Application of magnetotactic bacteria AMB-1 in improving health and delaying senescence
By using the magnetotropic bacteria AMB-1 to prepare drugs or health products, the technical problems of delaying aging are solved, and the health status of C. elegans is significantly improved, ROS accumulation is reduced, lipofuscin accumulation is reduced, and life span is prolonged.
Patent Information
- Application Number
- CN202510384097.9
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-03-28
- Publication Date
- 2025-08-01
AI Technical Summary
The application of magnetizing bacteria in the prior art in delaying aging has not been reported, and there is a lack of effective methods to improve health and delay aging.
Drugs or health care products are prepared using the magnetotaxis bacteria AMB-1, and by adding them to the culture medium of C. elegans as a food supplement, it reduces ROS levels, reduces lipofuscin accumulation, improves nematode survival rate, and prolongs life.
It significantly improves the health status of C. elegans, reduces ROS accumulation, reduces lipofuscin accumulation, prolongs the lifespan of nematodes, and has good biosafety and high efficiency.
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Figure CN120392826A_ABST
Abstract
Description
Technical Field
[0001] The present invention relates to the technical field of microbial applications, and in particular to an application of magnetotactic bacteria Magnetospirillummagneticum AMB-1 in improving health and delaying aging. Background Art
[0002] Aging is an inevitable part of life, characterized by a gradual decline in physiological functions and increased susceptibility to age-related diseases. Currently, the aging population is accelerating, bringing with it numerous challenges. The rapid growth of the elderly population has led to a surge in demand for medical and elderly care services, but resources are unevenly distributed. Urban elderly care beds are scarce, while rural facilities are underutilized, creating a supply-demand imbalance. Therefore, delaying aging has gradually become a major goal of modern biomedical research. Extending healthy lifespan not only delays the onset of age-related diseases such as Alzheimer's disease and cardiovascular disease, improving the quality of life for the elderly, but also reduces medical and social burdens and alleviates the pressure of elderly care.
[0003] In recent years, many efforts have been made to delay aging. In the early 1900s, it was discovered that by regulating the intestinal microbiota using host-friendly bacteria found in yogurt, health could be improved and aging could be delayed. Since then, probiotics have been widely studied as a microorganism that can benefit host health. For example, intestinal flora imbalance is closely related to aging-related diseases. Probiotics such as Bifidobacterium and Lactobacillus can maintain organ function and prolong life by inhibiting the proliferation of harmful bacteria, promoting the colonization of beneficial bacteria, reducing oxidative stress and inflammatory responses. For example, studies have shown that Lactobacillus paracasei HII01 can upregulate genes related to longevity in nematodes (such as skn-1, sir-2.1) and extend lifespan in a dose-dependent manner.
[0004] One of the core mechanisms of aging is the excessive accumulation of reactive oxygen species (ROS), which leads to an imbalance of oxidative stress, disrupting the structure and function of biological macromolecules (such as DNA, lipids, and proteins), and accelerating cell damage and apoptosis. Free radical theory suggests that ROS produced by mitochondrial metabolism attack DNA, proteins, and lipids, while the activity of antioxidant pathways (such as Keap1 / Nrf2) decreases with age, exacerbating the vicious cycle of oxidative stress. As a classic model organism for anti-aging research, Caenorhabditis elegans is widely used in the analysis of aging mechanisms and the screening of anti-aging substances due to its short life cycle (approximately 3 weeks), high genetic conservation (60%-80% of genes are homologous to humans), and ease of manipulation.
[0005] Magnetotactic bacteria are a type of bacteria that can move along magnetic field lines. As a special type of aquatic bacteria, they have a unique magnetosome structure with chain-like Fe3O4 / Fe3S4 nanoparticles inside the bacterial cells that can be observed under an electron microscope. Many studies have shown that iron oxide nanoparticles (IONPs, such as Fe3O4) have intrinsic peroxidase and catalase-like activities. This discovery reveals that ancient life forms may have relied on intracellular biomineralization of IONPs (proto-magnetosomes) to cope with reactive oxygen species (ROS) stress on early Earth. Additionally, some studies have shown that IONPs exhibit pH-dependent dual enzyme-like activities in the intracellular microenvironment: catalyzing the generation of hydroxyl radicals from H2O2 through peroxidase-like activity under acidic conditions, and catalyzing the generation of H2O and O2 from H2O2 through catalase-like activity under neutral and alkaline conditions. In strains such as AMB-1, the pH value of the magnetosome vesicles is close to neutral, indicating that Fe3O4 magnetosomes may mainly exhibit catalase-like activity in vivo. Compared with traditional antioxidant enzymes, IONPs show stronger stability under extreme temperature (4 - 90 °C) and pH (1 - 12) conditions, enabling them to maintain antioxidant functions in harsh environments. These properties support the hypothesis that IONPs served as key antioxidants in early life evolution.
[0006] Due to its special structure and magnetotactic and aerotactic properties, the removal of pollutants from wastewater, mainly including organic pollutants, radionuclides, and heavy metals, is one of the potential environmental applications of magnetotactic bacteria. In addition, there have been many studies on magnetotactic bacteria in the field of biomedicine. For example, using magnetotactic bacteria under a low-frequency and low-heat-generating oscillating magnetic field (sMF) can kill Staphylococcus aureus pathogens. Some studies have also shown that live magnetotactic bacteria can be internalized by human lung cancer cells A549, strongly inhibiting the proliferation of cancer cells through hyperthermia. Multiple studies have shown that magnetotactic bacteria have great potential in biomedicine research. However, whether magnetotactic bacteria have the function of delaying aging has not been reported. Summary of the Invention
[0007] The technical problem to be solved by the present invention is how to propose a new use of magnetotactic bacteria AMB-1 and a new method for improving health and delaying aging.
[0008] The present invention solves the above technical problems through the following technical means:
[0009] In the first aspect of the present invention, there is provided the use of magnetotactic bacteria AMB-1 in the preparation of a drug for improving the health and / or delaying the aging of Caenorhabditis elegans.
[0010] In the second aspect of the present invention, there is also provided the use of magnetotactic bacteria AMB-1 in one or more of the following:
[0011] (1) Use in the preparation of a product for reducing the level of ROS;
[0012] (2) Use in the preparation of a product for reducing the accumulation of lipofuscin;
[0013] (3) Use in the preparation of a product for increasing the survival rate of nematode animals.
[0014] Preferably, in (1) to (3), the product includes health products, drugs or feeds.
[0015] Preferably, the product is a drug, and the drug further includes a pharmaceutically acceptable carrier.
[0016] "Pharmaceutically acceptable" means a non-toxic material that does not reduce the activity of the active ingredient. Such pharmaceutically acceptable buffers, carriers or excipients are well known in the art (see Remington’s Pharmaceutical Sciences, 18th Edition, edited by A.R. Gennaro, Mack Publishing Company (1990) and handbook of Pharmaceutical Excipients, 3rd Edition, edited by A. Kibbe, Pharmaceutical Press (2000)).
[0017] Preferably, the pharmaceutically acceptable carrier is selected from one or more of diluents, excipients, fillers, binders, wetting agents, disintegrants, absorption promoters, surfactants, adsorption carriers and lubricants.
[0018] Preferably, the drug is made into a pharmaceutically acceptable dosage form.
[0019] Preferably, the pharmaceutically acceptable dosage form includes tablets, pills, ointments, oral liquids or granules.
[0020] Preferably, in (3), the nematode animals include but are not limited to Caenorhabditis elegans, Ascaris lumbricoides, Enterobius vermicularis, Wuchereria bancrofti, Trichinella spiralis.
[0021] The third aspect of the present invention provides a product for delaying the aging of Caenorhabditis elegans, and its active ingredient includes magnetotactic bacterium AMB-1 or its fermentation supernatant, lysate.
[0022] The fourth aspect of the present invention provides a method for improving the health and delaying the aging of Caenorhabditis elegans based on magnetotactic bacterium AMB-1. It is only necessary to add magnetotactic bacterium AMB-1 to the culture medium as a food supplement for Caenorhabditis elegans.
[0023] Preferably, the culture medium is MSGM medium.
[0024] The beneficial effects of the present invention are as follows:
[0025] 1. The present invention proposes a new use of magnetotactic bacterium AMB-1, specifically its application in the preparation of drugs for improving the health and / or delaying the aging of Caenorhabditis elegans. Through the cultivation of magnetotactic bacterium AMB-1, the cultivation of Caenorhabditis elegans, the detection of anti-aging effects, the detection of ROS, and the detection of lipofuscin accumulation, it is confirmed that magnetotactic bacterium AMB-1 can improve the health and delay the aging of organisms, which is relatively convenient, fast, and has high safety.
[0026] 2. Good biosafety. Multiple studies have shown that through cell experiments and nematode experiments at both the bacterial level and the magnetosome level, the good biosafety of magnetotactic bacterium AMB-1 has been demonstrated. It can significantly delay the aging of nematodes and reduce the accumulation of ROS in their bodies without toxic side effects.
[0027] 3. The anti-aging effect is significant. The results of nematode lifespan indicators show that Magnetospirillum magneticum AMB-1 can significantly improve the health of organisms and delay aging. Description of the Drawings
[0028] Figure 1 It is a comparison chart showing that Magnetospirillum magneticum AMB-1 in Example 1 can significantly improve the survival rate of Caenorhabditis elegans compared with Escherichia coli OP50;
[0029] Figure 2 It is a comparison chart showing that Magnetospirillum magneticum AMB-1 in Example 1 can significantly increase the average lifespan of Caenorhabditis elegans compared with Escherichia coli OP50;
[0030] Figure 3 It is a comparison chart showing that Magnetospirillum magneticum AMB-1 in Example 1 can significantly reduce the accumulation of ROS in the body of Caenorhabditis elegans 10 days after survival compared with Escherichia coli OP50;
[0031] Figure 4 It is a comparison chart showing that Magnetospirillum magneticum AMB-1 in Example 1 can significantly reduce the accumulation of lipofuscin in the body of Caenorhabditis elegans 10 days after survival compared with Escherichia coli OP50;
[0032] In the figure, * represents P < 0.05, and ** represents P < 0.01. Detailed Embodiments
[0033] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, rather than all of them. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts belong to the scope of protection of the present invention.
[0034] The test materials, reagents, etc. used in the following embodiments can be obtained from commercial channels without special instructions.
[0035] For those not specifying specific technologies or conditions in the embodiments, they can all be carried out according to the technologies or conditions described in the literature in the field or according to the product instructions. Without special instructions, the quantitative tests in the following embodiments are all set with more than three repeated experiments, and the results are averaged.
[0036] Succinic acid, Folic acid, Pyridoxine hydrochloride, Riboflavin, Biotin, Thiamine hydrochloride (VB1), Nicotinic acid, Pantothenic acid, Vitamin B12, p-Aminobenzoic acid, Thioctic acid, EDTA, Cobalt nitrate hexahydrate, Nickel chloride hexahydrate (NiCl2·6H2O), Peptone, and Yeast extract were all purchased from Sangon Biotech (Shanghai) Co., Ltd.; Sodium selenite anhydrous (Na2SeO3), Iron(III) chloride (FeCl3), and Quinic acid were all purchased from Shanghai Aladdin Biochemical Technology Co., Ltd.; Sodium tungstate dihydrate, Potassium dihydrogen phosphate, Magnesium sulfate heptahydrate (MgSO4·7H2O), Manganese sulfate monohydrate (MnSO4·H2O), Sodium chloride (NaCl), Iron(II) sulfate heptahydrate (FeSO4·7H2O), Calcium chloride (CaCl2), Zinc sulfate heptahydrate (ZnSO4·7H2O), Copper(II) sulfate pentahydrate (CuSO4·5H2O), Potassium alum (AlK(SO4)2), Boric acid (H3BO3), Sodium molybdate dihydrate (Na2MoO4·2H2O), and Sodium tungstate dihydrate (Na2WO4·2H2O) were all purchased from Sinopharm Chemical Reagent Co., Ltd.
[0037] Example 1:
[0038] Applying magnetotactic bacteria AMB-1 to improve the health and delay the aging of Caenorhabditis elegans specifically includes the following steps:
[0039] (1) Cultivation of magnetotactic bacteria Magnetospirillum magneticum AMB-1:
[0040] Bacterial activation: Take the magnetotactic bacterium AMB-1 (ATCC700264) out of the refrigerator at -80°C and place it in the refrigerator at -4°C for 8 min; after taking it out, let it stand at room temperature for 2 - 3 min, and immediately transfer it after the bacterial strain thaws; take 2 screw-cap test tubes (10 ml) containing 10 ml of sterilized MSGM medium, and add 800 μl of bacterial liquid to each; incubate statically at a constant temperature of 26°C for 10 days, and observe the growth status under an inverted microscope.
[0041] Bacterial transfer: Take out a 50-ml centrifuge tube containing 40 ml of sterilized MSGM medium stored at 4°C and let it stand until room temperature; select the bacterial liquid with better growth, take out 2 ml of bacterial liquid from the bottle with a pipette and transfer it into the centrifuge tube; mark the date and incubate statically at a constant temperature of 26°C for two days.
[0042] MSGM medium: 1.0 L of distilled water, 5.0 ml of inorganic salt mixture, 10.0 ml of vitamin mixture, 2.0 ml of ferric quinate, 0.68 g of potassium dihydrogen phosphate, 0.12 g of sodium nitrate, 0.74 g of succinic acid, 0.05 g of L-cysteine, 0.2 g of peptone, 0.1 g of yeast mixture. Add in the order of solid drugs, inorganic salt mixture, and vitamin mixture, adjust the pH to 6.75 with NaOH, and sterilize at 115°C under high-pressure steam for 30 min. After cooling, add the ferric quinate solution.
[0043] Ferric quinate stock solution: 1.0 L of distilled water, 1.9 g of quinic acid, 2.7 g of ferric chloride. Vitamin mixture: 1.0 L of distilled water, 2.0 mg of folic acid, 10.0 mg of vitamin B6, 5.0 mg of riboflavin, 2.0 mg of biotin, 5.0 mg of vitamin B1, 5.0 mg of nicotinic acid, 5.0 mg of pantothenic acid, 0.1 mg of vitamin B12, 5.0 mg of p-aminobenzoic acid, 5.0 mg of lipoic acid. Inorganic salt mixture: 1.0 L of distilled water, 0.5 g of EDTA, 0.1 g of ZnSO4·7H2O, 0.01 g of CuSO4·7H2O, 0.5 g of MnSO4·H2O, 0.01 g of Na2MoO4·H2O, 0.0001 g of Na2SeO3, 3.0 g of MgSO4·7H2O, 0.01 g of KAl(SO4)2, 0.1 g of FeSO4·7H2O, 0.1 g of Co(NO3)2·6H2O, 0.1 g of CaCl2, 0.01 g of NaWO4·2H2O, 0.02 g of NiCl2·6H2O. After preparing the ferric quinate solution, filter and sterilize it, and store it at 4°C.
[0044] (2) Nematode culture: Nematodes live in the solid agarose culture plate NGM prepared in the laboratory, and their food is the E. coli uracil auxotrophic strain OP50. The nematodes are cultured in a constant-temperature light-proof incubator at 20°C.
[0045] (3) Nematode synchronization: The purpose of nematode synchronization is to obtain nematodes at the same growth and development stage, so as to obtain comparable experimental results after treatment. Use M9 buffer (weigh 5 g of NaCl, 6 g of Na2HPO4 and 3 g of Na2HPO4, add ultrapure water and make up to 1000 mL, after autoclaving, cool to about 65 °C and then add 1 mL of 1 M MgSO4) to wash the adult worms during the egg-laying period from the NGM culture plate, suck them into a 1.5 mL centrifuge tube. After natural sedimentation, discard the supernatant. Each time, add 1 mL of M9 buffer to wash the worms, repeat the washing 3 times until the supernatant is clear. Aspirate the supernatant, then add 700 μL of M9 buffer, 200 μL of NaClO and 100 μL of 5 mol / L NaOH solution, invert the tube up and down to mix evenly, and gently shake until most of the worms in the tube are lysed. Centrifuge at 6000 rpm for 1 minute quickly, discard the supernatant, add 1 mL of M9 buffer to wash and centrifuge 3 times, discard the supernatant again. The precipitate at the bottom of the tube finally obtained is the developing fertilized eggs. Aspirate the precipitate and disperse it evenly in a culture dish (35 ml containing 3 ml of M9 buffer), and culture and incubate in an incubator at 20 °C for more than 14 hours to obtain larvae whose growth stage is stalled at the L1 stage due to the absence of OP50. These obtained L1-stage larvae can be inoculated onto NGM culture plates containing OP50 for continued culture or directly used for experimental treatment.
[0046] (4) Expose nematodes to Magnetospirillum magneticum AMB-1 throughout the entire life cycle and conduct nematode lifespan detection.
[0047] The specific operation is as follows: Spread Escherichia coli OP50 and Magnetospirillum magneticum AMB-1 evenly on a standard NGM plate to prepare a preparation plate. Transfer the synchronized L1-stage nematodes into separate preparation plates. After culturing for two days, pick the worms and transfer them to a new plate every other day, and record the number of dead nematodes every day until all the nematodes die.
[0048] (5) ROS detection: Wash the nematodes exposed for 10 days with M9 and put them into a 1.5 ml centrifuge tube, then wash twice with M9. Subsequently, add 10 μM DCFH-DA probe, place it in an incubator at 20 °C, and incubate for 3 h. Then wash twice with M9, observe and take pictures under a fluorescence microscope, and use Image J software to statistically analyze the average fluorescence intensity. The number of nematodes measured in each group should be ≥20.
[0049] (6) Detection of Lipofuscin: The nematodes after 10 days of exposure were rinsed with M9 and placed into a 1.5 ml centrifuge tube, then washed twice with M9, observed and photographed under a fluorescence microscope, and the average fluorescence intensity was statistically analyzed using the Image J software. The number of nematodes measured in each group should be ≥ 20.
[0050] Result analysis, as Figure 1 、 2 shown, the magnetotactic bacterium Magnetospirillum magneticum AMB-1 can significantly improve the survival rate of nematodes, delay aging; and increase the average lifespan of nematodes by 43.3%. It is proved that the magnetotactic bacterium AMB-1 has a good lifespan-extending effect on nematodes.
[0051] Result analysis, as Figure 3 、 4 shown, the magnetotactic bacterium Magnetospirillum magneticum AMB-1 can significantly reduce the level of ROS in nematodes after 10 days, with a 25.8% reduction. Lipofuscin is a fluorescent pigment that accumulates in senescent cells, mainly composed of oxidized lipids and proteins, and is commonly found in lysosomes. By detecting the accumulation of lipofuscin in nematodes after 10 days, it was found that the feeding of the magnetotactic bacterium AMB-1 reduced the accumulation of lipofuscin by 31.6%.
[0052] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that: they can still modify the technical solutions recorded in the foregoing embodiments, or perform equivalent replacements on some of the technical features; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the spirit and scope of the technical solutions of the various embodiments of the present invention.
Claims
1. Use of magnetotactic bacterium AMB-1 in the preparation of a drug for improving the health and / or delaying the aging of Caenorhabditis elegans.
2. Use of magnetotactic bacterium AMB-1 in one or more of the following: (1) Use in the preparation of a product for reducing the level of ROS; (2) Use in the preparation of a product for reducing the accumulation of lipofuscin; (3) Use in the preparation of a product for increasing the survival rate of nematode animals.
3. The application according to claim 2, wherein In (1)-(3), the product includes health products, drugs or feeds; in (3), the nematode animals include Caenorhabditis elegans, Ascaris lumbricoides, Enterobius vermicularis, Wuchereria bancrofti, Trichinella spiralis.
4. The application according to claim 3, characterized in that The product is a drug, and the drug further includes a pharmaceutically acceptable carrier.
5. The application according to claim 4, characterized in that, The pharmaceutically acceptable carrier is selected from one or more of diluents, excipients, fillers, binders, wetting agents, disintegrants, absorption promoters, surfactants, adsorption carriers and lubricants.
6. The application according to claim 4, characterized in that, The drug is made into a pharmaceutically acceptable dosage form.
7. The application according to claim 6, characterized in that, The pharmaceutically acceptable dosage form includes tablets, pills, ointments, oral liquids or granules.
8. A product for delaying the aging of Caenorhabditis elegans, characterized in that, Its active ingredient includes magnetotactic bacterium AMB-1 or its fermentation supernatant and lysate.
9. A method for improving the health and delaying the aging of Caenorhabditis elegans based on magnetotactic bacterium AMB-1, characterized in that, It is only necessary to add magnetotactic bacterium AMB-1 to the culture medium as a food supplement for Caenorhabditis elegans.
10. The method according to claim 9, characterized in that The culture medium is MSGM medium.