of CGMCC NO. 32691, Bifidobacterium longum infantis subspecies BR631 strain with a preservation number of CGMCC NO. 26243, Bifidobacterium bifidum BD-188 strain with a preservation number of CGMCC NO. 32484 and Lactobacillus plantarum PQQ-1 strain with a preservation number of CGMCC NO. 29303.

2. The use according to claim 1, characterized in that: The effective viable count of the lactic acid bacteria composition is equal to or greater than 5×10 9 CFU / g.

3. The use according to claim 1, characterized in that: The strain quantity ratio of the Bifidobacterium breve BBr-28 strain, the Bifidobacterium animalis lactis subspecies PS23 strain, the Bifidobacterium longum infantis subspecies BR631 strain, the Bifidobacterium bifidum BD-188 strain and the Lactobacillus plantarum PQQ-1 strain is (0.5-1.5): (1-2.5): (1-2.5): (1-2.5): (0.5-1.5).

4. The use according to claim 3, characterized in that: The strain quantity ratio of the Bifidobacterium breve BBr-28 strain, the Bifidobacterium animalis lactis subspecies PS23 strain, the Bifidobacterium longum infantis subspecies BR631 strain, the Bifidobacterium bifidum BD-188 strain and the Lactobacillus plantarum PQQ-1 strain is 1:2:2:2:

1.

5. The use according to claim 1, characterized in that: The composition for relieving emotions and promoting neuron growth is a food composition or a pharmaceutical composition.

6. The use according to claim 1, characterized in that: The composition comprises the lactic acid bacteria strain composition and a carrier.

7. The use according to claim 6, characterized in that: The carrier is a physiologically or pharmaceutically acceptable excipient or diluent.

8. The use according to claim 1, characterized in that: The composition is in the form of tablets, capsules or powders.

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