Microbial composition for regulating emotion, product and application thereof

Abstract

The invention provides a microorganism composition for regulating emotion, a product and application of the microorganism composition, and belongs to the technical field of nutritional compositions. The microbial composition comprises the following components: cannabidiol, lactobacillus helveticus, bifidobacterium longum and lactobacillus paracasei. The composition has the function of relieving anxiety and depression caused by pressure.

Description

Technical Field

[0001] The invention belongs to the technical field of nutritional compositions and relates to a mood regulating microbial composition, a product and an application thereof. Background Art

[0002] Cannabis sativa L. is an annual herbaceous plant of the Cannabaceae family and the Cannabis genus. Cannabis contains more than 100 terpene phenolic bioactive substances, namely cannabinoids. Among them, cannabidiol (CBD) is an important cannabinoid component.

[0003] The molecular formula of cannabidiol (CBD) is as follows:

[0004]

[0005] In recent years, as people continue to deepen their research on intestinal microbiota, the concept of the microbial-gut-brain axis has been proposed. Changes in the intestinal microbial community can lead to changes in the substances produced by intestinal microbial activities. After these substances are absorbed by the intestine, they may affect multiple extraintestinal systems including the central nervous system. An increasing number of studies have shown that probiotics have potential value in improving central nervous system-related diseases including anxiety, depression, and autism.

[0006] Patent CN111956542A provides a children's toothpaste made of hemp leaf extract that helps to fight irritability and depression. The contents of the components are: 35 to 45% glycerol, 35 to 45% water, 10 to 20% hydrated silica, 1 to 2% sodium lauryl sulfate, 0.01 to 1% flavor, 0.01 to 1% hydroxypropyl guar gum, 0.01 to 1% cellulose gum, 0.2 to 0.72% Lactobacillus paracasei, 0.015 to 0.01% hemp leaf extract, 0.01 to 1% xylitol, 0.01 to 1% hydroxyapatite, 0.01 to 1% trichlorogalactose, and 0.01 to 1% benzyl alcohol.

[0007] Patent CN117296922A provides a fermented milk for improving depression symptoms. The fermented milk is prepared from the following raw materials in parts by weight: 90-97 parts of raw milk, 1-5 parts of inulin, 1-5 parts of oligofructose, 0.1-2 parts of galacto-oligosaccharide, 0.1-2 parts of polydextrose, 0.1-2 parts of isomaltooligosaccharide, and 0.1-0.6 parts of xylo-oligosaccharide. The raw materials of the fermented milk also include composite bacterial powder; Bacillus, Lactobacillus plantarum, Lactobacillus paracasei, Bifidobacterium breve, Bifidobacterium lactis, and Bifidobacterium longum; the addition amount of each bacterial species in the composite bacterial powder is Lactobacillus gasseri (0.7-5)×10 8 CFU / 100g raw milk, Streptococcus thermophilus (0.1-9)×10 10CFU / 100g raw milk, Lactobacillus bulgaricus (0.1-9)×10 10 CFU / 100g raw milk, Lactobacillus acidophilus (0.5-7)×10 8 CFU / 100g raw milk, Lactobacillus plantarum (0.5-7)×10 8 CFU / 100g raw milk, Lactobacillus paracasei (0.1-9)×10 7 CFU / 100g raw milk, Bifidobacterium breve (1-5)×10 6 CFU / 100g raw milk, Bifidobacterium lactis (1-5)×10 6 CFU / 100g raw milk, Bifidobacterium longum (0.5-5)×10 6 CFU / 100g raw milk.

[0008] Patent CN117297098A discloses a probiotic composition for relieving depression and a preparation method thereof, in order to solve the technical problem that the long-term use of existing antidepressants can destroy intestinal flora, cause intestinal dysbiosis, and cause gastrointestinal reactions. It includes probiotics, prebiotics and nutrients. The probiotics include Bifidobacterium adolescentis, Bacillus coagulans, Bifidobacterium breve, Bifidobacterium infantis, Bifidobacterium longum, Lactobacillus casei, Lactobacillus helveticus, Lactobacillus paracasei, Lactobacillus plantarum, Lactobacillus rhamnosus and Lactobacillus salivarius. The prebiotics include oligofructose, lactitol, L-arabinose and inulin. The nutrients include vitamin B6, vitamin B9 and ginseng extract. The combined strain provided by the present invention has higher activity, and the effective combination of probiotics and nutrients can significantly improve the sleep, anxiety and diarrhea of ​​patients with depression.

[0009] However, none of the above-mentioned prior arts can provide a microbial composition containing cannabidiol that is better effective in improving anxiety and depression. Summary of the invention

[0010] Terms and Claims of the Present Invention:

[0011] 1. As used herein, the articles “a”, “an” and “the” include plural referents unless expressly limited to one or more referents otherwise.

[0012] 2. As used herein, numerical range: Unless otherwise expressly indicated, all ranges or ratios disclosed herein will be understood to include any and all sub-ranges or sub-ratios contained therein. For example, a range or ratio of 1 to 30 stated should be considered to be included between a minimum value of 1 and a maximum value of 30, and any sub-range or sub-ratio, integer, decimal, or sub-range or sub-ratio consisting of integers or decimals including the end points.

[0013] 3. As used herein, the terms "comprises," "including," "have," "have," "may," "contain" and variations thereof mean open-ended conjunctions or terms that do not exclude the possibility of additional compositions or structures.

[0014] In view of the problem that the prior art cannot provide a microbial composition containing cannabidiol that is better effective in improving anxiety and depression, the purpose of the present invention is to provide a mood-regulating microbial composition, product and application thereof.

[0015] To achieve the above-mentioned purpose of the invention, on the one hand, the present invention provides a microbial composition for regulating mood, comprising the following ingredients: cannabidiol, Lactobacillus helveticus, Bifidobacterium longum and Lactobacillus paracasei.

[0016] Preferably, the Lactobacillus helveticus is Lactobacillus helveticus R52.

[0017] Preferably, the Bifidobacterium longum is Bifidobacterium longum R175.

[0018] Preferably, the Lactobacillus paracasei is Lactobacillus paracasei R215.

[0019] Preferably, the ratio of the total viable count of the Lactobacillus helveticus, the Bifidobacterium longum and the Lactobacillus paracasei to the mass of the cannabidiol is 10 9 :0.1-0.4(CFU:mg).

[0020] More preferably, and as an embodiment of the present invention, the ratio of the total viable count of the Lactobacillus helveticus, the Bifidobacterium longum and the Lactobacillus paracasei to the mass of the cannabidiol is 10 9 :0.25(CFU:mg).

[0021] Preferably, the ratio of the viable counts of the Lactobacillus helveticus, the Bifidobacterium longum and the Lactobacillus paracasei is 1-10:1-10:1-10.

[0022] More preferably, and as an embodiment of the present invention, the ratio of the viable counts of the Lactobacillus helveticus, the Bifidobacterium longum and the Lactobacillus paracasei is 9:1:10.

[0023] In another aspect, the present invention provides a mood regulating product, the active ingredients of which include the above-mentioned microbial composition.

[0024] Preferably, the product is selected from at least one of food, nutritional supplement and medicine.

[0025] Preferably, the dosage form of the product is selected from at least one of granules, tablets, powders, pastes, solutions and lozenges.

[0026] In yet another aspect, the present invention provides use of the microbial composition in preparing a product for regulating mood.

[0027] Preferably, the emotion regulation is to relieve anxiety.

[0028] Preferably, the mood regulation is to relieve depression.

[0029] Preferably, the emotion regulation is to relieve the emotion caused by external pressure.

[0030] Compared with the prior art, the present invention has the following beneficial effects:

[0031] (1) The present invention provides a microbial composition for regulating mood with a simple formula, which is expected to be applied in the fields of food, nutritional supplements and medicines.

[0032] (2) Animal experiments show that the mood regulating composition provided by the present invention can significantly alleviate the anxiety and depression-like behaviors of mice. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1 This is the distribution diagram of the time mice spend in the middle area of ​​the open field.

[0034] Figure 2 This is the distribution diagram of the distance moved by mice in the open field.

[0035] Figure 3 Distribution of the number of times mice entered the middle area of ​​the open field.

[0036] Figure 4 is the distribution diagram of the time mice stayed in the open arms.

[0037] Figure 5 This is the distribution of the time mice spend immobile in the elevated maze.

[0038] Figure 6 This is the distribution diagram of the time mice stay in the bright box.

[0039] Figure 7 This is the time distribution diagram of the mice entering the dark room from the light room for the first time.

[0040] Figure 8 It is the distribution graph of the number of times mice pass through the light and dark boxes.

[0041] Fig. 9 This is the distribution diagram of the immobility time of mice during tail suspension. DETAILED DESCRIPTION

[0042] The following non-limiting examples can enable those skilled in the art to more fully understand the present invention, but do not limit the present invention in any way. The following content is merely an exemplary description of the scope of the present invention, and those skilled in the art can make various changes and modifications to the present invention based on the disclosed content, which should also fall within the scope of the present invention.

[0043] The present invention is further described below by way of specific examples. The various chemical reagents used in the examples of the present invention are obtained through conventional commercial channels unless otherwise specified. Unless otherwise specified, the contents described below are all weight contents. Unless otherwise specified, it is understood that the process is carried out at room temperature.

[0044] The raw material information used in the present invention is shown in the following table:

[0045]

[0046] In the following embodiments and comparative examples, Lactobacillus helveticus R52 used may be referred to as R52, Bifidobacterium longum R175 may be referred to as R175, Lactobacillus paracasei R215 may be referred to as R215, and cannabidiol may be referred to as CBD.

[0047] Example 1

[0048] Microbial composition, comprising:

[0049] 25mg cannabidiol, Lactobacillus helveticus R52, Bifidobacterium longum R175, Lacticaseibacillus paracaseiR215.

[0050] Among them, the total number of viable bacteria in R52, R175, and R215 is 10 11 CFU, the ratio of the number of live bacteria among the three is 9:1:10.

[0051] Effect evaluation

[0052] 1. Experimental Methods

[0053] 1.1. Experimental Materials

[0054] The following experiments were reviewed and approved by the Institutional Animal Care and Use Committee (IACUC) of the Institute of Brain Cognition and Brain Disorders (BCBDI), Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, with the ethics number SIAT-IACUC-20211222-NS-NTPZX-WLP-A2090-01.

[0055] Seven-week-old adult C57BL / 6J male mice used in the following experiments were purchased from Vitallife.

[0056] Experimental design

[0057] After one week of environmental adaptation, adult male C57BL / 6 mice were given samples of a single component or different formulations by long-term oral gavage for 5 weeks, and then at the 4th week of sampling, the mice in the model group, Example 1 group, CBD control group, and microbial control group were subjected to 1 week of stress induction, and the blank control group was not sampled and not subjected to stress induction; finally, behavioral tests (including anxiety-like behavioral tests and depression-like behavioral tests) were performed, and the mice were continuously sampled during the above-mentioned evaluation experiment.

[0058] The components or compositions involved in each group in the experiment are as follows:

[0059] Blank control group: intragastric administration of normal saline;

[0060] Model group: intragastric administration of normal saline;

[0061] Example 1 group: The microbial composition of Example 1 was administered intragastrically, wherein the dosage of CBD was 25 mg / kg mouse body weight / day; correspondingly, based on 20 g per mouse, it can be determined that the dosage of R52 was approximately 9×10 8 CFU per mouse per day, the dose of R175 is approximately 10 8 CFU per mouse per day, the dose of R215 is approximately 10 9 CFU per mouse per day, the dose of CBD is approximately 0.5 mg per mouse per day.

[0062] CBD control group: CBD was administered orally at a dose of 25 mg / kg mouse body weight / day.

[0063] Microbial control group: Oral administration of microorganisms. The dose of R52 was 9×10 8 CFU per mouse per day, the dose of R175 was 10 8 CFU per mouse per day, the dose of R215 was 10 9 CFU per mouse per day.

[0064] The stress response program is as follows:

[0065] Plantar stimulation was performed 10 times (current 0.5 mA, each time lasting 2 seconds, with an interval of 10 seconds); tail suspension was performed for 30 minutes; restraint was performed for 1 hour.

[0066] During the week of stress induction, the above stress regimen was performed once a day in a random order.

[0067] 1.3. Anxiety-like behavior test

[0068] 1.3.1. Open field test

[0069] The open field consists of a square box (35cm×35cm×30cm), which is divided into 16 areas (8.75cm×8.75cm), and the 4 areas in the middle will be defined as the middle area (17.5cm×17.5cm). Mice will be randomly placed in the middle area of ​​the open field. After 10 minutes of exploration, the time the mice stay in the middle area will be recorded and used as an indicator of anxiety. The longer the time spent in the middle area, the lower the anxiety level of the mice.

[0070] 1.3.2. Elevated maze test

[0071] The elevated maze apparatus is a cross-shaped maze with two open arms (35cm×5cm×15cm) and two closed arms (35cm×5cm). The middle area of ​​the cross maze is a fully open area (5cm×5cm). The entire cross maze is 50cm above the ground. At the beginning of the experiment, the mice were placed in the middle area of ​​the maze, facing the open arms. Then, the number of times the mice entered the open arms and closed arms within 7 minutes, the time they stayed in the open arms and closed arms, and the time they were still were automatically monitored. The time the mice stayed in the open arms was used as an anxiety indicator. The longer the mice stayed in the open arms, the lower their anxiety level.

[0072] 1.3.3. Light and dark box test

[0073] The size of the light-dark box device is 50cm×25cm×25cm, which contains two spaces, a light box and a dark box. The partition wall between the two boxes has a 7.5cm×7.5cm door hole for mice to shuttle through. The test time for each mouse is 5min. At the beginning of the test, the mouse is placed in the light box and allowed to explore freely in the device. The time the mouse stays in the light box, the number of times the light and dark boxes shuttle, and the time of the first entry into the dark room are monitored. The time spent in the light box is used as an anxiety indicator. The longer the mouse stays in the light box, the lower the anxiety level.

[0074] 1.4. Depression-like behavior test (tail suspension test)

[0075] The tail suspension test is widely used to assess despair behavior in rodents. Briefly, mice are suspended by tape about 1 cm from the tip of the tail 30 cm above the ground. The duration of the experiment is 6 min, and the immobility time (no struggling, little body movement) during the suspension process is automatically monitored during the last 5 min. The duration of immobility of mice will be used as an indicator of depression, and the longer the immobility time, the higher the degree of depression of the mice.

[0076] Statistical analysis

[0077] Prism software was used for statistical analysis. In the behavioral experimental results, each group was first checked for outliers, and then a significance analysis was performed after elimination. The blank control group and the model group used an unpaired t-test, while the model group and each embodiment group used a one-way analysis of variance.

[0078] 2. Experimental results

[0079] 2.1. Effects of stress on anxiety-like behavior in mice

[0080] 2.1.1. Open field test

[0081] The results of the open field test are as follows Figure 1-3 Compared with the CBD control group and the microbial control group, the microbial composition of Example 1 group has significantly better effects on increasing the time mice stay in the open field, increasing the distance mice move in the open field, and increasing the number of times mice enter the middle area of ​​the open field.

[0082] 2.1.2 Elevated maze test

[0083] The results of the elevated maze test are as follows Figure 4 , 5 Compared with the CBD control group and the microbial control group, the microbial composition of Example 1 group has a significantly better effect of increasing the time mice stay in the open arm and reducing the immobility time in the maze.

[0084] 2.1.3. Light-dark box experiment

[0085] The experimental results of the light-dark box experiment are as follows Figure 6-8 Compared with the CBD control group and the microbial control group, the microbial composition of Example 1 group has significantly better effects on increasing the time mice stay in the light box, increasing the time mice enter the dark room from the light room for the first time, and increasing the number of times mice pass through the light box.

[0086] 2.2. Effects of stress on depressive-like behavior in mice

[0087] The results of the mouse tail suspension experiment are as follows Fig. 9The results of the mouse tail suspension experiment showed that the microbial composition of Example 1 group had a significantly better effect of reducing the immobility time of mice compared with the CBD control group and the microbial control group.

[0088] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, rather than to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions of the technical solution of the present invention by ordinary technicians in this field do not deviate from the essence and scope of the technical solution of the present invention.

Claims

1. A microbial composition for regulating mood, characterized in that: Includes the following ingredients: Cannabidiol, Lactobacillus helveticus, Bifidobacterium longum, and Lactobacillus paracasei.

2. The microbial composition according to claim 1, characterized in that The Lactobacillus helveticus is Lactobacillus helveticus R52, the Bifidobacterium longum is Bifidobacterium longum R175, and the Lactobacillus paracasei is Lactobacillus paracasei R215.

3. The microbial composition according to claim 1, characterized in that The ratio of the total number of viable bacteria of Lactobacillus helveticus, Bifidobacterium longum and Lactobacillus paracasei to the mass of cannabidiol is 10 9 :0.1-0.4(CFU:mg).

4. The microbial composition according to claim 3, characterized in that The ratio of the total number of viable bacteria of Lactobacillus helveticus, Bifidobacterium longum and Lactobacillus paracasei to the mass of cannabidiol is 10 9 :0.25(CFU:mg).

5. The microbial composition according to any one of claims 1 to 4, characterized in that The ratio of the live bacterial counts of the Lactobacillus helveticus, the Bifidobacterium longum and the Lactobacillus paracasei is 1-10:1-10:1-10.

6. The microbial composition according to claim 5, characterized in that The ratio of the live bacterial counts of the Lactobacillus helveticus, the Bifidobacterium longum and the Lactobacillus paracasei is 9:1:

10.

7. A product for regulating emotions, characterized in that: The active ingredient comprises the microbial composition according to any one of claims 1 to 6.

8. The product according to claim 7, characterized in that The product is selected from at least one of food, nutritional supplements, and medicines; the dosage form of the product is selected from at least one of granules, tablets, powders, pastes, solutions, and lozenges.

9. Use of the microbial composition according to any one of claims 1 to 6 in the preparation of a product for regulating mood.

10. The use according to claim 9, characterized in that: The emotion regulation is selected from at least one of relieving anxiety, relieving depression, and relieving emotions caused by external pressure.

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