Strain of saccharomyces cerevisiae capable of relieving senescence and application of saccharomyces cerevisiae

Abstract

The invention discloses a strain of saccharomyces cerevisiae capable of relieving senescence and application of the saccharomyces cerevisiae, and belongs to the technical field of microorganisms and the technical field of medicines. Animal experiments prove that the Phocaecola dreei CCFM1429 has the effects of improving the weakness index of a mouse, increasing peroxide dismutase of the liver of the mouse, increasing the level of glutathione peroxidase (GSH-px), reducing an inflammatory factor IL-6, reducing an inflammatory factor TNF-alpha and improving the holding power of the mouse. The strain disclosed by the invention has a huge application prospect in preparation of products for relieving senescence.

Description

Technical Field

[0001] The invention relates to a strain of Bacteroides dorei capable of alleviating aging and application thereof, and belongs to the technical field of microorganisms and the technical field of medicine. Background Art

[0003] At the micro level, the characteristics of aging include cell aging, chronic inflammation, and intestinal flora disorders. Among them, a series of oxidative damage caused by chronic inflammation and cell aging is a difficult problem that has been waiting to be solved in the field of aging. At the macro level, aging causes many diseases such as Parkinson's disease and sarcopenia. A series of changes brought about by aging affect people's quality of life.

[0004] So far, the main challenge facing the existing technology is to find a safe, non-side-effect, and non-addictive treatment method that can regulate the body's health through intestinal bacteria and thus alleviate the symptoms associated with aging. It can regulate the body's inflammatory response level, oxidative stress, and alleviate common symptoms associated with aging such as muscle loss and reduced strength, thereby effectively improving the body's health level and alleviating aging. The existing technology has not yet found a microorganism that can effectively alleviate the level of inflammation, increased oxidative stress, and weakened muscle strength caused by aging. Summary of the invention

[0005] The present invention provides a strain of Phocaeicola dorei CCFM1429, and conducts animal test efficacy verification on the effect of alleviating aging.

[0006] The first object of the present invention is to provide a strain of Phocaeicola dorei CCFM1429, wherein the strain is deposited in Guangdong Province Microbiological Culture Collection Center with a deposit number of GDMCC No: 65155.

[0007] In one embodiment of the present invention, the Phocaeicola dorei CCFM1429 is isolated from human feces.

[0008] In one embodiment of the present invention, the growth characteristics of the Phocaeicola dorei CCFM1429 are as follows: the strain is an anaerobic bacterium, and after being inoculated into the culture medium, it is cultured at a culture temperature of 37° C. for 24 hours.

[0009] In one embodiment of the present invention, the culture medium is BHI medium.

[0010] In one embodiment of the present invention, the anti-aging effect of CCFM1429 was evaluated based on various biochemical indices of 20 experimental group mice (blank group, model group, spermidine group, Bacillus doreid CCFM1429 group, 5 mice in each group).

[0011] The second object of the present invention is to provide a microbial preparation containing the Bacillus multifiliis CCFM1429.

[0012] In one embodiment of the present invention, the microbial preparation contains one or more of the living cells, fermentation broth, lysate or extract of the Bacillus doreida CCFM1429.

[0013] The third object of the present invention is to provide a medicine containing the Bacillus multifiliis CCFM1429 or the microbial preparation.

[0014] In one embodiment of the present invention, the amount of Bacteroides doreniformis CCFM1429 added to the drug is not less than 2×10 8 CFU / mL or 2×10 8 CFU / g.

[0015] In one embodiment of the present invention, the dosage form of the drug includes powder, granules, capsules, tablets, pills or liquid preparations.

[0016] The fourth object of the present invention is to provide a method for preparing the microbial preparation, which is obtained by mixing the cultured Bacillus doreid CCFM1429 cells with a protective agent.

[0017] In one embodiment of the present invention, the protective agent includes one or more of skimmed milk powder, glycerol, maltodextrin, trehalose, and sodium L-glutamate.

[0018] The fifth object of the present invention is to provide the use of the Bacillus multifiliis CCFM1429 or the microbial preparation in the preparation of a drug for alleviating aging symptoms.

[0019] In one embodiment of the present invention, the delaying of aging includes reducing the level of oxidative stress, reducing the inflammatory response, reducing the frailty index, and increasing the level of acetylcholine in the brain.

[0020] In one embodiment of the present invention, the amount of Phocaeicola dorei CCFM1429 added to the drug is not less than 2×10 8 CFU / mL or 2×10 8 CFU / g.

[0021] In one embodiment of the present invention, the drug contains the Phocaeicoladorei CCFM1429, a drug carrier and / or a pharmaceutical excipient.

[0022] In one embodiment of the present invention, the drug carrier comprises microcapsules, microspheres, nanoparticles and / or liposomes.

[0023] In one embodiment of the present invention, the pharmaceutical excipients include one or more of fillers, binders, wetting agents, disintegrants, lubricants, and flavoring agents.

[0024] In one embodiment of the present invention, the drug is used to regulate intestinal flora, regulate oxidative stress response, reduce inflammatory response, reduce frailty index, bring health benefits and thus delay aging.

[0025] In one embodiment of the present invention, the drug is used for at least one of (a) to (h):

[0026] (a) reduce the frailty score index of aging individuals;

[0027] (b) reduce the level of MDA in liver tissue of aging individuals;

[0028] (c) increasing the level of SOD enzyme in liver tissue of aging individuals;

[0029] (d) increasing the level of CAT enzyme in liver tissue of aging individuals;

[0030] (e) increasing the level of GSH-px enzyme in liver tissue of aging individuals;

[0031] (f) increasing the level of acetylcholine in the brain tissue of aging individuals;

[0032] (g) reducing the expression level of pro-inflammatory factors in colon tissue of aging individuals; the pro-inflammatory factors include IL-6 and TNF-alpha;

[0033] (h) Improving grip strength in aging individuals.

[0034] Beneficial effects:

[0035] The present invention relates to a strain of Phocaeicola dorei CCFM1429 that can bring health benefits and thus alleviate aging. After the Phocaeicola dorei CCFM1429 provided by the present invention is applied to aged mice, the aging symptoms of the mice can be significantly alleviated, which is specifically embodied in:

[0036] (1) Significantly reduced the frailty score index of aging mice by 31.62%.

[0037] (2) Significantly increased the level of superoxide dismutase (SOD) in the liver of aged mice by 65.13%.

[0038] (3) Significantly increased the level of glutathione peroxidase (GSH-px) in the liver of aged mice by 109.40%

[0039] (4) Reduced the IL-6 level in the colon homogenate of aged mice by 25.43%.

[0040] (5) Reduced TNF-alpha levels in colon homogenates of aged mice by 21.20%.

[0041] (6) Improved the grip strength of aged mice by 19.89%.

[0042] Therefore, Phocaeicola dorei CCFM1429 has great application prospects in the preparation of drugs to alleviate aging. BRIEF DESCRIPTION OF THE DRAWINGS

[0043] Figure 1 : Frailty index of experimental mice in different groups;

[0044] Figure 2 : Superoxide dismutase (SOD) levels in the liver of experimental mice in different groups;

[0045] Figure 3 : Levels of glutathione peroxidase (GSH-px) in the liver of experimental mice in different groups;

[0046] Figure 4 :IL-6 levels in colon homogenates of experimental mice in different groups;

[0047] Figure 5 :TNF-alpha levels in the colon of different groups of experimental mice;

[0048] Figure 6 : The grip strength levels of experimental mice in different groups;

[0049] Figure 7 : Flowchart of animal experiments.

[0050] Deposit of biological materials:

[0051] A strain of Phocaeicola dorei CCFM1429, taxonomically named Phocaeicola dorei, was deposited in the Guangdong Provincial Microbial Culture Collection on September 20, 2024, with the deposit number GDMCC No: 65155. The deposit address is the 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, Institute of Microbiology, Guangdong Academy of Sciences. DETAILED DESCRIPTION

[0052] The C57BL / 6J male mice involved in the following examples were purchased from Beijing Weitong Lihua Animal Experiment Technology Co., Ltd., and the D-galactose and spermidine involved in the following test examples were purchased from Aladdin. The BHI culture medium involved in the following examples was purchased from Sinopharm. The Bacteroides dorei CCFM1429 involved in the following examples was deposited in the strain collection center of the Biotechnology Center of the School of Food Science and Engineering of Jiangnan University.

[0053] The method for detecting the number of viable bacteria involved in the following embodiments is based on the national standard "GB 4789.35-2016 National Food Safety Standard Food Microbiology Detection Lactic Acid Bacteria Detection".

[0054] The preparation method of the D-galactose solution involved in the following embodiments is as follows:

[0055] D-galactose solution: Weigh a certain amount of D-galactose, dissolve it in sterile saline to prepare D-galactose, and inject subcutaneously at 700 mg / kg body weight and 1000 mg / kg body weight.

[0056] Example 1: Screening and identification of Phocaeicola dorei CCFM1429

[0057] 1. Isolation and identification of Phocaeicola dorei CCFM1429

[0058] This strain was isolated from the feces of healthy adults, graded diluted with normal saline, spread on BHI solid medium, and incubated inverted in an anaerobic incubator at 37°C for 24 hours. Single colonies were picked and streaked on BHI medium for culture based on colony morphology.

[0059] After streaking the plate on the BHI solid culture medium, place it in an anaerobic incubator at 37°C and incubate it upside down for 48 hours. Pick a single colony and inoculate it into 5 mL of liquid MRS culture medium. After culturing it in an anaerobic incubator at 37°C for 24 hours, take 1.5 mL of the bacteria and centrifuge it at 6000 r / min for 3 minutes to remove the supernatant. Add 1 mL of 30% sterile glycerol for preservation. At the same time, keep a 1.5 mL portion of the bacteria, centrifuge it, remove the supernatant, and resuspend it in sterile water for strain identification.

[0060] The strain was sequenced and analyzed, and the sequence obtained by sequencing was compared with the nucleic acid sequence in Genbank. The result showed that the nucleic acid sequence similarity with that of Bacteroides dorei was greater than 99%. The strain was named Bacteroides dorei (Phocaeicola dorei) CCFM1429, and the Bacteroides dorei (Phocaeicola dorei) CCFM1429 has been preserved in Guangdong Provincial Microbial Culture Collection Center with a preservation number of GDMCC No: 65155.

[0061] 2. Morphology of Phocaeicola dorei CCFM1429

[0062] Phocaeicola dorei CCFM1429 was inoculated into BHI solid culture medium with L-cysteine ​​hydrochloride, and cultured anaerobically at 37°C for 48 hours. The colonies were observed and the bacteria were observed under a microscope. It was found that the colonies were milky white, smooth in surface, convex in the middle, and neat in edges.

[0063] 3. Preservation of Phocaeicola dorei CCFM1429

[0064] The strain was cultured in BHI medium, and Bacteroides dorei was cultured in an anaerobic incubator at 37°C for 24 hours for one generation. Single colonies on the solid medium of the second generation were selected and liquid cultured for the third generation under the same conditions as above. The tube containing the bacterial solution was centrifuged at 4°C and 8000g for 2 minutes, the supernatant was discarded, the precipitate was retained, and 1 mL of sterile 30% glycerol solution was added, and it was stored in a -80°C refrigerator after shaking.

[0065] Phocaeicola dorei CCFM1429 was inoculated into BHI liquid culture medium and cultured at 37°C for 48 h. The bacterial solution was then inoculated into fresh BHI liquid culture medium at an inoculum volume of 4% (v / v), and cultured under the same conditions for 36 h. The bacteria were centrifuged at 8000 r / min for 15 min, washed with 0.9% saline and centrifuged again at 8000 r / min for 10 min, the bacteria were collected, resuspended with 30% (v / v) glycerol solution, and resuspensions were prepared respectively, which were frozen at -80°C for use.

[0066] 4. Preparation of Phocaeicola dorei CCFM1429 bacterial suspension for intragastric administration

[0067] When Phocaeicola dorei CCFM1429 was used for oral administration to mice, the prepared resuspension was taken out from -80°C, centrifuged at 4°C, 8000r / min for 10min, the supernatant was discarded, the bacterial suspension was washed with 0.9% saline and resuspended to obtain a bacterial suspension for oral administration, and the bacterial concentration was: 2×10 8 CFU / mL.

[0068] Example 2: Effect of Phocaeicola dorei CCFM1429 on the Alleviation of Frailty Index in Aging Mice

[0069] Eight-week-old SPF-grade C57BL / 6J male mice were housed in the Experimental Animal Center of Jiangnan University, fed with ordinary feed, maintained at a constant temperature of 20-26°C, humidity of 40%-70%, noise level less than or equal to 60 dB, animal illumination of 15-20 LX, 12 h light, and 12 h dark (all animal experimental procedures were reviewed and approved by the Animal Welfare and Ethics Management Committee of Jiangnan University).

[0070] The experimental design is 20 weeks in total. The specific process is as follows: Figure 7 After entering the breeding facility, the mice were first adapted for one week. Galactose (CAS No. 59-23-4, Aladdin) was used to establish an aging model from 2 to 13 weeks. The control group mice were subcutaneously injected with sterile saline, and the other treatment groups were subcutaneously injected with 700 mg / kg BW / d of D-galactose dissolved in sterile saline. Because mice will develop a certain resistance to D-galactose, 1000 mg / kg BW / d of subcutaneous injection was continued from 14 to 20 weeks. At the same time, each group of mice was treated with the corresponding experiments. The control group and the model group were gavaged with 200 μL of sterile saline every day. The volume of injection liquid was the same for all groups.

[0071] The details are as follows:

[0072] Blank group: After entering the breeding facility, the mice were first adapted for one week. From 2 to 13 weeks, the mice in the blank group were subcutaneously injected with sterile saline; from 14 to 20 weeks, the mice were subcutaneously injected with sterile saline and gavaged with 200 μL of sterile saline daily.

[0073] Model group: After entering the breeding facility, mice were first adapted for one week. From week 2 to 13, the model group was subcutaneously injected with 700 mg / kg BW / d of D-galactose dissolved in sterile saline; from week 14 to 20, the mice were subcutaneously injected with 1000 mg / kg BW / d of D-galactose dissolved in sterile saline, and 200 μL of sterile saline was gavaged daily; food and water were freely available.

[0074] Experimental group: After entering the breeding facility, mice were first adapted for one week. From week 2 to 13, the model group was subcutaneously injected with 700 mg / kg BW / d of D-galactose dissolved in sterile saline; from week 14 to 20, the mice were subcutaneously injected with 1000 mg / kg BW / d of D-galactose dissolved in sterile saline, and were gavaged with 200 μL of Phocaeicola dorei CCFM1429 bacterial suspension (bacterial concentration was 2×10 8 CFU / mL); free access to food and water.

[0075] Drug group: After entering the breeding facility, mice were adapted for one week. From week 2 to 13, the model group was subcutaneously injected with 700 mg / kg BW / d of D-galactose dissolved in sterile saline; from week 14 to 20, the mice were subcutaneously injected with 1000 mg / kg BW / d of D-galactose dissolved in sterile saline, and were gavaged with spermidine 0.02 mg / kg body weight every day; they were allowed to eat and drink water freely.

[0076] Before killing the mice, the frailty of each aged mouse was scored, including the assessment of 13 variables, as shown in Table 1. The frailty index (FI) was calculated based on the scores of individual variables, and the specific scoring criteria were as follows: for each variable, 0 points represented no defect, 0.5 points represented mild defects, and 1 point represented severe defects. The total score was calculated for evaluation, and the total score was between 0 and 13.

[0077] Table 1: Mouse weakness index table

[0078]

[0079] Depend on Figure 1 It can be seen that the average frailty index of the blank group was 2.06, the average frailty index of the model group was 4.68, the average frailty index of the CCFM1429 group was 3.2, and the average frailty index of the spermidine group was 2. Compared with the model group, CCFM1429 reduced the frailty index of mice by 31.62%.

[0080] The above experimental results show that Phocaeicola dorei CCFM1429 can effectively reduce the frailty index of mice and improve the frail state of mice.

[0081] Example 3: Effect of Phocaeicola dorei CCFM1429 on the level of superoxide dismutase (SOD) in the liver of aged mice

[0082] The specific steps refer to Example 2. After the experiment, the mice were killed, the mouse livers were removed and placed in liquid nitrogen, stored in a -80°C refrigerator, and the tissues were rinsed with pre-cooled PBS (0.01M, pH=7.4) to remove residual blood and cut into pieces. Weigh about 1.2g of fresh tissue, add PBS at a weight-to-volume ratio of 1:9, add 1mL of universal protease inhibitor (Biyuntian) to every 100mL of this PBS, add an equal amount of grinding beads to each tissue, use a pre-cooled tissue grinder mold, and grind thoroughly on ice until no solids are completely eliminated. Further freeze-thaw repeatedly to fully lyse. Centrifuge the homogenate at 5000×g for 5-10 minutes, and take the supernatant and divide it into portions for later use. BCA method was used to measure protein concentration. 1.2 ml of protein standard solution (0.5 g bovine serum albumin, dissolved in distilled water and fixed to 100 ml, to make a 5 mg / ml solution, diluted ten times when used) was added to a tube of protein standard (30 mg BSA). After fully dissolved, a 25 mg / ml protein standard solution was prepared. Take an appropriate amount of 25 mg / ml protein standard and dilute to a final concentration of 0.5 mg / ml. According to the number of samples, 50 volumes of BCA reagent A (respectively weigh 10g BCA (1%), 20g Na2CO3·H2O (2%), 1.6g Na2C4H4O6·2H2O (0.16%), 4g NaOH (0.4%), 9.5g NaHCO3 (0.95%), add water to 1L, and adjust the pH to 11.25 with NaOH or solid NaHCO3) plus 1 volume of BCA reagent B (take 2g CuSO4·5H2O (4%), add distilled water to 50ml) (50:1) to prepare an appropriate amount of BCA working solution and mix thoroughly. Select an appropriate concentration within the standard concentration range of 0-1.5mg / mL to prepare a standard curve, and obtain the appropriate dilution multiple from the sample pre-experiment. In the formal experiment, add 20μL of sample or different concentrations of standard to each well, add 200μL BCA working solution to each well, and incubate at 37℃ in the dark for 30 minutes. The absorbance was measured at 562 nm using a microplate reader and the protein concentration of the sample was calculated.

[0083] The superoxide dismutase (SOD) content in mouse liver homogenate was detected according to the instructions of the liver superoxide dismutase (SOD) ELISA kit. The results are as follows Figure 2 shown.

[0084] The results show:

[0085] Depend on Figure 2It can be seen that the average value of liver superoxide dismutase (SOD) in the blank group was 27.90ng / mg protein, the average value of liver superoxide dismutase (SOD) in the model group was 16.254ng / mg protein, the average value of liver superoxide dismutase (SOD) in the CCFM1429 group was 26.84ng / mg protein, and the average value of liver superoxide dismutase (SOD) in the spermidine drug group was 29.51ng / mg protein. Compared with the model group, the liver SOD level of mice in the CCFM1429 group increased by 65.13% and recovered to the same level as the blank group.

[0086] The above experimental results show that Phocaeicola dorei CCFM1429 can effectively increase the level of superoxide dismutase (SOD) in mouse liver and improve the weak state of mice.

[0087] Example 4: Effect of Phocaeicola dorei CCFM1429 on the level of glutathione peroxidase (GSH-px) in the liver of aged mice

[0088] Specific implementation method Referring to Example 3, after obtaining mouse liver homogenate, the content of glutathione peroxidase (GSH-px) in the mouse liver homogenate was detected according to the instructions of the liver glutathione peroxidase (GSH-px) level ELISA kit. The results are as follows: Figure 3 shown.

[0089] The results show:

[0090] Depend on Figure 3 It can be seen that the average liver glutathione peroxidase (GSH-px) value of the blank group was 180.25U / mg protein, the average liver glutathione peroxidase (GSH-px) value of the model group was 77.07U / mg protein, the average liver glutathione peroxidase (GSH-px) value of the CCFM1429 group was 158.49U / mg protein, and the average liver glutathione peroxidase (GSH-px) value of the spermidine drug group was 161.38U / mg protein. Compared with the model group, the liver malondialdehyde (MDA) value of mice in the CCFM1429 group increased by 105.65%, which is equivalent to the regulatory effect of the drug.

[0091] The above experimental results show that Phocaeicola dorei CCFM1429 can effectively increase the level of glutathione peroxidase (GSH-px) in mouse liver and improve the weak state of mice.

[0092] Example 5: Effect of Phocaeicola dorei CCFM1429 on the reduction of IL-6 levels in colon homogenates of aged mice

[0093] Specific implementation method Referring to Example 2, after killing the mouse, the mouse colon was removed and placed in liquid nitrogen, stored in a -80°C refrigerator, and the tissue was rinsed with pre-cooled PBS (0.01M, pH=7.4) to remove residual blood and cut into pieces. Weigh about 1.2g of fresh weight tissue, add PBS at a weight-to-volume ratio of 1:9, add 1mL of universal protease inhibitor (Biyuntian) to every 100mL of this PBS, add an equal amount of grinding beads to each tissue, use a pre-cooled tissue grinder mold, and grind thoroughly on ice until no solids are completely gone. Further freeze and thaw repeatedly to fully lyse. Centrifuge the homogenate at 5000×g for 5-10 minutes, and take the supernatant and divide it into portions for later use. BCA method was used to measure protein concentration. 1.2 ml of protein standard solution (0.5 g bovine serum albumin, dissolved in distilled water and fixed to 100 ml, to make a 5 mg / ml solution, diluted ten times when used) was added to a tube of protein standard (30 mg BSA). After fully dissolved, a 25 mg / ml protein standard solution was prepared. Take an appropriate amount of 25 mg / ml protein standard and dilute to a final concentration of 0.5 mg / ml. According to the number of samples, 50 volumes of BCA reagent A (respectively weigh 10g BCA (1%), 20g Na2CO3·H2O (2%), 1.6g Na2C4H4O6·2H2O (0.16%), 4g NaOH (0.4%), 9.5g NaHCO3 (0.95%), add water to 1L, and adjust the pH to 11.25 with NaOH or solid NaHCO3) plus 1 volume of BCA reagent B (take 2g CuSO4·5H2O (4%), add distilled water to 50ml) (50:1) to prepare an appropriate amount of BCA working solution and mix thoroughly. Select an appropriate concentration within the standard concentration range of 0-1.5mg / mL to prepare a standard curve, and obtain the appropriate dilution multiple from the sample pre-experiment. In the formal experiment, add 20μL of sample or different concentrations of standard to each well, add 200μL BCA working solution to each well, and incubate at 37℃ in the dark for 30 minutes. The absorbance was measured at 562 nm using a microplate reader and the protein concentration of the sample was calculated.

[0094] The IL-6 content in the mouse colon homogenate was detected according to the instructions of the IL-6 ELISA kit. Figure 4 shown.

[0095] The results show:

[0096] Depend on Figure 4It can be seen that the average IL-6 in the colon homogenate of the blank group was 306.95pg / mg protein, the average IL-6 in the colon homogenate of the model group was 366.24pg / mg protein, the average IL-6 in the colon homogenate of the CCFM1429 group was 273.10pg / mg protein, and the average IL-6 in the colon homogenate of the spermidine group was 312.41pg / mg protein. Compared with the model group, the IL-6 value in the colon homogenate of mice in the CCFM1429 group decreased by 25.43%.

[0097] The above experimental results show that Phocaeicola dorei CCFM1429 can effectively reduce IL-6 in the colon homogenate of mice and improve the weak state of mice.

[0098] Example 6: Effect of Phocaeicola dorei CCFM1429 on the reduction of TNF-alpha levels in colon homogenates of aged mice

[0099] Specific implementation method Referring to Example 5, after obtaining the mouse colon homogenate, the TNF-alpha content in the mouse colon homogenate was detected according to the instructions of the TNF-alpha ELISA kit. The results are as follows: Figure 5 shown.

[0100] The results show:

[0101] Depend on Figure 5 It can be seen that the average TNF-alpha in the colon homogenate of the blank group was 281.00pg / mg protein, the average TNF-alpha in the colon homogenate of the model group was 324.56pg / mg protein, the average TNF-alpha in the colon homogenate of the CCFM1429 group was 255.77pg / mg protein, and the average TNF-alpha in the colon homogenate of the spermidine group was 267.28pg / mg protein. Compared with the model group, the TNF-alpha value in the colon homogenate of the mice in the CCFM1429 group decreased by 21.20%.

[0102] The above experimental results show that Phocaeicola dorei CCFM1429 can effectively reduce TNF-alpha in the colon homogenate of mice and improve the weak state of mice.

[0103] Example 7: Effect of Phocaeicola dorei CCFM1429 on the grip strength of aged mice

[0104] Specific implementation method Referring to Example 2, before killing the mouse, the mouse was placed flat on the T-frame of the grip strength meter (Jinan Yiyan Technology Co., Ltd.), the tail of the mouse was grasped and slowly pulled backwards, and the data was read. The results were as follows: Figure 6 shown.

[0105] The results show:

[0106] Depend on Figure 6 It can be seen that the average grasping force of mice in the blank group was 101.26g, the average grasping force of mice in the model group was 80.54g, the average grasping force of mice in the Dorenia spp. CCFM1429 group was 96.56g, and the average grasping force of mice in the spermidine drug group was 68.27g. Therefore, the effect of Dorenia spp. CCFM1429 on improving the grasping force of mice is better than that of drugs, and can restore the grasping force to a level equivalent to that of the blank group.

[0107] The above experimental results show that Phocaeicola dorei CCFM1429 can effectively increase the grip level of mice and improve the weak state of mice.

[0108] Example 8: Preparation of Phocaeicola dorei CCFM1429 microbial preparation

[0109] Preparation of liquid microbial preparation: Phocaeicola dorei CCFM1429 was cultured under anaerobic conditions for 24 h, the culture medium was removed by centrifugation, and the cells of Phocaeicola dorei CCFM1429 were resuspended in sterile saline to ensure that the content of Phocaeicola dorei CCFM1429 was not less than 2 × 10 8 CFU / mL, prepare bacterial suspension. If necessary, one or more of skimmed milk powder, glycerol, maltodextrin, trehalose, and sodium L-glutamate can be added to the bacterial suspension as a protective agent.

[0110] Preparation of solid microbial preparation: The cultured Bacillus multifiliis CCFM1429 cells are mixed with a protective agent and then freeze-dried and sprayed to obtain a solid microbial preparation. The protective agent includes one or more of skimmed milk powder, glycerol, maltodextrin, trehalose, and sodium L-glutamate, wherein the content of Bacillus multifiliis CCFM1429 is not less than 2×10 8 CFU / g.

[0111] Although the present invention has been disclosed as above in the form of a preferred embodiment, it is not intended to limit the present invention. Anyone familiar with this technology can make various changes and modifications without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention should be based on the definition of the claims.

Claims

1. A strain of Phocaeicola dorei CCFM1429, which was deposited in Guangdong Provincial Microbiological Culture Collection Center on September 20, 2024, with a deposit number of GDMCC No: 65155.

2. A microbial preparation containing the Bacillus multifiliis CCFM1429 according to claim 1.

3. The microbial preparation according to claim 2, characterized in that The microbial preparation contains one or more of the living cells, fermentation broth, lysate or extract of the Bacillus multifiliis CCFM1429.

4. A medicine containing the Bacillus multifiliis CCFM1429 according to claim 1, or the microbial preparation according to claim 2 or 3.

5. The drug according to claim 4, characterized in that In the drug, the added amount of Bacteroides dorenoides CCFM1429 is not less than 2×10 8 CFU / mL or 2×10 8 CFU / g.

6. The drug according to claim 4 or 5, characterized in that The dosage form of the drug includes powder, granules, capsules, tablets, pills or liquid preparations.

7. A method for preparing the microbial preparation according to claim 2 or 3, characterized in that: The cultured Bacillus doreid CCFM1429 cells are mixed with a protective agent to obtain the product.

8. The method according to claim 7, characterized in that The protective agent includes one or more of skimmed milk powder, glycerol, maltodextrin, trehalose, and sodium L-glutamate.

9. Use of the Bacillus doreniformis CCFM1429 described in claim 1, or the microbial preparation described in claim 2 or 3, in the preparation of a drug for alleviating aging symptoms.

10. The use according to claim 9, characterized in that: The drug is used for at least one of (a) to (h): (a) reduce the frailty score index of aging individuals; (b) reduce the level of MDA in liver tissue of aging individuals; (c) increasing the level of SOD enzyme in liver tissue of aging individuals; (d) increasing the level of CAT enzyme in liver tissue of aging individuals; (e) increasing the level of GSH-px enzyme in liver tissue of aging individuals; (f) increasing the level of acetylcholine in the brain tissue of aging individuals; (g) reducing the expression level of pro-inflammatory factors in colon tissue of aging individuals; the pro-inflammatory factors include IL-6 and TNF-alpha; (h) Improving grip strength in aging individuals.

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