Lactic acid bacteria strain composition with effect of improving sleep quality and application of lactic acid bacteria strain composition

Abstract

The invention relates to the technical field of probiotics, in particular to a lactic acid bacteria strain composition with an effect of improving sleep quality and application. 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 the physiological activity function of improving the sleep quality, so that the lactic acid bacteria strain composition can be used for preparing food or composition for improving the sleep quality, and the problem of poor sleep quality is solved.

Description

Technical Field

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

[0002] Sleep status not only affects physical health but also leads to premature aging. Therefore, sleep quality is an important factor for normal life functions. However, with the prevalence of 3C electronic products such as smartphones today, many people still use their phones before going to bed, such as browsing social messages, watching dramas, playing games, etc. These behaviors are likely to greatly reduce the sleep quality of users and also result in an increasing number of insomnia patients year by year. Therefore, the insomnia problem not only occurs in the elderly population, but there is a trend of younger people suffering from insomnia, and people aged 20 to 40 are the peak of insomnia.

[0003] Research on the health benefits of probiotics for humans is very extensive. Generally, 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). Various functional probiotics have also been disclosed in the prior art. However, the special effects of functional probiotics lie in the specificity of the strain rather than the species. That is, even for the same species, if they are different strains, they 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 with the effect of improving sleep quality to improve the sleep problem. Generally, lactic acid bacteria are considered safe. Therefore, finding a composition containing lactic acid bacteria strains with sleep quality-improving activity is the current goal that requires great effort. Summary of the Invention

[0005] The purpose of the present application is to provide a lactic acid bacteria strain composition having the effect of improving sleep quality and its application in view of the deficiencies of the prior art.

[0006] The above-mentioned invention purpose of the present application is achieved through the following technical solutions: One of the objectives of the present invention is to provide a lactic acid bacteria strain composition with the effect of improving sleep quality. The lactic acid bacteria strain composition includes Bifidobacterium breve BBr-28 strain with the preservation number of CGMCC NO.32485, Bifidobacterium animalis subsp. lactis PS23 strain with the preservation number of CGMCC NO.32691, Bifidobacterium longum subsp. infantis BR631 strain with the preservation number of CGMCC NO.26243, Bifidobacterium bifidum BD-188 strain with the preservation number of CGMCC NO.32484, and Lactiplantibacillus plantarum PQQ-1 strain with the preservation number of CGMCC NO.29303.

[0007] In the above technical solution, the strain quantity ratio of the Bifidobacterium breve BBr-28 strain, Bifidobacterium animalis subsp. lactis PS23 strain, Bifidobacterium bifidum BD-188 strain, and Lactiplantibacillus plantarum PQQ-1 strain is 1:(0.5 - 1.5):(0.5 - 1.5):(0.5 - 1.5):(0.5 - 1.5).

[0008] Another objective of the present invention is to provide the application of the above lactic acid bacteria strain composition in the preparation of a composition for improving sleep quality, and the composition is a food composition or a pharmaceutical composition.

[0009] In the above technical solution, the effective viable count of the lactic acid bacteria strain composition is equal to or greater than 5×10 9 CFU / g.

[0010] A third objective of the present invention is to provide a composition for improving sleep quality, and the composition includes the above lactic acid bacteria strain composition and a carrier.

[0011] In the above technical solution, the strain quantity ratio of the Bifidobacterium breve BBr-28 strain, Bifidobacterium animalis subsp. lactis PS23 strain, Bifidobacterium bifidum BD-188 strain, and Lactiplantibacillus plantarum PQQ-1 strain is 1:1:1:1:1.

[0012] In the above technical solution, in the composition, the effective viable count of all lactic acid bacteria strains 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 a lozenge, a capsule or a powder.

[0016] Advantages of the present application: The lactic acid bacteria strain composition provided by the present invention with the effect of improving sleep quality includes Bifidobacterium breve BBr-28 strain with the preservation number of CGMCC NO.32485, Bifidobacterium animalis subsp. lactis PS23 strain with the preservation number of CGMCC NO.32691, Bifidobacterium longum subsp. infantis BR631 strain with the preservation number of CGMCC NO.26243, Bifidobacterium bifidum BD-188 strain with the preservation number of CGMCC NO.32484, and Lactiplantibacillus plantarum PQQ-1 strain with the preservation number of CGMCC NO.29303. The present invention conducts experiments on the composition prepared from the bacterial powders of the above 5 lactic acid bacteria strains and a carrier. The sleep of the subjects is evaluated by the Pittsburgh Sleep Quality Index (PSQI). Before taking the compositions of the experimental group and the control group and 2 weeks after taking them, the blood of the subjects is collected to detect the change of serotonin, and the improvement of the sleep quality of the subjects is observed by a wearable medical watch sleep analyzer and a PSQI questionnaire. The results show that: 1) The experimental group containing five lactic acid bacteria strains is significantly better than the control group containing only a single lactic acid bacteria strain in increasing serotonin; 2) The experimental group containing five lactic acid bacteria strains is better than the control group containing only a single lactic acid bacteria strain in improving sleep quality; 3) Taking the composition of the experimental group can significantly improve the sleep quality of the subjects. This shows that the lactic acid bacteria strain composition of the present invention has the physiological activity function of improving sleep quality, and it can be used to prepare food or pharmaceutical compositions for improving sleep quality to solve the problem of sleep quality. Description of the Drawings

[0017] Figure 1 Test results of the change of serotonin in the subjects of Example 2 before and after taking the compositions of the experimental group and the control group respectively.

[0018] Figure 2Comparison results of the change in serotonin levels in the subjects of Example 2 before and after taking the compositions of the experimental group and the control group.

[0019] Figure 3 Test results of the proportion of deep sleep and light sleep in the subjects of Example 2 before and after taking the composition of the experimental group.

[0020] Figure 4 Test results of the proportion of deep sleep and light sleep in the subjects of Example 2 before and after taking the composition of the control group.

[0021] Figure 5 Comparison results of the change in the proportion of deep sleep and light sleep in the subjects of Example 2 after taking the composition of the experimental group.

[0022] Figure 6 Comparison results of the change in the proportion of deep sleep and light sleep in the subjects of Example 2 after taking the composition of the control group.

[0023] Figure 7 Change results of PSQI in the subjects of Example 2 before and after taking the composition of the experimental group. Detailed implementation methods

[0024] The microbial strain materials involved in the following examples are as follows: 1. Bifidobacterium breve BBr-28 strain Deposit date: 2024 / 11 / 04 Depositary institution: China General Microbiological Culture Collection Center Address of the depositary institution: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, China. Institute of Microbiology, Chinese Academy of Sciences, 100101 Deposit number: CGMCC No. 32485 Taxonomic nomenclature: Bifidobacterium breve 2. Bifidobacterium animalis subsp. lactis PS23 strain Deposit date: 2024 / 11 / 18 Depositary institution: China General Microbiological Culture Collection Center Address of the depositary institution: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, China. Institute of Microbiology, Chinese Academy of Sciences, 100101 Deposit number: CGMCC No. 32691 Taxonomic nomenclature: Bifidobacterium animalis subsp. lactis 3. Bifidobacterium longum subsp. infantis BR631 strain Deposit Date: December 25, 2022 Depositary Institution: China General Microbiological Culture Collection Center Address of Depositary Institution: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, China. Institute of Microbiology, Chinese Academy of Sciences, 100101 Deposit Number: CGMCC No.26243 Taxonomic Nomenclature: Bifidobacterium longum subsp. infantis 4. Bifidobacterium bifidum BD-188 strain Deposit Date: November 4, 2024 Depositary Institution: China General Microbiological Culture Collection Center Address of Depositary Institution: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, China. Institute of Microbiology, Chinese Academy of Sciences, 100101 Deposit Number: CGMCC NO.32484 Taxonomic Nomenclature: Bifidobacterium bifidum 5. Lactiplantibacillus plantarum PQQ-1 strain Deposit Date: December 12, 2023 Depositary Institution: China General Microbiological Culture Collection Center Address of Depositary Institution: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, China. Institute of Microbiology, Chinese Academy of Sciences, 100101 Deposit Number: CGMCC NO.29303 Taxonomic Nomenclature: Lactiplantibacillus plantarum The freeze-dried cultures of the above strains have been deposited in the China General Microbiological Culture Collection Center at No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, China. The detailed deposit information is shown in Table 1: Table 1. Deposit Information of the Lactic Acid Bacteria Strains of the Present Invention The following further elaborates on this application in conjunction with the accompanying drawings. Example 1: Research on the effects of probiotics on human health is extensive. Generally, 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). 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 for the same species, if they are different strains, they may still have different effects, or even opposite effects.

[0025] On this basis, the first embodiment of the present invention provides a lactic acid bacteria strain composition with the effect of improving sleep quality, including Bifidobacterium breve BBr-28 strain, Bifidobacterium animalis subsp. lactis PS23 strain, Bifidobacterium longum subsp. infantis BR631 strain, Bifidobacterium bifidum BD-188 strain, and Lactiplantibacillus plantarum PQQ-1 strain.

[0026] The strain ratio of Bifidobacterium breve BBr-28 strain, Bifidobacterium animalis subsp. lactis PS23 strain, Bifidobacterium longum subsp. infantis BR631 strain, Bifidobacterium bifidum BD-188 strain, and Lactiplantibacillus plantarum PQQ-1 strain is 1:(0.5-1.5):(0.5-1.5):(0.5-1.5):(0.5-1.5). Preferably, the ratio of the above five strains is 1:1:1:1:1.

[0027] The property characteristics of the lactic acid bacteria strains of the present invention: The taxonomic characteristics of the strains are confirmed according to the analysis results of 16S rRNA sequence analysis and API bacterial identification system. The morphological and general property characteristics of the lactic acid bacteria strains of the present invention are detailed in Table 2: Table 2. Morphological and general property characteristics of the lactic acid bacteria strains of the present invention The preparation method of the lactic acid bacteria strain composition of the present invention: Bifidobacterium breve strain BBr-28, Bifidobacterium animalis subsp. lactis strain PS23, Bifidobacterium longum subsp. infantis strain BR631, and Bifidobacterium bifidum strain BD-188 were isolated from breast milk; Lactiplantibacillus plantarum strain PQQ-1 was isolated from Tibetan snow lotus mushroom. The above five isolated lactic acid bacteria strains were stored in 20% glycerol at -80°C. When in use, each was inoculated into Difco TM MRS (De Man, Rogosa and Sharpe) medium at an inoculation amount of 1% (v / v) and cultured at 37°C for 20 hours. Then, the five resulting cultures were centrifuged at 4°C for 20 minutes, and then the supernatant was poured off. The resulting 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. Therefore, the above lactic acid bacteria strain composition has the use of improving sleep quality and can be used to prepare food or pharmaceutical compositions for improving sleep quality.

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

[0030] The carrier can be a physiologically acceptable excipient or diluent. Or, the carrier can be a pharmaceutically acceptable excipient or diluent. Specifically, the carrier can include oligosaccharides, sugar alcohols, sodium carboxymethylcellulose, ethylcellulose, galactooligosaccharides, stearic acid, magnesium stearate, potato starch, or a combination thereof. Further, the oligosaccharides can include fructooligosaccharides, isomaltooligosaccharides, xylooligosaccharides, stachyose, mannose, arabinose, or a combination thereof; the sugar alcohols can include erythritol, sorbitol, isomaltulose, 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 strain 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 strain composition is equal to or greater than 5×10 9 CFU / g.

[0032] The dosage form of the composition for improving sleep quality of the present invention can be tablets, capsules or powders for convenient oral use by users.

[0033] The preparation method of the composition for improving sleep quality of the present invention: Step 1. Prepare the lactic acid bacteria strain composition (the same as in Example 1). Bifidobacterium breve strain BBr-28, Bifidobacterium animalis subsp. lactis strain PS23, Bifidobacterium longum subsp. infantis strain BR631, and Bifidobacterium bifidum strain BD-188 were isolated from breast milk; Lactiplantibacillus plantarum strain PQQ-1 was isolated from Tibetan snow lotus mushroom. The above five isolated lactic acid bacteria strains were stored in 20% glycerol at -80°C. When in use, each strain was inoculated into Difco TM MRS (De Man, Rogosa and Sharpe) medium at an inoculation amount of 1% (v / v) and cultured at 37°C for 20 hours. Then, the five resulting cultures were centrifuged at 4°C for 20 minutes, and then the supernatant was removed, and the resulting 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 cell activity.

[0034] Step 2. Prepare the composition for improving sleep quality Mix the lactic acid bacteria bacterial powder in Step 1 with a carrier evenly 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 experiment and analysis of the composition of the present invention: The composition of the present invention was made into a capsule dosage form for convenient oral use by users. The 30 subjects participating in the test were divided into two groups, namely the experimental group and the control group, where: Experimental group: Take the composition capsule of the present invention 2 times a day, once between 2-3 pm and once 2 hours before bedtime, for 2 weeks.

[0036] Control group: The composition capsule taken only contains the bacterial powder of Bifidobacterium animalis subsp. lactis strain PS23, and the remaining administration method is the same as that of the experimental group.

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

[0038] Before taking the compositions of the experimental group and the control group and 2 weeks after taking them, the blood of the subjects was collected to detect the change of serotonin, and the improvement of the sleep quality of the subjects was observed using a wearable medical watch sleep analyzer and the PSQI questionnaire.

[0039] (1) Effects of taking the composition of the present invention on serotonin: Serotonin, also known as 5-hydroxytryptamine or 5-HT, is a monoamine neurotransmitter. Its functions include regulating mood, appetite, and sleep. Research indicates that when serotonin is insufficient, there is a tendency for the body to feel sluggish, mood swings, restlessness, insufficient melatonin, or difficulty falling asleep. Therefore, an increase in serotonin is helpful for sleep quality.

[0040] Before taking the compositions of the experimental group and the control group and 2 weeks after taking them, approximately 5 ml of blood from the subjects was collected 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 operation manual. After using HRP (horseradish peroxidase) to catalyze the color development of TMB (3,3',5,5'-Tetramethyl-benzidine), the absorbance was measured using an ELISA Reader at O.D 450 nm, and the serotonin concentration was converted by matching the standard curve. The test results are shown in Figure 1 and Figure 2 .

[0041] Figure 1 a shows the test results of the change in serotonin before and after the subjects took the composition of the experimental group; Figure 1 b shows the test results of the change in serotonin before and after the subjects took the composition of the control group. From Figure 1 a and Figure 1 b, it can be seen that 2 weeks after the subjects took the compositions of the experimental group and the control group, the serotonin in the blood increased, and there was a significant difference compared with before taking the composition (p < 0.001).

[0042] Figure 2 The results of comparing the change amounts of serotonin after the subjects took the compositions of the experimental group and the control group are presented as Mean ± SD. The statistics were calculated using a paired t-test, and p < 0.05 was considered to have a significant difference. As Figure 2 shown, by comparing the change 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) with the control group, the increase amount of serotonin in the subjects taking the composition of the experimental group was better than that of the control group, and there was a significant difference (p = 0.019). Therefore, the composition of the experimental group containing five lactic acid bacteria strains was significantly better than the control group containing only a single lactic acid bacteria strain in increasing serotonin.

[0043] 1. Effects of taking the composition of the present invention on deep sleep: A 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 strengthen declarative memory. In addition, during deep sleep, the pituitary gland secretes growth hormone, which helps body tissues grow and cells regenerate. To allow the brain and body sufficient time to repair, sufficient deep sleep is very important. Therefore, more deep sleep represents better sleep quality.

[0044] Before taking the compositions of the experimental group and the control group and 2 weeks after taking them, the sleep quality of the subjects was observed using a wearable medical watch sleep analyzer. The test results are shown in Figure 3 and Figure 4 .

[0045] Figure 3 a and Figure 3 b are the test results of the proportion of deep sleep and light sleep of the subjects before and after taking the composition of the experimental group, respectively. From Figure 3 a and Figure 3 b, 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 sleep quality of the subjects was indeed improved, and there was a significant difference. After taking the composition of the control group, the subjects also had the effect of improving sleep quality, and the results are as shown in Figure 4 a and Figure 4 b.

[0046] In addition, a comparison was made between the subjects who could improve sleep quality after taking the composition of the experimental group (i.e., the subjects with an increased proportion of deep sleep) and the subjects who could improve sleep quality after taking the composition of the control group. The results are as shown in Figure 5 and Figure 6 . From Figure 5 , it can be seen that after taking the composition of the experimental group, the increase in the proportion of deep sleep was better than that of the control group, and the decrease in the proportion of light sleep was also better than that of the control group, with a significant difference (p = 0.0471). Therefore, the composition of the experimental group containing five lactic acid bacteria strains is better than the control group containing only a single lactic acid bacteria strain in improving sleep quality.

[0047] 2. Effects of taking the composition of the present invention on PSQI: The Pittsburgh Sleep Quality Index (PSQI) has scores for seven aspects, namely sleep quality, sleep latency, sleep duration, sleep efficiency, sleep disturbances, use of sleeping pills, and daytime disability. The higher the PSQI score, the worse the sleep quality.

[0048] After 2 weeks of taking the composition of the experimental group of the present invention, the sleep quality of the subjects was evaluated again using the Pittsburgh Sleep Quality Index Questionnaire, and the results are as follows Figure 7 shown. From Figure 7 the results, it can be seen that compared with before taking the composition of the experimental group, after 2 weeks of taking the composition of the experimental group, the PSQI was significantly reduced (p = 0.002), indicating that taking the composition of the experimental group can significantly improve the sleep quality of the subjects.

[0049] 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 containing only a single lactic acid bacteria strain in the control group (control group), the composition containing 5 lactic acid bacteria strains (experimental group) can more significantly improve sleep quality. Thus, it can be seen that the combination of lactic acid bacteria strains of the present invention has unexpected effects.

[0050] The above embodiments are only used to illustrate the technical solutions of the present application, and are not intended to limit them; although the present application has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that they can still modify the technical solutions described in the foregoing embodiments, or perform equivalent replacements for some of the technical features; and these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the various embodiments of the present application, and should all be included within the protection scope of the present application.

Claims

1. A lactic acid bacteria strain composition having the effect of 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, 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 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 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: (0.5~1.5): (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, characterized in that: The composition is a food composition or a pharmaceutical composition.

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 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 1:1:1:1:

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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