Lactobacillus reuteri and application thereof

Abstract

The invention discloses a strain of lactobacillus reuteri YSLR01 and application of the strain of lactobacillus reuteri YSLR01 in preparation of anti-inflammatory, anti-oxidation, fat-reducing and anti-aging products. According to the invention, a YSLR01 strain with anti-inflammatory, anti-oxidation, fat-reducing and anti-aging effects is separated from excrement of healthy human beings, the strain is preserved in China General Microbiological Culture Collection Center on September 2, 2024, and the preservation number is CGMCC NO.31800. The invention further discloses a preparation method of the YSLR01 strain. Researches show that the YSLR01 strain can significantly reduce secretion of inflammatory factors in a lipopolysaccharide (LPS)-induced RAW264.7 cell inflammation model, improve the oxidation resistance of caenorhabditis elegans, enhance the body length, action ability and reproductive capacity of the caenorhabditis elegans, reduce fat deposition in the body and delay aging of the body.

Description

Technical Field

[0001] The present invention belongs to the technical field of probiotics, and more specifically, relates to Lactobacillus reuteri YSLR01 and its application in preparing anti-inflammatory, anti-oxidative, fat-reducing and anti-aging products. Background Art

[0002] Inflammation is a common defense measure. When the body is infected or the tissue is damaged, timely initiation of inflammatory response is beneficial to eliminate the infection source and repair tissue. Inflammatory response within a controllable range is beneficial. Excessive inflammation, such as septic shock, is harmful to the body. There is also a special inflammatory state - parainflammation. For example, the fat tissue of obese people shows chronic inflammation. This inflammation often starts with tissue dysfunction and is different from the inflammatory response caused by infection and tissue damage. It is usually characterized by changes in critical values ​​in the body. Obesity, type 2 diabetes, hypertension and a series of liver and intestinal diseases are all related to parainflammation.

[0003] Aging is a series of slow, complex and inevitable physiological processes that occur as the body ages, accompanied by the degeneration and loss of the overall function of cells, tissues and organs. In the normal aging process of the body, not only will many adverse changes occur, but also a series of changes at the genetic, cellular and epigenetic levels will occur, and the susceptibility to diseases will increase. Diseases such as diabetes, neurodegenerative diseases, immune diseases, cardiovascular diseases, and cancer are all related to aging.

[0004] Probiotics refer to living microorganisms that can have a beneficial effect on the host when ingested in sufficient doses. Studies have shown that probiotics can delay aging by improving intestinal flora, increasing the abundance of beneficial bacteria, inhibiting the colonization and reproduction of harmful bacteria and pathogens, reducing the damage of toxic and harmful products to the body, reducing inflammation, and improving the body's antioxidant and immune regulation capabilities. Currently reported probiotics with anti-aging effects include Lactobacillus and Bifidobacterium, such as Lactobacillus casei LTL1361, Lactobacillus paracasei NCC2461, Bifidobacterium infantis, Bifidobacterium longum, etc. However, most strains are strain-specific, and a strain has only a single function. Therefore, it is of broad and practical application value to continue to explore probiotic strains that have anti-inflammatory, antioxidant, fat-reducing, and anti-aging effects.

[0005] Lactobacillus reuteri (L. reuteri) is a lactic acid bacterium that exists in the intestines of almost all mammals, and is most commonly found in the proximal digestive tract of the host. It is a Gram-positive bacterium with slightly irregular, rounded-end Campylobacter. The bacterium has a strong adhesion ability to the intestinal mucosa, which can improve the distribution of intestinal flora, antagonize the colonization of harmful bacteria, and avoid intestinal diseases. In recent years, it has been increasingly used in the field of human health. According to research reports, L. reuteri can produce a non-protein broad-spectrum antibacterial substance called "reuterin", which is harmless to humans and animals and can widely inhibit the growth of Gram-positive bacteria, Gram-negative bacteria, yeasts, fungi and pathogens. In addition, L. reuteri can also improve glucose homeostasis and insulin sensitivity, which is beneficial to alleviate dyslipidemia. However, at present, the anti-inflammatory, antioxidant, fat-reducing and anti-aging effects of Lactobacillus reuteri are not significant. Summary of the invention

[0006] The technical problem to be solved by the present invention is to overcome the defects and shortcomings of the above-mentioned problems and to provide a Lactobacillus reuteri YSLR01 and its application in anti-inflammatory, anti-oxidation, fat reduction and anti-aging.

[0007] The second object of the present invention is to provide the use of the above-mentioned Lactobacillus reuteri in the preparation of anti-inflammatory products.

[0008] The third object of the present invention is to provide the use of the above-mentioned Lactobacillus reuteri in the preparation of antioxidant products.

[0009] The fourth object of the present invention is to provide the use of the above-mentioned Lactobacillus reuteri in the preparation of fat-reducing products.

[0010] The fifth object of the present invention is to provide the use of the above-mentioned Lactobacillus reuteri in the preparation of anti-aging products.

[0011] The above technical purpose of the present invention is achieved through the following technical scheme: the strain of Lactobacillus reuteri is deposited in the China General Microbiological Culture Collection Center (CGMCC), the preservation date is September 2, 2024, and the preservation number is CGMCC NO.31800.

[0012] Furthermore, the bacterial 16SrDNA gene sequence is shown in SEQ ID NO.1.

[0013] A strain of Lactobacillus reuteri is used as a probiotic in the preparation of anti-inflammatory, antioxidant, fat-reducing and anti-aging products.

[0014] Furthermore, the anti-inflammatory refers to inhibiting the inflammatory response in LPS-induced RAW264.7 cells; the antioxidant refers to enhancing the antioxidant capacity of the nematode body; the fat reduction refers to reducing fat deposition in Caenorhabditis elegans; and the anti-aging refers to extending the lifespan of nematodes and increasing their motility and fertility.

[0015] Furthermore, the LPS is lipopolysaccharide, which serves as an inducer in cellular inflammatory response.

[0016] Furthermore, the RAW264.7 cells are mouse macrophages, and the LPS-induced RAW264.7 cell inflammation model is established, which is the basis for screening probiotics with anti-inflammatory activity.

[0017] A product with anti-inflammatory, anti-oxidative, fat-reducing and anti-aging effects, comprising the above-mentioned YSLR01 strain and / or its bacterial liquid.

[0018] Furthermore, the product is a medicine, functional food, health product, feed or an additive thereof, and the product is in the form of powder, tablet, granule, capsule or liquid.

[0019] In summary, the present invention has the following beneficial effects: The Lactobacillus reuteri YSLR01 provided by the present invention can significantly reduce the secretion of inflammatory factors in the RAW264.7 cell inflammation model induced by lipopolysaccharide (LPS), enhance the antioxidant capacity of Caenorhabditis elegans, increase the body length, mobility and reproductive capacity of nematodes, reduce fat deposition in the body, and delay body aging. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 This is a graph showing the effect of different concentrations of probiotic YSLR01 supernatant on the survival rate of RAW264.7 cells; Figure 2 The content of inflammatory factors NO (A), IL-6 (B), and IL-1β (C) in the cell supernatant of the RAW264.7 inflammatory model induced by LPS was treated with the supernatant of probiotic YSLR01 within a relatively safe concentration range for 24 hours; Figure 3 This is a diagram showing the effect of AKK-PL001 on the life span of C. elegans N2; Figure 4 This is a diagram showing the changes in body length (A), mobility (B), and fertility (C) after feeding Caenorhabditis elegans with AKK-PL001; Figure 5 This is a graph showing the effect of feeding YSLR01 on mitochondrial membrane potential; Figure 6 This is a graph showing the effect of feeding YSLR01 on the fat content of nematodes; Figure 7The antioxidant properties of AKK-PL001 (A: MDA content; B: SOD activity; C: GSH content; D: CAT activity). DETAILED DESCRIPTION

[0021] The present invention is described in detail below in conjunction with the accompanying drawings and embodiments.

[0022] The reagents or formulations used in the following examples are: Mouse RAW264.7 cells were purchased from Saiye (Suzhou) Biotechnology Co., Ltd. The nitric oxide (NO) content detection kit was purchased from Bio-Technology Co., Ltd.; the mouse IL-6 and IL-1β ELISA kits were purchased from Ecosine Biotechnology (Taicang) Co., Ltd.

[0023] NGM solid medium: 3 g sodium chloride, 2.5 g tryptone, 17 g agar powder, 25 mL potassium phosphate buffer (1 mol / L), 1 mL CaCl2 solution (1 mol / L), 1 mL MgSO4 solution (1 mol / L), distilled water to 1000 mL, autoclave at 121 °C for 20 min. When the temperature drops to about 60 °C, add 1 mL cholesterol solution (5 mg / mL, dissolved in 95% ethanol aqueous solution) under sterile conditions, shake well, and pour on the plate.

[0024] MRS liquid culture medium: 5 g casein peptone, 10 g beef extract, 5 g yeast extract, 2 g diamine hydrogen citrate, 20 g glucose, 5 g sodium acetate, 2 g potassium dihydrogen phosphate, 0.58 g magnesium sulfate, 0.25 g manganese sulfate, then add 1 mL Tween-80, dilute to 1000 mL with distilled water, and sterilize at 121°C under high pressure for 20 min.

[0025] LB liquid medium: 5 g yeast powder, 10 g tryptone, 10 g NaCl, add distilled water to 1000 mL, and sterilize at 121 °C for 20 min.

[0026] Wild-type Caenorhabditis elegans N2 and Escherichia coli OP50 were purchased from Fujian Shangyuan Bioscience and Technology Co., Ltd. The kits for measuring malondialdehyde (MDA) content, glutathione (GSH) content and catalase (CAT) enzyme activity were purchased from Nanjing Jiancheng Biotechnology, and the kit for measuring superoxide dismutase (SOD) enzyme activity was purchased from Beyotime Biotechnology.

[0027] Example 1 Isolation and identification of strains Isolation of strains Sterile oxygen-free saline was used to dilute the stool samples in a gradient manner, and the concentration gradient was 10 -1 to 10 -6 Take 10 -4 , 10 -5 , 10 -6 Three gradients of bacterial liquid are applied to the specific screening medium. After evenly applied, the plate is inverted and placed in an anaerobic culture bag for culture. The strain growth time is about 24h-48h. Select and isolate a single colony suspected of being Lactobacillus reuteri from the colony, and transfer it to a new culture medium by streaking to continue culturing. Pick the flat, slightly gray single colony that has grown, use a 10μL pipette tip to gently scratch the surface of the colony, blow it evenly in 10μL pure water, then take 3μL of water and put it into a PCR tube, add universal primers, mix dye, and DD water, and perform PCR amplification.

[0028] Molecular Biology Identification (1) Pipette 3 μL of bacterial solution and prepare the PCR amplification system and run the program according to the following system (see Table 1 and Table 2) for PCR amplification.

[0029] Table 1: PCR reaction system Table 2: PCR reaction program (2) Prepare 1.5% agarose gel and add 10 μL Ultra GelRed (10000×) nucleic acid dye per 100 mL.

[0030] (3) Add 5 μL of the above PCR reaction solution to each well and use DL2000 marker as a band size control. The band size is around 1500 bp, which can be preliminarily determined to be Lactobacillus reuteri.

[0031] (4) The samples initially identified as Lactobacillus reuteri were further amplified using universal primers. The amplified PCR stock solution was sent to Qingke Biotechnology Co., Ltd. for sequencing to obtain the comparison results.

[0032] (5) After comparison, the sample identified as Lactobacillus reuteri was expanded and cultured in MRS liquid medium for 48 hours. 100 μL of the bacterial solution was aspirated, the bacterial solution was centrifuged and the supernatant was discarded. 100 μL of sterile water was added for re-dissolution, and PCR amplification was performed again. After amplification, it was sent to Qingke Biotechnology for sequencing. If it was identified as Lactobacillus reuteri, it was pure Lactobacillus reuteri. The bacterial solution was mixed with 60% glycerol solution in a 1:1 ratio and stored in a -80°C refrigerator.

[0033] Example 2 Effects of different concentrations of YSLR01 supernatant on the survival rate of RAW264.7 cells 1. Cell recovery After taking out the RAW264.7 cell cryopreservation tube from the liquid nitrogen tank, immediately put it into a water bath and shake it quickly to melt it in a short time. Use a pipette to slowly transfer the cell suspension to a centrifuge tube in the clean bench and centrifuge it at 1000rpm for 5 min. At the same time, add 7 mL of RPMI-1640 complete medium prepared with 1% triple antibody, 10% fetal bovine serum, and RPMI-1640 basal medium to the cell culture dish. After centrifugation, take out the centrifuge tube, discard the supernatant in the clean bench, add 1 mL of RPMI-1640 complete medium, resuspend the cells, and then transfer the cells to the culture dish, gently blow and aspirate, cover the culture dish lid and place it in a 37°C, 5% CO2 incubator for culture.

[0034] 2. Cell passaging After the cells are revived and placed in the incubator for 24 hours, take out the cells and observe them under a microscope. When the cell growth density is about 80% of the area at the bottom of the culture bottle, the cells can be passaged. When passaged, first pour out the old culture medium in the culture dish, then slowly rinse twice with PBS buffer solution, discard the PBS solution, add 1mL of trypsin digestion solution, let it stand for 2-3 minutes, and finally add 4mL of RPMI-1640 complete medium to terminate the digestion. Centrifuge at 1000 rpm for 5 minutes, then add 1mL of RPMI-1640 complete medium, resuspend the cells, transfer them to the culture dish, cover the culture dish, and place it in an incubator at 37°C and 5% CO2.

[0035] 3. CCK-8 cytotoxicity test (1) Cell plating: RAW264.7 cells with good growth status were plated at 5×10 4 / mL, 100 μL per well was inoculated into a 96-well cell culture plate, and the culture plate was placed in an incubator and cultured until the adhesion rate reached 80%-90%, and then cultured overnight; (2) Co-culture of probiotics and cells: RAW264.7 cells were treated with different concentrations of YSLR01 supernatant (25%, 12.5%, and 6.25%) using complete culture medium; four replicate wells were set up in each group and cultured in an incubator for 24 h; (3) Add CCK-8: Dilute the CCK-8 solution to 10% with complete culture medium, discard the old culture medium, add 100 µL of 10% CCK-8 solution to each well, place the well in an incubator and culture for 40 min. Measure the absorbance at 450 nm using a microplate reader.

[0036] 5. Test results The results are as follows Figure 1 As shown in the figure, the survival rate of RAW264.7 cells increased significantly with the decrease of the concentration of YSLR01 strain supernatant. The cytotoxicity of the strain supernatant was greater when the concentration was 25%, and the cell survival rate was high when the concentration was 12.5%, without cytotoxicity. Subsequently, the strain supernatant at a concentration of 12.5% ​​was selected to act on RAW264.7 cells.

[0037] Example 3 Effect of YSLR01 supernatant on NO, IL-6 and IL-1β levels in LPS-induced RAW264.7 cells 1. Test methods RAW264.7 cells in good growth condition were used at 2×10 6 / mL, 2mL per well was inoculated on a six-well cell culture plate. After the cells were firmly attached to the wall, the supernatant was discarded and the blank control group (CK), LPS-induced group, and YSLR01 supernatant treatment group were set. 2mL of complete medium was added to each well of the blank control group; 2mL of complete medium containing 2μg / mL LPS was added to each well of the LPS-induced group; and 2mL of complete medium containing 12.5% ​​and 2μg / mL LPS was added to each well of the YSLR01 supernatant group. After 24h, the cell supernatant was collected and the secretion of NO, IL-6, and IL-1β in the cell supernatant was determined using a kit.

[0038] 2. Test results The results are as follows Figure 2 As shown in the figure, compared with the blank control group (CK), LPS (2 μg / mL) stimulated RAW264.7 cells, which significantly increased the levels of inflammatory factors NO, IL-6, and IL-1β in the cell supernatant (p < 0.01). YSLR01 supernatant treatment significantly weakened the inflammatory effect of LPS. Figure 2 As shown in Figure A, YSLR01 supernatant treatment significantly attenuated the LPS-induced inflammatory level by 92.9%; Figure 2 As shown in Figure B, YSLR01 supernatant treatment significantly attenuated the LPS-induced inflammatory level by 85.8%; for IL-1β, Figure 2 As shown in C, YSLR01 supernatant treatment significantly reduced the inflammation by 19.3%, indicating that it has effective therapeutic ability against LPS-induced RAW264.7 cell inflammation.

[0039] Example 4 Lifespan test of Lactobacillus reuteri YSLR01 on wild-type Caenorhabditis elegans N2 (hereinafter referred to as nematodes) 1. Activation of bacteria Take 200 μL of Lactobacillus reuteri YSLR01 frozen at -80°C, inoculate it into 10 mL of MRS liquid culture medium, and culture it in a 37°C incubator for 24 to 48 hours to activate the strain.

[0040] 2. Cultivation of nematodes Escherichia coli OP50 (OD=0.5) was spread on NGM solid culture medium as nematode food, and then the nematodes were picked and placed on the surface of NGM solid culture medium, and cultured in a 20℃ biochemical incubator. During the culture period, the nematodes needed to be transferred to a fresh NGM plate coated with Escherichia coli OP50 every other day.

[0041] 3. Synchronization of nematodes Collect L4 nematodes into a centrifuge tube, add 1 mL of M9 buffer to wash away excess E. coli OP50, let stand and precipitate, remove the supernatant, and repeat the wash 3 times. Then add 1 mL of lysis buffer and shake thoroughly, remove the supernatant, and then centrifuge (3000 r / min, 1 min) with M9 buffer to wash 3 times, remove the supernatant and retain the eggs, transfer the eggs to a new NGM plate, and incubate in a 20 ℃ biochemical incubator. After about 16 to 18 hours, they will grow into L1 larvae.

[0042] 4. Determination of nematode lifespan Lactobacillus reuteri YSLR01 and Escherichia coli OP50 were cultured in LB liquid medium to the logarithmic phase, centrifuged at 8000 r / min at 4°C for 10 min, and Lactobacillus reuteri YSLR01 cells were collected as food. The cells fed only with Escherichia coli OP50 were used as a control and fed to nematodes to measure their lifespan.

[0043] 90 L1 larvae synchronized in the above steps were selected and placed on NGM plates containing Lactobacillus reuteri YSLR01 and Escherichia coli OP50, respectively, and cultured in a 20 ℃ biochemical incubator. All surviving nematodes were selected and transferred to fresh NGM plates coated with bacterial solution every other day to prevent newly hatched nematodes from affecting the experimental results. The experiment continued until all nematodes died, and the nematode lifespan curve was drawn.

[0044] 5. Test results The results are as follows Figure 3 As shown, compared with the average lifespan (10 d) and maximum lifespan (19 d) of nematodes fed with Escherichia coli OP50, the average lifespan of Lactobacillus reuteri YSLR01 was significantly extended by 31.2% (p<0.05), and the maximum lifespan was extended by 26 days, indicating that Lactobacillus reuteri YSLR01 can significantly extend the lifespan of nematodes.

[0045] Example 5 Effects of Lactobacillus reuteri YSLR01 on the body length, motility and fecundity of nematodes 1. Test methods (1) To verify whether the Lactobacillus reuteri YSLR01 strain affects the body length of nematodes, nematodes cultured on the third day were selected and soaked in a 70°C hot water bath for 30 min to make the nematodes stiff. The nematodes were then placed on a glass slide and their body lengths were observed and measured using an optical microscope. Each experiment was repeated for at least 10 nematodes.

[0046] (2) Exercise ability is one of the key indicators of health. Exercise status and exercise ability will gradually deteriorate and weaken as the body ages. The nematodes were cultured with the Lactobacillus reuteri YSLR01 strain in the same manner as in Example 1. The control group was fed with Escherichia coli OP50 alone. On the third day, 20 nematodes were randomly selected and placed on a fresh NGM plate. After the nematodes moved freely for 30 seconds, the frequency of body bending and swinging within 20 seconds was observed using a stereo microscope. One body bend refers to the body bending from one direction to another and then returning to the original direction.

[0047] (3) The anti-aging drugs or strains to be screened should not damage the reproductive capacity of the body as a prerequisite. Reproductive capacity is one of the most critical indicators of health. Therefore, the effect of Lactobacillus reuteri YSLR01 on the reproductive capacity of nematodes was verified by using the number of offspring as an indicator. According to the method described in Example 4, the nematodes were cultured with the strain to be tested until the L4 stage, and one nematode was randomly selected and placed on the NGM plate coated with Lactobacillus reuteri YSLR01 and Escherichia coli OP50, with 5 parallels in each group. During the nematode egg-laying period, the egg-laying adults were transferred to a new plate every 1 day, and the old plate continued to be placed in a 20°C biochemical incubator until the eggs hatched, and the total number of hatched larvae on the plate was counted until the nematode eggs hatched.

[0048] 2. Experimental results Compared with the E. coli OP50 control group, the body length of nematodes in the Lactobacillus reuteri YSLR01 group increased by 12.0% and 12.69% on the 3rd and 12th days, respectively ( Figure 4 A in the figure, p<0.05), the body bending and swinging frequencies increased by 22.0% and 34.0% ( Figure 4 B in the figure, p<0.05), the total egg laying increased by 19.8% ( Figure 4 C in (p<0.05). This indicates that feeding Lactobacillus reuteri YSLR01 can significantly enhance the body length, mobility and reproductive capacity of nematodes, and has good anti-aging properties.

[0049] Example 6 Effects of Lactobacillus reuteri YSLR01 on fat content and mitochondrial membrane potential in nematodes 1. Test methods (1) Lactobacillus reuteri YSLR01 was used to culture nematodes. The culture method was the same as that in Example 4. Escherichia coli OP50 was used as the control group. After continuing to culture for 3 days at the L4 stage, 20 nematodes were randomly selected to measure the nematode fat by Oil Red O staining. The nematodes were rinsed 3 times with M9 buffer, the throat was sealed with 25 mM levamisole hydrochloride, 200 μL of 4% paraformaldehyde solution was added and allowed to stand for 20 minutes, the solution was removed and then frozen and thawed with liquid nitrogen 3 times, and then stained with 60% isopropanol Oil Red O. After 5 hours, the excess dye was rinsed with M9 buffer and observed with an optical microscope.

[0050] (2) Caenorhabditis elegans was cultured with Lactobacillus reuteri YSLR01 in the same manner as in Example 4. Escherichia coli OP50 was used as a control group. After culturing for 3 days at the L4 stage, 20 nematodes were randomly selected and the mitochondrial membrane potential was measured using JC-1 dye. The nematodes were incubated with the freshly prepared JC-1 dye solution in the dark for 2 hours. The dye on the surface of the nematodes was washed off with M9 buffer, and red fluorescence (emission wavelength of about 590 nm) and green fluorescence (emission wavelength of 529 nm) were observed under a fluorescence microscope, photographed and recorded, and quantitatively calculated using Image J software.

[0051] 2. Test results Compared with the E. coli OP50 control group, the fat content of nematodes fed with Lactobacillus reuteri YSLR01 was significantly reduced ( Figure 5 A in the figure). Using Image J software to analyze the red light optical density of fat, it was found that the red light optical density of nematode fat was significantly reduced after the intervention of Lactobacillus reuteri YSLR01, decreasing by 26.1% ( Figure 5 B, p<0.05). JC-1 staining showed increased red fluorescence and decreased green fluorescence compared with OP50 ( Figure 6 A in the figure), and the red-green fluorescence ratio increased by 63.2% ( Figure 6 B in (p<0.05), indicating that Lactobacillus reuteri YSLR01 significantly reduced fat deposition in nematodes, thereby affecting nematode fat metabolism, slowing down the decline in mitochondrial membrane potential, and reducing mitochondrial oxidative damage.

[0052] Example 7 Effect of Lactobacillus reuteri YSLR01 on the Antioxidant Capacity of Nematodes 1. Test methods The nematodes were cultured using Lactobacillus reuteri YSLR01 strain, and the culture method was the same as in Example 4. Escherichia coli OP50 was fed alone as the control group. The nematodes were collected after 3 days of continuous culture in the L4 stage, washed 3 times with M9 buffer, and then the nematodes were allowed to settle naturally. The nematodes were placed in a homogenizer, ground on ice, and centrifuged at 4 ° C, 4000 r / min to obtain the supernatant, which was the protein homogenate. The malondialdehyde (MDA) content, superoxide dismutase (SOD) enzyme activity, glutathione (GSH) content, and catalase (CAT) enzyme activity were measured according to the operating instructions of the kit.

[0053] 2. Test results Compared with the control group of Escherichia coli OP50, the MDA content in nematodes fed with Lactobacillus reuteri YSLR01 was significantly reduced by 36.4% ( Figure 7 A in the experiment, p<0.05), SOD activity was significantly increased by 59.7% ( Figure 7 B, p<0.05), GSH content increased significantly by 33.6% ( Figure 7 C in the experiment, p<0.05), CAT activity was significantly increased by 57.1% ( Figure 7 The above results indicate that Lactobacillus reuteri YSLR01 can enhance the antioxidant capacity of nematodes by increasing SOD activity, CAT activity and GSH content and reducing MDA content in the body.

[0054] In summary, the YSLR01 strain provided by the present invention can significantly reduce the secretion of inflammatory factors in the RAW264.7 cell inflammation model induced by lipopolysaccharide (LPS), enhance the antioxidant capacity of Caenorhabditis elegans, increase the body length, mobility and reproductive capacity of nematodes, reduce body fat deposition, and delay body aging. This indicates that the strain can be used as a probiotic to prepare or develop products with anti-inflammatory, antioxidant, fat-reducing and anti-aging effects.

[0055] The above is only a preferred embodiment of the present invention, and the protection scope of the present invention is not limited to the above embodiments. All technical solutions under the concept of the present invention belong to the protection scope of the present invention. It should be pointed out that for ordinary technicians in this technical field, some improvements and modifications without departing from the principle of the present invention should also be regarded as the protection scope of the present invention.

Claims

1. A strain of Lactobacillus reuteri YSLR01, which is deposited in the China General Microbiological Culture Collection Center (CGMCC) on September 2, 2024, with a deposit number of CGMCC NO.31800.

2. A Lactobacillus reuteri according to claim 1, characterized in that: The bacterial 16SrDNA gene sequence is shown in SEQ ID NO.

1.

3. Use of a Lactobacillus reuteri strain according to claim 1 as a probiotic in the preparation of anti-inflammatory, antioxidant, fat-reducing and anti-aging products.

4. The use according to claim 3, characterized in that: The anti-inflammatory refers to inhibiting the inflammatory response in RAW264.7 cells induced by LPS; the antioxidant refers to enhancing the antioxidant capacity of the nematode body; the fat reduction refers to reducing fat deposition in Caenorhabditis elegans; and the anti-aging refers to extending the lifespan of nematodes and increasing their motility and fertility.

5. The use according to claim 4, characterized in that: The LPS is lipopolysaccharide, which acts as an inducer in cellular inflammatory response.

6. The use according to claim 4, characterized in that: The RAW264.7 cells are mouse macrophages, and the LPS-induced RAW264.7 cell inflammation model is established, which is the basis for screening probiotics with anti-inflammatory activity.

7. A product with anti-inflammatory, anti-oxidant, fat-reducing and anti-aging effects, characterized in that: Containing the YSLR01 strain and / or its bacterial solution as described in claim 1.

8. The product according to claim 7, characterized in that: The product is a medicine, functional food, health product, feed or its additive, and its product form is powder, tablet, granule, capsule or liquid.

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