Lactobacillus johnsonii A21065 capable of resisting oxidation, prolonging life and delaying Parkinson's symptom as well as product and application of lactobacillus johnsonii A21065

Abstract

The invention discloses lactobacillus johnsonii A21065 capable of resisting oxidation, prolonging life and delaying Parkinson's disease as well as a product and application of the lactobacillus johnsonii A21065, and belongs to the technical field of functional strains. The preservation number of the lactobacillus johnsonii A21065 is GDMCC (China General Microbiological Culture Collection Center) No: 65767. In a caenorhabditis elegans model, the lactobacillus johnsonii A21065 shows that the lactobacillus johnsonii A21065 has better effects of improving the oxidation resistance of nematodes, resisting environmental stress and prolonging the life, also shows a good effect in in-vitro experiments of resisting artificial gastrointestinal fluid, resisting cholate and the like, meanwhile, the lactobacillus johnsonii A21065 also has an effect of improving the cognition of a Parkinson nematode model, and the lactobacillus johnsonii A21065 has a good application prospect. The invention has important significance for preventing and / or treating Parkinson's disease.

Description

Technical Field

[0001] The present invention relates to the technical field of functional strains, and particularly relates to a Lactobacillus johnsonii A21065 with antioxidant, life extension, and Parkinson's symptom delaying effects, as well as its products and applications. Background Art

[0002] The gut microbiota is the most complex and diverse microbial community in the animal digestive tract. They can not only maintain the structure and physiological functions of the host digestive tract, but also promote the metabolism of nutrients, adjust the immune system, and control the growth of pathogenic bacteria. Among them, Lactobacillus johnsonii is one of the important probiotic species in the human intestine. It is a Gram-positive bacterium, belonging to facultative homofermentative lactic acid bacteria, facultative anaerobic, growing poorly under aerobic conditions but growing well under low oxygen pressure. Lactobacillus johnsonii was initially isolated from human blood, is the dominant species of Lactobacillus in the infant intestine, is also commonly present in the adult intestine and excreted with feces, and as an important component of the normal intestinal microbial system, it accompanies the host throughout life and is of great significance for maintaining the balance of the intestinal microecology. The main biological activities of Lactobacillus johnsonii include enhancing the body's immunity, alleviating intestinal inflammation, inhibiting intestinal pathogenic microorganisms, and regulating the balance of the intestinal flora, etc. Research shows that Lactobacillus johnsonii is a probiotic that can improve memory impairment through the brain-gut axis. At the same time, it can also restore the morphology and function of liver mitochondria, reduce liver lipids, and improve whole-body glucose metabolism.

[0003] Due to the high safety of this strain, the National Health Commission of China has included Lactobacillus johnsonii in the "List of Strains Allowed to be Used in Foods", allowing its application in foods, which lays a foundation for the development and application of Lactobacillus johnsonii.

[0004] A strain of Lactobacillus johnsonii isolated from the feces of centenarians in the present invention has a good effect of extending lifespan, anti-aging, and antioxidant on nematodes, and can provide a reference for the development of Lactobacillus johnsonii into health foods or drugs for anti-aging, life extension, etc., aiming to expand the application of Lactobacillus johnsonii. Summary of the Invention

[0005] The present invention aims to at least solve the above technical problems in the prior art. For this reason, the object of the present invention is to provide a Lactobacillus johnsonii A21065 with antioxidant, life extension, and Parkinson's symptom delaying effects, as well as its products and applications.

[0006] In order to achieve the above object, the technical solutions adopted by the present invention are:

[0007] In the first aspect of the present invention, a Lactobacillus johnsonii is provided. The Lactobacillus johnsonii is Lactobacillus johnsonii A21065, taxonomically named Lactobacillus johnsonii, and was deposited at the Guangdong Microbial Culture Collection Center (GDMCC) on January 13, 2025 (the deposit address is: 5th Floor, Building 59, No. 100 Compound, Xianlie Middle Road, Guangzhou), with the deposit number GDMCC No: 65767.

[0008] In some embodiments of the present invention, the colony morphology of Lactobacillus johnsonii A21065 is circular, convex, entire, and colorless colonies.

[0009] In some embodiments of the present invention, the 16S sequence of Lactobacillus johnsonii A21065 is as shown in SEQ ID NO: 3.

[0010] In some embodiments of the present invention, the screening and isolation method of Lactobacillus johnsonii A21065 is as follows: Take fecal samples from centenarians in Guangxi Zhuang Autonomous Region and inoculate them into MRS solid medium, culture in an incubator for 48 hours, pick colonies with circular, convex, entire, and colorless colony morphology from the plate and culture them in at least a small amount of MRS liquid medium. After PCR amplification and identification, Lactobacillus johnsonii A21065 is obtained.

[0011] In some embodiments of the present invention, the primers used for PCR amplification and identification are as shown in SEQ ID NO: 1 and SEQ ID NO: 2.

[0012] In some embodiments of the present invention, the amplification system for PCR amplification and identification is:

[0013] Component Content 10 μM forward primer 16S-F 1 μL 10 μM reverse primer 16S-R 1 μL PCR amplification solution 12.5 μL DNA template 1 μL <![CDATA[ddH2O]]> Make up to 25 μL.

[0014] In some embodiments of the present invention, the amplification program for PCR amplification and identification is 95°C for 3 min; 95°C for 40 s, 56°C for 30 s, 72°C for 1 min, cycle 30 times; 72°C for 8 min.

[0015] In the second aspect of the present invention, a culture of Lactobacillus johnsonii A21065 is provided.

[0016] In some embodiments of the present invention, the culture includes the supernatant obtained after culturing Lactobacillus johnsonii A21065.

[0017] In the third aspect of the present invention, a bacterial agent is provided.

[0018] In some embodiments of the present invention, the bacterial agent comprises Lactobacillus johnsonii A21065 described in the above aspects and / or a culture of Lactobacillus johnsonii A21065.

[0019] In some embodiments of the present invention, the bacterial agent is a solid bacterial agent or a liquid bacterial agent.

[0020] The fourth aspect of the present invention provides the use of any one or more of Lactobacillus johnsonii A21065 described in the above aspects and / or a culture of Lactobacillus johnsonii A21065 and / or the bacterial agent in the preparation of a medicament for preventing or treating Parkinson's disease.

[0021] In some embodiments of the present invention, the dosage form of the medicament includes powder, granule, capsule, tablet, pill or oral liquid.

[0022] The fifth aspect of the present invention provides the use of any one or more of Lactobacillus johnsonii A21065 described in the above aspects and / or a culture of Lactobacillus johnsonii A21065 and / or the bacterial agent in at least one of the following (I) to (III);

[0023] (I) Preparing a product for prolonging life;

[0024] (II) Preparing an anti-aging product;

[0025] (III) Preparing an antioxidant product.

[0026] The sixth aspect of the present invention provides a product containing any one or more of Lactobacillus johnsonii A21065, a culture of Lactobacillus johnsonii A21065 and the bacterial agent described in the above aspects.

[0027] In some embodiments of the present invention, the product includes food, food additives, feed, feed additives, medicines and cosmetics.

[0028] In some embodiments of the present invention, the product further contains excipients.

[0029] In some embodiments of the present invention, the excipients include food additives, pharmaceutically acceptable adjuvants or cosmetic excipients.

[0030] In some embodiments of the present invention, the form of Lactobacillus johnsonii in the product includes freeze-dried powder, bacterial liquid and granular inoculum.

[0031] The beneficial effects of the present invention are:

[0032] The present invention provides Lactobacillus johnsonii A21065 and its products and applications with antioxidant, life extension, and Parkinson's symptom delaying effects. Lactobacillus johnsonii A21065 can improve the locomotor ability of Parkinson's nematodes and reduce the aggregation of α-synuclein, and can be used to delay the onset of Parkinson's disease symptoms. At the same time, Lactobacillus johnsonii A21065 also shows good effects in experiments such as resistance to artificial gastrointestinal fluid, bile salt resistance, anti-aging, life extension, and antioxidant. Therefore, this strain can be used as a probiotic agent for preparing products that regulate the intestinal flora, which is of great significance for the prevention and / or treatment of Parkinson's disease. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1 It is the phylogenetic tree of Lactobacillus johnsonii A21065.

[0034] Figure 2 It is the antioxidant effect of Lactobacillus johnsonii A21065.

[0035] Figure 3 It is the heat stress effect of Lactobacillus johnsonii A21065.

[0036] Figure 4 It is the effect of Lactobacillus johnsonii A21065 on the lifespan of Caenorhabditis elegans.

[0037] Figure 5 It is the effect of Lactobacillus johnsonii A21065 on the body bend of nematodes.

[0038] Figure 6 It is the colony of A21065 grown on Columbia blood agar.

[0039] Figure 7 It is the effect of Lactobacillus johnsonii A21065 on the body bend experiment of NL5901.

[0040] Figure 8 It is the effect of Lactobacillus johnsonii A21065 on the head swing experiment of NL5901.

[0041] Figure 9 It is the effect of Lactobacillus johnsonii A21065 on the aggregation of α-syn in NL5901. DETAILED DESCRIPTION OF THE INVENTION

[0042] The content of the present invention will be further described in detail through specific examples below. The raw materials, reagents, or devices used in the examples and comparative examples can be obtained from conventional commercial channels or can be obtained by existing technical methods without special instructions. Unless otherwise specified, the test or testing methods are conventional methods in the art.

[0043] Example 1 Isolation and purification of Lactobacillus johnsonii A21065

[0044] This embodiment provides a method for the isolation and purification of Lactobacillus johnsonii A21065, and the specific experimental steps are as follows.

[0045] (1) Take 100 μL of fecal samples from centenarians in Guangxi Zhuang Autonomous Region, dilute them step by step using phosphate buffer solution (PBS), and spread them on MRS medium plates, then culture them in an incubator at 37 °C for 48 h;

[0046] (2) After the culture is completed, pick colonies with round, convex, entire and colorless colony morphologies on the plates for purification culture. For the purified cultured strains, use an inoculation loop to inoculate them one by one into sterile MRS broth medium on a sterile operating table, place them in a shaker, and culture them at 37 °C for 48 h;

[0047] (3) Add PCR amplification solution (2×Taq PCR StarMix with Loading Dye, Beijing Kangrun Chengye Biotechnology Co., Ltd.) to the cultured PCR tubes, and add 16S primers for PCR amplification (the amplification system is shown in Table 1 below);

[0048] Among them, the 16S primers are:

[0049] Forward primer 16S-F: 5’-AGAGTTTGATCCTGGCTCAG-3’ (SEQ ID NO:1);

[0050] Reverse primer 16S-R: 5’-TACGGCTACCTTGTTACGACTT-3’ (SEQ ID NO:2);

[0051] Table 1 PCR amplification system for Lactobacillus johnsonii

[0052] Component Content 10 μM forward primer 16S-F 1 μL 10 μM reverse primer 16S-R 1 μL PCR amplification solution 12.5 μL DNA template 1 μL <![CDATA[ddH2O]]> Make up to 25 μL

[0053] The amplification program is: 95 °C for 3 min; 95 °C for 40 s, 56 °C for 30 s, 72 °C for 1 min, cycle 30 times; 72 °C for 8 min;

[0054] Send the amplification product to Sangon Biotech (Shanghai) Co., Ltd. for sequencing and identification.

[0055] After sequencing and identification, it is confirmed to be Lactobacillus johnsonii, and its 16S sequence is:

[0056]

[0057] (4) The confirmed culture medium was purified by streak plate method twice. Single colonies after purification were picked for repeated culture to obtain bacterial liquid. The bacterial liquid was mixed with 40% glycerol preservation solution in a volume ratio of 1:1 in a strain cryopreservation tube and placed in a -80 °C refrigerator for storage;

[0058] (5) The 16S sequences obtained in (3) were used to construct a phylogenetic tree by the neighbor-joining method on the MEGA-X 10.2.2 software (https: / / www.megasoftware.net / ).

[0059] Sequence alignment was performed on NCBI and it was found that the 16S rDNA sequence of the strain had the highest homologous similarity with that of Lactobacillus johnsonii of the genus Lactobacillus, and the alignment similarity was 99.86%. The strain was identified as Lactobacillus johnsonii.

[0060] The construction result of the phylogenetic tree is as Figure 1 shown, further confirming that the strain isolated and purified in this example is Lactobacillus johnsonii, and it was named Lactobacillus johnsonii A21065.

[0061] The obtained Lactobacillus johnsonii A21065 was sent to the Guangdong Provincial Culture Collection Center of Microorganisms (GDMCC) for preservation. Its taxonomic name is: Lactobacillus johnsonii, the preservation date is: January 13, 2025, the preservation address is: 5th Floor, Building 59, No. 100 Compound, Xianlie Middle Road, Guangzhou, and the preservation number is: GDMCC No: 65767.

[0062] Example 2 Antioxidant effect of Lactobacillus johnsonii A21065

[0063] This example provides a test for the antioxidant ability of Lactobacillus johnsonii A21065. The specific experimental steps are as follows.

[0064] (1) Lactobacillus johnsonii A21065 was cultured in a conventional MRS liquid medium and placed in a shaker at 37 °C for 48 hours;

[0065] (2) Escherichia coli OP50 was cultured in LB liquid medium under the following conditions: temperature 37 °C, time 20 hours, rotation speed 200 rpm;

[0066] (3) Bacterial liquids of Lactobacillus johnsonii A21065 and Escherichia coli OP50 were respectively plated on nematode growth medium (NGM) and cultured overnight;

[0067] The formula of the NGM medium is as follows: 3 g of sodium chloride, 17 g of agar powder, 2.5 g of peptone, 975 mL of deionized water. After fully mixing the above components evenly, seal them with tin foil. After autoclaving at high pressure for 20 min, cool them in a water bath to 55 °C. Under aseptic conditions, add the following sterilized solutions: 1 mL of 1 M CaCl2, 1 mL of 1 M MgSO4, 25 mL of 1 M KPO4 buffer (108.3 g of KH2PO4 and 35.6 g of K2HPO4, dissolved in 1000 mL of distilled water, adjust the pH value to 6.0), 1 mL of 5 mg / mL cholesterol (dissolved in 95% ethanol);

[0068] (4) After centrifuging the thawed wild-type nematodes (Caenorhabditis elegans, kindly provided by Researcher Wang Bin of Guangxi Academy of Sciences), add them separately onto the NGM medium with Escherichia coli OP50, and place them in an incubator at 20 °C for 3 days;

[0069] (5) Suspend the nematodes cultured in (4) using M9 buffer (Na2HPO4: 6.0 g; KH2PO4: 3.0 g; NaCl: 5.0 g; MgSO4·7H2O: 0.25 g; dissolved in 1 L of distilled water, autoclave at high temperature and high pressure), and aspirate them into a culture tube. Add lysis solution (5 mol / L NaOH and 5% sodium hypochlorite solution by volume ratio) to each tube and lyse for 6 minutes;

[0070] (6) Then centrifuge the lysed liquid in (5) at 3500 r / min for 1 min, discard the supernatant, wash it repeatedly 4 times with M9 buffer, centrifuge again and discard the supernatant. Transfer the precipitate onto the NGM medium and culture it overnight at 20 °C to obtain L1-stage nematode larvae;

[0071] (7) After rinsing and centrifuging once with M9 buffer, transfer the nematode larvae onto the NGM plate containing Escherichia coli OP50 and culture them at 20 °C for 28 - 30 hours to obtain synchronized nematodes, that is, L4-stage nematodes;

[0072] (8) Randomly divide the synchronized L4-stage hermaphrodite nematodes with comparable size and vitality into 2 groups, namely the control group (fed with Escherichia coli OP50) and the experimental group (fed with experimental strain A21065), and culture them in an incubator at 20 °C. Mark the day when L4-stage nematodes are picked as day 0. On the 5th day, transfer the nematodes to the NGM medium containing 4.25 mM H2O2 (30 nematodes per group). At intervals of 1 hour, count the number of dead nematodes under a microscope according to the nematode death standard until all nematodes die.

[0073] The experimental results are as Figure 2 shown. Lactobacillus johnsonii A21065 can enhance the antioxidant capacity of nematodes by 80%.

[0074] Example 3: Anti-thermal stress effect of Lactobacillus johnsonii A21065 on Caenorhabditis elegans

[0075] This example provides a test on the anti-thermal stress ability of Lactobacillus johnsonii A21065 against Caenorhabditis elegans. The specific experimental steps are as follows.

[0076] (1) Prepare L4 synchronized nematodes, A21065, and OP50 bacterial solutions according to the experimental steps in Example 2;

[0077] (2) Randomly divide the synchronized nematodes into two groups, namely the control group (OP50) and the experimental group (Lactobacillus johnsonii A21065). Each strain of bacteria corresponds to 1 plate, with 50 nematodes on each plate. The day when L4-stage nematodes are picked is recorded as day 0. After culturing for 5 days respectively, transfer the nematodes to a 37 °C incubator for heat shock (30 nematodes per dish). At 1-hour intervals, count the number of surviving nematodes under a microscope (stimulate the head and body of the nematodes multiple times with a gold foil needle tip, and those with a response are considered alive, while those without a response are considered dead) and record.

[0078] The experimental results are as Figure 3 shown. Lactobacillus johnsonii A21065 can improve the anti-thermal stress ability of nematodes by 28.57%, and can improve the ability of the host to cope with environmental stress.

[0079] Example 4: Effect of Lactobacillus johnsonii A21065 on the lifespan of Caenorhabditis elegans

[0080] This example provides a test on the effect of Lactobacillus johnsonii A21065 on the lifespan of Caenorhabditis elegans. The specific experimental steps are as follows.

[0081] (1) Prepare L4 synchronized nematodes, A21065, and OP50 bacterial solutions according to the experimental steps in Example 2;

[0082] (2) Randomly divide the synchronized nematodes into two groups, namely the control group (OP50) and the experimental group (Lactobacillus johnsonii A21065), with 3 plates in each group and 50 nematodes on each plate. The day when L4-stage nematodes are picked is recorded as day 0. During the experiment, the NGM medium is changed every 2 days for the nematodes and the lifespan is counted until all nematodes die. Abnormally dead nematodes are not included in the statistics.

[0083] The experimental results are as Figure 4 shown. Lactobacillus johnsonii A21065 can extend the lifespan of nematodes by 18.75%.

[0084] Example 5: Anti-aging test of Lactobacillus johnsonii A21065

[0085] This example provides a test on the anti-aging ability of Lactobacillus johnsonii A21065. The specific experimental steps are as follows.

[0086] (1) Prepare L4 synchronized nematodes, A21065, and OP50 bacterial solution according to the experimental steps of Example 2;

[0087] (2) Randomly divide the synchronized nematodes into two groups, with each strain of bacteria corresponding to one plate, 50 nematodes per plate. They are respectively the control group (OP50) and the experimental group (Lactobacillus johnsonii A21065). The day when L4-stage nematodes are picked is recorded as day 0. Randomly pick 30 nematodes on the 5th, 10th, and 15th days respectively, and use a stereomicroscope (Motic) to measure their body bending rate. Put the test nematodes into a blank NGM culture plate to crawl freely for 30 s to adapt before starting the detection. When the nematodes show spontaneous and / or regular sine / cosine movements, one swing is recorded as one time, and record for 30 s. Subsequently, use GraphPad Prism 9 for statistics and analysis by t-test. P < 0.05 indicates significant difference, and P < 0.01 indicates extremely significant difference.

[0088] The experimental results are as Figure 5 shown. In the body bending test, Lactobacillus johnsonii A21065 can improve the locomotor ability of nematodes on the 5th, 10th, and 15th days of nematode culture. The improvement effects are 19.29%, 24.09%, and 129.29% respectively, indicating that A21065 can improve the locomotor effect of nematodes in the early, middle, and late stages of nematode culture.

[0089] Example 6 Hemolysis test of Lactobacillus johnsonii A21065

[0090] This example provides a hemolysis test of Lactobacillus johnsonii A21065. The specific experimental steps are as follows.

[0091] Activate and culture the Lactobacillus johnsonii A21065 obtained in Example 1 on a Columbia agar plate containing 5% sheep blood, and then place it in an anaerobic workstation at 37 °C for 48 hours to observe whether there is a hemolysis zone around the colony.

[0092] The experimental results are as Figure 6 shown. There is no hemolysis zone around the colony of Lactobacillus johnsonii A21065, indicating that Lactobacillus johnsonii A21065 has high safety.

[0093] Example 7 In vitro experiment on the resistance of Lactobacillus johnsonii A21065 to artificial gastrointestinal fluids

[0094] This example provides an in vitro experiment on the resistance of Lactobacillus johnsonii A21065 to artificial gastrointestinal fluids. The specific experimental steps are as follows.

[0095] (1) Take the Lactobacillus johnsonii A21065 obtained in Example 1 for overnight culture and activation, and inoculate the bacterial solution into artificial gastric juice (Regen Biotech) with pH = 3 at an inoculation amount of 10% by volume;

[0096] (2) 100 μL of the solution was taken at 0 h, 1 h, and 3 h respectively and spread on MRS plates, and the number of single colonies was counted after anaerobic culture at 37 °C for 48 h.

[0097] (3) Using the bacterial solution treated with gastric juice for 3 h as the initial bacterial solution concentration for the intestinal juice experiment, the bacterial solution was inoculated into artificial intestinal juice (Regen Biotech) with a pH of 8. 100 μL of the solution was taken at 2 h, 4 h, 6 h, and 8 h respectively and spread on MRS plates, and the number of single colonies was counted after anaerobic culture at 37 °C for 48 h. The survival rate of Lactobacillus johnsonii A21065 in artificial gastrointestinal juice was calculated with the 0 h of gastric juice as the initial concentration based on the number of single colonies in the above experiment.

[0098] The experimental results showed that the survival rate of Lactobacillus johnsonii A21065 in artificial gastric juice for 3 h was 593.75%, the survival rate in intestinal juice for 6 h was 21.43%, and the survival rate in intestinal juice for 8 h was 14.29%, indicating that Lactobacillus johnsonii A21065 had a certain colonization ability in the gastrointestinal tract without cryoprotectant.

[0099] Example 8 In vitro experiment of Lactobacillus johnsonii A21065 against bile salts

[0100] This example provides an in vitro experiment on the bile salt tolerance of Lactobacillus johnsonii A21065. The specific experimental steps are as follows.

[0101] The Lactobacillus johnsonii A21065 obtained in Example 1 was taken for overnight culture and activation. According to an inoculation amount of 10% by volume, the bacterial solution was inoculated into MRS liquid medium containing 0.1 g / L of bile salts. 100 μL of the solution was taken at 0 h, 1 h, 2 h, 3 h, and 4 h respectively and spread on MRS plates, and the number of single colonies was counted after anaerobic culture at 37 °C for 48 h.

[0102] The experimental results showed that the survival rate of Lactobacillus johnsonii A21065 in bile salts for 4 h was 118.18%, indicating that A21065 could survive and even proliferate in 0.1 g / L of bile salts.

[0103] Example 9 Effect of Lactobacillus johnsonii A21065 on the locomotor ability of Caenorhabditis elegans with Parkinson's disease

[0104] This example provides a detection of the effect of Lactobacillus johnsonii A21065 on Caenorhabditis elegans. NL5901 is a human α-synuclein (α-syn) transgenic nematode (purchased from Caenorhabditis Genetics Center, CGC). When there is α-syn aggregation in the nematode, green fluorescent protein (GFP) can be observed in the nematode. The specific experimental steps are as follows.

[0105] (1) Synchronized L4-stage NL5901 nematodes, A21065, and OP50 bacterial solution were cultured according to the experimental steps of Example 2.

[0106] (2) The synchronized nematodes were randomly divided into two groups. The bacterial solutions of OP50 and A21065 were respectively spread on blank NGM plates, divided into a control group (OP50) and an experimental group (Lactobacillus johnsonii A21065). 50 L4-stage nematodes were picked into each plate and cultured at 20 °C. The day of inoculation was counted as day 0. Body bending and head swinging experiments were carried out on the 5th and 8th days respectively. The method of the head swinging experiment was as follows: The frequency of head swinging of nematodes within 30 s was counted one by one in an NGM plate containing M9 buffer. A swing of the nematode's head from one side to the other and back to the original position was counted as one swing. The method of the body bending experiment was as follows: The frequency of body bending of nematodes within 30 s was counted on a blank NGM plate. A swing of the nematode's head from one side to the other and back to the original position was counted as one swing. Subsequently, statistical analysis was performed using GraphPad Prism 9, and a t-test was used for analysis. P < 0.05 indicated a significant difference, and P < 0.01 indicated a highly significant difference.

[0107] The experimental results are shown in Figure 7 and Figure 8 As shown, after feeding Lactobacillus johnsonii A21065, the body bending ( Figure 7 ) and head swinging ( Figure 8 ) of NL5901 were significantly improved, indicating that A21065 can improve the locomotor ability of NL5901.

[0108] Example 10 Effect of Lactobacillus johnsonii A21065 on the expression level of α-syn in Parkinson's nematodes

[0109] This example provides a detection of the effect of Lactobacillus johnsonii A21065 on the expression level of α-syn in Parkinson's nematodes. Under the condition of a shooting magnification of 20×, the aggregated expression level of α-syn near the gonopore was selected for comparison. The specific experimental steps are as follows.

[0110] (1) Synchronized L4-stage NL5901 nematodes, A21065, and OP50 bacterial solution were cultured according to the experimental steps of Example 2.

[0111] (2) The Parkinson's nematode model NL5901 was selected to photograph and detect the expression level of α-syn under a fluorescence microscope. Synchronized L4-stage nematodes NL5901 were selected. The control group was fed OP50, and the experimental group was fed Lactobacillus johnsonii A21065. Subsequently, they were cultured at 20 °C. On the 5th day, the nematodes were anesthetized with 2% tetramisole hydrochloride. After all nematodes were paralyzed, they were placed under the green channel of a fluorescence microscope for photography. The shooting magnification was 20×, and the shooting conditions should be consistent for subsequent analysis.

[0112] The experimental results are as Figure 9 shown. The A21065 group has an obvious effect of reducing the aggregation of α-syn, proving that A21065 has the potential to delay the onset of Parkinson's symptoms.

[0113] The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications made without departing from the spirit and principle of the present invention shall be equivalent replacement methods and are all included in the protection scope of the present invention.

Claims

1. A Lactobacillus johnsonii, characterized in that, The Lactobacillus johnsonii is Lactobacillus johnsonii A21065, taxonomically named Lactobacillus johnsonii, and was deposited at the Guangdong Microbial Culture Collection Center on January 13, 2025, with the deposit number GDMCC No: 65767.

2. The culture of the Lactobacillus johnsonii according to claim 1.

3. A bacterial agent, characterized in that, The bacterial agent comprises the Lactobacillus johnsonii according to claim 1 and / or the culture according to claim 2.

4. The microbial agent according to claim 3, wherein The bacterial agent is a solid bacterial agent or a liquid bacterial agent.

5. Use of any one or more of the Lactobacillus johnsonii according to claim 1, the culture of the Lactobacillus johnsonii according to claim 2, and the bacterial agent according to claim 3 or 4 in the preparation of a drug for preventing or treating Parkinson's disease.

6. The application according to claim 5, wherein The dosage form of the drug includes powder, granule, capsule, tablet, pill or oral liquid.

7. Use of any one or more of the Lactobacillus johnsonii according to claim 1, the culture of the Lactobacillus johnsonii according to claim 2, and the bacterial agent according to claim 3 or 4 in at least one of the following (I) to (III); (I) Preparation of a product for prolonging lifespan; (II) Preparation of an anti-aging product; (III) Preparation of an antioxidant product.

8. A product containing any one or more of the Lactobacillus johnsonii described in claim 1, the culture of Lactobacillus johnsonii described in claim 2, and the bacterial agent described in claim 3 or 4, characterized in that The product includes food, food additive, feed, feed additive, medicine and cosmetics.

9. The product according to claim 8, wherein The product further contains excipients; the excipients include food additives, pharmaceutically acceptable adjuvants or cosmetic excipients.

10. The product according to claim 8, characterized in that, The form of the Lactobacillus johnsonii in the product includes freeze-dried powder, bacterial liquid and granular inoculum.

Images