Application of lactococcus lactis in product for improving sleep

Abstract

The invention relates to the field of microorganisms, and discloses application of lactococcus lactis in a product for improving sleep. The lactococcus lactis is named as WHH2078, is separated from yoghurt butter collected from a country in Sankarst city in Tibet Autonomous Region in China, is preserved in China General Microbiological Culture Collection Center on February 25, 2021, has a preservation number of CGMCCNO.21831, and is classified and named as Lactococcus lactis. The Lactococcus lactis is named as Lactococcus lactis, and the Lactococcus lactis is named as Lactococcus lactis. The lactococcus lactis has the effects of shortening the sleep latency, prolonging the total sleep time, improving 5-hydroxytryptamine and gamma-aminobutyric acid of the brain, serum and colon, lowering the corticosterone level, regulating back the diversity of intestinal microorganisms alpha, regulating up the abundance of intestinal acidobacilli and lowering the abundance of actinomycetes, desulfurization bacilli and verruca microflora. The sleep disorder can be effectively improved.

Description

Technical Field

[0001] The invention relates to the technical field of microorganisms, and in particular to application of Lactococcus lactis in a product for improving sleep. Background Art

[0002] With the development of modern society and the accelerated pace of people's lives, improving sleep quality and seeking safe and effective means of aiding sleep have become urgent health issues. Among the many means of improving sleep, drug therapy is a common way. However, although traditional sleep aids such as benzodiazepines, non-benzodiazepines and antihistamines can effectively relieve insomnia symptoms, their long-term use is prone to dependence and drug resistance, and may be accompanied by a series of side effects, such as daytime sleepiness, inattention, memory loss, etc. Therefore, exploring safe, side-effect-free natural sleep aids has gradually become a research hotspot.

[0003] Studies have found that the imbalance of intestinal flora is closely related to a variety of psychiatric and neurological diseases such as depression, anxiety, and sleep disorders. By regulating the intestinal microbiota, new therapeutic strategies may be provided to improve these diseases. Lactic acid bacteria are an important part of the intestinal flora of healthy individuals. Lactococcus lactis, as a common lactic acid bacteria, is widely used in the food fermentation industry. Patent CN202110743748.0 authorizes a strain of Lactococcus lactis WHH2078 with the effect of improving depression. This strain has good adhesion properties, acid and bile resistance, and intestinal barrier protection function. It can significantly lower serum corticosterone levels, relieve depression / anxiety-like behaviors by adjusting the depression-related neural factors in the body, and has a significant effect on improving depression.

[0004] So far, there are no reports indicating that Lactococcus lactis WHH2078 has the effect of improving sleep. Furthermore, within the genus (Lactococcus) to which this strain belongs, no strain has been found that can improve sleep through the combined effects of shortening the latency to fall asleep, prolonging the total sleep duration, increasing 5-hydroxytryptamine and γ-aminobutyric acid in the brain, serum and colon, downregulating corticosterone levels, adjusting the alpha diversity of intestinal microorganisms, upregulating the abundance of intestinal acidobacteria, and downregulating the abundance of actinomycetes, desulfurizing bacteria and verrucomicrobia. Summary of the invention

[0005] In order to solve the technical problems that the above-mentioned drugs for improving sleep quality have low safety and side effects, the present invention provides an application of Lactococcus lactis in a product for improving sleep. Lactococcus lactis WHH2078 has significant physiological functions, can effectively alleviate sleep problems, and is highly safe without side effects.

[0006] The specific technical scheme of the present invention is: an application of Lactococcus lactis in a product for improving sleep, wherein the Lactococcus lactis is named WHH2078, which has been deposited in the General Microbiological Center of the China Microbiological Culture Collection Administration Committee on February 25, 2021, at No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, China, with a deposit number of CGMCC NO.21831. The strain has been disclosed in another patent, and the patent information is as follows: the patent name is "A strain of Lactococcus lactis with the effect of improving depression and its application", the publication number is CN113512514A, and the publication date is October 19, 2021.

[0007] The lactococcus lactis WHH2078 of the present invention can simultaneously shorten the latency period of falling asleep, prolong the total sleep time, increase the 5-hydroxytryptamine and γ-aminobutyric acid in the brain, serum and colon, downregulate the level of corticosterone, adjust the alpha diversity of intestinal microorganisms, upregulate the abundance of intestinal acidobacteria, and downregulate the abundance of actinomycetes, desulfurization bacteria and verrucomicrobia, thereby improving sleep. Specifically: The lactococcus lactis WHH2078 of the present invention can effectively shorten the sleep latency of insomnia mice induced by PCPA (para-chlorophenylalanine) and prolong the total sleep time. The dosage is 1×10 9 CFU / d had a significant effect. Compared with the model group, the sleep latency was shortened by 49.63% and the total sleep duration was prolonged by 38.49%, which was equivalent to the effect of melatonin (0.5 mg / kg). Lactococcus lactis WHH2078 can effectively shorten the sleep latency and prolong the total sleep duration of insomnia mice induced by caffeine. 9 CFU / d had a significant effect. Compared with the model group, the sleep latency was shortened by 20.18% and the total sleep time was prolonged by 64.05%, which was equivalent to that of the drug melatonin (0.5 mg / kg).

[0008] The lactococcus lactis WHH2078 of the present invention can effectively improve the levels of 5-HT (5-hydroxytryptamine) and GABA (gamma-aminobutyric acid) in the central nervous system of insomnia mice induced by PCPA, and the dosage is 1×10 9 CFU / d had a significant effect, with the central nervous system 5-HT level rising by 39.74% to normal levels and GABA level rising by 31.87% to normal levels, which was equivalent to the effect of melatonin (0.5 mg / kg). Lactococcus lactis WHH2078 can effectively increase the central nervous system 5-HT and GABA levels in mice with insomnia induced by caffeine. 9CFU / d had a significant effect, with central 5-HT levels rising 40.00% to normal levels and GABA levels upregulated 30.51% to normal levels, which were equivalent to the effects of the drug melatonin (0.5 mg / kg).

[0009] The lactococcus lactis WHH2078 of the present invention can effectively down-regulate the serum corticosterone level of insomnia mice induced by PCPA, and the dosage is 1×10 9 CFU / d had a significant effect, and the serum corticosterone level of PCPA-induced insomnia mice was reduced by 33.79% to normal levels, which was equivalent to the effect of melatonin (0.5 mg / kg). Lactococcus lactis WHH2078 can effectively downregulate the serum corticosterone level of caffeine-induced insomnia mice. The dose of 1×10 9 CFU / d had a significant effect, reducing serum corticosterone levels by 33.60% to normal levels, which was equivalent to the effect of the drug melatonin (0.5 mg / kg).

[0010] After being colonized in the intestine, the lactococcus lactis WHH2078 of the present invention affects the body's activities through its own metabolic activity, the interaction with the intestinal flora, and the influence on the synthesis of substances in the intestine, thereby playing a role in improving sleep. For example, WHH2078 can increase the alpha diversity of intestinal microorganisms, up-regulate the abundance of intestinal acid bacteria, down-regulate the abundance of actinomycetes, desulfurization bacteria and verrucomicrobia, and adjust the levels of 5-HT and GABA in the colon of insomniac mice, thereby improving the sleep quality of insomniac mice.

[0011] Preferably, the product is used to shorten the sleep onset latency, prolong the total sleep duration, increase 5-hydroxytryptamine and gamma-aminobutyric acid in the brain, serum and colon, and downregulate corticosterone levels.

[0012] Preferably, the product is used to adjust the alpha diversity of intestinal microorganisms, upregulate the abundance of intestinal acidobacteria and downregulate the abundance of actinobacteria, desulfurobacteria and verrucomicrobia.

[0013] Preferably, the product is a health product or a medicine.

[0014] Preferably, the product is a capsule, a tablet or a powder.

[0015] Preferably, the product is an oral product.

[0016] Lactococcus lactis WHH2078 can be taken orally to improve sleep and relieve stress.

[0017] The invention discloses an application of Lactococcus lactis in preparing a product for shortening the sleep-onset latency period. The Lactococcus lactis is named as WHH2078 and has a preservation number of CGMCC NO.21831.

[0018] The invention discloses an application of Lactococcus lactis in preparing a product for prolonging the total sleep time. The Lactococcus lactis is named as WHH2078 and has a preservation number of CGMCC NO.21831.

[0019] The invention discloses an application of Lactococcus lactis in preparing a product for regulating the level of sleep-related substances in a body. The Lactococcus lactis is named WHH2078 and has a preservation number of CGMCC NO.21831. The regulator is used for increasing 5-hydroxytryptamine and gamma-aminobutyric acid and down-regulating corticosterone.

[0020] Preferably, the 5-hydroxytryptamine is the 5-hydroxytryptamine in the brain, serum and colon; and the γ-aminobutyric acid is the γ-aminobutyric acid in the brain, serum and colon.

[0021] The invention discloses an application of Lactococcus lactis in preparing a product for regulating intestinal microorganisms. The Lactococcus lactis is named as WHH2078 and has a preservation number of CGMCC NO.21831. The intestinal microorganisms are intestinal microorganisms related to improving sleep.

[0022] Preferably, the product is used to restore intestinal microbial alpha diversity.

[0023] Preferably, the product is used to up-regulate the abundance of the phylum Acidobacteria, or to down-regulate the abundance of the phylum Actinobacteria, the phylum Desulfobacteria and the phylum Verrucomicrobia.

[0024] Compared with the prior art, the present invention has the following advantages: The Lactococcus lactis WHH2078 of the present invention has the effects of simultaneously shortening the latency period to fall asleep, prolonging the total sleep time, increasing 5-hydroxytryptamine and γ-aminobutyric acid in the brain, serum and colon, downregulating the level of corticosterone, adjusting the alpha diversity of intestinal microorganisms, upregulating the abundance of intestinal acidobacteria and downregulating the abundance of actinomycetes, desulfurizing bacteria and verrucomicrobia, and can effectively improve sleep disorders. DETAILED DESCRIPTION

[0025] The present invention will be further described below in conjunction with the embodiments.

[0026] A use of Lactococcus lactis in a product for improving sleep, wherein the Lactococcus lactis is named WHH2078 and was deposited at the General Microbiological Center of the China Culture Collection Administration on February 25, 2021, at No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, China, with a deposit number of CGMCC NO.21831.

[0027] As a specific embodiment, the product is used to shorten the latency to fall asleep, prolong the total sleep time, increase the central nervous system 5-hydroxytryptamine and γ-aminobutyric acid, and down-regulate the level of corticosterone. The product is used to shorten the latency to fall asleep, prolong the total sleep time, increase the 5-hydroxytryptamine and γ-aminobutyric acid in the brain, serum and colon, and down-regulate the level of corticosterone. The product is used to adjust the alpha diversity of intestinal microorganisms, increase the abundance of intestinal acid bacteria, and down-regulate the abundance of actinomycetes, desulfurization bacteria and verrucomicrobia. The product is a health product or a medicine. The product is a capsule, tablet or powder. The product is an oral product. The product is used for humans or animals.

[0028] The invention discloses an application of Lactococcus lactis in preparing a product for shortening the sleep-onset latency period. The Lactococcus lactis is named as WHH2078 and has a preservation number of CGMCC NO.21831.

[0029] The invention discloses an application of Lactococcus lactis in preparing a product for prolonging the total sleep time. The Lactococcus lactis is named as WHH2078 and has a preservation number of CGMCC NO.21831.

[0030] The invention discloses an application of Lactococcus lactis in preparing a product for regulating the level of sleep-related substances in a body. The Lactococcus lactis is named WHH2078 and has a preservation number of CGMCC NO.21831. The regulator is used for increasing 5-hydroxytryptamine and gamma-aminobutyric acid and down-regulating corticosterone.

[0031] As a specific embodiment, the 5-hydroxytryptamine is the 5-hydroxytryptamine in the brain, serum and colon; the γ-aminobutyric acid is the γ-aminobutyric acid in the brain, serum and colon.

[0032] The invention discloses an application of Lactococcus lactis in preparing a product for regulating intestinal microorganisms. The Lactococcus lactis is named as WHH2078 and has a preservation number of CGMCC NO.21831. The intestinal microorganisms are intestinal microorganisms related to improving sleep.

[0033] As a specific embodiment, the product is used to adjust the alpha diversity of intestinal microorganisms. The product is used to increase the abundance of intestinal acid bacteria, or to decrease the abundance of actinobacteria, desulfurobacteria and verrucomicrobia.

[0034] The present invention is described below by specific examples. It should be understood that these examples are only used to illustrate the present invention and are not used to limit the scope of the present invention. Without departing from the spirit and scope of the inventive concept, changes and advantages that can be thought of by those skilled in the art are included in the present invention, and the attached claims and any equivalents thereof are the protection scope of the present invention.

[0035] Unless otherwise defined, all technical terms and scientific terms used in the present invention have the same meanings as those commonly understood by ordinary technicians in the field to which the present disclosure belongs. Unless otherwise specified, the raw materials and equipment used in the present invention are conventional raw materials and equipment in the field and can be obtained from conventional commercial channels; unless otherwise specified, the methods used in the present invention are conventional methods in the field.

[0036] Example 1 This example uses the PCPA insomnia mouse model to test the sleep-improving effect of Lactococcus lactis WHH2078.

[0037] This example uses 40 healthy SPF male BALB / c mice, which are 8 weeks old and weigh 20±0.2g. The mice are raised in a clean environment, and the environmental conditions are controlled to be room temperature 26±0.5°C, humidity 50-60%, and light-dark alternation time (12 hours light / 12 hours dark cycle). During the raising period, the mice can freely eat and drink water.

[0038] Forty mice were randomly divided into WHH2078 group, drug group, control group and model group, with 10 mice in each group. The mice in the WHH2078 group were intragastrically gavaged with 200 μL of Lactococcus lactis WHH2078 strain solution every day, with the number of viable bacteria being 1×10 9 CFU / d; the drug group was gavaged with 200 μL melatonin (manufacturer: Sigma-Aldrich, model: M5250) solution every day, the melatonin concentration was 0.5 mg / kg, dissolved in 4.0 ml sterile water; the control group and model group mice were gavaged with 200 μL sterile saline every day. Starting from the first day of gavage treatment, mice in each group were gavaged on the 1st to 14th days of the test period; on the 15th and 16th days of the test period, the model group and WHH2078 group mice were intraperitoneally injected with 200 μL PCPA (manufacturer is Sigma-Aldrich, model C6506) solution, the concentration of which is 350 mg / kg, dissolved in 10.0 ml sterile water, and injected continuously for 2 days; on the 17th day of the test period, all mice were intraperitoneally injected with 200 μL sodium pentobarbital (manufacturer is Henan Putian Tongchuang Biotechnology Co., Ltd., model 230106) solution, the concentration of which is 42 mg / kg, dissolved in 8.0 ml sterile water, and the sleep quality test was performed, and the sleep latency and total sleep duration of the mice were recorded. After the test, the serum corticosterone of the mice was detected, and the brain tissue of the mice was collected for the detection of central 5-HT and GABA levels.

[0039] The sleep quality test results of mice in each group are shown in Table 1.

[0040] Table 1 Example 1 Mouse sleep quality test results Group Sleep latency (s) Total sleep time(s) Control group <![CDATA[456.54±18.32 b ]]> <![CDATA[1632.42±28.57 a ]]> Model Group <![CDATA[922.29±17.66 a ]]> <![CDATA[987.43±21.62 b ]]> Drug group <![CDATA[484.78±7.43 b ]]> <![CDATA[1565.26±22.34 a ]]> WHH2078 Group <![CDATA[464.56±8.21 b ]]> <![CDATA[1605.34±21.66 a ]]> a, b: significant difference, P<0.05.

[0041] As shown in Table 1, compared with the control group, the sleep latency of the mice in the model group induced by PCPA was significantly prolonged, and the total sleep time was significantly shortened (P<0.05), indicating that the model was successfully established; compared with the model group, the sleep latency of the mice in the WHH2078 group was significantly shortened, and the total sleep time was significantly prolonged (P<0.05). At the same time, the test results of the WHH2078 group were similar to those of the drug group, indicating that the administration concentration of Lactococcus lactis WHH2078 was 1×10 9 CFU / d has the ability to improve the sleep quality of PCPA-induced insomnia mice, and its effect is comparable to that of melatonin at a concentration of 0.5 mg / kg.

[0042] The results of serum corticosterone, brain 5-HT and GABA levels in each group of mice are shown in Table 2.

[0043] Table 2 Results of sleep-related biological indicators of mice in Example 1 a, b: significant difference, P<0.05.

[0044] From Table 2, we can see that: (1) The central 5-HT concentration of the insomnia mice induced by PCPA in the model group was significantly lower than that in the control group (P<0.05), indicating that the model was successfully established. The central 5-HT concentration of the mice in the WHH2078 group was significantly higher than that in the insomnia group (P<0.05) and similar to that in the drug group, indicating that the administration concentration of Lactococcus lactis WHH2078 was 1×10 9 CFU / d has the ability to upregulate the level of 5-HT in the brain of mice with insomnia induced by PCPA, and its effect is comparable to that of the drug melatonin at a concentration of 0.5 mg / kg.

[0045] (2) The central GABA concentration of the insomnia mice induced by PCPA in the model group was significantly lower than that in the control group (P<0.05), indicating that the model was successfully established. The central GABA concentration of the mice in the WHH2078 group was significantly higher than that in the insomnia group (P<0.05) and was similar to that in the drug group, indicating that the administration concentration of Lactococcus lactis WHH2078 was 1×10 9 CFU / d has the ability to upregulate GABA levels in the brain of mice with insomnia induced by PCPA, and its effect is comparable to that of the drug melatonin at a concentration of 0.5 mg / kg.

[0046] (3) The serum corticosterone concentration of the insomnia mice induced by PCPA in the model group was significantly higher than that in the control group (P<0.05), indicating that the model was successfully established. The serum corticosterone concentration of the mice in the WHH2078 group was significantly lower than that in the insomnia group (P<0.05) and was similar to that in the drug group, indicating that the administration concentration of Lactococcus lactis WHH2078 was 1×10 9 CFU / d has the ability to correct the serum corticosterone concentration level of PCPA-induced insomnia mice, and its effect is comparable to that of melatonin at a concentration of 0.5 mg / kg.

[0047] In summary, the Lactococcus lactis WHH2078 strain was administered at a concentration of 1×10 9 CFU / d can significantly improve the sleep disorders of PCPA-induced insomnia mice, including shortening the latency to fall asleep and prolonging the total sleep time, and adjusting the levels of brain 5-HT, GABA and blood corticosterone. It has an effect equivalent to that of the drug melatonin at a concentration of 0.5 mg / kg, and is a strain with the function of improving sleep.

[0048] Example 2 This example uses a caffeine insomnia mouse model to test the sleep-improving effect of Lactococcus lactis WHH2078.

[0049] This example uses 40 healthy SPF male BALB / c mice, which are 8 weeks old and weigh 20±0.2g. The mice are raised in a clean environment, and the environmental conditions are controlled to be room temperature 26±0.5°C, humidity 50-60%, and light-dark alternation time (12 hours light / 12 hours dark cycle). During the raising period, the mice can freely eat and drink water.

[0050] Forty mice were randomly divided into WHH2078 group, drug group, control group and model group, with 10 mice in each group. The mice in the WHH2078 group were intragastrically gavaged with 200 μL of Lactococcus lactis WHH2078 strain solution every day, with the number of viable bacteria being 1×10 9CFU / d; the drug group was gavaged with 200 μL melatonin (manufacturer: Sigma-Aldrich, model: M5250) solution every day, the melatonin concentration was 0.5 mg / kg, dissolved in 4.0 ml sterile water; the control group and model group mice were gavaged with 200 μL sterile saline every day. Starting from the first day of gavage treatment, mice in each group were gavaged on the 1st to 21st days of the test period; on the 22nd day of the test period, 200 μL of caffeine (manufacturer: Sigma-Aldrich, model 205548) solution was injected intraperitoneally into the model group and WHH2078 group mice, with a concentration of 15 mg / kg, dissolved in 10.0 ml of sterile water, and 30 minutes later, 200 μL of sodium pentobarbital (manufacturer: Henan Putian Tongchuang Biotechnology Co., Ltd., model 230106) solution was injected intraperitoneally into all mice, with a concentration of 42 mg / kg, dissolved in 8.0 ml of sterile water, and a sleep quality test was performed to record the sleep latency and total sleep duration of the mice. After the test, the serum corticosterone of the mice was detected, and the brain tissue of the mice was collected for the detection of central 5-HT and GABA levels.

[0051] The sleep quality test results of mice in each group are shown in Table 1.

[0052] Table 3 Example 2 Mouse Sleep Quality Test Results Group Sleep latency (s) Total sleep time(s) Control group <![CDATA[273.46±9.78 c ]]> <![CDATA[1948.56±14.66 a ]]> Model Group <![CDATA[400.77±9.90 a ]]> <![CDATA[560.99±21.33 c ]]> Drug group <![CDATA[310.65±9.89 b ]]> <![CDATA[1448.12±21.48 b ]]> WHH2078 Group <![CDATA[319.90±10.65 b ]]> <![CDATA[1560.67±23.44 b ]]> a, b: significant difference, P<0.05.

[0053] As shown in Table 3, compared with the control group, the caffeine-induced model group had a significantly prolonged sleep latency and a significantly shortened total sleep time (P<0.05), indicating that the model was successfully established. Compared with the model group, the WHH2078 group had a significantly shortened sleep latency and a significantly prolonged total sleep time (P<0.05). At the same time, the test results of the WHH2078 group were similar to those of the drug group, indicating that the administration concentration of Lactococcus lactis WHH2078 was 1×10 9 CFU / d has the ability to improve the sleep quality of caffeine-induced insomnia mice, and its effect is comparable to that of melatonin at a concentration of 0.5 mg / kg.

[0054] The results of the serum corticosterone, brain 5-HT and GABA levels of mice in each group are shown in Table 4.

[0055] Table 4 Results of sleep-related biological indicators in mice in Example 2 a, b: significant difference, P<0.05.

[0056] From Table 4, we can see that: (1) The central 5-HT concentration of the caffeine-induced insomnia mice in the model group was significantly lower than that in the control group (P<0.05), indicating that the model was successfully established. The central 5-HT concentration of the WHH2078 group was significantly higher than that of the insomnia group (P<0.05) and similar to that of the drug group, indicating that the administration concentration of Lactococcus lactis WHH2078 was 1×10 9 CFU / d has the ability to upregulate the level of 5-HT in the brain of caffeine-induced insomnia mice, and its effect is comparable to that of melatonin at a concentration of 0.5 mg / kg.

[0057] (2) The central GABA concentration of the caffeine-induced insomnia mice in the model group was significantly lower than that in the control group (P<0.05), indicating that the model was successfully established. The central GABA concentration of the WHH2078 group was significantly higher than that of the insomnia group (P<0.05) and similar to that of the drug group, indicating that the administration concentration of Lactococcus lactis WHH2078 was 1×10 9 CFU / d has the ability to upregulate GABA levels in the brain of caffeine-induced insomnia mice, and its effect is comparable to that of melatonin at a concentration of 0.5 mg / kg.

[0058] (3) The serum corticosterone concentration of the caffeine-induced insomnia mice in the model group was significantly higher than that in the control group (P<0.05), indicating that the model was successfully established. The serum corticosterone concentration of the WHH2078 group was significantly lower than that in the insomnia group (P<0.05) and was similar to that in the drug group, indicating that the administration concentration of Lactococcus lactis WHH2078 was 1×10 9 CFU / d has the ability to adjust the serum corticosterone concentration level of mice with caffeine-induced insomnia, and its effect is comparable to that of melatonin at a concentration of 0.5 mg / kg.

[0059] In summary, the Lactococcus lactis WHH2078 strain was administered at a concentration of 1×10 9 CFU / d can significantly improve sleep disorders in caffeine-induced insomnia mice, including shortening the latency to fall asleep and prolonging the total sleep time, and adjusting the levels of brain 5-HT, GABA and blood corticosterone. Its effect is equivalent to that of the drug melatonin at a concentration of 0.5 mg / kg. It is a strain with the function of improving sleep.

[0060] Example 3 This example uses the PCPA insomnia mouse model to test the sleep-improving effect of Lactococcus lactis WHH2078.

[0061] This example uses 30 healthy SPF male BALB / c mice, which are 8 weeks old and weigh 20±0.2g. The mice are raised in a clean environment, and the environmental conditions are controlled to be room temperature 26±0.5°C, humidity 50-60%, and light-dark alternation time (12 hours light / 12 hours dark cycle). During the raising period, the mice can freely eat and drink water.

[0062] Thirty mice were randomly divided into WHH2078 group, control group and model group, with 10 mice in each group. The mice in the WHH2078 group were intragastrically gavaged with 200 μL of Lactococcus lactis WHH2078 strain solution every day, with the number of viable bacteria being 1×10 9 CFU / d; mice in the control group and model group were gavaged with 200 μL sterile saline every day. Starting from the first day of gavage treatment, mice in each group were gavaged on the 1st to 14th days of the test period; on the 15th and 16th days of the test period, mice in the model group and WHH2078 group were intraperitoneally injected with 200 μL PCPA (manufacturer: Sigma-Aldrich, model C6506) solution, with a concentration of 350 mg / kg, dissolved in 10.0 ml sterile water, and injected continuously for 2 days; on the 17th day of the test period, all mice were intraperitoneally injected with 200 μL sodium pentobarbital (manufacturer: Henan Putian Tongchuang Biotechnology Co., Ltd., model 230106) solution, with a concentration of 42 mg / kg, dissolved in 8.0 ml sterile water. Brain tissue, colon tissue and serum samples of mice were collected for the detection of 5-HT and GABA levels, and fecal samples of mice were collected to detect changes in intestinal microbial composition.

[0063] The results of 5-HT levels in mice in each group are shown in Table 5.

[0064] Table 5 Example 3 Mouse 5-HT level detection results b: Significant difference, P<0.05.

[0065] From Table 5, we can see that: The concentration of 5-HT in the central nervous system of the insomnia mice induced by PCPA in the model group was significantly lower than that in the control group (P<0.05), indicating that the model was successfully established. The concentration of 5-HT in the central nervous system, blood, and intestine of the WHH2078 group was significantly higher than that of the insomnia group (P<0.05), indicating that the administration concentration of Lactococcus lactis WHH2078 was 1×10 9 CFU / d can upregulate the level of 5-HT in the brain of mice with PCPA-induced insomnia by promoting the gut-brain axis.

[0066] The results of GABA levels in mice in each group are shown in Table 6.

[0067] Table 6 Example 3 Mouse GABA level detection results b: Significant difference, P<0.05.

[0068] From Table 6, we can see that: The central GABA concentration of the insomnia mice induced by PCPA in the model group was significantly lower than that in the control group (P<0.05), indicating that the model was successfully established. The central, blood, and intestinal GABA concentrations of the WHH2078 group were significantly higher than those of the insomnia group (P<0.05), indicating that the administration of Lactococcus lactis WHH2078 at a concentration of 1×10 9 CFU / d can upregulate the concentration of GABA in the brain of mice with PCPA-induced insomnia by promoting the gut-brain axis.

[0069] The results of intestinal microbial alpha diversity of mice in each group are shown in Table 7.

[0070] Table 7 Example 3 Mouse intestinal microbial alpha diversity detection results Group Shannon Index Chao1 Index Control group <![CDATA[3.38±0.45 a ]]> <![CDATA[3.60±0.28 a ]]> Model Group <![CDATA[1.87±0.34 b ]]> <![CDATA[2.07±0.67 b ]]> WHH2078 Group <![CDATA[3.63±0.22 a ]]> <![CDATA[3.78±0.43 a ]]> b: Significant difference, P<0.05.

[0071] From Table 7, we can see that: The alpha diversity of intestinal microorganisms in the model group, including Shannon index and Chao1 index, was significantly lower than that in the control group (P<0.05). The alpha diversity index of intestinal microorganisms in the WHH2078 group was significantly higher than that in the insomnia group (P<0.05), indicating that the administration concentration of Lactococcus lactis WHH2078 was 1×10 9 CFU / d has the ability to restore the alpha diversity of intestinal microorganisms in PCPA-induced insomnia mice.

[0072] The relative abundance results of the intestinal microbial composition at the phylum level of each group of mice are shown in Table 8.

[0073] Table 8 Relative abundance test results of mouse intestinal microbial composition at the phylum level in Example 3 b: Significant difference, P<0.05.

[0074] From Table 8, we can see that: The relative abundance of Actinobacteria, Desulfobacteria, and Verrucomicrobia in the intestinal tract of mice with insomnia induced by PCPA in the model group was significantly higher than that in the control group (P<0.05). The relative abundance of Actinobacteria, Desulfobacteria, and Verrucomicrobia in the intestinal tract of mice in the WHH2078 group was significantly lower than that in the insomnia group (P<0.05), and the relative abundance of Acidobacteria was significantly lower than that in the insomnia group (P<0.05), indicating that the administration concentration of Lactococcus lactis WHH2078 was 1×10 9 CFU / d has the ability to improve the dysregulation of relative abundance of intestinal microbial phyla in PCPA-induced insomnia mice.

[0075] The raw materials and equipment used in the present invention, unless otherwise specified, are all commonly used raw materials and equipment in the art; the methods used in the present invention, unless otherwise specified, are all conventional methods in the art.

[0076] The above description is only a preferred embodiment of the present invention and does not limit the present invention in any way. Any simple modification, change and equivalent transformation made to the above embodiment based on the technical essence of the present invention still falls within the protection scope of the technical solution of the present invention.

Claims

1. An application of Lactococcus lactis in a product for improving sleep, characterized in that: The lactococcus lactis is named as WHH2078, and its deposit number is CGMCC NO.21831.

2. The use according to claim 1, characterized in that: The product is used to shorten the latency to fall asleep, prolong the total sleep duration, increase 5-hydroxytryptamine and gamma-aminobutyric acid in the brain, serum and colon, and downregulate corticosterone levels.

3. The use according to claim 1 or 2, characterized in that: The product is used to adjust the alpha diversity of intestinal microorganisms, upregulate the abundance of intestinal acidobacteria, and downregulate the abundance of actinobacteria, desulfurobacteria and verrucomicrobia.

4. A use of Lactococcus lactis in preparing a product for shortening the sleep latency period, characterized in that: The lactococcus lactis is named as WHH2078, and its deposit number is CGMCC NO.21831.

5. A use of Lactococcus lactis in preparing a product for prolonging the total sleep time, characterized in that: The lactococcus lactis is named as WHH2078, and its deposit number is CGMCC NO.21831.

6. A use of Lactococcus lactis in preparing a product for regulating the level of sleep-related substances in the body, characterized in that: The lactococcus lactis is named as WHH2078 and its deposit number is CGMCC NO.21831; the regulator is used for increasing 5-hydroxytryptamine and gamma-aminobutyric acid and down-regulating corticosterone.

7. The use according to claim 6, characterized in that: The 5-HT is the 5-HT in the brain, serum and colon; the γ-aminobutyric acid is the γ-aminobutyric acid in the brain, serum and colon.

8. Use of Lactococcus lactis in preparing a product for regulating intestinal microorganisms, characterized in that: The lactococcus lactis is named as WHH2078, and its preservation number is CGMCC NO.21831; and the intestinal microorganisms are intestinal microorganisms related to improving sleep.

9. The use according to claim 8, characterized in that: The product is used to restore intestinal microbial alpha diversity.

10. The use according to claim 8, characterized in that: The product is used to up-regulate the abundance of the phylum Acidobacteria, or to down-regulate the abundance of the phylum Actinobacteria, the phylum Desulfobacteria and the phylum Verrucomicrobia.

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