Anti-aging pharmaceutical composition and its use for delaying aging

Abstract

The anti-aging pharmaceutical composition of the present invention comprises: (a) an isolated lactic acid bacteria strain or its fermentate, and (b) an excipient, diluent or carrier; wherein the isolated lactic acid bacteria strain comprises: Bifidobacterium bifidum VDD088 strain, Bifidobacterium breve Bv-889 strain, and Bifidobacterium longum BLI-02 strain. In addition, the present invention further provides the use of the above anti-aging pharmaceutical composition for preparing a pharmaceutical composition for delaying aging.

Description

Technical Field

[0001] The present invention relates to an anti-aging pharmaceutical composition, and particularly to an anti-aging pharmaceutical composition containing lactic acid bacteria or its fermented product. The present invention further relates to the use of the above-mentioned pharmaceutical composition for delaying aging. Background Art

[0002] Aging is a universal, progressive, cumulative, and deleterious physiological decline phenomenon, but due to individual differences, it cannot be described by a single or simple model. Research has found that human aging is closely related to reactive oxygen species (ROS) in the body. Reactive oxygen species can cause damage to molecules such as DNA, proteins, and lipids in body cells, resulting in the gradual loss of physiological functions, leading to diseases and organ aging. Many studies have also pointed out that oxidants are related to cardiovascular diseases, cancers, and aging. Many literatures have further pointed out that reactive oxygen species are one of the main factors causing organ aging. Accordingly, searching for new antioxidant substances to delay aging is one of the main topics in the current anti-aging research field.

[0003] Many literature studies have confirmed that resveratrol has excellent antioxidant properties, and its anti-aging function has also been found in animal experiments. However, resveratrol still has potential side effects. Clinically, it has been found that taking high doses of resveratrol (2.5 to 5.0 g per day) can cause symptoms such as stomach nausea, flatulence, abdominal discomfort, and diarrhea. Resveratrol may also have an anticoagulant effect, which may interfere with enzymes related to drug metabolism in the liver (such as cytochrome P450 or CYP1A2), thereby interfering with drug effects, or may cause unknown risks. Therefore, it is necessary to consult a doctor or pharmacist before taking. It has also been found in animal experiments that it has a slight estrogen effect, so patients sensitive to estrogen should avoid taking it, such as breast cancer patients, ovarian cancer patients, or uterine cancer patients.

[0004] Generally speaking, the activation and increased expression of the intracellular p53 / p21 signaling pathway will induce cell cycle arrest, leading to cell senescence. According to the research paper published in Nature Aging volume 1, pages 962 - 973 (2021), after intermittently clearing senescent cells with highly expressed P21 gene in aging mice, the physical functions of the mice were significantly restored. In other words, reducing the expression of the P21 gene in senescent cells helps to delay aging.

[0005] Probiotics are important symbiotic bacteria in the human body. Depending on the strain and species, they have different physiological functions, such as: regulating the gut microbiota, digestive tract function, regulating blood sugar, enhancing immunity, antioxidant and anti-fatigue effects, etc. In addition, some strains even have multiple different functions. For example, Lactobacillus rhamnosus GG (LGG) is the most studied strain at present, and it has multiple different functions, such as: balancing and improving gastrointestinal function, enhancing the body's own immune ability, preventing and treating diarrhea, reducing respiratory tract infections, expelling toxins and preventing dental caries, etc. The discovery of these functional strains is of great help in improving human health. In addition, in recent years, some probiotic strains have also been found to have antioxidant functions.

[0006] The topics of elderly health and elderly care have been issues that countries around the world have attached importance to in recent years. Therefore, it is important to develop a safe and long-term anti-aging supplement. Generally speaking, lactic acid bacteria are quite safe to consume. Finding lactic acid bacteria strains with anti-aging functions is one of the current urgent goals. Summary of the Invention

[0007] This invention is completed based on the fact that the pharmaceutical composition containing multiple lactic acid bacteria strains or their fermentates has been confirmed to have anti-aging physiological activity through cell experiments and animal experiments.

[0008] This invention provides an anti-aging pharmaceutical composition, which comprises: (a) isolated lactic acid bacteria strains or their fermentates, and (b) excipients, diluents or carriers; wherein the isolated lactic acid bacteria strains include: Bifidobacterium bifidum VDD088 strain deposited with the China General Microbiological Culture Collection Center under the deposit number CGMCC15211, Bifidobacterium breve Bv-889 strain deposited with the China General Microbiological Culture Collection Center under the deposit number CGMCC 16145, and Bifidobacterium longum BLI-02 strain deposited with the China General Microbiological Culture Collection Center under the deposit number CGMCC 15212.

[0009] The present invention further provides a use of an anti-aging pharmaceutical composition for preparing a pharmaceutical composition for delaying aging, wherein the anti-aging pharmaceutical composition comprises: (a) an isolated lactic acid bacteria strain or its fermentate, and (b) an excipient, a diluent or a carrier; and the isolated lactic acid bacteria strain comprises: Bifidobacterium bifidum VDD088 strain deposited with the China General Microbiological Culture Collection Center under the deposit number CGMCC 15211, Bifidobacterium breve Bv-889 strain deposited with the China General Microbiological Culture Collection Center under the deposit number CGMCC 16145, and Bifidobacterium longum BLI-02 strain deposited with the China General Microbiological Culture Collection Center under the deposit number CGMCC 15212. Description of the Drawings

[0010] Figure 1 It is a result graph of free radical scavenging efficiency analysis, showing the influence of different strains on free radical scavenging.

[0011] Figure 2 It is a result graph of FRAP analysis, showing the influence of different strains on iron ion reduction.

[0012] Figure 3 It is a result graph of P21 gene expression, showing the influence of different samples on the P21 gene expression level in mouse 3T3-L1 cells.

[0013] Figure 4 It is a result graph of anti-aging analysis, showing the influence of different samples on the content of oxidized protein carbonyl in the posterior brain of mice.

[0014] Figure 5 It is a result graph of anti-aging analysis, showing the influence of different samples on the catalase activity in the heart of mice.

[0015] Figure 6 It is a result graph of anti-aging analysis, showing the influence of different samples on the content of 8-oxo-2'-deoxyguanosine in the mitochondria of the liver of mice. Detailed Description of the Invention

[0016] To make the above and / or other objects, effects, features of the present invention more obvious and understandable, the following preferred embodiments are specifically described in detail as follows:

[0017] The lactic acid bacteria strains described in the present invention are deposited in the China Center for Type Culture Collection, located at Wuhan University, China, or deposited in the China General Microbiological Culture Collection Center, located at No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. The detailed deposit information is shown in Table 1:

[0018] Table 1. Deposit Information of Lactic Acid Bacteria Strains

[0019]

[0020] It is found in this article that the deposited Bifidobacterium bifidum VDD088 strain, Bifidobacterium breve Bv-889 strain, Bifidobacterium longum BLI-02 strain, Lactobacillus plantarum PL-02 strain, Bifidobacterium animalis subsp. lactis CP-9 strain, and Bifidobacterium longum subsp. longum OLP-01 strain listed in Table 1 have anti-aging physiological activities. Therefore, all the listed strains or their fermentates can be used for the purpose of delaying aging.

[0021] One embodiment of the present invention provides an anti-aging pharmaceutical composition, which comprises: (a) an isolated lactic acid bacteria strain or its fermentate, and (b) an excipient, diluent or carrier; characterized in that the isolated lactic acid bacteria strain comprises: the Bifidobacterium bifidum VDD088 strain deposited at the China General Microbiological Culture Collection Center with the deposit number CGMCC 15211, the Bifidobacterium breve Bv-889 strain deposited at the China General Microbiological Culture Collection Center with the deposit number CGMCC 16145, and the Bifidobacterium longum BLI-02 strain deposited at the China General Microbiological Culture Collection Center with the deposit number CGMCC 15212.

[0022] To further enhance the anti-aging efficacy of the pharmaceutical composition, the isolated lactic acid bacteria strain further comprises: the Bifidobacterium animalis CP-9 strain deposited at the China Center for Type Culture Collection with the deposit number CCTCC M2014588 and the Lactobacillus plantarum PL-02 strain deposited at the China General Microbiological Culture Collection Center with the deposit number CGMCC 20485.

[0023] To further enhance the anti-aging efficacy of the pharmaceutical composition, the isolated lactic acid bacteria strain further comprises: the Bifidobacterium longum subsp. longum OLP-01 strain deposited at the China General Microbiological Culture Collection Center with the deposit number CGMCC 17345.

[0024] In addition, the excipient, diluent or carrier can be a pharmaceutically acceptable excipient, diluent or carrier. The pharmaceutical composition can be in an oral dosage form or a topical skin dosage form. The oral dosage form can be tablets, capsules, solutions or powders, etc.

[0025] Furthermore, the lactic acid bacteria strain can be an active strain or an inactivated strain alone. The total number of strains is preferably 10 6 CFU or more, more preferably 10 9CFU or more. In addition, the fermented product may include the fermented liquid of inactivated strains, the fermented liquid with bacteria removed, or the dried powder of any of these. For example, the fermented liquid may be a fermentation supernatant or a whey fermented liquid, etc. Based on the total weight of the pharmaceutical composition, the content of the dried powder of the fermented liquid is preferably 0.5 wt% or more; alternatively, the content of the fermented liquid of inactivated strains or the fermented liquid with bacteria removed is preferably 2.5 wt% or more.

[0026] Another embodiment of the present invention provides a use of the above anti-aging pharmaceutical composition, which is for preparing a pharmaceutical composition for delaying aging. Specifically, the prepared pharmaceutical composition can be administered to an individual with a need for delaying aging to achieve the effect of delaying aging by scavenging free radicals, reducing the expression level of the P21 gene, reducing the protein carbonyl content, enhancing the catalase activity and / or reducing the content of 8-oxo-2'-deoxyguanosine in the individual. In addition, the prepared pharmaceutical composition is preferably administered to the individual at a total number of strains of 10 6 to 10 10 CFU per kilogram of body weight per day of the individual to achieve the purpose of delaying aging, and more preferably at a total number of strains of 8.3x10 7 to 3.4x10 8 CFU per kilogram of body weight per day of the individual.

[0027] <Example 1: Morphology and General Properties of Lactic Acid Bacteria Strains>

[0028] The taxonomic characteristics of the lactic acid bacteria strains were confirmed by 16S rDNA sequence analysis and API bacterial identification system analysis. The characteristics of the lactic acid bacteria strains in terms of morphology and general properties are shown in Table 2:

[0029] Table 2. Morphological and General Property Characteristics of Lactic Acid Bacteria Strains

[0030]

[0031]

[0032]

[0033] <Example 2: Collection of Lactic Acid Bacteria Strains>

[0034] The lactic acid bacteria strains were stored in 20% glycerol at -80°C. Before use, they were activated twice for 24 hours at 37°C in MRS broth (DIFCO) containing 0.05% cysteine. The lactic acid bacteria strains used were Bifidobacterium bifidum VDD088 strain, Bifidobacterium breve Bv-889 strain, Bifidobacterium longum BLI-02 strain, Lactobacillus plantarum PL-02 strain, Bifidobacterium animalis subsp. lactis CP-9 strain, and Bifidobacterium longum subsp. longum OLP-01 strain, all of which were sourced from the human intestine. In one embodiment, the liquid medium of the lactic acid bacteria strains contains at least one of 5 to 30 wt% milk and 1 to 10 wt% soybean powder; in one embodiment, the liquid medium of the lactic acid bacteria strains contains at least one of a carbon source and a nitrogen source. The carbon source includes glucose, fructose, lactose, sucrose, maltose, galactose, mannose, trehalose, starch, molasses, potato starch, corn starch, malt extract, maltodextrin, or any combination thereof, and the nitrogen source includes (NH 4 ) 2 SO 4 、(NH 4 ) 3 PO 4 、NH 4 NO 3 、NH 4 Cl, casein amino acids, urea, peptone, polypeptone, tryptone, meat extract, yeast extract, yeast powder, milk, soybean powder, whey, or any combination thereof; in one embodiment, based on the total weight of the liquid medium, glucose and maltodextrin together account for 2 to 5 wt%.

[0035] The ferment produced by the fermentation of lactic acid bacteria strains is obtained by fermenting a fermentation broth using at least one of the above lactic acid bacteria strains, followed by centrifugation, filtration, heat sterilization, and finally purification. According to actual needs, the ferment can be further dried into a lactic acid bacteria ferment powder, and the powder dosage form or the formed aqueous solution dosage form can be stored at room temperature.

[0036] <Example 3: Free Radical Scavenging Activity Analysis>

[0037] DPPH (di(phenyl)-(2,4,6-trinitrophenyl)iminoazanium) is a stable free radical molecule, and the DPPH free radical methanol solution has the highest absorption value at a wavelength of 517 nm. When the DPPH free radical reacts with an antioxidant, the antioxidant provides hydrogen ions to scavenge the free radical, causing the DPPH free radical to lose its original blue-violet color and resulting in a decrease in absorbance.

[0038] The lactic acid bacteria strains and their fermentation broths to be tested (the total number of strains is approximately 2x10 9CFU, OD about 2.0) was mixed with 0.2 mM DPPH methanol solution in a ratio of 1:1. Additionally, Streptococcus thermophilus strain L-243 (total number of strains about 2x10 9 CFU, OD about 2.0) without antioxidant activity was used as the negative control group, and secondary water was used as the blank group and the same mixing method was adopted. After mixing evenly, the reaction was carried out for 30 minutes at room temperature in a dark room. Then, after centrifuging at 12,000 rpm for 2 minutes at 4°C, 200 μl of the reaction solution was taken into a 96-well plate and its OD 517 value was measured. The calculation formula for the free radical scavenging efficiency is as follows:

[0039]

[0040] Among them, OD sample represents the OD 517 value of the sample to be measured, and OD blank represents the OD 517 value of the blank group.

[0041] As Figure 1 shown, compared with other lactic acid bacteria strains, Bifidobacterium breve strain Bv-889, Bifidobacterium longum subsp. strain OLP-01, Bifidobacterium longum strain BLI-02, Lactobacillus plantarum strain PL-02, Bifidobacterium animalis strain CP-9, and Bifidobacterium bifidum strain VDD088 have relatively high free radical scavenging efficiency.

[0042] <Example 4: Ferric Ion Reducing Power Analysis>

[0043] FRAP is the abbreviation of ferric ion reducing antioxidant power (or ferric reducing ability of plasma), which is called "ferric ion reduction / antioxidant capacity method" in Chinese. It is a method for measuring the antioxidant capacity of a sample by generating a blue-violet complex of ferrous ion and tripyridyltriazine (TPTZ) under low pH conditions. This method is widely used in the analysis of the antioxidant capacity of foods and health products.

[0044] Specifically, the principle of FRAP is that under acidic conditions, antioxidants can reduce Fe 3+ -TPTZ to blue-violet Fe 2+-TPTZ, and then the absorbance was measured at a wavelength of 593 nm as an index of total antioxidant capacity. In addition, acidic conditions can inhibit some endogenous interferences. Since the total concentration of iron ions or ferrous ions in the sample to be tested is usually less than 10 μM, the iron ions or ferrous ions in the sample will not significantly interfere with the test results. In addition, since the iron ions or ferrous ions in the reaction system will chelate with TPTZ, the trace metal ion chelating agents contained in the sample to be tested usually will not significantly affect the detection results.

[0045] As Figure 2 shown, compared with other lactic acid bacteria strains, Bifidobacterium breve Bv-889 strain, Bifidobacterium longum subsp. longum OLP-01 strain, Bifidobacterium longum BLI-02 strain, Lactobacillus plantarum PL-02 strain, Bifidobacterium animalis CP-9 strain, and Bifidobacterium bifidum VDD088 strain have relatively high iron ion reducing ability, that is, these strains have relatively high antioxidant ability.

[0046] <Example 5: P21 Gene Expression Analysis>

[0047] Referring to the literature Probiotics Antimicrob Proteins. 2020 Jun; 12(2):563-576, mouse 3T3-L1 cells were used as a model and the cells were senesced under H 2 O 2 oxidative stress. In addition, the cells were treated with the sample to be tested and the expression of the target gene P21 was observed.

[0048] Mouse 3T3-L1 cells purchased from the Food Industry Research and Development Institute were cultured in Dulbecco’s modified Eagle’s medium (DMEM) containing 10% FBS at a culture temperature of 37°C and a CO 2 concentration of 5%. After subculturing for 3 to 5 generations, the experiment was carried out.

[0049] The cells were seeded in a 6-well culture plate with 3x10 4 cells per well, and continuously cultured for several days until the cells adhered and reached 80% confluence. H at a concentration of 150 μM 2 O 2 was used as a cell oxidant and added to the wells to co-culture with the cells for 3 hours. After that, it was washed with PBS and then DMEM was added. The above steps were carried out once every 24 hours for a total of three times. The sample to be tested was added to co-culture with the cells at the same time as H 2 O 2 was added. The sample to be tested was added to co-culture with the cells at the same time as H 2 O 2After co-culturing with cells for 3 hours, first wash with PBS, and then add the same test sample to co-culture with the cells. Finally, extract the RNA of the cells and perform real-time quantitative polymerase chain reaction (Q-PCR) to analyze the expression level of the P21 gene. The primer pair for the P21 gene is the forward primer 5’-tgcatccgtttcacccaacc-3’ and the reverse primer 5’-tcatttttccaaagtgctattcagg-3’. The primer pair for the internal control group GADPH gene is the forward primer 5’-gaaggtgaaggtcggagt-3’ and the reverse primer 5’-gaagatggtgatggatttc-3’.

[0050] As Figure 3 shown, under H 2 O 2 oxidative stress, the relative expression levels of the P21 gene in cells treated with Bifidobacterium bifidum strain VDD088 (total number of each strain is 2x10 9 CFU), cells treated with Bifidobacterium animalis strain CP-9 (total number of each strain is 2x10 9 CFU), cells treated with Bifidobacterium breve strain Bv-889 (total number of each strain is 2x10 9 CFU), cells treated with Bifidobacterium longum strain BLI-02 (total number of each strain is 2x10 9 CFU), and cells treated with Lactobacillus plantarum strain PL-02 (total number of each strain is 2x10 9 CFU) are all lower than those in cells not treated with any test sample, indicating that the above-mentioned lactic acid bacteria strains have the ability to delay aging.

[0051] Again, as Figure 3 shown, under H 2 O 2 oxidative stress, the relative expression level of the P21 gene in cells co-treated with Bifidobacterium bifidum strain VDD088, Bifidobacterium breve strain Bv-889, and Bifidobacterium longum strain BLI-02 (total number of each strain is 2x10 9 CFU, and the ratio of different types of strains is 1:1:1) is less than that in cells treated only with Bifidobacterium bifidum strain VDD088, cells treated only with Bifidobacterium breve strain Bv-889, and cells treated only with Bifidobacterium longum strain BLI-02, indicating that the combination of Bifidobacterium bifidum strain VDD088, Bifidobacterium breve strain Bv-889, and Bifidobacterium longum strain BLI-02 shows a synergistic effect on delaying aging.

[0052] Once again, asFigure 3 As shown, under H 2 O 2 oxidative stress, the relative expression level of the P21 gene in cells co-treated with Bifidobacterium bifidum strain VDD088, Bifidobacterium animalis strain CP-9, Bifidobacterium breve strain Bv-889, Bifidobacterium longum strain BLI-02, and Lactobacillus plantarum strain PL-02 (the total number of each strain is 2x10 9 CFU, and the ratio of the number of different strains is 1:1:1:1:1) is less than that in cells treated only with Bifidobacterium bifidum strain VDD088, only with Bifidobacterium animalis strain CP-9, only with Bifidobacterium breve strain Bv-889, only with Bifidobacterium longum strain BLI-02, and only with Lactobacillus plantarum strain PL-02, indicating that the combination of Bifidobacterium bifidum strain VDD088, Bifidobacterium animalis strain CP-9, Bifidobacterium breve strain Bv-889, Bifidobacterium longum strain BLI-02, and Lactobacillus plantarum strain PL-02 shows a synergistic effect in delaying aging.

[0053] Another example is Figure 3 as shown, under H 2 O 2 oxidative stress, the relative expression level of the P21 gene in cells co-treated with Bifidobacterium bifidum strain VDD088, Bifidobacterium animalis strain CP-9, Bifidobacterium breve strain Bv-889, Bifidobacterium longum strain BLI-02, Lactobacillus plantarum strain PL-02 (the total number of each strain is 2x10 9 CFU, and the ratio of the number of different strains is 1:1:1:1:1) and their fermentation broth is less than that in cells co-treated with Bifidobacterium bifidum strain VDD088, Bifidobacterium animalis strain CP-9, Bifidobacterium breve strain Bv-889, Bifidobacterium longum strain BLI-02, Lactobacillus plantarum strain PL-02 (the total number of each strain is 2x10 9 CFU, and the ratio of the number of different strains is 1:1:1:1:1), indicating that the fermentation broth of lactic acid bacteria strains has the ability to delay aging.

[0054] <Example 6: Anti-aging analysis of mice>

[0055] The content of protein carbonyl, the activity of catalase (CAT), and the content of mitochondrial 8-hydroxy-2'-deoxyguanosine (8OHdG) are important indicators of tissue aging. The anti-aging properties of the test samples in mice were analyzed using these three indicators below.

[0056] Ten-month-old mice were divided into six groups of four mice each. The dose of lactic acid bacteria was based on the initial estimation method announced by the US Food and Drug Administration in 2005; the "blank control group" was fed only a normal diet; the "resveratrol group" was fed a normal diet and an additional 20.5 mg of resveratrol per kilogram of body weight per day for the mice; the "low-dose lactic acid bacteria group" was fed a normal diet and an additional 1.03x10 9 CFU of lactic acid bacteria strain mixture; the "low-dose lactic acid bacteria and ferment group" was fed a normal diet and an additional 1.03x10 9 CFU of lactic acid bacteria strain mixture and 20.5 mg of lactic acid bacteria ferment; the "high-dose lactic acid bacteria group" was fed a normal diet and an additional 4.1x10 9 CFU of lactic acid bacteria strain mixture; the "high-dose lactic acid bacteria and ferment group" was fed a normal diet and an additional 4.1x10 9 CFU of lactic acid bacteria strain mixture and 20.5 mg of lactic acid bacteria ferment; among them, the "lactic acid bacteria strain mixture" used included Bifidobacterium bifidum strain VDD088, Bifidobacterium breve strain Bv-889, Bifidobacterium longum strain BLI-02, Lactobacillus plantarum strain PL-02, Bifidobacterium animalis strain CP-9, and Bifidobacterium longum subsp. longum strain OLP-01, and the ratio of the number of different strains was 1:1:1:1:1:1. After continuous feeding for six months, the brains, hearts, and livers of the mice were removed, and the aging conditions of the mice in each group were evaluated. The experimental procedures such as the determination of the carbonyl content of oxidized proteins, the determination of catalase activity, and the determination of the 8-oxo-2'-deoxyguanosine content in mitochondria are detailed below:

[0057] 1. After continuous feeding for six months, the mice were sacrificed first, and then the tissues such as the brain, heart, and liver were removed with surgical instruments and placed in microcentrifuge tubes respectively. 200 μl of 0.4 M perchloric acid was added and homogenized with an ultrasonic homogenizer.

[0058] 2. The homogenates of each mouse organ were operated according to the experimental standard procedures provided by the Protein Carbonyl Colorimetric Assay Kit (Cayman Chemical, model: 10005020), Catalase Assay Kit (Cayman Chemical, model: 707002), and 8-OHdG Check ELISA kit (JaICA, model: KOG200S / E) to detect the content of organ aging indicators.

[0059] As Figures 4 to 6As shown, the mice fed with high-dose lactic acid bacteria combined with fermented products had the lowest content of protein carbonyl, an oxidative substance, in the brain, the highest catalase activity in the heart, and the lowest content of 8-oxo-2'-deoxyguanosine in the liver mitochondria, indicating that the combination of lactic acid bacteria strains and their fermentation broth has the ability to delay aging.

[0060] The preservation information of the biological materials involved in the present invention is as follows:

[0061] 1. CN, China, China General Microbiological Culture Collection Center, January 15, 2018, CGMCC 15211;

[0062] 2. CN, China, China General Microbiological Culture Collection Center, July 23, 2018, CGMCC 16145;

[0063] 3. CN, China, China General Microbiological Culture Collection Center, January 15, 2018, CGMCC 15212;

[0064] 4. CN, China, China General Microbiological Culture Collection Center, August 6, 2020, CGMCC 20485;

[0065] 5. CN, China, China Center for Type Culture Collection, November 24, 2014, CCTCC M2014588;

[0066] 6. CN, China, China General Microbiological Culture Collection Center, March 18, 2019, CGMCC 17345.

[0067] However, the above are only the preferred embodiments of the present invention, and the protection scope of the present invention cannot be limited thereby; therefore, all simple equivalent changes and modifications made according to the protection scope of the patent application of the present invention and the content of the invention specification still fall within the protection scope of the present invention.

Claims

1. An anti-aging pharmaceutical composition, comprising: (a) an isolated lactic acid bacteria strain, the isolated lactic acid bacteria strain consisting of: consisting of: Bifidobacterium bifidum VDD088 strain deposited with the China General Microbiological Culture Collection Center under the deposit number CGMCC No. 15211, Bifidobacterium breve Bv-889 strain deposited with the China General Microbiological Culture Collection Center under the deposit number CGMCC No. 16145, Bifidobacterium longum subsp. infantis BLI-02 strain deposited with the China General Microbiological Culture Collection Center under the deposit number CGMCC No. 15212, Bifidobacterium animalis subsp. lactis CP-9 strain deposited with the China Center for Type Culture Collection under the deposit number CCTCC No: M2014588, and Lactobacillus plantarum PL-02 strain deposited with the China General Microbiological Culture Collection Center under the deposit number CGMCC No. 20485; and (b) an excipient or a carrier.

2. An anti-aging pharmaceutical composition, comprising: (a) an isolated lactic acid bacteria strain, the isolated lactic acid bacteria strain consisting of: consisting of: Bifidobacterium bifidum strain VDD088 deposited with the China General Microbiological Culture Collection Center under deposit number CGMCC No. 15211, Bifidobacterium breve strain Bv-889 deposited with the China General Microbiological Culture Collection Center under deposit number CGMCC No. 16145, Bifidobacterium longum subsp. infantis strain BLI-02 deposited with the China General Microbiological Culture Collection Center under deposit number CGMCC No. 15212, Bifidobacterium animalis subsp. lactis strain CP-9 deposited with the China Center for Type Culture Collection under deposit number CCTCC No: M2014588, Lactobacillus plantarum strain PL-02 deposited with the China General Microbiological Culture Collection Center under deposit number CGMCC No. 20485, and Bifidobacterium longum subsp. longum strain OLP-01 deposited with the China General Microbiological Culture Collection Center under deposit number CGMCC No. 17345; and (b)excipient or carrier.

3. The pharmaceutical composition according to any one of claims 1 to 2, characterized in that the isolated lactic acid bacteria strain is an active strain.

4. The pharmaceutical composition according to claim 1, characterized in that the strain quantity ratio among the Bifidobacterium bifidum VDD088 strain, the Bifidobacterium breve Bv-889 strain, the Bifidobacterium longum subsp. infantis BLI-02 strain, the Bifidobacterium animalis subsp. lactis CP-9 strain, and the Lactobacillus plantarum PL-02 strain is 1:1:1:1:

1.

5. Use of a pharmaceutical composition according to claim 1 or 2, characterized in that for preparing an anti-aging pharmaceutical composition for administration to an individual in need of anti-aging to reduce the expression level of the P21 gene in the individual.

6. The use according to claim 5, characterized in that the isolated lactic acid bacteria strain is an active strain.

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