Lactic acid bacteria strain composition for improving sleep quality and application

Abstract

The invention relates to the technical field of probiotics, in particular to a lactic acid bacteria strain composition for improving sleep quality and application, and 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 and a cheese paracasei BR176 strain. The lactic acid bacteria strain composition has the physiological activity function of improving the sleep quality, so that the lactic acid bacteria strain composition can be used for preparing the composition for improving the sleep quality so as to improve the sleep quality.

Description

Technical Field

[0001] The present invention relates to the technical field of probiotics, and in particular to a lactic acid bacteria strain composition for improving sleep quality and an application thereof. Background Art

[0002] Sleeping conditions not only affect physical health, but also lead to premature aging. Therefore, sleep quality is an important factor in whether one can have normal life functions. However, with the popularity of 3C electronic products such as smart phones, many people still use their phones before going to bed, such as browsing social messages, watching TV series, playing games, etc. These behaviors can easily reduce the user's sleep quality and lead to an increasing number of insomniacs every year. Therefore, insomnia problems do not only occur in the elderly population, but have a trend of becoming younger, and the peak of insomnia is among people aged 20 to 40.

[0003] 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). The prior art also discloses various functional probiotics, but the special effects of 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.

[0004] In view of this, it is urgent to develop a safe and long-term edible nutritional supplement that can improve sleep quality to improve sleep problems. Lactic acid bacteria are generally considered safe, so finding a composition containing lactic acid bacteria strains that have the activity of improving sleep quality is a goal that needs to be worked on at present. Summary of the invention

[0005] The purpose of the present application is to address the deficiencies of the prior art and to provide a lactic acid bacteria strain composition and application for improving sleep quality.

[0006] The above invention objectives of the present application are achieved through the following technical solutions: One of the purposes of the present invention is to provide a lactic acid bacteria strain composition for improving sleep quality. The lactic acid bacteria strain composition includes Bifidobacterium breve BBr-28 strain with a preservation number of CGMCC NO.32485, Bifidobacterium animalis subsp. lactis PS23 strain with a preservation number of CGMCC NO.32691, Bifidobacterium longum subsp. infantis BR631 strain with a preservation number of CGMCC NO.26243, and Lacticaseibacillus paracasei BR176 strain with a preservation number of CGMCC NO.21563.

[0007] In the above technical solution, 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 and the Casei Bacillus paracasei BR176 strain is 1:(0.5~1.5):(0.5~1.5):(0.5~1.5).

[0008] A second object of the present invention is to provide an application of the lactic acid bacteria composition in the preparation of a composition for improving sleep quality.

[0009] 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.

[0010] A third object of the present invention is to provide a composition for improving sleep quality, wherein the composition comprises the above-mentioned lactic acid bacteria strain composition and a carrier.

[0011] In the above technical solution, the strain amount ratio of the Bifidobacterium breve BBr-28 strain, the Bifidobacterium animalis subspecies lactis PS23 strain, the Bifidobacterium longum subspecies infantis BR631 strain and the Casei Bacillus paracasei BR176 strain is 1:1:1:1.

[0012] In the above technical solution, the effective viable count of all lactic acid bacteria strains in the composition is equal to or greater than 5×10 9 CFU / g.

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

[0014] In the above technical solution, the carrier is a pharmaceutically acceptable excipient or diluent.

[0015] In the above technical solution, the dosage form of the composition is tablet, capsule or powder.

[0016] Beneficial effects of this application: The invention provides a lactic acid bacteria strain composition for improving sleep quality, comprising 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, and a Lacticaseibacillus paracasei BR176 strain with a preservation number of CGMCC NO.21563. The present invention experiments on the composition made of the above-mentioned lactic acid bacteria strain composition and the carrier, and evaluates the sleep of the subjects with the Pittsburgh Sleep Quality Index (PSQI). Before taking the composition of the experimental group and the control group and after taking it for 2 weeks, the blood of the subjects is collected to detect the changes in serotonin, and the wearable medical watch sleep analyzer and the PSQI questionnaire are used to observe the improvement of the sleep quality of the subjects. The results show that 1) the increase in serotonin in the experimental group containing four lactic acid bacteria strains is significantly better than that in the control group containing only a single lactic acid bacteria strain; 2) the improvement of sleep quality in the experimental group containing four lactic acid bacteria strains is better than that in the control group containing only a single lactic acid bacteria strain; 3) taking the composition of the experimental group can significantly improve the sleep quality of the subjects. It can be shown that the lactic acid bacteria strain composition of the present invention has a physiologically active function of improving sleep quality, which can be used to prepare a composition for improving sleep quality to improve the problem of sleep quality. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 The test results are for the changes in serotonin levels of the subjects of Example 2 before and after taking the compositions of the experimental group and the control group, respectively.

[0018] Figure 2 The results are compared for the changes in serotonin levels of the subjects in Example 2 before and after taking the compositions of the experimental group and the control group, respectively.

[0019] Figure 3 The test results of the deep sleep and light sleep ratios of the subjects in Example 2 before and after taking the composition of the experimental group are shown.

[0020] Figure 4These are the test results of the percentage of deep sleep and light sleep of the subjects in Example 2 before and after taking the composition of the control group.

[0021] Figure 5 The comparison results of the changes in the proportions of deep sleep and light sleep after the subjects of Example 2 took the composition of the experimental group.

[0022] Figure 6 The results are compared for the changes in the percentages of deep sleep and light sleep after the subjects of Example 2 took the composition of the control group.

[0023] Figure 7 The results of changes in PSQI for the subjects of Example 2 before and after taking the composition of the experimental group. DETAILED DESCRIPTION

[0024] 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. Bacillus paracasei BR176 strain Storage date: 2020 / 12 / 28 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.21563 Classification name: Lacticaseibacillus paracasei 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 The present application is further described in detail below in conjunction with the accompanying drawings. Embodiment 1: 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.

[0025] On this basis, the first embodiment of the present invention proposes a lactic acid bacteria strain composition for improving sleep quality, including Bifidobacterium breve BBr-28 strain, Bifidobacterium animalis lactis subsp. PS23 strain, Bifidobacterium longum subsp. infantis BR631 strain and Bacillus paracasei BR176 strain.

[0026] The strain ratio of Bifidobacterium BBr-28, Bifidobacterium animalis subsp. lactis PS23, Bifidobacterium longum subsp. infantis BR631 and Bacillus paracasei BR176 is 1:(0.5-1.5):(0.5-1.5):(0.5-1.5). Preferably, the strain ratio of the above four strains is 1:1:1:1.

[0027] 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 preparation method of the lactic acid bacteria composition of the present invention: Bifidobacterium breve BBr-28, Bifidobacterium animalis subsp. lactis PS23, and Bifidobacterium longum subsp. infantis BR631 were isolated from breast milk; and Casei paracasei BR176 was isolated from infant feces. The above four 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 four 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.

[0028] The above lactic acid bacteria strain composition has the physiological activity function of improving sleep quality, so the above lactic acid bacteria strain composition has the purpose of improving sleep quality and can be used to prepare a composition for improving sleep quality.

[0029] Embodiment 2: The second embodiment of the present invention provides a composition for improving sleep quality, comprising a lactic acid bacteria strain composition having an active effect of improving sleep quality and a carrier. The lactic acid bacteria strain composition is the same as that in Example 1.

[0030] The carrier may be a physiologically acceptable excipient or diluent. Alternatively, the carrier may be a pharmaceutically acceptable excipient or diluent. Specifically, the carrier may include oligosaccharides, sugar alcohols, sodium carboxymethyl cellulose, ethyl cellulose, galacto-oligosaccharides, stearic acid, magnesium stearate, potato starch or a combination thereof. Further, the oligosaccharides may include fructooligosaccharides, isomaltooligosaccharides, xylo-oligosaccharides, stachyose, mannose, arabinose or a combination thereof; the sugar alcohols may include erythritol, sorbitol, isomalt, xylitol, mannitol or a combination thereof.

[0031] In the composition for improving sleep quality of the present invention, the effective viable count of the lactic acid bacteria composition is equal to or greater than 1×10 6 CFU / g, as a preferred embodiment, the effective viable count of the lactic acid bacteria composition is equal to or greater than 5×10 9 CFU / g.

[0032] The composition for improving sleep quality of the present invention can be in the form of tablets, capsules or powders, so as to facilitate oral administration by users.

[0033] The preparation method of the composition for improving sleep quality of the present invention: Step 1: preparing a lactic acid bacteria strain composition (same as in Example 1) Bifidobacterium breve BBr-28, Bifidobacterium animalis subsp. lactis PS23, and Bifidobacterium longum subsp. infantis BR631 were isolated from breast milk; and Casei paracasei BR176 was isolated from infant feces. The above four 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 four 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.

[0034] Step 2: Prepare a composition for improving sleep quality The lactic acid bacteria powder of step 1 is mixed evenly with the carrier to obtain the composition of the present invention, wherein the effective viable count of the lactic acid bacteria strain is equal to or greater than 5×10 9 CFU / g.

[0035] Physiological activity experiments and analysis of the composition of the present invention: The composition of the present invention is prepared into a capsule dosage form for convenient oral use by users. The 30 subjects participating in the experiment were divided into two groups, namely an experimental group and a control group, wherein: Experimental group: The patients took the capsules of the composition of the present invention twice a day, once between 2 and 3 pm and once 2 hours before going to bed, for a period of 2 weeks.

[0036] Control group: The taken composition capsules only contained bacterial powder of Bifidobacterium animalis subsp. lactis PS23 strain, and the rest of the taking method was the same as that of the experimental group.

[0037] The subjects were evaluated for sleep using the Pittsburgh Sleep Quality Index (PSQI) questionnaire. PSQI was proposed by American scholars Buysse et al. It has seven scores. The higher the PSQI score, the worse the sleep quality. When PQSI>5, it is assessed as having sleep disorders.

[0038] Before taking the composition of the experimental group and the control group and after taking it for 2 weeks, the subjects' blood was collected to detect changes in serotonin, and the improvement of the subjects' sleep quality was observed using a wearable medical watch sleep analyzer and a PSQI questionnaire.

[0039] (I) Effects of taking the composition of the present invention on serotonin: Serotonin, also known as 5-hydroxytryptamine or serotonin (5-HT), is a monoamine neurotransmitter that has functions including regulating mood, appetite and sleep. Studies have shown that when serotonin is insufficient, it is easy to feel sluggish, moody, restless, lack of melatonin or sleeplessness. Therefore, the increase of serotonin helps the quality of sleep.

[0040] Before taking the experimental group and the control group's composition and after taking it for 2 weeks, about 5 ml of blood was collected from each subject 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 absorbance was measured using an ELISA Reader OD 450nm, and the serotonin concentration was converted using a standard curve. The test results are shown in Figure 1 and Figure 2 .

[0041] Figure 1a is the test result of the changes of serotonin before and after the subjects took the composition of the experimental 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 experimental group and the control group composition for 2 weeks, the serotonin in the blood increased, and there was a significant difference compared with before taking the composition.

[0042] Figure 2 The results are the comparison of the changes in serotonin levels after the subjects took the composition of the experimental group and the control group. The values ​​are presented as Mean±SD. The statistics were calculated using paired t-test, and p<0.05 was considered to be significantly different. Figure 2 As shown, the changes in the amount of serotonin in the subjects before and after taking the composition of the experimental group (i.e., the difference in serotonin before and after taking the composition) were compared with those of the control group. The increase in the amount of serotonin in the subjects taking the composition of the experimental group was better than that of the control group, and the difference was significant (p=0.0075).

[0043] Therefore, the composition of the experimental group containing four lactic acid bacteria strains was significantly better than the control group containing only a single lactic acid bacteria strain in increasing serotonin.

[0044] 1. Effects of taking the composition of the present invention on deep sleep: The sleep cycle includes light sleep, deep sleep and rapid eye movement (REM) sleep. Deep sleep helps the brain rest, repair, and restore physical strength, and also helps to strengthen declarative memory. In addition, during the deep sleep stage, the pituitary gland will secrete growth hormone, which helps the body's tissue growth and cell regeneration. In order to allow the brain and body enough time to repair, sufficient deep sleep is very important. Therefore, more deep sleep means better sleep quality.

[0045] Before taking the composition of the experimental group and the control group and after taking it for 2 weeks, the sleep quality of the subjects was observed using a wearable medical watch sleep analyzer. Figure 3 and Figure 4 .

[0046] Figure 3 a and Figure 3 b are the test results of the deep sleep and light sleep ratios before and after the subjects took the experimental group's composition. Figure 3 a and Figure 3b It can be seen that after taking the composition of the experimental group, the proportion of light sleep of the subjects decreased, while the proportion of deep sleep increased. Therefore, after taking the composition of the experimental group, the subjects can indeed improve their sleep quality, and there is a significant difference.

[0047] Depend on Figure 4 a and Figure 4 b It can be seen that the subjects also had the effect of improving sleep quality after taking the composition of the control group.

[0048] In addition, the subjects who had improved sleep quality after taking the composition of the experimental group (i.e., the subjects whose deep sleep ratio increased) were compared with the subjects who had improved sleep quality after taking the composition of the control group. The results are as follows Figure 5 and Figure 6 As shown. Figure 5 It can be seen that the increase in the deep sleep percentage of the experimental group after taking the composition is better than that of the control group, and the reduction in the shallow sleep percentage is also better than that of the control group, with significant differences (p=0.0201). Therefore, the composition of the experimental group containing four lactic acid bacteria strains is better than the control group containing only a single lactic acid bacteria strain in improving sleep quality.

[0049] 2. Effect of taking the composition of the present invention on PSQI: The Pittsburgh Sleep Quality Index (PSQI) has seven scores, namely sleep quality, sleep latency, sleep duration, sleep efficiency, sleep disturbance, use of sleeping pills, and daytime activity disability. The higher the PSQI score, the worse the sleep quality.

[0050] After taking the composition of the present invention for 2 weeks, the sleep quality of the subjects was evaluated again using the Pittsburgh Sleep Quality Index Questionnaire. The results are as follows: Figure 7 As shown. Figure 7 It can be seen from the results that, compared with before taking the composition of the experimental group, the PSQI was significantly reduced (p=0.006) after taking the composition of the experimental group for 2 weeks, indicating that taking the composition of the experimental group can significantly improve the sleep quality of the subjects.

[0051] In summary, the composition of the present invention has the physiological activity function of improving sleep quality. It should be noted that compared with the composition of the control group containing only a single lactic acid bacteria strain (control group), the composition containing 4 lactic acid bacteria strains (experimental group) can more significantly improve sleep quality, which shows that the lactic acid bacteria strain combination of the present invention has unexpected effects.

[0052] 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. A lactic acid bacteria strain composition for improving sleep quality, characterized in that: The lactic acid bacteria strain composition includes 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, and Bacillus paracasei BR176 strain with a preservation number of CGMCC NO. 21563.

2. The lactic acid bacteria strain composition according to claim 1, characterized in that: The strain quantity ratio of the Bifidobacterium breve BBr-28 strain, the Bifidobacterium animalis subspecies lactis PS23 strain, the Bifidobacterium longum subspecies infantis BR631 strain and the Bacillus paracasei BR176 strain is 1: (0.5-1.5): (0.5-1.5): (0.5-1.5).

3. Use of the lactic acid bacteria strain composition according to claim 1 or 2 in the preparation of a composition for improving sleep quality.

4. The use according to claim 3, characterized in that: The effective viable count of the lactic acid bacteria composition is equal to or greater than 5×10 9 CFU / g.

5. A composition for improving sleep quality, characterized in that: The composition comprises the lactic acid bacteria strain composition according to claim 1 or 2 and a carrier.

6. The composition according to claim 5, characterized in that: The strain quantity ratio of the Bifidobacterium breve BBr-28 strain, the Bifidobacterium animalis subspecies lactis PS23 strain, the Bifidobacterium longum subspecies infantis BR631 strain and the Casei bacillus paracasei BR176 strain is 1:1:1:

1.

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

8. The composition according to claim 5, characterized in that: The carrier is a physiologically acceptable excipient or diluent.

9. The composition according to claim 5, characterized in that: The carrier is a pharmaceutically acceptable excipient or diluent.

10. The composition according to claim 5, characterized in that: The composition is in the form of tablets, capsules or powders.

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