Application of pediococcus pentosaceus strain A20001 in preparation of product for improving memory concentration and anxiety behavior

Abstract

The invention provides an application of a pediococcus pentosaceus strain A20001 in preparation of a product for improving memory concentration and anxiety and depression behaviors, and belongs to the technical field of biological medicines. Animal experiments show that the pediococcus pentosaceus strain A20001 has the functions of resisting anxiety behavior, resisting depression, improving cognitive function and improving memory concentration. Therefore, the invention provides the application of the pediococcus pentosaceus strain A20001 in the preparation of the medicine for preventing and / or treating the anxiety disorder, the depression or the cognitive impairment and improving the memory, and further provides the application of the pediococcus pentosaceus strain A20001 in the preparation of the medicine for preventing and / or treating the anxiety disorder, the depression or the cognitive impairment.

Description

Technical Field

[0001] The invention belongs to the technical field of biological medicines, and specifically relates to an application of a Pediococcus pentosaceus strain A20001 in preparing a product for improving memory, concentration and anxiety behavior. Background Art

[0002] There are trillions of microorganisms in the human intestine, which play an important role in digestion, immunity and overall health. Damage to any part may lead to diseases and other problems. According to current research reports, the intestine is interconnected with various organs in the host body, which is called the "axis". Researchers have proposed the concept of the microbial "gut-brain" axis to further explore the detailed mechanism. Many research evidences show that the homeostasis of intestinal flora has a profound impact on maintaining host health, including anxiety and cognitive dysfunction. Anxiety is characterized by persistent and excessive worry and fear, which are often inconsistent with the reality and difficult to control. Cognitive dysfunction (cognitive impairment) refers to the impairment of an individual's function in the cognitive field, including decreased ability in memory, attention, language, visual-spatial skills, etc. This disorder may be caused by a variety of factors, including brain damage, neurological diseases (such as Alzheimer's disease), mental illness (such as depression, anxiety), etc. The severity of cognitive dysfunction can range from mild to severe, affecting people's work and life. Anxiety disorder and cognitive dysfunction are both neurological diseases, but due to the complexity of the pathogenesis, differences between individuals, environmental differences and other factors, the existing clinical treatment drugs have limited effects and are scarce, resulting in low cure rates for the two diseases.

[0003] In recent years, studies have reported that the central nervous system can be regulated by intestinal microbial flora, specifically through the long-range effects on the large intestine through probiotics or probiotic metabolites. Currently, there are reports of a group of biological preparations composed of probiotics, postbiotics, prebiotics or synbiotics, which can produce and deliver neuroactive substances such as γ-aminobutyric acid and 5-hydroxytryptamine, and act through the microbial gut-brain axis to improve disease symptoms. However, there are currently no reports on the use of Pediococcus pentosaceus in the field of anti-anxiety and improving cognitive functions such as concentration and memory. Summary of the invention

[0004] In view of this, the object of the present invention is to provide the use of Pediococcus pentosaceus strain A20001 in the preparation of products for improving memory, concentration and anxiety behavior.

[0005] The invention provides an application of a Pediococcus pentosaceus strain A20001 in preparing a medicine for preventing and / or treating anxiety or depression. The preservation number of the Pediococcus pentosaceus strain A20001 is GDMCC No: 64674.

[0006] Preferably, the anxiety disorder includes at least one of the following anxiety behaviors and complications: acute anxiety, chronic anxiety, neurological dysfunction, cardiovascular and cerebrovascular diseases, and digestive system diseases.

[0007] The invention provides an application of a Pediococcus pentosaceus strain A20001 in preparing a medicine for preventing and / or treating cognitive dysfunction and / or improving memory. The preservation number of the Pediococcus pentosaceus strain A20001 is GDMCC No: 64674.

[0008] Preferably, the cognitive dysfunction includes at least one of the following behavioral disorders: memory impairment, aphasia, visuospatial disorder, executive dysfunction, calculation disorder, apraxia and agnosia.

[0009] Preferably, the Pediococcus pentosaceus strain A20001 comprises at least one of the following cultures: bacterial cells of the Pediococcus pentosaceus strain A20001, metabolites of the Pediococcus pentosaceus strain A20001, and fermentation broth of the Pediococcus pentosaceus strain A20001.

[0010] Preferably, the Pediococcus pentosaceus strain A20001 also exists in the form of a probiotic.

[0011] Preferably, the probiotic comprises at least one of the following: tablets, drops, capsules and powders.

[0012] Preferably, the live bacterial concentration of Pediococcus pentosaceus strain A20001 in the probiotic is not less than 2×10 9 CFU / mL or 2×10 9 CFU / g.

[0013] Preferably, the viable bacterial concentration of the Pediococcus pentosaceus strain A20001 is 5×10 9 CFU / mL~100×10 9 CFU / mL or 5×10 9 CFU / g~100×10 9 CFU / g.

[0014] Preferably, the drug comprises at least one of the following oral dosage forms: tablets, powders, capsules and oral liquids.

[0015] The present invention provides an application of Pediococcus pentosaceus strain A20001 in the preparation of a drug for preventing and / or treating anxiety or depression, and the deposit number of the Pediococcus pentosaceus strain A20001 is GDMCC No: 64674. The present invention establishes a chronic unpredictable mild stress model in mice, and uses a sucrose preference test, a tail suspension test, and a forced swimming test to evaluate the antidepressant efficacy of the Pediococcus pentosaceus strain A20001. The results of the sucrose preference experiment show that compared with the model group, the sugar water preference value of the mice in the strain A20001 group is significantly increased (p < 0.0001), indicating that the strain A20001 of the present invention has the function of improving the anhedonia of depressed mice. The results of the tail suspension experiment show that compared with the model group, the immobility time of the mice in the strain A20001 group is significantly shortened (p < 0.001). At the same time, the results of the forced swimming experiment show that compared with the model group, the immobility time of the mice in the A20001 group is significantly shortened (p < 0.001). This shows that the strain A20001 can effectively alleviate the depressive-like behavior of the model mice. At the same time, the present invention uses an open field test and an elevated plus maze test to evaluate the efficacy of the anti-anxiety behavior of the Pediococcus pentosaceus strain A20001. The results of the open field test showed that compared with the model group, the time that the mice in the A20001 group entered the center of the open field was significantly increased; the results of the elevated plus maze test showed that compared with the model group, the number of times the mice in the A20001 group entered the open arm and the percentage of time in the open arm were significantly increased, and the time in the closed arm was significantly shortened. This shows that the Pediococcus pentosaceus strain A20001 can effectively alleviate the anxiety behavior of the model mice, providing a new means for the treatment of anxiety disorders.

[0016] The present invention provides an application of Pediococcus pentosaceus strain A20001 in the preparation of a drug for preventing and / or treating cognitive dysfunction and / or improving memory, and the deposit number of the Pediococcus pentosaceus strain A20001 is GDMCC No: 64674. The present invention uses a novel object recognition experiment to evaluate the efficacy of the Pediococcus pentosaceus strain A20001 in improving cognitive function and improving learning and memory. The results of the novel object recognition experiment showed that compared with the model group, the percentage of time that the mice in the A20001 group explored new objects was significantly increased, indicating that the A20001 strain can effectively improve the cognitive behavior of model mice and improve the memory of mice. The technical solution of the present invention provides a new means for the treatment of cognitive dysfunction and related neurological diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 The results show that strain A20001 significantly increases the body weight of CUMS mice;

[0018] Figure 2 The results show that strain A20001 significantly increases sucrose preference in CUMS mice;

[0019] Figure 3 The results show that strain A20001 significantly shortens the tail suspension immobility time of CUMS mice;

[0020] Figure 4 The results show that strain A20001 significantly shortened the forced swimming immobility time of CUMS mice;

[0021] Figure 5 The results show that strain A20001 significantly increases the exploratory ability of CUMS mice in an open field.

[0022] Figure 6 The strain A20001 significantly increased the time and number of times CUMS mice spent in the open arms;

[0023] Figure 7 The results showed that strain A20001 significantly increased the time CUMS mice spent exploring new objects;

[0024] Figure 8 The results show that strain A20001 significantly increases the serum BDNF level in mice.

[0025] Biomaterial Deposit Information

[0026] Pediococcus pentosaceus strain A20001 was deposited in Guangdong Provincial Microbiological Culture Collection Center on May 23, 2024, with the abbreviation of GDMCC. The address is 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, and the deposit number is GDMCC No: 64674.

[0027] Lactobacillus plantarum strain A21337 was deposited in Guangdong Provincial Microbiological Culture Collection Center on May 23, 2024. The unit is abbreviated as GDMCC. The address is 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou. The deposit number is GDMCC No: 64677. DETAILED DESCRIPTION

[0028] The invention provides an application of a Pediococcus pentosaceus strain A20001 in preparing a medicine for preventing and / or treating anxiety or depression. The preservation number of the Pediococcus pentosaceus strain A20001 is GDMCC No: 64674.

[0029] In the present invention, the Pediococcus pentosaceus strain A20001 is isolated from the feces of healthy centenarians in Guangxi. The strain A20001 has the characteristics of fast growth, entering the logarithmic growth phase within 2 hours of inoculation, and reaching the stable phase at 14 hours. The strain grows stably and does not show a decline phase within 24 hours.

[0030] In the present invention, the pentosaceous Pediococcus strain A20001 preferably includes at least one of the following cultures: the thalli of the pentosaceous Pediococcus strain A20001, the metabolites of the pentosaceous Pediococcus strain A20001 and the fermentation broth of the pentosaceous Pediococcus strain A20001. The method for preparing the fermentation broth of the pentosaceous Pediococcus strain A20001 preferably inoculates the activated pentosaceous Pediococcus strain A20001 in MRS liquid culture to obtain a seed solution; inoculates the seed solution in MRS liquid culture for expansion culture, and inoculates the pentosaceous Pediococcus strain A20001 bacterial solution obtained by the expansion culture in MRS liquid culture again for fermentation culture to obtain a fermentation broth. The activation method of the strain A20001 is preferably that the strain A20001 is inoculated on an MRS plate for streak culture, and a single colony is picked. The temperature of the streak culture is preferably 36-38°C, more preferably 37°C. The inoculation amount during the streak culture is preferably 0.5%-2%, more preferably 1%. The method for preparing seed liquid of the strain A20001 is preferably to inoculate the above-picked single colony into MRS liquid culture medium for cultivation to obtain seed liquid. The culture temperature is preferably 36-38°C, more preferably 37°C. The culture time is preferably 2-4h, more preferably 3h. During the culture, the inoculation amount is preferably 3%-6%, more preferably 4%-5%. The temperature of the expanded culture is 36-38°C, more preferably 37°C. The inoculation amount during the expanded culture is preferably 5%-10%, more preferably 8%. The present invention does not impose any special restrictions on the formula of the MRS liquid culture medium and the MRS plate, and the MRS liquid culture medium and the MRS plate formula known in the art can be used. For example, the MRS liquid culture medium formula is 20g / L glucose, 10g / L beef extract, 10g / L peptone, 5g / L yeast powder, 2g / L dipotassium hydrogen phosphate, 2g / L diammonium citrate, 5g / L sodium acetate, 0.58g / L magnesium sulfate, 0.25g / L manganese sulfate, 0.1% Tween-80, and the pH value is 6.6-6.8. The difference between the MRS plate and the MRS liquid culture medium is that 1.5%-2% agar is added additionally. The method for preparing the bacterial body or metabolite of the pentosaceae strain A20001 is to separate the fermentation broth prepared above into solid and liquid, collect the precipitate as the bacterial body of the pentosaceae strain A20001, and collect the liquid as the metabolite of the pentosaceae strain A20001.

[0031] In the present invention, the Pediococcus pentosaceus strain A20001 preferably also exists in the form of a probiotic. The probiotic refers to supplementing the beneficial bacteria in the intestine so that the beneficial bacteria have sufficient quantity and exert unique biological effects, thereby achieving the purpose of health care or disease prevention and treatment. The probiotic preferably includes at least one of the following: tablets, drops, capsules and powders. The powder is processed by low-temperature freeze-drying technology to ensure the activity of the probiotics. The auxiliary materials of the powder preferably include a freeze-drying agent, such as skim milk powder, etc. The powder is suitable for warm water and is suitable for infants. The tablet is prepared by mixing the Pediococcus pentosaceus strain A20001 and auxiliary materials and pressing them into tablets, which are suitable for chewing and taking. The auxiliary materials preferably include sweeteners or cocoa powder, etc. The capsule preferably contains the Pediococcus pentosaceus strain A20001 alone or mixed with auxiliary materials as a capsule core in the capsule shell. The drops are prepared by mixing Pediococcus pentosaceus strain A20001 with edible oil to form drops that isolate air and moisture. When taking, the dropper port is opened to squeeze the encapsulated oil into the mouth. The live bacteria concentration of Pediococcus pentosaceus strain A20001 in the probiotic is preferably not less than 1×10 10 CFU / mL or 1×10 10 CFU / g, which can be 10×10 10 CFU / mL~100×10 10 CFU / mL or 10×10 10 CFU / g~100×10 10 CFU / g, or 15×10 10 CFU / mL, 20×10 10 CFU / mL, 10×10 10 CFU / mL, 30×10 10 CFU / mL, 40×10 10 CFU / mL, 50×10 10 CFU / mL, 60×10 10 CFU / mL, 70×10 10 CFU / mL or 15×10 10 CFU / g, 20×10 10 CFU / g, 10×10 10 CFU / g, 30×10 10 CFU / g, 40×10 10 CFU / g, 50×10 10 CFU / g, 60×10 10 CFU / g, 70×10 10 CFU / g.

[0032] In the present invention, the drug preferably includes at least one of the following oral dosage forms: tablets, powders, capsules and oral liquids. The present invention has no particular limitation on the preparation method of the drug, and any drug preparation method known in the art can be used.

[0033] In the present invention, the anxiety disorder preferably includes at least one of the following anxiety behaviors and complications: acute anxiety, chronic anxiety, neurological dysfunction, cardiovascular and cerebrovascular diseases, and digestive system diseases. The acute anxiety is mainly manifested as acute panic attacks, and patients often suddenly feel anxious, panicked or excited, which leads to hallucinations and delusions, and sometimes mild confusion of consciousness. The acute anxiety attack can generally last for a few minutes or hours, and the course of the disease is generally not long, and it will gradually tend to ease after a period of time. Acute anxiety patients almost always have more or less sleep disorders, most of which are manifested as difficulty in falling asleep, easy to wake up after falling asleep, often accompanied by nightmares, restless when waking up, and feeling very scared after waking up. The chronic anxiety refers to the anxiety that can last for a long time, and its anxiety level also fluctuates from time to time. Chronic anxiety in the elderly is usually manifested as being more sensitive and irritable at ordinary times, and being upset and inattentive when there is something unsatisfactory in life, and sometimes sulking and losing temper. The neurological dysfunction includes dizziness, vertigo, insomnia, nightmares, insomnia at night or excessive sleepiness during the day, confusion of right and wrong, tension, oppression, cold sweats or a lot of sweat, fear of cold and wind. The cardiovascular and cerebrovascular diseases include the impact of anxiety on blood pressure. Patients with common cerebrovascular diseases in clinical practice are not nervous. Once anxious and nervous, their blood pressure rises, which leads to cerebral hemorrhage and brings great risks. The indigestion is that long-term anxiety can cause colitis, stomach pain, chronic gastritis, and patients often have loss of appetite and irritability. Some patients will overeat, causing symptoms such as gastrointestinal indigestion.

[0034] In one embodiment of the present invention, an open field test and an elevated plus maze test were conducted to evaluate the efficacy of Pediococcus pentosaceus strain A20001 or its preparation in anti-anxiety behavior. The results of the open field test showed that compared with the model group, the time that the mice in the A20001 group entered the center of the open field was significantly increased (p < 0.001). The results of the elevated plus maze test showed that compared with the model group, the number of times the mice in the A20001 group entered the open arm and the percentage of time in the open arm were significantly increased (p < 0.0001), and the time in the closed arm was significantly shortened (p < 0.0001).

[0035] In the present invention, the depression includes single-episode depression, bipolar depression and persistent depression (chronic depression). The single-episode depression refers to a patient who experiences only one significant depressive episode, which usually lasts for several months to about a year and then enters a remission period. Symptoms may include: persistent low mood and depression, loss of interest and pleasure in daily activities, lack of energy, fatigue and physical exhaustion, sleep problems (such as insomnia or excessive sleep), decreased attention and concentration, reduced self-worth, self-blame and inferiority, negative thinking, negative concepts and pessimistic attitudes, and possible anxiety, guilt and despair. The bipolar depression, also known as manic depression, is a mood disorder in which patients alternate between depression and mania. Manic phase symptoms may include: high mood, excitement and impulse, excessive self-confidence, impulsive behavior and high-risk decision-making, distraction, easy distraction and multitasking, and possible delusions or hallucinations. The persistent depression (chronic depression) refers to a depressive symptom duration of up to two years or longer. Symptoms may be mild, but persist, affecting daily life and function. Common symptoms include: depressed mood, loss of interest and pleasure, decreased energy or fatigue, changes in appetite, decreased libido, sleep disorders (such as insomnia or excessive sleep), decreased attention and concentration, low self-esteem, and pessimism about the future.

[0036] In the present invention, a chronic unpredictable mild stress model of mice was established, and the antidepressant efficacy of Pediococcus pentosaceus strain A20001 was evaluated using sucrose preference test, tail suspension test, and forced swimming test. The results of the sucrose preference experiment showed that compared with the model group, the sugar water preference value of the strain A20001 group mice was significantly increased, indicating that the strain A20001 of the present invention has the function of improving the anhedonia of depressed mice. The results of the tail suspension experiment showed that compared with the model group, the immobility time of the A20001 group mice was significantly shortened. At the same time, the results of the forced swimming experiment showed that compared with the model group, the immobility time of the A20001 group mice was significantly shortened. It shows that the A20001 strain can effectively alleviate the depressive-like behavior of the model mice.

[0037] The invention provides an application of a Pediococcus pentosaceus strain A20001 in preparing a medicine for preventing and / or treating cognitive dysfunction and / or improving memory. The preservation number of the Pediococcus pentosaceus strain A20001 is GDMCC No: 64674.

[0038] In the present invention, the Pediococcus pentosaceus strain A20001 is the same as the above technical solution and will not be described in detail here.

[0039] In the present invention, the cognitive dysfunction preferably includes at least one of the following behavioral disorders: memory impairment, aphasia, visuospatial disorder, executive dysfunction, calculation disorder, apraxia and agnosia.

[0040] In one embodiment of the present invention, a novel object recognition experiment was conducted to evaluate the effect of Pediococcus pentosaceus strain A20001 on the improvement of cognitive function, especially memory, of model mice. The experimental results showed that compared with the model group, the percentage of time spent exploring novel objects in the strain A20001 group of mice was significantly increased (p < 0.0001), indicating that strain A20001 can effectively improve the cognitive behavior of model mice and improve the memory of mice.

[0041] In another embodiment of the present invention, in order to further explore the molecular mechanism of the anti-anxiety and cognitive and memory functions of Pediococcus pentosaceus strain A20001, the serum BDNF of mice in the model group and the drug-treated group was detected, and the results showed that the BDNF content of mice in the strain A20001 group was significantly increased compared with the model group. Brain-derived neurotrophic factor (BDNF) is a protein that promotes the growth and survival of neurons, participates in cognitive processes such as synaptic plasticity, learning and memory, and plays an important role in the development and functional maintenance of the nervous system. BDNF regulates cognitive function. The present invention increases the level of BDNF in model mice through strain A20001, regulates the activity of neurons, and then promotes the expression and secretion of BDNF, increases cognitive processes such as anti-anxiety behavior and learning and memory, and enhances neural network function.

[0042] The following is a detailed description of the application of a Pediococcus pentosaceus strain A20001 provided by the present invention in the preparation of products for improving memory, concentration and anxiety behavior in conjunction with the examples, but they should not be construed as limiting the scope of protection of the present invention.

[0043] Example 1

[0044] Preparation method of probiotic containing Pediococcus pentosaceus strain A20001

[0045] The frozen strain A20001 was inoculated on an MRS plate at a 1% inoculation rate and streaked at 37°C, and a single colony was picked to obtain an activated strain A20001. The single colony picked above was inoculated in an MRS liquid culture medium and cultured at 37°C for 2 hours to obtain a seed solution. The seed solution was inoculated in an MRS liquid culture medium at a 5% inoculation rate and expanded at 37°C to a logarithmic phase to obtain a bacterial solution containing the Pediococcus pentosaceus strain A20001, and glycerol accounting for 40% of the volume of the bacterial solution was added to obtain a probiotic agent containing the Pediococcus pentosaceus strain A20001.

[0046] The MRS liquid culture medium contains 20 g / L glucose, 10 g / L beef extract, 10 g / L peptone, 5 g / L yeast powder, 2 g / L dipotassium hydrogen phosphate, 2 g / L diammonium citrate, 5 g / L sodium acetate, 0.58 g / L magnesium sulfate, 0.25 g / L manganese sulfate, and 0.1% Tween-80, and the pH value is 6.6 to 6.8. The difference between the MRS plate and the MRS liquid culture medium is that 1.5% to 2% agar is added additionally.

[0047] Example 2

[0048] 1. Establishment of a chronic unpredictable mild stress model in mice

[0049] 48 healthy 6-week-old SPF male C57BL / 6J mice, weighing 18-22g, were purchased from Sibeifu (Beijing) Biotechnology Co., Ltd., and the experimental animal production license is SCXK (Beijing) 2019-0010. The mice were kept in a constant temperature and humidity barrier environment (temperature 22±1℃, humidity 55±10%), with a light-dark alternation time of 12h:12h, and free food and water. After a week of adaptation, the mice were randomly grouped according to their weight, and the grouping is shown in Table 1. The following 1-3 different stimulations were used every day: restraint for 4h, crowding for 4h, tail clamping for 1min, ice water swimming for 5min, wet bedding for 24h, lighting all night for 24h, continuous darkness for 24h, no bedding for 24h, fasting for 24h, water deprivation for 24h, cage tilting for 24h, noise stimulation for 10min, foreign body stimulation for 4h; stimulation was performed at irregular intervals every day, and the model was continuously established for 4 weeks, and the mice were weighed once a week. After the modeling, the mice were subjected to behavioral tests to evaluate whether the depression model was successfully established. Specifically, the antidepressant effect of strain A20001 was evaluated by sucrose preference test (SPT), tail suspension test (TST), and forced swim test (FST). The anti-anxiety effect of strain A20001 was evaluated by open field test (OFT) and elevated plus maze test (EPM). The novel object recognition test (NOR) was used to evaluate the improvement of cognitive impairment and learning and memory function of strain A20001. At the same time, strain A21337 was used as a control.

[0050] Table 1 Treatment methods of mice in each group

[0051] Group Number deal with Normal group 12 Daily intragastric administration of normal saline, 0.2 ml / rat CUMS Group 12 CUMS model was established, and normal saline was gavaged daily, 0.2 ml / rat A20001 Group 12 <![CDATA[Modeling with CUMS, intragastric administration of A20001 at 0.2 ml / rat per day, 1×10 10 CFU / mL]]> A21337 Group 12 <![CDATA[CUMS model establishment, intragastric administration of A21337 at 0.2 ml / rat per day, 1×10 10 CFU / mL]]>

[0052] 2. Mouse Weight Statistics

[0053] The mice were modeled according to the above modeling method, and the weight of the mice was counted once a week.

[0054] The results are as follows Figure 1 The experimental results showed that compared with the normal group, the weight of mice in the CUMS model group decreased significantly (p < 0.0001). Compared with the model group, the weight loss trend of mice after oral administration of strain A20001 was significantly slowed down (p < 0.01), and was better than the control A21337 strain group, indicating that oral administration of strain A20001 can increase the weight of depressed mice caused by CUMS.

[0055] 3. Sucrose preference experiment

[0056] Before the formal test, all mice were first trained with sugar water for 3 days, that is, they were given a bottle of drinking water containing 1% sucrose and a bottle of regular drinking water, and the positions of the two bottles were swapped from time to time. After the training period, all mice were fasted and deprived of water, and a formal sugar water test was conducted. Two bottles of drinking water containing 1% sucrose and regular drinking water of the same volume were prepared. After 24 hours, the sugar water consumption (g) of each mouse was counted, and the sugar water preference value was calculated according to Formula I:

[0057] Sugar water preference value = (mass of sugar water consumed / total mass of liquid consumed) × 100% Formula I.

[0058] The results are as follows Figure 2 As shown, compared with the normal group, the sugar preference value of mice in the CUMS model group was significantly reduced (p < 0.0001), indicating that the model was successfully constructed. In addition, after intragastric administration of strain A20001, the sugar preference value of mice was significantly increased (p < 0.0001), indicating that the strain A20001 of the present invention has the function of improving the anhedonia of depressed mice.

[0059] 4. Tail suspension test

[0060] The mouse tail was fixed with tape at the 1 / 3 of the back, and hung on a stand. The timing and video recording were started immediately, and the video recording was stopped after 6 minutes of observation. The immobility time of the mouse in the last 4 minutes was counted using Smart 3.0 analysis software. Immobility refers to the state in which the animal gives up active struggle and is completely immobile.

[0061] The results are as follows Figure 3 As shown in the figure, compared with the normal group, the immobility time of mice in the CUMS model group was significantly increased (p < 0.001), indicating that the model was successfully constructed. Compared with the model group, the immobility time of mice in the strain A20001 group was significantly shortened (p < 0.001). This shows that strain A20001 can effectively alleviate the depressive-like behavior of mice caused by CUMS.

[0062] 5. Forced swimming test

[0063] The mice were placed in a transparent experimental bucket (filled with sterile water about 30 cm high) for a 6-minute swimming test. The Smart 3.0 analysis software was used for real-time video recording and analysis, and the immobility time of the mice in the last 4 minutes (immobility, the judgment standard was immobility of the limbs or only slight movement of the hind limbs) was calculated.

[0064] The results are as follows Figure 4 As shown, compared with the normal group, the immobility time of mice in the CUMS model group was significantly increased (p < 0.0001), indicating that the model was successfully constructed. Compared with the model group, the immobility time of mice in the strain A20001 group was significantly shortened (p < 0.0001), indicating that it can effectively alleviate the depressive-like behavior of mice caused by CUMS.

[0065] 6. Open field experiment

[0066] The mouse was placed in a corner of the open field, and the timing and recording were started immediately. The recording was stopped after 10 minutes of observation. When replacing the animal, in order to avoid affecting the next test result, the surface needs to be wiped with 75% alcohol to remove the residual information of the previous animal (such as urine, feces, odor, etc.). Smart 3.0 analysis software was used for real-time recording and analysis, and the time (s) that the mouse stayed in the center area of ​​the open field was counted.

[0067] The results are as follows Figure 5 As shown in the figure, compared with the normal group, the time for mice in the CUMS model group to enter the center of the open field was significantly shortened (p < 0.01). Compared with the model group, the time for mice in the strain A20001 group to enter the center of the open field was significantly increased (p < 0.001), and was better than the control strain A21337. This shows that strain A20001 can effectively alleviate the anxiety behavior of mice accompanied by depression-like symptoms caused by CUMS.

[0068] 7. Elevated plus maze test

[0069] Place the mouse in the central area of ​​the elevated plus maze (the mouse faces the open arm), start timing and recording immediately, and stop recording after 5 minutes of observation. When replacing the animal, in order to avoid affecting the next test result, it is necessary to wipe the surface with 75% alcohol to remove the residual information of the previous animal (such as urine, feces, odor, etc.). Use Smart 3.0 analysis software for real-time recording and analysis, and count the number of times the mouse enters the open arm, and the percentage of time in the open arm and the closed arm (%).

[0070] The results are as follows Figure 6As shown, compared with the normal group, the number of times mice in the CUMS model group entered the open arm and the percentage of time in the open arm were significantly reduced (p < 0.05). Compared with the model group, the number of times mice in the strain A20001 group entered the open arm and the percentage of time in the open arm were significantly increased (p < 0.0001), and the time in the closed arm was significantly shortened (p < 0.0001), and both were significantly better than the control strain A21337, indicating that strain A20001 can effectively alleviate the anxiety behavior of mice accompanied by depression-like symptoms caused by CUMS.

[0071] 8. New object recognition experiment

[0072] The novel object recognition test (NOR) is mainly used to evaluate the cognitive function, especially memory, of rodents (such as mice and rats). The core of this test is the difference in the animals' exploration behavior towards new and old objects, which is used to evaluate their memory and cognitive abilities.

[0073] Before the formal test, the mice were trained for 2 days. On the first day, the mice were allowed to freely explore and adapt in the open field for 10 minutes. On the second day, two identical objects A were placed in diagonal positions, and the mice were allowed to explore freely for 5 minutes. One hour later, one of the objects A was replaced with another new object B, and the mice were allowed to explore freely for 5 minutes. It should be noted that each time the animal is replaced, in order to avoid affecting the results of the next test, it is necessary to wipe the open field and the surface of the object with 75% alcohol to remove the residual information of the previous animal (such as urine, feces, odor, etc.). In the formal experiment, object A was retained in the open field, and object B was replaced with object C. Each mouse was placed in the same direction and allowed to explore freely for 5 minutes. Smart 3.0 analysis software was used for real-time video recording and analysis, and the percentage (%) of time the mouse explored the new object C was counted.

[0074] The results are as follows Figure 7 As shown in the figure, compared with the normal group, the percentage of time spent exploring new objects in the CUMS model group was significantly shortened (p < 0.01). Compared with the model group, the percentage of time spent exploring new objects in the strain A20001 group was significantly increased (p < 0.0001), and was superior to the control strain A21337 group, indicating that strain A20001 can effectively improve the cognitive behavior of CUMS depressed mice and improve the memory of mice.

[0075] Example 3

[0076] Detection of mouse serum BDNF levels

[0077] The mice were divided into groups according to the method of Example 2 and treated in the same way. An appropriate amount of mouse serum was taken from each group of mice and tested according to the instructions of the mouse BDNF ELISA kit (provided by Lianke Bio). The results of mouse serum BDNF test are shown in Figure 2. Figure 8 As shown, compared with the normal group, the BDNF level in the serum of mice in the CUMS model group was significantly decreased (p < 0.0001). Compared with the model group, the BDNF content of mice in the strain A20001 group was significantly increased (p < 0.001), and was significantly better than that of the control strain A21337 group.

[0078] In summary, the Pediococcus pentosaceus strain A20001 may alleviate the depressive-like behavior of mice by increasing the level of BDNF, and the effect is significantly better than the control strain A21337, effectively improving the cognitive behavior of CUMS depressed mice and improving the memory of mice.

[0079] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. Use of a Pediococcus pentosaceus strain A20001 in the preparation of a drug for preventing and / or treating anxiety or depression, wherein the Pediococcus pentosaceus strain A20001 has a deposit number of GDMCC No: 64674.

2. The use according to claim 1, characterized in that: The anxiety disorder includes at least one of the following anxiety behaviors and complications: acute anxiety, chronic anxiety, neurological dysfunction, cardiovascular and cerebrovascular diseases, and digestive system diseases.

3. Use of Pediococcus pentosaceus strain A20001 in the preparation of a drug for preventing and / or treating cognitive dysfunction and / or improving memory, wherein the deposit number of the Pediococcus pentosaceus strain A20001 is GDMCC No: 64674.

4. The use according to claim 3, characterized in that: The cognitive dysfunction includes at least one of the following behavioral disorders: memory impairment, aphasia, visuospatial disorder, executive dysfunction, calculation disorder, apraxia and agnosia.

5. The use according to any one of claims 1 to 4, characterized in that: The Pediococcus pentosaceus strain A20001 includes at least one of the following cultures: bacterial bodies of the Pediococcus pentosaceus strain A20001, metabolites of the Pediococcus pentosaceus strain A20001, and fermentation broth of the Pediococcus pentosaceus strain A20001.

6. The use according to claim 5, characterized in that: The Pediococcus pentosaceus strain A20001 also includes being present in the form of a probiotic.

7. The use according to claim 6, characterized in that: The probiotics include at least one of the following: tablets, drops, capsules and powders.

8. The use according to claim 6, characterized in that: The live bacterial concentration of Pediococcus pentosaceus strain A20001 in the probiotic is not less than 1×10 10 CFU / mL or 1×10 10 CFU / g.

9. The use according to claim 8, characterized in that: The live bacterial concentration of the Pediococcus pentosaceus strain A20001 is 5×10 10 CFU / mL~100×10 10 CFU / mL or 10×10 9 CFU / g~100×10 10 CFU / g.

10. The use according to any one of claims 1 to 4, characterized in that: The drug comprises at least one of the following oral dosage forms: tablets, powders, capsules and oral liquids.

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