Application of lactobacillus plantarum strain A21337 in preparation of product for resisting anxiety and improving memory and concentration

Abstract

The invention provides an application of a lactobacillus plantarum strain A21337 in preparation of a product for resisting anxiety and improving memory and concentration, and belongs to the technical field of biological medicines. Animal experiments show that the plant lactobacillus strain A21337 has the effects of resisting anxiety, resisting depression, improving cognitive impairment function and improving memory concentration, and a new means is provided for preventing and / or treating nerve diseases related to anxiety disorder, depression and cognitive impairment.

Description

Technical Field

[0001] The invention belongs to the technical field of biological medicines, and particularly relates to an application of a Lactobacillus plantarum strain A21337 in preparing products for resisting anxiety and improving memory and concentration. Background Art

[0002] Lactobacillus plantarum is a well-known intestinal probiotic. It is reported that the intestinal probiotic effects of Lactobacillus plantarum include enhancing immunity, improving gastrointestinal function, exerting antioxidant function, lowering blood sugar and blood pressure, enhancing intestinal mucosal barrier function to maintain intestinal flora balance, degrading uric acid, inhibiting harmful bacteria and improving nutrient absorption, and alleviating alcoholic liver damage. It can be seen that Lactobacillus plantarum plays an important role in promoting human health and preventing diseases. However, it should be noted that different strains may have different characteristics and effects.

[0003] Currently, there are no reports on the use of Lactobacillus plantarum in improving anxiety and enhancing cognitive functions such as memory. Summary of the invention

[0004] In view of this, the object of the present invention is to provide a use of Lactobacillus plantarum strain A21337 in the preparation of products for resisting anxiety and improving memory and concentration.

[0005] The invention provides an application of a Lactobacillus plantarum strain A21337 in preparing a medicine for preventing and / or treating anxiety or depression. The deposit number of the Lactobacillus plantarum strain A21337 is GDMCC No: 64677.

[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 Lactobacillus plantarum strain A21337 in preparing a medicine for preventing and / or treating cognitive dysfunction and / or improving memory. The deposit number of the Lactobacillus plantarum strain A21337 is GDMCC No: 64677.

[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 Lactobacillus plantarum strain A21337 comprises at least one of the following cultures: cells of the Lactobacillus plantarum strain A21337, metabolites of the Lactobacillus plantarum strain A21337 and fermentation broth of the Lactobacillus plantarum strain A21337.

[0010] Preferably, the Lactobacillus plantarum strain A21337 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 bacteria concentration of Lactobacillus plantarum strain A21337 in the probiotic is not less than 1×10 10 CFU / mL or 1×10 10 CFU / g.

[0013] Preferably, the viable bacterial concentration of the Lactobacillus plantarum strain A21337 is 5×10 10 CFU / mL~100×10 10 CFU / mL or 5×10 10 CFU / g~100×10 10 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 Lactobacillus plantarum strain A21337 in preparing a drug for preventing and / or treating anxiety or depression, and the deposit number of the Lactobacillus plantarum strain A21337 is GDMCC No: 64677. The present invention establishes a chronic unpredictable mild stress model in mice, and uses a sucrose preference experiment, a tail suspension experiment, and a forced swimming experiment to evaluate the antidepressant efficacy of Lactobacillus plantarum strain A21337. 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 A21337 group is significantly increased (p < 0.0001), indicating that the strain A21337 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 A21337 group is significantly shortened (p < 0.01). 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 A21337 group is significantly shortened (p < 0.0001). This shows that the strain A21337 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 anti-anxiety behavior efficacy of plant lactobacillus strain A21337. The results of the open field test showed that compared with the model group, the time for mice in the A21337 group to enter 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 mice in the A21337 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 plant lactobacillus strain A21337 can effectively alleviate the anxiety and depression behaviors of model mice, providing a new means for the treatment of depression and anxiety.

[0016] The present invention provides an application of Lactobacillus plantarum strain A21337 in the preparation of a drug for preventing and / or treating cognitive dysfunction and / or improving memory, and the deposit number of the Lactobacillus plantarum strain A21337 is GDMCC No: 64677. The present invention uses a novel object recognition experiment to evaluate the efficacy of Lactobacillus plantarum strain A21337 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 A21337 group mice explored new objects was significantly increased, indicating that the A21337 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 This is the result of the effect of strain A21337 on the body weight of CUMS mice;

[0018] Figure 2 The test results show that strain A21337 significantly improves sucrose preference in CUMS mice;

[0019] Figure 3 The test results show that strain A21337 significantly shortens the tail suspension immobility time of CUMS mice;

[0020] Figure 4 The strain A21337 significantly shortened the immobility time of forced swimming in CUMS mice.

[0021] Figure 5 The test results show that strain A21337 significantly increases the exploratory ability of CUMS mice in an open field;

[0022] Figure 6 The results of the strain A21337 in the elevated plus maze test;

[0023] Figure 7 The strain A21337 significantly increased the time CUMS mice spent exploring new objects;

[0024] Figure 8 The test results show that strain A21337 significantly increases the serum 5-HT level in mice;

[0025] Fig. 9 The strain A21337 significantly increased the level of BDNF in mouse serum;

[0026] Fig.10 The strain A21337 significantly increased the BDNF level in the mouse hippocampus.

[0027] Biomaterial Deposit Information

[0028] 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

[0029] The invention provides an application of a Lactobacillus plantarum strain A21337 in preparing a medicine for preventing and / or treating anxiety or depression. The deposit number of the Lactobacillus plantarum strain A21337 is GDMCC No: 64677.

[0030] In the present invention, the Lactobacillus plantarum strain A21337 is isolated from a sample of intestinal microbial flora of centenarians. Morphological identification and 16S rDNA sequencing show that the strain A21337 belongs to Lactobacillus plantarum.

[0031] In the present invention, the plant lactobacillus strain A21337 preferably includes at least one of the following cultures: the bacterial body of plant lactobacillus strain A21337, the metabolites of plant lactobacillus strain A21337 and the fermentation liquid of plant lactobacillus strain A21337. The preparation method of the fermentation liquid of the plant lactobacillus strain A21337 preferably inoculates the activated plant lactobacillus strain A21337 in MRS liquid culture to obtain seed liquid; inoculates the seed liquid in MRS liquid culture for expansion culture, and inoculates the plant lactobacillus strain A21337 bacterial liquid obtained by the expansion culture in MRS liquid culture again for fermentation culture to obtain fermentation liquid. The activation method of the strain A21337 is preferably that the strain A21337 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 strain A21337 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 flat plate, and the MRS liquid culture medium and the MRS flat plate formula known in the art can be used. For example, the MRS liquid culture medium formula is 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, 0.1% Tween-80, and the pH value is 6.6-6.8. The difference between the MRS flat plate and the MRS liquid culture medium is that 1.5%-2% agar is added additionally. The method for preparing the cell or metabolite of the plant lactobacillus strain A21337 is to separate the fermentation liquid prepared above into solid and liquid, collect the cell of the plant lactobacillus strain A21337 as the precipitate, and collect the metabolite of the plant lactobacillus strain A21337 as the liquid.

[0032] In the present invention, the plant lactobacillus strain A21337 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 plant lactobacillus strain A21337 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 packs the plant lactobacillus strain A21337 alone or mixed with auxiliary materials as a capsule core in the capsule shell. The drop is prepared by mixing Lactobacillus plantarum strain A21337 with edible oil to form a drop that is isolated from air and moisture. When taken, the drop opening is opened to squeeze the encapsulated oily substance into the mouth. The live bacteria concentration of Lactobacillus plantarum strain A21337 in the probiotic is preferably not less than 2×10 10 CFU / mL or 2×10 10 CFU / g, can be 5×10 10 CFU / mL~100×10 10 CFU / mL or 5×10 10 CFU / g~100×10 10 CFU / g, or 10×10 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 10×10 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.

[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 carried out to evaluate the efficacy of plant lactobacillus strain A21337 or its preparation in anti-anxiety behavior. The results of the open field test showed that compared with the model group, the time for mice in the strain A21337 group to enter 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 strain A21337 group of mice entered the open arm and the percentage of time in the open arm increased significantly, and the time in the closed arm was significantly shortened. This shows that the plant lactobacillus strain A21337 has the effect of improving the anxiety behavior of model mice.

[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 plant lactobacillus strain A21337 was evaluated using sucrose preference experiment, tail suspension experiment and forced swimming experiment. The results of the sucrose preference experiment showed that the sugar water preference value of strain A21337 group mice was significantly increased compared with the model group, indicating that strain A21337 of the present invention has the function of improving the anhedonia of depressed mice. The results of the tail suspension experiment showed that the immobility time of strain A21337 group mice was significantly shortened compared with the model group. At the same time, the results of the forced swimming experiment showed that the immobility time of strain A21337 group mice was significantly shortened compared with the model group. This shows that strain A21337 can effectively alleviate the depressive-like behavior of model mice.

[0037] The invention provides an application of a Lactobacillus plantarum strain A21337 in preparing a medicine for preventing and / or treating cognitive dysfunction and / or improving memory. The deposit number of the Lactobacillus plantarum strain A21337 is GDMCC No: 64677.

[0038] In the present invention, the source of the Lactobacillus plantarum strain A21337 and the type of probiotics are the same as those in the above technical solution, which 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 Lactobacillus plantarum strain A21337 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 new objects in the strain A21337 group of mice increased significantly, indicating that strain A21337 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 plant lactobacillus strain A21337 in anti-anxiety and improving cognitive ability and memory function, 5-hydroxytryptamine (5-HT) and brain-derived neurotrophic factor (BDNF) in the serum of mice in the model group and the drug-treated group were detected respectively. The results showed that compared with the model group, the 5-HT content of mice in the strain A21337 group increased significantly. At the same time, compared with the model group, the BDNF content of mice in the strain A21337 group increased significantly. 5-hydroxytryptamine (5-HT) is an important neurotransmitter in the body, widely distributed in the central nervous system, and participates in regulating emotions, sleep, appetite and behavior, etc. Brain-derived neurotrophic factor (BDNF) is a protein that promotes neuron growth and survival, participates in cognitive processes such as synaptic plasticity, learning and memory, and plays an important role in the development and function maintenance of the nervous system. 5-HT and BDNF synergistically regulate cognitive function. The present invention improves the levels of 5-HT and BDNF in model mice through strain A21337, so that 5-HT regulates the activity of neurons by binding to its receptor, thereby promoting the expression and secretion of BDNF, increasing cognitive processes such as anti-anxiety behavior and learning and memory, and enhancing neural network function.

[0042] The following is a detailed description of the application of a Lactobacillus plantarum strain A21337 provided by the present invention in the preparation of products for anti-anxiety and improving memory and concentration, 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] A preparation method of probiotic agent of Lactobacillus plantarum strain A21337

[0045] The frozen strain A21337 is inoculated on an MRS plate at an inoculum size of 1% and streaked at 37°C, and a single colony is picked to obtain an activated strain A21337. The single colony picked above is inoculated in an MRS liquid culture medium at 37°C and cultured for 2h to obtain a seed solution. The seed solution is inoculated in an MRS liquid culture medium at an inoculum size of 5% and expanded and cultured at 37°C to a logarithmic phase to obtain a bacterial solution containing plant lactobacillus strain A21337, an aqueous solution containing 12% skim milk is added as a protective agent at a volume ratio of 1:1, and vacuum freeze-dried to obtain a probiotic containing plant lactobacillus strain A21337.

[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 the Chronic Unpredictable Mild Stress (CUMS) Model in Mice

[0049] Thirty-six healthy 6-week-old SPF male C57BL / 6J mice, weighing 18-22 g, were purchased from Sibeifu (Beijing) Biotechnology Co., Ltd., and the experimental animal production license is SCXK (Beijing) 2019-0010. The mice were housed 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. Every day, 1 to 3 different stimulations were used: 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, and foreign body stimulation for 4h; the stimulation was irregular 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 modeling was successful. The antidepressant behavioral efficacy of Lactobacillus plantarum strain A21337 was evaluated using the sucrose preference test (SPT), tail suspension test (TST), and forced swim test (FST). The anti-anxiety behavioral efficacy of Lactobacillus plantarum strain A21337 was evaluated using the open field test (OFT) and elevated plus maze test (EPM). The novel object recognition test (NOR) was used to evaluate the efficacy of Lactobacillus plantarum strain A21337 in improving cognitive dysfunction and memory and concentration.

[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 A21337 Group 12 <![CDATA[Modeling by CUMS, 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. The results are as follows: Figure 1 shown.

[0054] 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, there was no significant change in the weight loss trend of mice after oral administration of A21337, indicating that oral administration of A21337 had no effect on the weight of depressed mice.

[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 A21337, the sugar preference value of mice was significantly increased (p < 0.0001), indicating that the strain A21337 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 established. Compared with the model group, the immobility time of mice in the A21337 group was significantly shortened (p < 0.01), indicating that the A21337 strain 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 4As shown in the figure, 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 established. Compared with the model group, the immobility time of mice in the A21337 group was significantly shortened (p < 0.0001). This shows that the A21337 strain 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 that the mice in the CUMS model group entered the center of the open field was significantly shortened (p < 0.01). Compared with the model group, the time that the mice in the A21337 group entered the center of the open field was significantly increased (p < 0.05). This shows that the A21337 strain 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 6 As shown in the figure, compared with the normal group, the number of times the mice in the CUMS model group entered the open arm and the percentage of time in the open arm were significantly shortened (p < 0.05). Compared with the model group, the number of times the mice in the A21337 group entered the open arm was significantly increased (p < 0.01) and the percentage of time in the open arm was significantly increased (p < 0.0001), and the time in the closed arm was significantly shortened (p < 0.0001), indicating that the A21337 strain 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 A21337 group was significantly increased (p < 0.001), indicating that strain A21337 can effectively improve the cognitive behavior of CUMS depressed mice and improve the memory of mice.

[0075] Example 3

[0076] Detection of serum 5-HT and BDNF levels in mice

[0077] The three groups of mice were treated according to the method of Example 2, and appropriate amounts of serum were taken from each group of mice, and the detection was performed according to the instructions of the mouse 5-HT ELISA kit (provided by Jianglai Bio) and the mouse BDNF ELISA kit (provided by Lianke Bio). In addition, hippocampal tissue was isolated from each group of mice and detected according to the instructions of the total RNA extraction kit (provided by Promeg Bio), the single-stranded synthesis kit (provided by Quanshijin), and the 2X SYBR Green qPCR MiX (provided by Si Kejie). The primers used for qPCR are shown in Table 2.

[0078] Table 2 Primer sequences for qPCR detection

[0079]

[0080] The results of mouse serum 5-HT test are as follows Figure 8As shown, compared with the normal group, the 5-HT level in the serum of mice in the CUMS model group was significantly decreased (p < 0.001). Compared with the model group, the 5-HT content in mice in the strain A21337 group was significantly increased (p < 0.001).

[0081] The results of mouse serum BDNF detection are as follows Fig. 9 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 in mice in the strain A21337 group was significantly increased (p < 0.01).

[0082] The results of BDNF detection in mouse hippocampus are as follows Fig.10 As shown in the figure, compared with the normal group, the BDNF level in the hippocampus of the mice in the CUMS model group was significantly decreased (p<0.001). Compared with the model group, the BDNF content in the mice in the strain A21337 group was significantly increased (p<0.01).

[0083] In summary, strain A21337 may alleviate the depressive-like behavior of mice by increasing the levels of 5-HT and BDNF, and can effectively improve the cognitive behavior of CUMS depressed mice and improve the memory of mice.

[0084] 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 plant lactobacillus strain A21337 in the preparation of a medicament for preventing and / or treating anxiety or depression, wherein the deposit number of the plant lactobacillus strain A21337 is GDMCC No: 64677.

2. The application 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 a plant lactobacillus (Pediococcus pentosaceus) strain A21337 in the preparation of a drug for preventing and / or treating cognitive dysfunction and / or improving memory, wherein the deposit number of the plant lactobacillus (Pediococcus pentosaceus) strain A21337 is GDMCC No: 64677.

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 Lactobacillus plantarum strain A21337 comprises at least one of the following cultures: bacterial bodies of the Lactobacillus plantarum strain A21337, metabolites of the Lactobacillus plantarum strain A21337 and fermentation liquid of the Lactobacillus plantarum strain A21337.

6. The use according to claim 5, characterized in that: The Lactobacillus plantarum strain A21337 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 bacteria concentration of Lactobacillus plantarum strain A21337 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 bacteria concentration of the plant lactobacillus strain A21337 is 5×10 10 CFU / mL~100×10 10 CFU / mL or 5×10 10 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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