Microecological preparation capable of relieving aging and application thereof

Abstract

The invention belongs to the technical field of biological medicine, and discloses a microecological preparation capable of relieving senescence and application of the microecological preparation. Tests on D-galactose-induced aging mice show that the microecological preparation can effectively reduce the weakness index of the aging mice, maintain muscle health, improve cognitive ability decline caused by aging, improve the antioxidant ability of liver tissues (reduce the MDA content and increase the activity of SOD, CAT and GSH-Px), improve the anti-aging ability of the mice, improve the anti-aging ability of the mice, improve the anti-aging ability of the mice, improve the anti-aging ability of the mice, and improve the anti-aging ability of the mice and the anti-aging ability of the mice and the anti-aging ability of the mice. The levels of proinflammatory factors TNF-alpha, IL-1beta and IL-6 in serum are reduced, and the level of an anti-inflammatory factor IL-10 in the serum is increased. Therefore, the microecological preparation disclosed by the invention can effectively relieve senescence and has a relatively great application prospect in senescence relieving products.

Description

Technical Field

[0001] The present invention belongs to the field of biotechnology, and in particular relates to a microecological preparation capable of alleviating aging and an application thereof. Background Art

[0002] During aging, the intestinal microbiome exhibits significant dysbiosis, such as changes in microbial diversity, alterations in key microorganisms, and decreased levels of beneficial metabolites derived from microorganisms. These changes can lead to inflammation, weakness, and even the risk of death. Current research suggests that interventions targeting the intestinal microbiome may reverse age-related cognitive impairment, sarcopenia, and other phenotypes. For example, patent CN108576823A discloses that synbiotics can effectively promote intestinal health, inhibit the production of pathogenic bacteria and toxins, protect the intestinal mucosa, maintain the biofilm barrier, and reduce the production of free radicals, thereby delaying aging. Patent CN112708581A discloses a probiotic preparation of Bifidobacterium adolescentis DH162, which can exert anti-aging effects through mechanisms such as enhancing the body's antioxidant capacity, improving immunity, regulating the intestinal flora, and improving neurotransmitters. Patent CN118773037A discloses Bifidobacterium bifidum NKUFB3-12, which can alleviate chronic inflammation, cognitive impairment, and immune aging caused by aging. However, there is currently no microecological preparation that can reduce the frailty index, increase grip strength, increase muscle content, improve muscle tissue morphology, relieve liver oxidative stress, reduce the expression level of inflammatory factors, and increase the expression level of anti-inflammatory factors. Summary of the Invention

[0003] To solve the above problems, the present invention provides a microecological preparation capable of alleviating aging and its application.

[0004] The technical solutions of the present invention are as follows:

[0005] The present invention provides a microecological preparation capable of alleviating aging, wherein the microecological preparation comprises the above-mentioned Bifidobacterium bifidum and fucoidan.

[0006] Preferably, the content of the Bifidobacterium bifidum CCFM1424 strain is ≥1×10 9 CFU / mL, and the concentration of fucoidan should not be less than 30mg / kg BW / day.

[0007] The application of the above-mentioned microecological preparation in the preparation of products for alleviating aging.

[0008] Preferably, the alleviating aging includes any of the following functions:

[0009] (1) Reduce the frailty index of elderly individuals;

[0010] (2) increase grip strength in elderly individuals;

[0011] (3) Improve muscle mass in aging mice;

[0012] (4) Improve muscle tissue morphology in aging individuals;

[0013] (5) Can significantly alleviate cognitive decline caused by aging;

[0014] (6) significantly alleviate oxidative stress in the liver of elderly individuals, wherein the antioxidant index includes at least one of MDA, SOD, CAT, or GSH-Px;

[0015] (7) reducing the expression level of inflammatory factors in the serum of elderly individuals, wherein the inflammatory factors include at least one of IL-1β, IL-6, or TNF-α;

[0016] (8) increasing the expression level of anti-inflammatory factors in the serum of elderly individuals, wherein the anti-inflammatory factor is IL-10;

[0017] (9) Alleviate the intestinal microbiome disorder caused by aging.

[0018] Beneficial effects:

[0019] The beneficial effects of the microecological preparation of the present invention are as follows:

[0020] (1) Reduce the frailty index of elderly individuals;

[0021] (2) increase grip strength in elderly individuals;

[0022] (3) increase muscle mass in aging mice;

[0023] (4) Improve muscle tissue morphology in aging individuals;

[0024] (5) Cognitive function of aging mice improved;

[0025] (6) Reduce the MDA concentration in liver tissue of aging mice;

[0026] (7) Increase SOD activity in liver tissue of aging mice;

[0027] (8) Increase CAT activity in liver tissue of aging mice;

[0028] (9) Increase GSH-Px activity in liver tissue of aging mice;

[0029] (10) Reduce the concentration of inflammatory factor IL-1β in the serum of aged mice;

[0030] (11) Reduce the concentration of inflammatory factor TNF-α in the serum of aging mice;

[0031] (12) Reduce the concentration of inflammatory factor IL-6 in the serum of aging mice;

[0032] (13) Increase the concentration of anti-inflammatory factor IL-10 in the serum of aging mice.

[0033] (14) Regulate the intestinal microbiome disorder caused by aging, reduce the F / B value, and restore the intestinal microbiome to a healthy state.

[0034] Therefore, it has great application prospects in the preparation of products that alleviate aging.

[0035] Preservation Instructions

[0036] Bifidobacterium bifidum CCFM1424 strain

[0037] Storage address: 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, Guangdong Province

[0038] Deposit date: September 20, 2024

[0039] Species name: Bifidobacterium bifidum

[0040] Latin name: Bifidobacterium bifidum

[0041] Strain ID: CCFM1424

[0042] Depository: Guangdong Provincial Microbial Culture Collection Center

[0043] Depository abbreviation: GDMCC

[0044] GDMCC registration number: GDMCC No:65151 BRIEF DESCRIPTION OF THE DRAWINGS

[0045] The present invention will be further described below with reference to the accompanying drawings and embodiments.

[0046] Figure 1 Flowchart of animal experiments.

[0047] Figure 2 This is a graph showing the frailty index scores of experimental mice in different groups.

[0048] Figure 3 The graph shows the forelimb grip strength results of experimental mice in different groups.

[0049] Figure 4 The graph shows the muscle content of experimental mice in different groups.

[0050] Figure 5 These are muscle tissue sections of experimental mice in different groups.

[0051] Figure 6 The performance of different groups of experimental mice in the water maze behavioral test is shown in the figure.

[0052] (A) Escape latency in the water maze test for 4 consecutive days;

[0053] (B) Heat map of representative swimming trajectories of the spatial exploration test on day 5;

[0054] (C) Escape latency in the space exploration test on day 5;

[0055] (D) is the number of times the platform was crossed during the space exploration experiment on the fifth day;

[0056] (E) Cumulative duration in the target quadrant during the spatial exploration test on day 5.

[0057] Figure 7 The antioxidant index diagram of the liver tissue of different groups of experimental mice.

[0058] (A) is the MDA concentration;

[0059] (B) is SOD activity;

[0060] (C) is CAT activity;

[0061] (D) GSH-Px activity.

[0062] Figure 8 The following is a graph showing the levels of inflammatory factors and anti-inflammatory factors in the serum of experimental mice in different groups.

[0063] (A) is the level of TNF-α;

[0064] (B) is the IL-1β level;

[0065] (C) is the IL-6 level;

[0066] (D) IL-10 levels.

[0067] Figure 9 Figure 2 shows the regulatory effects of different interventions on the intestinal flora of aging mice.

[0068] (A) is α-diversity;

[0069] (B) is β-diversity;

[0070] (C) shows the relative abundance changes of Proteobacteria;

[0071] (D) is the F / B value.

[0072] In the above figures: *, **, *** indicate that the p-values ​​compared with the blank group are less than 0.05, 0.01, and 0.001, respectively. DETAILED DESCRIPTION

[0073] In order to better understand the present invention, the present invention is further described in detail below with reference to the embodiments and drawings. However, those skilled in the art will understand that the following embodiments are not limitations on the scope of protection of the present invention, and any changes and modifications made on the basis of the present invention are within the scope of protection of the present invention.

[0074] Unless otherwise specified, the experimental methods used in the following examples are conventional methods.

[0075] Unless otherwise specified, the materials and reagents used in the following examples can be obtained from commercial sources.

[0076] C57BL / 6 male mice used in the following examples were purchased from Beijing Weitonglihua Laboratory Animal Technology Co., Ltd. Elisa assay kits for TNF-α, IL-1β, IL-6, and IL-10 were purchased from Nanjing Senbeijia Biotechnology Co., Ltd. SOD, CAT, GSH-Px, and MDA assay kits were purchased from Nanjing Jiancheng. D-galactose used in the following examples was purchased from Sinopharm Group Co., Ltd.

[0077] The preparation method of the D-galactose solution involved in the following examples is as follows:

[0078] Each mouse was subcutaneously injected with 1000 mg / kg BW D-galactose solution, which was mixed with normal saline.

[0079] The culture medium involved in the following examples is as follows:

[0080] MRS liquid medium ( / L): peptone 10 g, beef extract 10 g, yeast powder 5 g, anhydrous glucose 20 g, anhydrous sodium acetate 2 g, magnesium sulfate (MgSO4·7H2O) 0.5 g, manganese sulfate (MnSO4·H2O) 0.25 g, diammonium hydrogen citrate 2 g, dipotassium hydrogen phosphate (K2HPO4·3H2O) 2.6 g, Tween 80 1 mL, pH 7.2-7.4.

[0081] MRS solid culture medium formula ( / L): peptone 10 g, beef extract 10 g, yeast powder 5 g, anhydrous glucose 20 g, anhydrous sodium acetate 2 g, magnesium sulfate (MgSO4·7H2O) 0.5 g, manganese sulfate (MnSO4·H2O) 0.25 g, diammonium hydrogen citrate 2 g, dipotassium hydrogen phosphate (K2HPO4·3H2O) 2.6 g, Tween 80 1 mL, agar 15 g, pH 7.2-7.4.

[0082] The present invention will be further described below with reference to specific embodiments.

[0083] Example 1 Effect on Frailty Index of Aging Mice

[0084] Eight-week-old SPF-grade C57BL / 6J male mice were divided into six groups, namely, a blank group, a model group, an experimental group (CCFM1424 group, a fucoidan group, a fucoidan + CCFM1424 group (referred to as the proecological preparation group)), and a positive drug control group (referred to as the arginine group); 6 mice in each group were housed in the Experimental Animal Center of Jiangnan University, fed with ordinary feed, and maintained at a constant temperature of 21-26°C, a humidity of 40-70%, a noise level of less than or equal to 60 dB, and an animal illumination of 15-20 LX (all animal experimental procedures were reviewed and approved by the Animal Welfare and Ethics Management Committee of Jiangnan University).

[0085] The specific procedures of animal experiments are shown in Figure 1 As shown. The experiment was divided into 8 weeks, with 1 week for adaptation, 7 weeks for modeling, and 7 weeks for intervention. Starting from the second week, mice in the model group, experimental group, and control group were subcutaneously injected with D-galactose (dose 1000 mg / kg BW / d) every day for modeling. Mice in the blank group were subcutaneously injected with the same amount of sterile saline every day. The body weight of all mice was recorded every week, and the dose of D-galactose was adjusted accordingly. At the same time, intervention was carried out in the second week. The number of viable bacteria in the CCFM1424 group was 1×10 9 CFU / mL of CCFM1424 bacterial suspension, the fucoidan group was gavaged with 0.2 mL of fucoidan at a concentration of 30 mg / kg BW / day, and the proecological preparation group was gavaged with 0.2 mL of solution containing fucoidan at a concentration of 30 mg / kg BW / day and 1×10 9 CFU / mL of CCFM1424 were used. The positive drug control group was gavaged with 0.2 mL of arginine at a concentration of 0.4 mg / kg BW / d, while the model and blank groups were gavaged with 0.2 mL of sterile saline for a total of 7 weeks. During this period, all groups had free access to water and food.

[0086] Before sacrifice, each aged mouse was scored for clinical signs, including assessment of 27 variables, as shown in Table 1. A frailty index (FI) was calculated based on the scores of individual variables. For each variable, a score of 0 indicates no impairment, 0.5 indicates mild impairment, and 1 indicates severe impairment. All scores were summed and divided by the total number of variables assessed, 27, to determine the FI score for each animal (FI values ​​range from 0 to 1).

[0087] Table 1 Clinical signs scoring table of mouse aging

[0088]

[0089] The results are as follows Figure 2 As shown, the aging score, characterized by the FI index, was higher in the model group (0.37±0.02) compared to the blank group (0.12±0.04), an increase of 198.33%. Mice in the model group exhibit varying degrees of hair loss, beard loss, hair graying, and behavioral changes, common symptoms of aging in C57BL / 6J mice. After treatment with CCFM1424, fucoidan, fucoidan + CCFM1424, and arginine, the frailty index of aging mice significantly decreased (p<0.001) by 44.69%, 41.34%, 69.83%, and 45.53%, respectively, compared to the model group. The frailty index decreased most significantly after treatment with fucoidan + CCFM1424, maintaining a comparable level to the blank group. These mice, with the exception of a few mice lacking whiskers, had intact body hair compared to the other groups. There were no obvious lesions around the eyes, on the body, or on the body, and no behavioral abnormalities.

[0090] Example 2 Effect on the Forelimb Grip Strength of Aging Mice

[0091] The animal experiment design was the same as in Example 1.

[0092] At week 8, a grip strength test was conducted on mice: the mice were placed flat on the grip strength tester grid plate, and the tail of the mouse was grasped and slowly pulled backwards. The data was read and repeated five times, and the maximum value was recorded. At the end of the experiment, the equipment was wiped with 75% (v / v) ethanol to eliminate the influence of odor. The results are as follows: Figure 3 shown.

[0093] The results showed that the grip strength of the blank group, model group, CCFM1424 group, fucoidan group, fucoidan + CCFM1424 group, and arginine group were 149.23 ± 3.42 N, 97.2 ± 6.14 N, 123.03 ± 8.25 N, 111.13 ± 5.1 N, 130.13 ± 9.25 N, and 122.07 ± 11.32 N, respectively. Compared with the blank group, the forelimb grip strength of mice in the model group was significantly reduced (p < 0.001), decreasing by 35%. The forelimb grip strength of mice in the CCFM1424 group, fucoidan + CCFM1424 group, and arginine group was significantly increased compared with the model group (p < 0.05), with the increase in the fucoidan + CCFM1424 group being greater than that in the other intervention groups.

[0094] Example 3 Effect on Muscle Content in Aging Mice

[0095] The animal experiment design was the same as in Example 1.

[0096] After the experiment, the mice were dissected in accordance with ethical requirements, and the gastrocnemius, soleus, and tibialis anterior muscles of the right calf were weighed and the total weight was calculated to calculate the muscle content of the mice = total muscle weight / mouse body weight × 100%. Figure 4 shown.

[0097] Decreased muscle mass and increased fat mass are the main causes of strength loss in aging. As shown in the figure, the model group showed a significant decrease in muscle mass, a 37% decrease, compared to the blank group. After intervention in both the experimental and positive drug control groups, muscle mass increased to varying degrees in the aging mice, with the fucoidan + CCFM1424 group showing the most significant increase (p<0.05), a 66% increase.

[0098] Example 4 Effects on Muscle Morphology in Aging Mice

[0099] The animal experiment design was the same as in Example 1.

[0100] After the experiment, the mice were dissected according to ethical requirements. The tibialis anterior muscles of the mice were fixed in environmentally friendly GD fixative, embedded in paraffin, and sliced ​​5 mm thick. The slices were stained with hematoxylin and eosin. The stained slices were observed using a pathology section scanner and images were randomly collected. Figure 5 shown.

[0101] Muscle slices Figure 5 As shown in the results, the muscle fiber structure of the blank group mice was tightly arranged and regularly shaped, with polygonal cross-sections and small gaps between muscle fibers; the muscle fibers of the model group mice were disorderly arranged, with irregular fiber shapes and increased gaps between fibers in some areas; after intervention with CCFM1424, fucoidan, fucoidan + CCFM1424 and arginine, the pathological symptoms of the muscle tissue of aging mice were improved to varying degrees. The fucoidan + CCFM1424 group showed that the muscle fiber arrangement was relatively restored to normal, with a more regular shape, reduced gaps between muscle fibers, and the muscle fibers in some areas restored to a tight arrangement, which to a certain extent improved the muscle lesions of the model group.

[0102] Example 5 Effects on Spatial Learning and Cognitive Ability of Aging Mice

[0103] The animal experiment design was the same as in Example 1.

[0104] The Morris water maze experiment was carried out one week before the mice were sacrificed (week 8). The water maze experiment included a 4-day positioning navigation experiment and a 1-day spatial exploration experiment. In the positioning navigation experiment, each mouse was trained 4 times a day, entering the water from the four quadrants respectively, with an interval of 2 hours between each trial. The time it took for each mouse to find the hidden platform within 60 seconds (escape latency) was recorded. If the mouse could not find the platform within 60 seconds, the escape latency was recorded as 60 seconds. At the same time, the mouse was guided to the platform and stood on the platform for observation for 10 seconds. On the fifth day, the hidden platform was removed for the spatial exploration experiment. Each mouse was placed in the water from the third quadrant, and the number of times the mouse crossed the center of the target quadrant (where the platform was previously located) within 60 seconds was recorded. During the experiment, the overhead camera equipped with the equipment was used to record the number of times the experimental mice crossed the platform and the swimming trajectory.

[0105] The results are as follows Figure 6 As shown in (A), after 4 consecutive days of positioning navigation training, the escape latency of mice in each group shortened with the extension of training time. Compared with the model group, the escape latency of the fucoidan + CCFM1424 group on the 4th day was significantly decreased (p < 0.05). The spatial exploration test was carried out on the 5th day, and the results are shown in Figure 6(BE). The escape latency, number of times the mice crossed the platform, and time they stayed in the target quadrant of the model group were 45.4±13.02s, 0.33±0.58 times, and 7.42±4.28%, respectively; the escape latency, number of times the mice crossed the platform, and time they stayed in the target quadrant of the blank group were 11.93±6.59s, 4.33±0.58 times, and 25.91±3.04%, respectively; the escape latency, number of times the mice crossed the platform, and time they stayed in the target quadrant of the CCFM1424 group were 22.2±4.2s, 1.33±0.58 times, and 16.61±1.33%, respectively; the escape latency, number of times the mice crossed the platform, and time they stayed in the target quadrant of the fucoidan group were 22.2±4.2s, 1.33±0.58 times, and 16.61±1.33%, respectively. The escape latency, the number of times the mice crossed the platform and the time they stayed in the target quadrant were 17.67±4.11s, 1.67±1.15 times and 19.16±0.51% respectively; the escape latency, the number of times the mice crossed the platform and the time they stayed in the target quadrant of the mice in the fucoidan+CCFM1424 group were 8.73±4.35s, 4.00±1.00 times and 27.13±3.00% respectively; the escape latency, the number of times the mice crossed the platform and the time they stayed in the target quadrant of the mice in the arginine group were 17.27±3.70s, 3.00±1.00 times and 22.48±1.07% respectively. The results show that compared to the blank group, the model group had a significantly increased escape latency (p<0.01), a significantly decreased number of platform crossings (p<0.01), and a significantly decreased time spent in the target quadrant (p<0.001). After intervention, these indicators improved in both the experimental and positive drug control groups, with the most significant improvement seen in the fucoidan + CCFM1424 group. This suggests that the fucoidan + CCFM1424 combination can improve the decline in spatial learning and memory function caused by aging.

[0106] Example 6 Effects of oxidative stress on liver tissue of aging mice

[0107] The animal experiment design was the same as in Example 1.

[0108] After the experiment, the mice were dissected in accordance with ethical requirements, and the mouse liver tissue was weighed and mixed with physiological saline at a ratio of 1:9. The tissue was crushed using a tissue crusher at 65 Hz for 30 s (cycled 5-8 times), and finally centrifuged at 3000×g for 10 min. The supernatant was taken and the activity of brain tissue SOD (item number: A001-3-2, purchased from Nanjing Jiancheng), CAT (item number: A007-1-1, purchased from Nanjing Jiancheng) and GSH-Px (item number: A005-1-2, purchased from Nanjing Jiancheng) and the concentration of MDA (item number: A003-1-2, purchased from Nanjing Jiancheng) were detected by kits. The protein concentration of the supernatant was determined using a BCA protein concentration assay kit. The results are shown in Figure 2. Figure 5-6 .

[0109] The content of MDA can not only indirectly reflect the degree of tissue oxidative damage, but also be used to test whether the D-galactose-induced aging model is successful. Figure 7 (A) shows that the MDA content in liver tissue of the model group was 4.6±0.70 nmol / mg protein, while that of the blank group was 1.94±0.10 nmol / mg protein. The MDA content in the model group increased by 138% compared with the blank group, indicating the success of D-galactose aging modeling. Compared with the model group, the MDA content in the experimental groups (CCFM1424 group, fucoidan group, and fucoidan + CCFM1424 group) decreased by 51%, 47%, and 58%, respectively. The fucoidan + CCFM1424 group had a more significant effect in reducing MDA content (p<0.01). The MDA content in the positive drug control group decreased by 45%.

[0110] like Figure 7 As shown in (B), SOD activity in liver tissue was 192.95±3.54 U / mg protein in the blank group, 154.48±4.88 U / mg protein in the model group, 189.3±11.22 U / mg protein in the CCFM1424 group, 195.57±13.24 U / mg protein in the fucoidan group, 222.46±12.15 U / mg protein in the fucoidan + CCFM1424 group, and 196.34±10.59 U / mg protein in the arginine group. Compared with the SOD activity in the blank group, the SOD activity in the model group decreased by 20%. Compared with the SOD activity of the model group, the SOD activity of the CCFM1424 group, fucoidan group, fucoidan + CCFM1424 group and arginine group increased by 23%, 27%, 44% and 27%, respectively, among which the increase in the fucoidan + CCFM1424 group was the most significant (p<0.001).

[0111] like Figure 7 As shown in (C), CAT activity in liver tissue was 23.54±2.64 U / mg protein in the blank group, 16.1±1.96 U / mg protein in the model group, 22.17±0.88 U / mg protein in the CCFM1424 group, 22.83±2 U / mg protein in the fucoidan group, 26.39±2.53 U / mg protein in the fucoidan + CCFM1424 group, and 24.18±3.18 U / mg protein in the arginine group. Compared with the blank group, CAT activity in the model group decreased by 32%. Compared with the model group, the other experimental groups and the positive drug control group all significantly reduced CAT activity in mouse liver tissue, with the fucoidan + CCFM1424 group showing the greatest effect in reducing CAT activity in mouse liver tissue (p<0.01).

[0112] like Figure 7As shown in (D), GSH-Px activity in liver tissue was 313.15±18.26 U / mg protein in the blank group, 250.68±10.78 U / mg protein in the model group, 343.26±15.49 U / mg protein in the CCFM1424 group, 308.97±11.25 U / mg protein in the fucoidan group, 377.17±11.99 U / mg protein in the fucoidan + CCFM1424 group, and 277.55±13.63 U / mg protein in the arginine group. Compared with the GSH-Px activity in the blank group, the GSH-Px activity in the model group decreased by 20%. Compared with the model group, the GSH-Px activities of CCFM1424 group, fucoidan group, fucoidan+CCFM1424 group and arginine group increased by 37%, 23%, 50% and 11%, respectively, among which the increase in fucoidan+CCFM1424 group was the most significant (p<0.001).

[0113] Example 7 Effects on Cytokines in Serum of Aging Mice

[0114] The animal experiment design was the same as in Example 1.

[0115] After the experiment, in accordance with ethical requirements, the mice were eyeballed and blood was collected and dissected. The blood was allowed to stand for 2 hours and then centrifuged at 3000 r / min for 15 minutes. The mouse serum was collected and the levels of cytokines TNF-α (catalog number: SBJ-M0030-96T, purchased from Nanjing Senbega Biotechnology Co., Ltd.), IL-1β (catalog number: SBJ-M0027-96T, purchased from Nanjing Senbega Biotechnology Co., Ltd.), IL-6 (catalog number: SBJ-M0657-96T, purchased from Nanjing Senbega Biotechnology Co., Ltd.), and IL-10 (catalog number: SBJ-M0073-96T, purchased from Nanjing Senbega Biotechnology Co., Ltd.) in the mouse serum were measured using a kit (catalog number 439807, purchased from Nanjing Senbega Biotechnology Co., Ltd.). The results are shown in the table. Figure 8 shown.

[0116] Serum inflammatory factor IL-1β concentration is shown in Figure 8 A. The serum IL-1β concentration in the blank group was 50.96±1.28 ng / L, while that in the model group was 74.5±4.14 ng / L, a 46% increase. IL-1β concentrations in the CCFM1424, fucoidan, fucoidan + CCFM1424, and arginine intervention groups were 50.99±2.09 ng / L, 55.33±3.26 ng / L, 48.42±1.26 ng / L, and 50.63±3.24 ng / L, respectively, significantly lower than those in the model group (p<0.001), with decreases of 32%, 26%, 35%, and 32%, respectively. The fucoidan + CCFM1424 group showed the greatest reduction.

[0117] Serum inflammatory factor IL-6 concentration is shown in Figure 8 As shown in Figure 2, the serum IL-6 concentration in the blank group was 23.49±2.23 pg / mL, while that in the model group was 31.57±3.35 pg / mL, a 34% increase. The IL-6 concentrations in CCFM1424, fucoidan, fucoidan + CCFM1424, and arginine interventions were 24.95±1.7 pg / mL, 22.02±2.86 pg / mL, 20.48±1.1 pg / mL, and 23.84±0.57 pg / mL, respectively, significantly lower than those in the model group (p<0.05), representing decreases of 21%, 30%, 35%, and 34%, respectively. The fucoidan + CCFM1424 group showed the greatest decrease in serum IL-6 concentration.

[0118] Serum inflammatory factor TNF-α concentration is shown in Figure 8 C. The serum TNF-α concentration in the blank group was 200.29±10.26 ng / L, while that in the model group was 268.98±10.77 ng / L, a 46% increase. The TNF-α concentrations in CCFM1424, fucoidan, fucoidan + CCFM1424, and arginine interventions were 229.79±15.81 ng / L, 219.89±22.18 ng / L, 179.69±10.91 ng / L, and 190.19±8.93 ng / L, respectively, significantly lower than those in the model group (p<0.05), with decreases of 15%, 18%, 33%, and 29%, respectively. Among them, the fucoidan + CCFM1424 group had the most significant effect in reducing serum TNF-α levels in mice (p<0.001), surpassing both the CCFM1424 and fucoidan groups.

[0119] Serum anti-inflammatory factor IL-10 concentration is shown in Figure 8 D. The serum IL-10 concentration in the blank group was 332.18±8.78 pg / mL, while that in the model group was 270.58±1.67 pg / mL, a decrease of 19%. The IL-10 concentrations in the CCFM1424, fucoidan, fucoidan + CCFM1424, and arginine intervention groups were 283.11±2.88 pg / mL, 299.38±9.61 pg / mL, 332.45±11.21 pg / mL, and 287.11±1.6 pg / mL, respectively, all increasing compared with the model group. Significant increases were observed in the fucoidan (p<0.01) and fucoidan + CCFM1424 groups (p<0.001), increasing by 11% and 23%, respectively.

[0120] Example 8 Effects on Intestinal Flora in Aging Mice

[0121] The animal experiment design was the same as in Example 1.

[0122] Before the end of the experiment, mouse feces were collected and total DNA was extracted from the mouse feces samples using the MP feces kit. The specific operation steps are as follows, mainly referring to the kit instructions. V3-V4 region PCR amplification was performed with primers (341F: 5'-CCTAYGGGRBGCASCAG-3', 806R: 5'-GGACTACNNGGGTATCTAAT-3'). After amplification, the target band was recovered according to the QIAquick Gel Extraction Kit instructions after gel electrophoresis detection. The concentration and purity of the gel-recovered DNA were measured and recorded using a NanoDrop 2000 ultramicrophotometer. The library was constructed according to the TurSeq DNA LTSample Preparation Kit and its instructions, and finally the sequence was sequenced on an Illumina Miseq sequencer according to the MiSeq Regent Kit and its instructions. The QIIME2 analysis pipeline was used to process the sequence data, and the Lianchuan Cloud website (https: / / www.omicstudio.cn / tool) was used for principal coordinates analysis (PCoA).

[0123] Alpha diversity and beta diversity are used to evaluate the changes in the diversity of fecal microbiota ( Figure 9 ). Among them, α diversity is characterized by observed_features, and the results show that ( Figure 9 A), observed_features in the model group decreased significantly (p<0.05), while after intervention with fucoidan + CCFM1424, observed_features increased significantly to normal levels (p<0.05). β diversity was evaluated using NMDS, and the results showed that ( Figure 9 B), there were significant differences among the blank group, model group, fucoidan + CCFM1424 group and arginine group (p = 0.001). From the perspective of the relative abundance changes at the phylum level, the relative abundance of Proteobacteria in the model group was significantly higher than that in the blank group (p < 0.05, Figure 9 C), Proteobacteria include a variety of pathogens including Escherichia coli, Salmonella, and Vibrio cholerae. After the intervention of fucoidan + CCFM1424, the relative abundance of Proteobacteria decreased significantly (p<0.05), indicating that fucoidan + CCFM1424 can reduce the relative abundance of harmful intestinal bacteria and restore the health of intestinal flora. The ratio of Firmicutes to Bacteroidetes (F / B value) in the model group was significantly higher than that in the blank group (p<0.05, Figure 9D), while after fucoidan + CCFM1424 intervention, the F / B value was significantly decreased (p<0.05), indicating that fucoidan + CCFM1424 is beneficial to the health of intestinal microbiota.

[0124] In summary, the microecological preparation provided by the present invention has a good effect in alleviating aging.

[0125] Although the specific embodiments of the present invention are described above, those skilled in the art should understand that the specific embodiments described are merely illustrative and are not intended to limit the scope of the present invention. Equivalent modifications and changes made by those skilled in the art in accordance with the spirit of the present invention should be included within the scope of protection of the claims of the present invention.

Claims

1. A microecological preparation that can alleviate aging, characterized in that: The microecological preparation comprises Bifidobacterium bifidum and fucoidan.

2. The microecological preparation according to claim 1, characterized in that The Bifidobacterium bifidum is Bifidobacterium bifidum CCFM1424, which is deposited in Guangdong Provincial Microbiological Culture Collection Center with a deposit number of GDMCC No: 65151.

3. The microecological preparation according to claim 1, characterized in that The content of the Bifidobacterium bifidum CCFM1424 strain is ≥1×10 9 CFU / mL.

4. The microecological preparation according to claim 1, characterized in that The concentration of the fucoidan is not less than 30 mg / kg BW / day.

5. Use of the probiotic preparation according to claim 1 in the preparation of products for alleviating aging.

6. The use of the composition for preparing an anti-aging product according to claim 1, characterized in that: The aging alleviation function includes any of the following: (1) Reduce the frailty index of elderly individuals; (2) increase grip strength in elderly individuals; (3) Improve muscle mass in aging mice; (4) Improve muscle tissue morphology in aging individuals; (5) Can significantly alleviate cognitive decline caused by aging; (6) significantly alleviate oxidative stress in the liver of elderly individuals, wherein the antioxidant index includes at least one of MDA, SOD, CAT, or GSH-Px; (7) reducing the expression level of inflammatory factors in the serum of elderly individuals, wherein the inflammatory factors include at least one of IL-1β, IL-6, or TNF-α; (8) increasing the expression level of anti-inflammatory factors in the serum of elderly individuals, wherein the anti-inflammatory factor is IL-10; (9) Alleviate the intestinal microbiome disorder caused by aging.

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