CN117603861A - Lactobacillus gasseri and application thereof in relieving anxiety or improving sleep - Google Patents

Lactobacillus gasseri and application thereof in relieving anxiety or improving sleep Download PDF

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CN117603861A
CN117603861A CN202311569598.1A CN202311569598A CN117603861A CN 117603861 A CN117603861 A CN 117603861A CN 202311569598 A CN202311569598 A CN 202311569598A CN 117603861 A CN117603861 A CN 117603861A
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lactobacillus gasseri
prosci
sleep
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CN117603861B (en
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刘晓军
刘磊
张建军
张凌宇
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Beijing Scitop Bio Tech Co ltd
Jinhua Yinhe Biotechnology Co ltd
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Jinhua Yinhe Biotechnology Co ltd
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Abstract

The invention discloses a lactobacillus gasseri and application thereof in relieving anxiety or improving sleep, wherein the lactobacillus gasseri ProSci-108 is preserved in China general microbiological culture Collection center (CGMCC) No.25448. The lactobacillus gasseri ProSci-108 can improve sleeping, anxiety stress or bad emotion of human body to a certain extent; therefore, the administration of the product containing the lactobacillus gasseri ProSci-108 can help to improve the sleep problems such as insomnia, night waking, difficulty in falling asleep and the like caused by anxiety and stress, promote deep sleep, improve sleep quality and regulate bad emotion; the lactobacillus gasseri ProSci-108 has great application prospect in preparing products for relieving anxiety, improving sleep or regulating emotion.

Description

Lactobacillus gasseri and application thereof in relieving anxiety or improving sleep
Technical Field
The invention relates to the field of microorganisms, in particular to lactobacillus gasseri and application thereof in relieving anxiety or improving sleep.
Background
Sleep is an important physiological phenomenon, and is not only a basic autonomic nerve function regulating process, but also an important link of physical energy recovery, memory integration, consolidation and other processes. Sleep disorders refer to abnormalities in sleep quality and quantity, as well as a series of clinical symptoms that occur during sleep. Long-term sleep disorders can cause damage to personal physical, psychological and social functions, not only affecting the quality of life of the individual, but also inducing psychotic disorders, cardiovascular and cerebrovascular disorders, and the like.
Modern fast-paced life and high-intensity work bring more and more pressure to current people, sleep disorder problems are easy to occur, and even the emotion is influenced by poor long-term sleep quality, the unstable emotion can easily further stress people, and then anxiety, depression and other symptoms occur, so that the aim of adjusting the emotion is very necessary by relieving pressure and improving the sleep quality.
At present, barbiturates, benzodiazepines, non-benzodiazepines and the like are generally used for clinically treating sleep disorders, and the substances have the advantages of quick response, clear action mechanism and good curative effect, but side effects such as residual effect, forgetting effect, drug withdrawal effect and the like are easy to generate after long-term administration, and in addition, the situation that the improving effect is not obvious and the like can occur when traditional Chinese medicines are adopted in the process of relieving pressure and improving sleep, so that the problem of anxiety and sleep improvement is effectively solved in the prior art.
In recent years, probiotics are increasingly paid attention to the effect of regulating the ecology of intestinal flora and relieving abnormal emotion, but the effect of relieving anxiety, improving sleep or regulating the health function of emotion of the existing disclosed lactobacillus gasseri is not found.
Disclosure of Invention
Accordingly, the present invention has been made keeping in mind the above problems occurring in the prior art, and has not been accomplished by the present invention, which has been accomplished by the present invention, to provide a strain of lactobacillus gasseri that can achieve the effects described above.
A Lactobacillus gasseri Lactobacillus gasseri ProSci-108, the Lactobacillus gasseri ProSci-108 has been deposited in China general microbiological culture Collection center with the collection number of CGMCC No.25448, the collection address of China national academy of sciences of China, no.3, the university of North Chen West Lu 1, the Korean area of Beijing, and the date of deposit of the strain of 2022, and the strain is detected to be a viable strain.
The acquisition process of the lactobacillus gasseri ProSci-108 is as follows:
taking inner Mongolia natural fermentation goat milk as a sample, sucking 0.5mL of the sample, adding the sample into 5mL of MRS liquid culture medium, and carrying out aerobic culture at 37 ℃ for 24 hours to obtain an enriched culture solution; absorbing 0.5mL of the enrichment culture solution and adding the enrichment culture solution into 4.5mL of sterile physiological saline to obtain 10 -1 Diluent and then aspirate 0.5mL 10 -1 The diluted solution was dissolved in 4.5mL of physiological saline to give 10 -2 Dilution ofAccording to the operation, 10 is obtained in turn -3 、10 -4 、10 -5 、10 -6 A dilution liquid; 100 mu L of gradient dilution is absorbed and coated on MRS solid culture medium, 10 -4 、10 -5 、10 -6 1 plate per gradient, aerobically cultured at 37 ℃ for 48 hours to obtain colonies; selecting a colony with typical characteristics of lactobacillus gasseri on an MRS solid culture medium according to the shape, size, edge, transparency and the like of the colony, picking the colony by an inoculating loop, streaking the colony on the MRS solid culture medium, and carrying out aerobic culture for 48 hours at 37 ℃ to obtain a purified single colony; the purified single colony is picked and inoculated into 5mL MRS liquid culture medium respectively, and the bacterial liquid is obtained by aerobic culture for 24 hours at 37 ℃; after numbering each strain corresponding to each bacterial liquid, gram staining, strain identification, physiological and biochemical experiments and genome identification analysis are carried out by referring to the steps described in textbook "microbiology" (Shen Ping, chen Xiangdong main code), and a strain having typical characteristics of lactobacillus gasseri is selected to obtain a strain ProSci-108.
Wherein, the strain identification process is as follows:
taking bacterial strain ProSci-108, extracting genome of bacterial strain ProSci-108, and using a primer pair 27F of 27F/1492R,SEQ ID NO.2 with sequences shown in SEQ ID NO.2 and SEQ ID NO.3 respectively: 5'-AGAGTTTGATCCTGGCTCAG-3', SEQ ID NO.3 primer pair 1492R:5'-AAGGAGGTGATCCAGCC-3' the genome of the extracted strain ProSci-108 is used as a template for amplification to obtain the 16S rRNA of the strain ProSci-108, and the 16S rDNA sequence of the strain ProSci-108 is shown as SEQ ID NO. 1: AAAATGAGAGTTTGATCCTGGCTCAGGACGAACGCTGGCGGCGTGCCTAATACATGCAAGTCGAGCGAGCTTGCCTAGATGAATTTGGTGCTTGCACCAGATGAAACTAGATACAAGCGAGCGGCGGACGGGTGAGTAACACGTGGGTAACCTGCCCAAGAGACTGGGATAACACCTGGAAACAGATGCTAATACCGGATAACAACACTAGACGCATGTCTAGAGTTTAAAAGATGGTTCTGCTATCACTCTTGGATGGACCTGCGGTGCATTAGCTAGTTGGTAAGGTAACGGCTTACCAAGGCAATGATGCATAGCCGAGTTGAGAGACTGATCGGCCACATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCACAATGGACGCAAGTCTGATGGAGCAACGCCGCGTGAGTGAAGAAGGGTTTCGGCTCGTAAAGCTCTGTTGGTAGTGAAGAAAGATAGAGGTAGTAACTGGCCTTTATTTGACGGTAATTACTTAGAAAGTCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGATTTATTGGGCGTAAAGCGAGTGCAGGCGGTTCAATAAGTCTGATGTGAAAGCCTTCGGCTCAACCGGAGAATTGCATCAGAAACTGTTGAACTTGAGTGCAGAAGAGGAGAGTGGAACTCCATGTGTAGCGGTGGAATGCGTAGATATATGGAAGAACACCAGTGGCGAAGGCGGCTCTCTGGTCTGCAACTGACGCTGAGGCTCGAAAGCATGGGTAGCGAACAGGATTAGATACCCTGGTAGTCCATGCCGTAAACGATGAGTGCTAAGTGTTGGGAGGTTTCCGCCTCTCAGTGCTGCAGCTAACGCATTAAGCACTCCGCCTGGGGAGTACGACCGCAAGGTTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGA AGCAACGCGAAGAACCTTACCAGGTCTTGACATCCAGTGCAAACCTAAGAGATTAGGTGTTCCCTTCGGGGACGCTGAGACAGGTGGTGCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGTCATTAGTTGCCATCATTAAGTTGGGCACTCTAATGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAGTCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGACGGTACAACGAGAAGCGAACCTGCGAAGGCAAGCGGATCTCTGAAAGCCGTTCTCAGTTCGGACTGTAGGCTGCAACTCGCCTACACGAAGCTGGAATCGCTAGTAATCGCGGATCAGCACGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGAGAGTCTGTAACACCCAAAGCCGGTGGGATAACCTTTATAGGAGTCAGCCGTCTAAGGTAGGACAGATGATTAGGGTGAAGTCGTAACAAGGTAGCCGTAGGAGAACCTGCGGCTGGATCACCTCCTTT;
the nucleic acid sequence of the 16S rDNA of the strain ProSci-108 was aligned in the Blastn program of NCBI, and the result shows that the strain was Lactobacillus gasseri, designated as Lactobacillus gasseri (Lactobacillus gasseri) ProSci-108.
The invention also provides the application of the lactobacillus gasseri ProSci-108 in preparing any one of the following (1) - (3),
(1) An anxiety-reducing product;
(2) A sleep improving product;
(3) And (3) a mood-regulating product.
The product is food, health product or medicine.
The food is yogurt, yogurt block, solid beverage, and probiotic beverage.
The medicine is in the form of powder, tablet or capsule.
A product for alleviating anxiety, improving sleep or regulating mood comprising a lactobacillus gasseri ProSci-108 as described above.
The total viable count of the lactobacillus gasseri ProSci-108 in the product is not lowAt 3X 10 10 CFU/mL or 3X 10 10 CFU/g。
When the product is a medicine, the product also comprises chemical medicines.
The chemical is barbital sodium.
The raw material of the lactobacillus gasseri ProSci-108 in the product comprises any one or more of a culture, a fermentation product and freeze-dried powder of the strain.
The technical scheme of the invention has the following advantages:
the lactobacillus gasseri ProSci-108 provided by the invention can improve sleeping, anxiety stress or bad emotion of human body to a certain extent. Therefore, the administration of the lactobacillus gasseri ProSci-108 can help to improve the sleep problems such as insomnia, night waking, difficulty in falling asleep and the like caused by anxiety and stress, promote deep sleep, improve sleep quality and regulate bad emotion, and has great application prospect in preparing products for relieving anxiety, improving sleep or regulating emotion.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are needed in the description of the embodiments or the prior art will be briefly described, and it is obvious that the drawings in the description below are some embodiments of the present invention, and other drawings can be obtained according to the drawings without inventive effort for a person skilled in the art.
Fig. 1 is a statistical graph of PSQI scores of 4 groups of subjects before and after intervention in experimental example 1;
FIG. 2 is a statistical plot of the time to sleep for 4 groups of subjects prior to intervention in Experimental example 1;
FIG. 3 is a statistical plot of the time to fall asleep of 4 groups of subjects in the intervention of experimental example 1;
FIG. 4 is a statistical chart of the fall asleep time of the group of 4 subjects following the dry period in experimental example 1;
FIG. 5 is a statistical plot of the mid-night wake up of the 4 groups of subjects prior to intervention in Experimental example 1;
FIG. 6 is a statistical plot of the mid-night wake up of 4 groups of subjects in the intervention of Experimental example 1;
FIG. 7 is a statistical chart showing the wake-up of the dry and wet 4 groups of subjects in experimental example 1 at midnight;
fig. 8 is a statistical chart of sleep duration of 4 groups of subjects before intervention in experimental example 1;
FIG. 9 is a statistical chart of sleep duration of 4 groups of subjects in the intervention of experimental example 1;
FIG. 10 is a statistical chart of sleep duration of the 4 groups of subjects after the dry prognosis in experimental example 1;
FIG. 11 is a statistical diagram of mental states of 4 groups of subjects prior to intervention in experimental example 1;
FIG. 12 is a statistical diagram of the mental states of the 4 groups of subjects with the prognosis of the subject in experiment example 1.
Detailed Description
The following examples are provided for a better understanding of the present invention and are not limited to the preferred embodiments described herein, but are not intended to limit the scope of the invention, any product which is the same or similar to the present invention, whether in light of the present teachings or in combination with other prior art features, falls within the scope of the present invention.
The specific experimental procedures or conditions are not noted in the examples and may be followed by the operations or conditions of conventional experimental procedures described in the literature in this field. The reagents or apparatus used were conventional reagent products commercially available without the manufacturer's knowledge.
Example 1
The preparation method of the probiotic bacterial powder of the lactobacillus gasseri ProSci-108 comprises the following steps:
1) Activation of strains: inoculating the strain (Lactobacillus gasseri ProSci-108) frozen and preserved at-40deg.C into MRS liquid culture medium sterilized at 121deg.C for 15min, anaerobic culturing at 37deg.C for 18-24 hr, and subculturing for 1-2 times to obtain activated strain.
2) Preparation of optimized culture medium: preparing the components of the optimized culture medium according to a proportion, uniformly mixing, adjusting the pH value to 6.5, and sterilizing for 15min at 121 ℃;
the optimized medium composition was as follows: 23.5Kg of sucrose, 12.0Kg of lactose and soy protein15.0Kg peptone, 5.0Kg yeast powder, 12Kg yeast peptone, na 2 HPO 3 21.0Kg, 2.0Kg of citric acid, mgSO 4 ·7H 2 O0.6Kg，MnSO 4 ·5H 2 O0.3 Kg, tween-80 1.0Kg, L-cysteine hydrochloride 0.3Kg, distilled water to 1000L.
3) Preparing seed liquid: and (3) correspondingly inoculating each strain activated in the step (1) into the optimized culture medium prepared in the step (2), and stopping obtaining seed liquid when anaerobic culture is performed at 37 ℃ until the pH value is 4.5-4.8.
4) Inoculating and fermenting: inoculating the seed liquid obtained in the step 3) into the optimized culture medium prepared in the step 2) according to the proportion of 1 per mill, and fermenting for 18h under controlled fermentation conditions;
the fermentation conditions are controlled as follows: culturing at 30deg.C in the early stage of fermentation, and naturally fermenting until pH is 5.0; then adjusting the fermentation temperature to 37 ℃ for constant temperature culture, and controlling the pH value to be 6.0 by adding a neutralizing agent NaOH, and keeping the anaerobic fermentation to reach the total fermentation time of 18 hours. Wherein the anaerobic condition is realized by a method of introducing nitrogen once every two hours.
5) Terminating fermentation: stopping fermentation when the acid production of thallus is stopped (whether the acid production is stopped or not can be judged according to the fact that the pH is not reduced when the addition of the neutralizer is stopped), obtaining high-density fermentation liquor of the strain, wherein the viable count of the high-density fermentation liquor reaches 2 multiplied by 10 10 cfu/ml or more.
6) Freeze drying
a. And (3) concentration of thalli: concentrating the high-density fermentation broth by 12000g centrifugation;
b. and (3) adding a protective agent: adding 3-8 times of volume multiple of protective agent solution into the bacterial concentrate; the composition of the protectant solution is as follows: 10-15Kg of skim milk, 8-12Kg of lactose, 1-2Kg of vitamin C, 1-2Kg of sodium glutamate and distilled water to 1000L.
c. And (3) drying: freeze drying the bacterial suspension added with the protective agent to obtain freeze-dried bacterial powder of strain bacteria, wherein the total number of viable bacteria in the bacterial powder reaches 5.0X10 11 cfu/g or more.
Example 2
In this example, a composite powder of Lactobacillus gasseri ProSci-108 was prepared by the following steps:
streaking the strain on an MRS solid culture medium, and culturing for 48 hours at 37 ℃ to obtain a single colony; single colony is selected and inoculated in MRS liquid culture medium, and is cultivated again at 37 ℃ for continuous activation for two generations to obtain activating solution; inoculating the activating solution into an optimized culture medium according to the inoculum size of 2% (v/v), and continuously culturing at 37 ℃ for 20 hours to obtain bacterial solution; centrifuging 12000g of bacterial liquid for 10min to obtain bacterial mud; washing the bacterial mud with normal saline, and then placing the bacterial mud in a protective agent to obtain bacterial suspension; finally, freeze-drying the bacterial suspension to obtain bacterial powder, the number of the viable bacteria of which is 1 multiplied by 10 11 CFU/g。
In this embodiment, the following steps are first 1: adding maltodextrin into the bacterial powder in proportion of 1, and mixing to obtain the bacterial powder with viable count of 5×10 10 Powder of CFU/g; the addition amount of maltodextrin in the powder is 1-20g, and other auxiliary materials are 1-50g, and the composite powder is obtained after uniform mixing. The other adjuvants can be one or more of galactooligosaccharide, inulin, stachyose, erythritol, maltitol or resistant dextrin.
The composite powder of the lactobacillus gasseri ProSci-108 is obtained by mixing the preparation method, and can be applied to the fields of foods, medicines or health care products.
Example 3
In this example, a sample tablet of Lactobacillus gasseri ProSci-108 was prepared as follows:
streaking the strain on an MRS solid culture medium, and culturing for 48 hours at 37 ℃ to obtain a single colony; single colony is selected and inoculated in MRS liquid culture medium, and is cultivated again at 37 ℃ for continuous activation for two generations to obtain activating solution; inoculating the activating solution into a culture medium according to the inoculum size of 2% (v/v), and continuously culturing at 37 ℃ for 20 hours to obtain bacterial solution; centrifuging 12000g of bacterial liquid for 10min to obtain bacterial mud; washing the bacterial mud with normal saline, and then placing the bacterial mud in a protective agent to obtain bacterial suspension; finally, freeze-drying the bacterial suspension to obtain bacterial powder, the number of the viable bacteria of which is 1 multiplied by 10 11 CFU/g。
The fungus powder is prepared according to the following proportion of 1:1 proportion of maltodextrin, and diluting until the viable count is 5×10 10 CFU/g, maltodextrin adding amount is 1-30g, other auxiliary materials are weighed 1-40g, and tablet samples are obtained after uniform mixing and tabletting. The other is provided withThe adjuvant can be fructo-oligosaccharide, inulin, raffinose, xylitol or maltitol.
The lactobacillus gasseri ProSci-108 compound tablet sample is obtained by mixing the preparation method, and can be applied to the fields of food, medicines or health care products.
Example 4
In this example, a fermented milk of Lactobacillus gasseri ProSci-108 was prepared by the following steps:
streaking the strain on an MRS solid culture medium, and culturing for 48 hours at 37 ℃ to obtain a single colony; single colony is selected and inoculated in MRS liquid culture medium, and is cultivated again at 37 ℃ for continuous activation for two generations to obtain activating solution; inoculating the activating solution into a culture medium according to the inoculum size of 2% (v/v), and continuously culturing at 37 ℃ for 20 hours to obtain bacterial solution; centrifuging 12000g of bacterial liquid for 10min to obtain bacterial mud; washing the bacterial mud with normal saline, and then placing the bacterial mud in a protective agent to obtain bacterial suspension; finally, freeze-drying the bacterial suspension to obtain bacterial powder, the number of the viable bacteria of which is 1 multiplied by 10 11 CFU/g。
Adding sucrose into pure milk at a certain proportion, heating to 65deg.C, homogenizing under 18-20 MPa, maintaining at 95deg.C, sterilizing for 5min, and cooling; to a sample temperature of 37℃at 0.03% by weight and 5X 10 6 Inoculating basic starter and lactobacillus gasseri ProSci-108 respectively with CFU/mL inoculum size, and fermenting at 37deg.C; after fermentation, the mixture is refrigerated at 4 ℃ for 24 hours for after-ripening to obtain a fermented milk sample, and the fermented milk sample can be applied to the field of functional foods.
Experimental example 1
The bacterial powder prepared in the example 1 is used for sleep quality evaluation to judge the effects of regulating emotion, relieving stress and improving anxiety, and the specific process is as follows:
design of experiments
The Pittsburgh sleep quality index scale (pittsburgh sleep quality index, PSQI) is a self-evaluation scale for sleep quality evaluation, and comprises 7 components including subjective sleep evaluation, sleep time, sleep duration, sleep efficiency, sleep disorder, use condition of hypnotic drugs, daytime functions and the like, each component is scored according to 0-3, each score is accumulated as PSQI total score, the total score range is 0-21 score, and the higher the PSQI total score is, the worse the sleep quality is indicated. The sleeping quality of 0-5 minutes is good, the sleeping quality of 6-10 minutes is good, the sleeping quality of 11-15 minutes is general, and the sleeping quality of 16-21 minutes is poor.
1. Group of subjects: the subject should be a population suffering from sleep disorder with 18 years old or older (not limited to men and women) and a PSQI score of 8 or more as the observation subject.
2. Inclusion criteria: only if the subject meets all of the following criteria, it is suitable for the study.
1) Either male or female, the age being over 18 years old;
2) PSQI score 8 or more for people with sleep disorder;
3) Antibiotics were not used 1 month prior to group entry;
4) No history of chronic gastrointestinal disease;
5) Agreeing to participate in the study, and signing written informed consent;
6) The newly discovered or the existing patients with sleep problems can have no other complications and are not treated by sleep medicines;
3. exclusion criteria: the subject cannot be enrolled if they meet any of the following:
1) Abnormal functions of heart, liver and kidney;
2) Patients suffering from severe psychotic disorders;
3) Those with history of allergy or high allergic constitution of lactobacillus and its products;
4) Patients who received other medications 1 month prior to group entry;
4) Alcoholics;
5) Other sleep-aid products cannot be taken in compliance with the test guidelines or halfway through the test period.
4. Diagnostic criteria:
diagnostic criteria for insomnia, including difficulty in falling asleep, difficulty in sleep maintenance, early waking, daytime dysfunction, etc., are adopted in the third edition of the Chinese mental disorder classification and diagnostic criteria (mental disorder classification), and occur more than 3 times a week for more than 1 month, and are not caused by other diseases.
5. Sample size: 200 cases are divided into 4 groups randomly according to the ratio of 1:1:1, namely a probiotic group, a placebo group, a probiotic compound drug (barbital sodium) group and a drug group (barbital sodium), wherein 50 persons are in each group, and the 4 groups of people have no statistical significance on the difference of gender and age and are comparable.
6. The main curative effect index is as follows:
the sleep disorder-related symptoms are mainly counted, wherein the sleep disorder-related symptoms comprise sleep time, the number of awakenings in the middle of the night, the sleeping condition again and the total sleeping duration.
7. The composition and the intervention mode of the product are as follows:
probiotic group: lactobacillus gasseri ProSci-108 (300 hundred million cfu/d) +maltodextrin, 2 g/bag; placebo group: maltodextrin, 2 g/bag; probiotic composite drug group: lactobacillus gasseri ProSci-108 (300 hundred million cfu/d) +barbital sodium (50 mg/d) +maltodextrin, 2 g/bag; drug group: barbituric sodium (50 mg/d) +maltodextrin, 2 g/bag, the four above groups of products give the same package, color and smell.
8. Intervention dose and administration mode: 1 bag per day, and is taken after supper with warm water (below 37deg.C) for 30 days.
9. Statistical analysis
The 0d, 15d and 30d human subjects PSQI and sleep symptom system meters were collected. Count data is expressed as a rate (%), and metering data is expressed as mean ± standard deviation (±s). Check level α=0.05.
(II) results of experiments
As shown in fig. 1-4 and in table 1 below.
Wherein, fig. 1 is a graph of PSQI score statistics of 4 groups of subjects before, during and after the intervention, and as can be seen from fig. 1: the PSQI scores of the 4 groups before intervention were respectively probiotic group (14.25±0.52, n=50), placebo group (13.70±0.55, n=50), probiotic composite drug group (14.11±0.16, n=50) and drug group (13.59±0.23, n=50) and no significant difference (P > 0.05) was compared, and the population test could be continued. After 30d intervention, PSQI score of the probiotic group is obviously reduced, the 15 th d is reduced by 29.47%, and after the intervention is finished, the PSQI score is reduced by 43.72%; there was no significant difference in the placebo group PSQI scores before and after intervention, there was a certain decrease in both other two groups of PSQI scores after intervention and a 48.72% decrease in the PSQI scores after probiotic-drug intervention.
Fig. 2 to 4 are graphs showing the statistics of the falling asleep time of the 4 groups of the subjects before, during and after the intervention, fig. 2 shows the statistics of the falling asleep time of the subject before the intervention (0 d), fig. 3 shows the statistics of the falling asleep time of the subject during the intervention (15 d), and fig. 4 shows the statistics of the falling asleep time of the subject after the intervention (30 d). Statistics is carried out on the proportion of people who can fall asleep within 30min before and after intervention, and the result shows that the proportion of people who can fall asleep within 30min by the probiotics group is increased to 54.0% from the original 22.0% compared with the proportion of people before the intervention after 15d, and is increased to 72.0% after the intervention is finished; the ratio of the number of people falling asleep in 30min before and after intervention is increased from original 24.0% to 25.0%, no significant difference exists, the ratio of the number of people falling asleep in 30min before and after intervention is also increased to a certain extent, the difference is significant (P < 0.05), and the ratio of the number of people falling asleep in 30min after the probiotics and the drug group are matched.
Fig. 5 to 7 are graphs showing the statistics of the mid-night wake-up condition of the 4 groups of the subjects before, during and after the intervention, fig. 5 shows the statistics of the mid-night wake-up condition of the 4 groups of the subjects before (0 d) the intervention, fig. 6 shows the statistics of the mid-night wake-up condition of the 15d group of the subjects during the intervention, and fig. 7 shows the statistics of the mid-night wake-up condition of the 30d group of the subjects after the intervention. The results show that after 30d of intervention, compared with the prior intervention, the number of times of awakening the probiotics group in the middle night is obviously reduced, the number of people who feel the sky bright is increased from 10.0% to 43.0%, and the number of people who wake in the middle night and cannot fall asleep very quickly is reduced from 60.0% to 14.0%; the number of awakenings before and after the intervention of the placebo group is not significantly different, and the number of people who feel bright is not significantly changed before and after the intervention; the remaining two groups also showed a trend of increasing the proportion of people sleeping to the sky, the probiotic-matched drug group had a proportion of people sleeping to the sky increased from 15% to 48% and 33% and the drug group increased from 14% to 22% compared to before the intervention.
Fig. 8 to 10 are graphs of statistics of total sleeping time lengths of 4 groups of tested persons before, during and after the intervention, fig. 8 is a graph of statistics of total sleeping time lengths of the tested persons before the intervention (0 d), fig. 9 is a graph of statistics of total sleeping time lengths of the tested persons during the intervention (15 d), and fig. 10 is a graph of statistics of total sleeping time lengths of the tested persons after the intervention (30 d). The result shows that after 30d of intervention, compared with the prior intervention, the change of the sleeping time length of the probiotics group has obvious difference (P < 0.05), the number of people with the total sleeping time length of more than 7h is increased from original 12.0% to 31.0%, the number of people with the sleeping time length of 3-5h is reduced by 29%, and the obvious descending trend is shown in the middle of the intervention; the sleeping time length before and after the intervention of the placebo group has no obvious difference, the total sleeping time length of the other two groups has obvious change before and after the intervention, and the total sleeping time length of the probiotics matched with the drug group treatment after the intervention is over 7 hours is increased to 29.0 percent from the original 12.0 percent; the total sleep time after the intervention is over 7 hours in the drug group treatment is also 13% higher than that in the probiotic treatment group.
Fig. 11 to 12 are diagrams of statistics of mental states of 4 groups of subjects before and after the intervention, fig. 11 is statistics of mental states of the subjects after the waking of the group of subjects before the intervention (0 d), and fig. 12 is statistics of mental states of the group of subjects after the intervention (30 d). The results show that the difference of the mental state change of the population of the probiotics group after the intervention is over (P < 0.05), the number of people who are full after sleeping is increased by 23%, and the number of people who feel not sleeping or feel more tired is reduced; the ratio of the mental states of the population before and after intervention of the placebo group is still unchanged, the ratio of the population with full energy of the other two groups is in an ascending trend, and the change of the probiotics matched with the drug treatment group is more obvious.
The demographics of the 4 groups of subjects with improved effects on the application of the different intervention groups to the adjustment of mood, relief of stress and improvement of anxiety after the intervention process to the end of the intervention are shown in table 1 below.
TABLE 1
As can be seen from table 1, the taking effect of all groups of subjects is counted at the beginning of taking 10d, and the result shows that 12 people feel mood is relieved when the probiotics group is intervened 10d, mood depression and dysphoria are improved, and 14 people mood is relieved when the probiotics compound medicine group is used; after 20d of intervention, 30 people feel relaxed, no more irritability, pessimistic aversion and the like, and 32 people feel obviously improved; intervention is performed for 30d, 46 people have stable mood, the mood is smooth, the attention is more concentrated, the working efficiency is improved, only 4 people can not be relieved, and 44 people also have pleasant mood, and the pressure and anxiety are obviously relieved; in addition, in the whole intervention process, the drug group has a certain treatment effect on the aspect of conditioning emotion of the tested person group, but the number of people with improved emotion is relatively small compared with the two groups; the placebo group had no apparent effect of intervention on the population tested throughout the intervention time, and no variability.
From the results of fig. 1-12 and table 1, it can be seen that the probiotic group, the placebo group, the probiotic compound drug group and the drug group are compared with each other for improving effect, PSQI score, sleep time, wake-up condition at midnight, total sleep time and mental state after wake-up of the people suffering from sleep disorder, and the results show that the sleep quality of the patients can be better improved by taking the compound probiotic, and the emotion of the patients is obviously better after the sleep quality is improved.
It is apparent that the above examples are given by way of illustration only and are not limiting of the embodiments. Other variations or modifications of the above teachings will be apparent to those of ordinary skill in the art. It is not necessary here nor is it exhaustive of all embodiments. While still being apparent from variations or modifications that may be made by those skilled in the art are within the scope of the invention.

Claims (10)

1. The lactobacillus gasseri ProSci-108 is characterized in that the lactobacillus gasseri ProSci-108 is preserved in China general microbiological culture Collection center with the preservation number of CGMCC No.25448.
2. Use of Lactobacillus gasseri ProSci-108 for the preparation of any one of the following products (1) - (3),
(1) An anxiety-reducing product;
(2) A sleep improving product;
(3) And (3) a mood-regulating product.
3. Use according to claim 2, characterized in that the product is a food, a health product or a pharmaceutical product.
4. Use according to claim 3, characterized in that the food product is a yoghurt, a yoghurt block, a solid beverage, a probiotic beverage.
5. The use according to claim 3, wherein the pharmaceutical product is in the form of a powder, a tablet or a capsule.
6. A product for alleviating anxiety, improving sleep or regulating mood comprising a lactobacillus gasseri ProSci-108 according to claim 1.
7. The product according to claim 6, wherein the total viable count of Lactobacillus gasseri ProSci-108 in the product is not less than 3X 10 10 CFU/mL or 3X 10 10 CFU/g。
8. Use according to any of claims 6-7, wherein when the product is a pharmaceutical product, the product further comprises a chemical.
9. The use according to claim 8, wherein the chemical is barbital sodium.
10. An ingested product according to claim 6, wherein a source of lactobacillus gasseri ProSci-108 in the product comprises any one or more of a culture, a ferment and a lyophilized powder of the strain.
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