Application of Guangxi Akkermansia strain or its probiotic agent in the preparation of products for prolonging life, resisting aging and delaying Parkinson's disease

Abstract

The present invention provides an application of Akkermansia guangxiensis sp. strain in the preparation of products for prolonging life, resisting aging, and delaying Parkinson's disease, belonging to the technical field of microbial products. The application of Akkermansia guangxiensis sp. strain or a probiotic containing Akkermansia guangxiensis sp. strain in the preparation of products for resisting aging, prolonging life and / or delaying Parkinson's disease, wherein the deposit number of the Akkermansia guangxiensis sp. strain includes GDMCC No: 62888 and / or GDMCC No: 62889. The present invention uses Caenorhabditis elegans as an animal model, and studies have shown that two strains of Akkermansia can slow down the aging of nematodes by prolonging life, improving motor ability, and improving the stress injury ability of nematodes, and have broad prospects for being developed into health foods or medicines for prolonging life and resisting aging.

Description

Technical Field

[0001] The invention belongs to the technical field of microbial products, and specifically relates to the application of Guangxi Akkermansia strains or probiotic agents thereof in the preparation of products for prolonging life, resisting aging and delaying Parkinson's disease. Background Art

[0002] Aging is an inevitable physiological process that is time-dependent and determined by the interaction of multiple factors. The hallmarks of aging are the functional decline of tissues, cells, and organs and the increased risk of death. Aging is inevitable, but delaying aging is possible. As the world's population ages, the economic burden of aging-related diseases will soar rapidly, and aging-related research has received increasing attention. More than a century ago, Nobel Prize winner Metchnikoff first emphasized the importance of intestinal microorganisms to human health and aging, and that the intestinal flora will also change with the aging process. With the in-depth study of the intestinal flora, Akkermansia has also quickly attracted a lot of attention and research resources in the academic community because of its "beneficial" role in many diseases, and has even been directly dubbed the "universal bacteria" and "star bacteria".

[0003] At present, the models used in aging research include mice, fruit flies, nematodes, zebrafish and non-human primates. Specifically, among nematodes, Caenorhabditis elegans is widely used in aging research because of its clear genetic background, simple individual structure, transparent body, easy observation, short lifespan and easy control of growth conditions. It is the "top student" among model organisms.

[0004] The Akkermansia guangxiensis sp. strains with the deposit numbers GDMCC No: 62888 and GDMCC No: 62889 are two functional microbial strains isolated and screened from the intestines of centenarians. They have excellent antioxidant, tumor cell growth inhibition and weight loss and lipid-lowering effects. However, there are no reports on the anti-aging effects of these two strains and Akkermansia. Summary of the invention

[0005] In view of this, the object of the present invention is to provide the use of Guangxi Akkermansia strain or its probiotic agent in the preparation of anti-aging products.

[0006] The present invention provides an application of an Akkermansia guangxiensis sp. strain or a probiotic containing the Akkermansia guangxiensis sp. strain in preparing an anti-aging product. The Akkermansia guangxiensis sp. strain comprises the Akkermansia guangxiensis sp. strain N21116 with a deposit number of GDMCC No: 62888 and / or the Akkermansia guangxiensis sp. strain N21169 with a deposit number of GDMCC No: 62889.

[0007] Preferably, the anti-aging content includes improving at least one of the following abilities: motor function and stress resistance.

[0008] Preferably, the stress resistance includes heat stress resistance and / or oxidative stress resistance.

[0009] Preferably, the enhanced oxidative stress resistance includes the ability to resist hydrogen peroxide stimulation.

[0010] The invention provides use of an Akkermansia guangxiensis sp. strain or a probiotic containing the Akkermansia guangxiensis sp. strain in preparing a product for prolonging life. The Akkermansia guangxiensis strain comprises the Akkermansia guangxiensis strain N21116 with a deposit number of GDMCC No: 62888 and / or the Akkermansia guangxiensis strain N21169 with a deposit number of GDMCC No: 62889.

[0011] The invention provides an application of an Akkermansia guangxiensis sp. strain or a probiotic containing the Akkermansia guangxiensis sp. strain in preparing a product for delaying Parkinson's disease. The Akkermansia guangxiensis sp. strain comprises the Akkermansia guangxiensis sp. strain N21116 with a preservation number of GDMCC No: 62888 and / or the Akkermansia guangxiensis sp. strain N21169 with a preservation number of GDMCC No: 62889.

[0012] Preferably, said delaying Parkinson's disease comprises delaying Parkinson's disease symptoms.

[0013] Preferably, the delaying of Parkinson's disease symptoms includes reducing the frequency of head shaking and / or restoring the patient's basic behavior.

[0014] Preferably, the product comprises at least one of the following: a medicine, a feed additive and a feed.

[0015] Preferably, the probiotic containing the Guangxi Akkermansia strain is a freeze-dried powder of the Guangxi Akkermansia strain.

[0016] The present invention provides an application of an Akkermansia guangxiensis sp. strain or a probiotic containing the Akkermansia guangxiensis sp. strain in preparing an anti-aging product. The Akkermansia guangxiensis sp. strain comprises the Akkermansia guangxiensis sp. strain N21116 with a deposit number of GDMCC No: 62888 and / or the Akkermansia guangxiensis sp. strain N21169 with a deposit number of GDMCC No: 62889. The present invention uses Caenorhabditis elegans as an experimental object, and uses two strains of Akkermansia isolated from the intestines of centenarians as active ingredients, and detects three aging-related indicators: motor function, heat stress and oxidative stress, to explore its function in delaying aging. The experiment shows that strain N21116 and strain N21169 can significantly increase the head swing frequency of nematodes in the middle stage (equivalent to human middle age), indicating that both strains can improve the vitality of nematodes; the results of the heat stress experiment show that compared with the control group, the survival time of nematodes in the strain N21116 and N21169 groups after heat shock is prolonged, and the difference is significant; the results of the hydrogen peroxide stimulation experiment show that under the stimulation of 3.375mM H2O2, compared with the control group, the N21116 group can increase the antioxidant capacity of nematodes by 33.33%. It can be seen that the Guangxi Akkermansia strain or probiotics containing the Guangxi Akkermansia strain provide new evidence for the anti-aging research of Akkermansia and new ideas for the research and development of anti-aging probiotic products.

[0017] The invention provides use of an Akkermansia guangxiensis sp. strain or a probiotic containing the Akkermansia guangxiensis sp. strain in preparing a product for prolonging life. The Akkermansia guangxiensis strain comprises the Akkermansia guangxiensis strain N21116 with a deposit number of GDMCC No: 62888 and / or the Akkermansia guangxiensis strain N21169 with a deposit number of GDMCC No: 62889. The present invention uses Caenorhabditis elegans as the experimental object, and uses two strains of Akkermansia isolated from the intestines of centenarians as the active ingredients to detect the life span of the synchronized nematodes and explore its function in extending life span. The experiment shows that compared with the control group, the nematodes significantly extended their life span during the 10th to 20th day after feeding with N21116 and N21169. The average life spans of the OP50 control group, N21116, and N21169 experimental groups were 17, 21, and 21 days, respectively. Compared with the control group, N21116 extended the life span of the nematodes by 23.53%, and N21169 extended the life span of the nematodes by 23.53%. It can be seen that the application provided by the present invention provides a new idea for the development or preparation of products related to extending life span.

[0018] The invention provides use of an Akkermansia guangxiensis sp. strain or a probiotic containing the Akkermansia guangxiensis sp. strain in preparing a product for delaying Parkinson's disease. The Akkermansia guangxiensis sp. strain comprises the Akkermansia guangxiensis sp. strain N21116 with a preservation number of GDMCC No: 62888 and / or the Akkermansia guangxiensis sp. strain N21169 with a preservation number of GDMCC No: 62889. The present invention uses a Caenorhabditis elegans disease model suffering from Parkinson's disease as an experimental object, and uses two strains of Akkermansia isolated from the intestines of centenarians as active ingredients, statistically analyzes the clinical phenotype of the nematode disease model, and explores its function in the prevention and treatment of Parkinson's disease. The experiment shows that compared with the control group, the model group has a higher head swing frequency in M9 buffer, which is significant, while the head swing frequency of the nematodes in the N21116 and N21169 groups is comparable to that of the OP50 control group, which is not statistically significant. It can be seen that the feeding of the strain can effectively slow down the frequency of the patient's head swing. At the same time, the improvement of basic behavior is also statistically analyzed. The body bending frequency of the nematode model group and the OP50 control group shows an opposite trend. The nematodes in the model group move faster when there is no food, proving that the dopamine neurons of the nematodes in the model group may be damaged, and after feeding the N21116 and N21169 strains, the nematodes recover the basic behavior. It can be seen that the experimental results prove that the N21116 and N21169 strains have the function of alleviating the symptoms of Parkinson's disease, providing new ideas for the development or preparation of life-extending related products. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 This is the phylogenetic tree of Akkermansia guangxiensis N21116 and N21169;

[0020] Figure 2 This is the ANI analysis result of Akkermansia in Guangxi;

[0021] Figure 3 is the lifespan curve of Caenorhabditis elegans;

[0022] Figure 4 The results for head shaking in Caenorhabditis elegans;

[0023] Figure 5 The results of the heat stress test of Caenorhabditis elegans are shown in Figure 1.

[0024] Figure 6 The results of the test on the anti-oxidative stress ability of Caenorhabditis elegans;

[0025] Figure 7 The results of the head shaking experiment on the nematode Caenorhabditis elegans with Parkinson's disease;

[0026] Figure 8The results of a basic mitigation experiment on Parkinson's disease in Caenorhabditis elegans.

[0027] Biodeposit Survival Certificate

[0028] Akkermansia guangxiensis sp. strain N21116 is deposited in Guangdong Microbiological Culture Collection Center, abbreviated as GDMCC, with the address at 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, Guangdong Institute of Microbiology, deposit date December 2, 2022, and deposit number GDMCC No: 62888.

[0029] Akkermansia guangxiensis sp. strain N21169 is deposited in the Guangdong Provincial Microbiological Culture Collection Center, abbreviated as GDMCC, with the address at 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, Guangdong Institute of Microbiology, deposit date December 2, 2022, and the deposit number is GDMCC No: 62889. DETAILED DESCRIPTION

[0030] The present invention provides an application of an Akkermansia guangxiensis sp. strain or a probiotic containing the Akkermansia guangxiensis sp. strain in preparing an anti-aging product. The Akkermansia guangxiensis sp. strain comprises the Akkermansia guangxiensis sp. strain N21116 with a deposit number of GDMCC No: 62888 and / or the Akkermansia guangxiensis sp. strain N21169 with a deposit number of GDMCC No: 62889.

[0031] In the present invention, Guangxi Akkermansia strain N21116 with a deposit number of GDMCC No: 62888 and / or Guangxi Akkermansia strain N21169 with a deposit number of GDMCC No: 62889 are separated from the feces of centenarians and disclosed in the patent with publication number CN116200312A. The Guangxi Akkermansia strain N21116 with a deposit number of GDMCC No: 62888 and / or Guangxi Akkermansia strain N21169 with a deposit number of GDMCC No: 62889 have good resistance to gastric and intestinal juices, good safety, can be colonized in the human gastrointestinal tract for a long time, and can meet the requirements of probiotics.

[0032] In the present invention, the probiotic containing the Guangxi Akkermansia strain is preferably a freeze-dried powder of the Guangxi Akkermansia strain. The preparation method of the freeze-dried powder of the Guangxi Akkermansia strain preferably comprises the following steps:

[0033] amplifying and culturing the Akkermansia guangxiensis strain to obtain a culture solution;

[0034] The bacterial cells are separated from the culture solution, mixed with a freeze-drying protective agent, and then freeze-dried to obtain freeze-dried powder.

[0035] In the present invention, the method of amplification and culture is preferably to culture Guangxi Akkermansia strain N21116 or N21169 anaerobically at 37°C for 72 hours in a synthetic liquid medium. The method of isolating the bacteria is preferably centrifugation. The speed of the centrifugation is preferably 7500-8500rpm, more preferably 8000rpm, and the centrifugation time is preferably 8-12min, more preferably 10min. The centrifugation temperature is preferably 4°C. The mass ratio of the bacteria and the lyophilization protective agent is preferably 1:0.5-3, more preferably 1:1.3. The lyophilization protective agent preferably includes skim milk, trehalose, sorbitol, and vitamin C.

[0036] In the present invention, the anti-aging content preferably includes improving at least one of the following abilities: motor function and stress resistance. The motor ability is a reflection of the aging degree of nematodes. The older the nematodes are, the worse their motor ability is. In the embodiment of the present invention, the head swing frequency of nematodes is used as an indicator to characterize the vitality of nematodes. The experimental results show that both N21116 and N21169 can significantly improve the head swing frequency of nematodes in the middle stage (equivalent to human middle age). The stress resistance preferably includes heat stress resistance and / or oxidative stress resistance. The heat stress resistance refers to the length of survival time after heat stimulation. The longer the survival time, the stronger the heat stress resistance of nematodes. The experimental results of the embodiment of the present invention show that compared with the OP50 control group, the survival time of nematodes fed N21116 and N21169 after heat shock was prolonged. Compared with the control group, the N21116 and N21169 groups can respectively improve the heat stress resistance of nematodes by 83.33% and 66.67%. The oxidative stress capacity preferably refers to the survival capacity of nematodes growing in a medium containing hydrogen peroxide, and a higher survival rate indicates that the nematodes have a stronger ability to resist oxidative stress. In the present embodiment, under the stimulation of 3.375 mM H2O2, compared with the control group, the N21116 group can improve the antioxidant capacity of nematodes by 33.33%; N21169 has no obvious effect.

[0037] The invention provides use of an Akkermansia guangxiensis sp. strain or a probiotic containing the Akkermansia guangxiensis sp. strain in preparing a product for prolonging life. The Akkermansia guangxiensis strain comprises the Akkermansia guangxiensis strain N21116 with a deposit number of GDMCC No: 62888 and / or the Akkermansia guangxiensis strain N21169 with a deposit number of GDMCC No: 62889.

[0038] In the examples of the present invention, compared with the OP50 control group, the lifespan of nematodes after feeding with N21116 and N21169 was significantly extended during the 10th to 20th day. The average lifespans of the OP50 control group, N21116, and N21169 experimental groups were 17, 21, and 21 days, respectively. Relative to the control group, N21116 extended the lifespan of nematodes by 23.53%, and N21169 extended the lifespan of nematodes by 23.53%.

[0039] The invention provides use of an Akkermansia guangxiensis sp. strain or a probiotic containing the Akkermansia guangxiensis sp. strain in preparing a product for delaying Parkinson's disease. The Akkermansia guangxiensis sp. strain comprises the Akkermansia guangxiensis sp. strain N21116 with a preservation number of GDMCC No: 62888 and / or the Akkermansia guangxiensis sp. strain N21169 with a preservation number of GDMCC No: 62889.

[0040] In the present invention, the delaying of Parkinson's disease preferably includes delaying the symptoms of Parkinson's disease. The delaying of the symptoms of Parkinson's disease preferably includes reducing the frequency of head shaking and / or restoring the patient's basic behavior. The results of the embodiments of the present invention show that strains N21116 and / or N21169 can delay the symptoms of Parkinson's disease.

[0041] In the present invention, the product preferably includes at least one of the following: medicine, feed additive and feed. The present invention has no special restrictions on the preparation method of the medicine, feed additive and feed, and the preparation method known in the art can be used.

[0042] The application of the Guangxi Akkermansia strain or its probiotic agent provided by the present invention in the preparation of products for prolonging life, resisting aging, and delaying Parkinson's disease is described in detail below in conjunction with the examples, but they should not be construed as limiting the scope of protection of the present invention.

[0043] 1. Materials and Reagents:

[0044] Escherichia coli OP50 and wild-type Caenorhabditis elegans N2 were kindly donated by Researcher Wang Bin from Guangxi Academy of Sciences.

[0045] Disodium hydrogen phosphate, potassium dihydrogen phosphate, sodium chloride, dipotassium hydrogen phosphate, sodium hydroxide, Sinopharm Chemical Reagent Co., Ltd.; agar and Triton X-100, Beijing Solebold Technology Co., Ltd.; calcium chloride and cholesterol, Sangon Biotech (Shanghai) Co., Ltd.; peptone, Thermo Fisher Scientific (China) Co., Ltd.; magnesium sulfate, Tianjin Bodi Chemical Co., Ltd.; tryptone and yeast extract, Weibaiao (Beijing) Biotechnology Co., Ltd.; BHI medium, Qingdao Haibo Biotechnology Co., Ltd.; sodium hypochlorite, Tianjin Damao Chemical Reagent Factory; isopropanol, Tianjin Fuyu Fine Chemical Co., Ltd.; paraformaldehyde, Shanghai MacLean Biochemical Technology Co., Ltd.; hydrogen peroxide, Nanguo Pharmaceutical Co., Ltd.

[0046] 2. Source of instruments and equipment

[0047] Vertical pressure steam sterilizer, Shanghai Boxun Medical Biological Instrument Co., Ltd.; electronic balance, Shenyang Longteng Electronics Co., Ltd.; HPX-II-250, Shanghai Yuejin Medical Instrument Co., Ltd.; BSC-1500II B2-X, Jinan Xinbeisi Biotechnology Co., Ltd.; stereo microscope, Motic Industrial Group Co., Ltd.; PCR amplification instrument, Suzhou Dongsheng Xingye Science Co., Ltd.; biochemical incubator, Wuhan Ruihua Instrument Equipment Co., Ltd.

[0048] Example 1

[0049] Isolation and identification of Akkermansia N21116 and N21169 from Guangxi

[0050] The separation and purification steps of Akkermansia Guangxi in this embodiment are as follows:

[0051] Take an appropriate amount of stool samples from two centenarians in Guangxi, dilute them with PBS, evenly spread them on mucin solid culture medium, and place them in an anaerobic workstation. Culture them at 37°C for two days under the gas conditions of 90% (v / v) N2 and 10% CO2. After the culture, pick the grayish-white colonies with round, raised, smooth, and neat edges on the plate and identify them using PCR amplification technology.

[0052] The PCR identification method of Akkermansia guangxiensis in this embodiment is:

[0053] Use a sterile toothpick to pick up colonies that meet the above Akkermansia morphological characteristics, transfer them to 2× PCR premix, and add upstream and downstream specific primers and sterile water in sequence (the specific PCR amplification system is shown in Table 1). The thermal cycle parameters are set as follows: 95°C pre-denaturation for 3 min; 95°C denaturation for 40 s, 56°C annealing for 30 s, 72°C extension for 1 min, 35 cycles; 72°C final extension for 10 min.

[0054] Table 1 Akkermansia PCR amplification system

[0055]

[0056]

[0057] Among them, the Akkermansia specific amplification primers used for identification are:

[0058] Upstream primer F: 5′-CAGCACGTGAAGGTGGGGAC-3′ (SEQ ID NO: 1);

[0059] Downstream primer R: 5'-CCTTGCGGTTGGCTTCAGAT-3' (SEQ ID NO: 2).

[0060] The amplified products were detected by agarose gel electrophoresis, and when the length of the amplified fragment was consistent with expectations, the isolated strain could be preliminarily determined to be a suspected Akkermansia strain.

[0061] The strains identified as suspected Akkermansia by PCR amplification were further subjected to 16S rRNA molecular identification.

[0062] The 16S rRNA molecular identification system was the same as in Table 1. The thermal cycle parameters were as follows: pre-denaturation at 95°C for 2 min; denaturation at 95°C for 40 s, annealing at 56°C for 30 s, extension at 72°C for 90 s, 30 cycles; and final extension at 72°C for 10 min.

[0063] Among them, the specific amplification primers used for 16S rRNA molecule identification are:

[0064] Upstream primer 16S-F: 5'-AGAGTTTGATCCTGGCTCAG-3' (SEQ ID NO: 3); downstream primer 16S-R: 5'-TACGGCTACCTTGTTACGACTT-3' (SEQ ID NO: 4).

[0065] The amplified products were sent to Shanghai Bioengineering for sequencing.

[0066] Among them, the 16S sequence of suspected Akkermansia strain 1 (number N21169) obtained by sequencing is shown in SEQ ID NO: 5. The 16S sequence of suspected Akkermansia strain 2 (number N21116) is shown in SEQ ID NO: 6. Using the NCBIBLAST tool for sequence retrieval, it was found that the 16S rRNA sequence similarities of the two strains with the Akkermansia muciniphila ATCC BAA-835 standard strain were 95.45% and 95.53%, respectively, which shows that these two strains may be new species of Akkermansia that have not been discovered.

[0067]

[0068]

[0069] To further determine the evolutionary relationship of the above strains, the whole genome of the two suspected strains was sequenced, and ANI analysis was performed on them compared with the only two current members of the genus Akkermansia.

[0070] The results are as follows Figure 1 and Figure 2 shown.

[0071] The results showed that the ANI values ​​of N21116 and Akkermansia muciniphila ATCC BAA-835T of the genus Akkermansia and Akkermansia saccharophila PyT isolated from pythons were 68.95% and 68.34%, respectively, and the ANI values ​​of N21169 and Akkermansia muciniphila ATCC BAA-835T of the genus Akkermansia and Akkermansia saccharophila PyT isolated from pythons were 68.83% and 68.49%, respectively, meeting the ANI evaluation criteria (95%-96%) for the classification of new species. In addition, the sequence identity between N21116 and N21169 was 98.59%. It can be seen that the two belong to the same species.

[0072] Considering that the two strains of microorganisms were isolated from Guangxi, the inventors temporarily named them Akkermansia guangxiensis sp., and their Latin names were temporarily determined as Akkermansia guangxiensis sp.nov.

[0073] The phylogenetic tree based on the 16S rRNA sequences of the two strains of Akkermansia spp. isolated in this example is shown in Figure 1 , which revealed that it belongs to the phylum Verrucomicrobia, class Verrucomicrobiales, order Verrucomicrobiales, family Akkermansiaceae, genus Akkermansia.

[0074] The two strains of Akkermansia (N21116 and N21169) obtained in the above examples were sent to the Guangdong Microbiological Culture Collection Center (GDMCC, the collection address is: 5th Floor, Building 59, No. 100, Xianlie Middle Road, Guangzhou) for preservation on December 2, 2022. The taxonomic names are all: Akkermansia sp., and the collection numbers are: GDMCC No: 62888 and GDMCC No: 62889.

[0075] Example 2

[0076] Bacteria strains, nematode culture, and nematode synchronization

[0077] E. coli OP50 was cultured in LB liquid medium. After overnight culture, an appropriate amount was plated on nematode growth medium (NGM) and cultured overnight. Thawed nematodes were centrifuged and supernatant was removed. Then, the thawed nematodes were added to NGM medium with OP50 and cultured in a biochemical incubator at 20°C. N21116 and Na21169 were cultured in BHI liquid medium at 37°C for 3 days for use.

[0078] Adult worms were suspended in M9 buffer and pipetted into culture tubes. Lysis buffer (5N NaOH and 5% sodium hypochlorite solution) was added to each tube of nematodes. Lysis was performed for 6 minutes. Centrifugation was performed at 3500 r / min for 1 minute. The supernatant was discarded. The tubes were washed with M9 buffer for 4 times, the supernatant was discarded, and the tubes were transferred to NGM culture medium. The tubes were cultured at 20°C overnight to obtain L1 larvae. After washing and centrifugation with M9 buffer once, the nematodes were added to NGM culture medium with OP50 and cultured at 20°C for 28-30 hours to obtain synchronized L4 nematodes.

[0079] Example 3

[0080] Lifespan experiment

[0081] The synchronized nematodes were randomly divided into 3 groups, each with 3 plates and 50 nematodes per plate, which were the control group (fed with OP50) and the experimental group (fed with N21116 or N21169). The day of picking the L4 nematodes was recorded as day 0. During the experiment, the NGM medium of the nematodes was replaced every 2 days and the life span was counted until all the nematodes died. The nematodes that died abnormally were not included in the statistics.

[0082] The results are shown in Table 2 and Figure 3 Compared with the control group, the life span of nematodes was significantly extended during the 10th to 20th day after feeding with N21116 and N21169. The average life spans of the OP50 control group, N21116, and N21169 experimental groups were 17, 21, and 21 days, respectively. Compared with the control group, N21116 extended the life span of nematodes by 23.53%, and N21169 extended the life span of nematodes by 23.53%.

[0083] Table 2 Life test results

[0084]

[0085] Example 4

[0086] Head shaking frequency

[0087] The nematodes were cultured according to the method of Example 2, and 30 nematodes were randomly selected and their head swing rates were measured using a stereo microscope (Motic) on the 10th day of culture. The test nematodes were placed in M9 buffer and swam freely for 30 seconds to adapt to the test. When the head of the nematode showed spontaneous and / or regular sine / cosine motion, one swing was counted as one time, and the record was recorded for 30 seconds.

[0088] Results Figure 4 The frequency of head shaking of nematodes is a reflection of the vitality of nematodes. The test was conducted in M9 buffer. Figure 4 It can be seen that compared with the OP50 control group, both N21116 and N21169 can significantly increase the head swing frequency of middle-aged nematodes (equivalent to middle-aged humans).

[0089] Example 5

[0090] Heat stress tolerance in nematodes

[0091] Nematodes were cultured according to the method of Example 2. The 5th antenna nematodes were transferred to a 37°C incubator for heat shock (30 nematodes / dish). The number of surviving nematodes was counted under a microscope every hour. The head and body of the nematodes were stimulated with a gold foil needle tip several times. A response was considered alive, and no response was considered dead. The survival of the nematodes was recorded.

[0092] The results are shown in Table 3 and Figure 5 Compared with the control group, the survival time of nematodes fed with N21116 and N21169 was prolonged after heat shock. Compared with the OP50 control group, the N21116 and N21169 groups could improve the heat stress resistance of nematodes by 50% and 50%, respectively.

[0093] Table 3 Results of heat stress tolerance of nematodes

[0094]

[0095] Example 6

[0096] Antioxidant stress capacity of nematodes

[0097] The nematodes were cultured according to the method in Example 2, and the 5th antenna nematodes were transferred to NGM medium containing 3.375 mM H2O2, with each strain corresponding to one plate and 50 nematodes per plate. The death of C. elegans was observed and recorded every 1 hour.

[0098] Results Figure 6 Under the stimulation of 3.375 mM H2O2, compared with the OP50 control group, both the N21116 group and the N21169 group could enhance the antioxidant capacity of nematodes by 40%.

[0099] Example 7

[0100] Establishment of Parkinson's disease model and the effects of N21116 and N21169 on this nematode model

[0101] After synchronization of the nematodes, the L1 stage nematodes were washed with M9 buffer, and then treated with 6-hydroxydopamine (a drug commonly used to establish a Parkinson's disease model) to a final concentration of 50 mM. The nematodes were induced in the dark for 1.5 h. After washing twice with M9 buffer, the nematodes were added to plates coated with OP50, N21116, and N21169. After being placed at 25°C for 48 h, the head swing frequency of the nematodes within 30 s and the body bending frequency on the sterile plate and the plate with bacteria were counted. The number of nematodes in each group was more than 10.

[0102] In addition, we also used these nematodes to conduct a basic slowing experiment. First, 10 nematodes from each group were selected and crawled on a sterile plate and a plate with bacteria, respectively. After the nematodes adapted, the body bending frequency of each group of nematodes within 30 seconds was counted.

[0103] One of the clinical manifestations of Parkinson's disease is abnormal behavior. Figure 7 As shown, compared with the control group, the head swing frequency of the model group in M9 buffer was higher, which was significantly higher, while the head swing frequency of nematodes in the N21116 and N21169 groups was comparable to that of the control group, which was not statistically significant.

[0104] In the basic behavior of nematodes, the speed of movement is affected by the environment. When there is sufficient food, the dopamine neurons of the nematodes will send signals, causing the movement to slow down. On the contrary, in an environment where food is scarce, the nematodes will release serotonin to inhibit the nematodes from slowing down, causing the nematodes to move faster. Figure 8 As shown, the body bending frequency of the nematode model group and the OP50 control group showed opposite trends. The nematodes in the model group moved faster when there was no food, proving that the dopamine neurons of the nematodes in the model group may have been damaged. After feeding the N21116 and N21169 strains, the nematodes recovered their basal behavior. Figure 7 and Figure 8 Both of them proved that nematodes could alleviate the symptoms of Parkinson's disease after eating N21116 and N21169 strains.

[0105] From the results of the above embodiments, it can be seen that the two strains of Akkermansia selected in this application, using Caenorhabditis elegans as an animal experimental model, conducted a series of screening experiments through lifespan experiments and detection of three aging-related indicators: motor function, heat stress and oxidative stress, and found that strains N21116 and N21169 can significantly prolong the lifespan of nematodes, improve the ability of nematodes to resist heat stress, and improve the head swing frequency and anti-oxidative stress ability of Caenorhabditis elegans; in addition, the research results also show that N21116 and N21169 can delay the symptoms of Parkinson's disease. Therefore, these two strains of Akkermansia can slow down the aging of nematodes by prolonging lifespan, improving motor ability, and improving the ability of nematodes to resist stress damage, and have broad prospects for developing drugs such as prolonging lifespan and anti-aging.

[0106] 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. Akkermansia guangxiensis ( Akkermansia guangxiensis sp.) strain or a probiotic containing the Guangxi Akkermansia strain in the preparation of anti-aging products, wherein the Guangxi Akkermansia strain is the Guangxi Akkermansia strain N21116 with a deposit number of GDMCC No: 62888 and / or the Guangxi Akkermansia strain N21169 with a deposit number of GDMCC No: 62889; The aging refers to a decline in motor function and / or stress resistance; The anti-stress ability is anti-heat stress ability and / or anti-oxidative stress ability; The anti-oxidative stress ability is the ability to resist the stimulation of hydrogen peroxide.

2. Akkermansia guangxiensis ( Akkermansia guangxiensis sp.) strain or a probiotic containing the Guangxi Akkermansia strain in the preparation of a product that prolongs life, wherein the Guangxi Akkermansia strain is the Guangxi Akkermansia strain N21116 with a deposit number of GDMCC No: 62888 and / or the Guangxi Akkermansia strain N21169 with a deposit number of GDMCC No: 62889.

3. Akkermansia guangxiensis ( Akkermansia guangxiensis sp.) strain or a probiotic containing the Guangxi Akkermansia strain in the preparation of a product for delaying Parkinson's disease, wherein the Guangxi Akkermansia strain is the Guangxi Akkermansia strain N21116 with a deposit number of GDMCC No: 62888 and / or the Guangxi Akkermansia strain N21169 with a deposit number of GDMCC No: 62889; The delaying of Parkinson's disease symptoms is to reduce the frequency of head shaking and / or restore the patient's basic behavior.

4. The use according to any one of claims 1 to 3, characterized in that: The product described is a drug.

5. The use according to any one of claims 1 to 3, characterized in that: The probiotic containing the Guangxi Akkermansia strain is freeze-dried powder of the Guangxi Akkermansia strain.

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