Dietary composition for preventing and treating senile sarcopenia and application thereof

Abstract

The invention discloses a dietary composition for preventing and treating senile sarcopenia and application thereof, and relates to the technical field of dietary nutrition and health. The dietary composition disclosed by the invention comprises lactobacillus paracasei LC86, lactobacillus rhamnosus LRa05, lactobacillus plantarum Lp05 and plant protein, and the components are combined according to the proportion of (8.3-50) * 10 < 9 > CFU of lactobacillus paracasei LC86, (8.3-50) * 10 < 9 > CFU of lactobacillus rhamnosus LRa05 and (8.3-50) * 10 < 9 > CFU of lactobacillus plantarum Lp05 in every 5g of plant protein. The dietary composition improves the biological value of vegetable protein, does not introduce the fat intake risk of animal protein, improves the symptoms of the senile sarcopenia, and has a remarkable effect.

Description

Technical Field

[0001] This invention relates to the field of dietary nutrition and health technology, and more specifically to a dietary composition for preventing and treating sarcopenia in the elderly and its application. Background Technology

[0002] With the accelerating aging of society, the elderly face more and more complex health problems than younger people, leading to increased care needs and making their disease burden the highest globally. Sarcopenia, an age-related progressive skeletal muscle disease characterized by the accelerated loss of skeletal muscle mass, strength, and / or function, is a significant public health issue in aging societies. Its insidious progression and gradual worsening are important risk factors for mobility impairments, falls, frailty, and reduced quality of life in the elderly.

[0003] Currently, there are no specific drugs for sarcopenia in clinical practice, and the main recommendations are physical exercise and a reasonable diet. However, physical exercise has high environmental requirements and is somewhat difficult for the elderly due to their special physiological conditions, while improving diet seems to have more practical application value.

[0004] Studies have shown that increasing dietary protein intake can prevent sarcopenia to some extent. However, plant protein is not a high-quality protein and its actual effect on improving sarcopenia is not good. Animal protein may increase fat intake, which may increase the risk of dyslipidemia in the elderly.

[0005] Therefore, how to address the risks of fat intake from animal protein and the biological value of plant protein in order to prevent and treat sarcopenia in the elderly is a problem that urgently needs to be solved by those skilled in the art. Summary of the Invention

[0006] In view of this, the present invention provides a dietary composition for preventing and treating sarcopenia in the elderly and its application.

[0007] To achieve the above objectives, the present invention adopts the following technical solution:

[0008] A dietary composition for preventing and treating sarcopenia in the elderly includes *Lactobacillus paracasei* LC86, *Lactobacillus rhamnosus* LRa05, *Lactobacillus plantarum* Lp05, and plant protein, at a ratio of (8.3~50) × 10 per 5 grams of plant protein. 9 CFU Lactobacillus paracasei LC86, (8.3~50) × 10 9 CFU Lactobacillus rhamnosus LRa05 and (8.3~50)×10 9 The ratio of CFU to Lactobacillus plantarum Lp05.

[0009] Preferably, the dietary composition for preventing and treating sarcopenia in the elderly is formulated with a plant protein ratio of 50 × 1 ... 9 CFU Lactobacillus paracasei LC86, 50×10 9 CFU Lactobacillus rhamnosus LRa05 and 50×10 9 The ratio of CFU to Lactobacillus plantarum Lp05.

[0010] Another object of the present invention is to provide the application of the above-mentioned dietary composition for preventing and treating sarcopenia in the preparation of products for preventing and treating sarcopenia.

[0011] Beneficial Effects: The dietary composition of this invention for preventing and treating sarcopenia in the elderly synergistically combines *Lactobacillus paracasei* LC86, *Lactobacillus rhamnosus* LRa05, *Lactobacillus plantarum* Lp05, and plant protein, significantly improving the absolute grip strength and body weight-normalized grip strength of aged rats, while also significantly increasing the gastrocnemius and quadriceps muscle indices. Furthermore, this dietary formula significantly increased serum SOD levels and decreased serum MDA, IL-6, and TNF-α levels in aged rats. In targeted metabolomics and high-throughput sequencing, this dietary composition significantly increased the content of branched-chain amino acids and short-chain fatty acids such as acetic acid, propionic acid, and butyric acid, while also improving the gut microbiota structure. Mechanistically, this dietary composition significantly increased the protein expression levels of AMPK-α1 and p70 S6K, suggesting that it can improve sarcopenia by enhancing the body's protein synthesis rate. This dietary composition improves the biological value of plant protein without introducing the risk of fat intake associated with animal protein, significantly improving the symptoms of sarcopenia in the elderly. Attached Figure Description

[0012] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on the provided drawings without creative effort.

[0013] Figure 1 This is a flowchart of the experimental design for this invention.

[0014] Figure 2 This invention investigates the effects of dietary interventions on muscle function and muscle mass in rats.

[0015] Figure 3 This invention investigates the effects of dietary interventions on blood biochemical indicators in rats.

[0016] Figure 4 This invention investigates the effects of dietary interventions on short-chain fatty acids in rat feces.

[0017] Figure 5 This invention illustrates the effects of dietary interventions on the diversity of gut microbiota in rats. Specifically, A represents the effect of dietary intervention on gut α-diversity (ACE), B represents the effect of dietary intervention on gut α-diversity (Shannon), C represents the effect of dietary intervention on gut health index, and D represents the effect of dietary intervention on gut β-diversity.

[0018] Figure 6 This invention investigates the effects of dietary interventions on the composition and structure of the gut microbiota in rats.

[0019] Figure 7 This invention compares the abundance of differentially expressed microbiota between the NC and MC groups during dietary intervention.

[0020] Figure 8 This invention compares the abundance of differentially expressed microbiota between the MC group and the high-dose group during dietary intervention.

[0021] Figure 9 This invention provides a comparative analysis of the LDA of gut microbiota in dietary interventions.

[0022] Figure 10 This invention investigates the effects of dietary interventions on the serum amino acid profiles of rats in various groups.

[0023] Figure 11 This invention investigates the effects of dietary interventions on the expression levels of proteins related to muscle synthesis in rats. Detailed Implementation

[0024] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0025] This invention discloses a dietary composition for preventing and treating sarcopenia in the elderly. The raw materials used, including *Lactobacillus paracasei* LC86 lyophilized powder, *Lactobacillus rhamnosus* LRa05 lyophilized powder, *Lactobacillus plantarum* Lp05 lyophilized powder, and plant protein, are all commercially available. For example, *Lactobacillus paracasei* LC86 lyophilized powder, *Lactobacillus rhamnosus* LRa05 lyophilized powder, and *Lactobacillus plantarum* Lp05 lyophilized powder can be purchased from Microcare Probiotics (Suzhou) Co., Ltd. (specification 1.0 × 10⁻⁶). 11 The plant protein (CFU / g) and soy protein are available from Tianmeijian Biotechnology (Beijing) Co., Ltd. Unless otherwise specified, the methods involved in this invention are conventional methods and will not be described in detail here.

[0026] Example 1

[0027] Lactobacillus paracasei LC86, Lactobacillus rhamnosus LRa05, Lactobacillus plantarum Lp05, and plant protein were administered at a ratio of 8.3 × 10⁻⁶. 9 CFU, 8.3×10 9 CFU, 8.3×10 9 CFU and 5.0g were combined (as the baseline formula, i.e., the low-dose group, represented as A+B, where A represents 5.0g of plant protein and B represents 8.3×10 of each of the three probiotics). 9 CFU (Continuous Fuel Cells) are used for subsequent experiments.

[0028] Example 2

[0029] Lactobacillus paracasei LC86, Lactobacillus rhamnosus LRa05, Lactobacillus plantarum Lp05, and plant protein were mixed at a concentration of 16 × 10⁻⁶. 9 CFU, 16×10 9 CFU, 16×10 9 CFU and 5.0g were combined (i.e., the medium-dose group, denoted as A+2B) for subsequent experiments.

[0030] Example 3

[0031] Lactobacillus paracasei LC86, Lactobacillus rhamnosus LRa05, Lactobacillus plantarum Lp05, and plant protein were mixed at 50×10⁻⁶. 9 CFU, 50×10 9 CFU, 50×10 9 CFU and 5.0g were combined (i.e., the high-dose group, denoted as A+6B) for subsequent experiments.

[0032] Example 4

[0033] 1. Laboratory animals and their husbandry methods

[0034] All animal experiments in this invention comply with national regulations and have been approved by the Experimental Animal Ethics Committee of Hangzhou Medical College (Approval No.: 2023-037).

[0035] Experimental animals: 6-week-old SPF-grade Sprague Dawley (SD) rats were purchased from Zhejiang Provincial Experimental Animal Center (License No.: SCXK (Zhejiang) 2024-0002).

[0036] SD rats were raised in a controlled environment until they were 18 months old, with free access to food and water, a 12-hour light / dark cycle, a temperature of 20-22℃, and a humidity of 40±5%.

[0037] 2. Experimental Design (see Appendix) Figure 1 (n=8)

[0038] NC group (adolescent control group): 6-month-old non-aged control + normal feed + physiological saline;

[0039] MC group (aged model control group): 18-month-old aged rats + normal diet + physiological saline;

[0040] Low group (low-dose intervention group): 18-month-old aged rats + normal diet + dietary composition of Example 1, wherein the amount of dietary composition of Example 1 added is (A+B) / kg / day;

[0041] Mid group (medium-dose intervention group): 18-month-old aged rats + normal diet + dietary composition of Example 2, wherein the amount of dietary composition of Example 2 added is (A+2B) / kg / day;

[0042] High group (high-dose intervention group): 18-month-old aged rats + normal diet + dietary composition of Example 3, wherein the amount of dietary composition of Example 3 added is (A+6B) / kg / day.

[0043] The probiotic dosage in the intervention group of this invention was based on previous studies, with low, medium, and high doses equivalent to 5 times, 10 times, and 30 times the recommended dose for a 60kg adult, respectively; the amount of soy protein used was 10 times the acceptable range of plant protein (30g / d) for the elderly population recommended in the 2023 edition of the "Chinese Dietary Reference Intakes".

[0044] Rats’ body weight and food intake were recorded regularly during the experiment, and the intervention lasted for 12 weeks.

[0045] 3. Indicator Measurement

[0046] (1) Measurement of animal gripping force

[0047] The grip strength of the whole limb was measured using the YLS-13A rat grip strength tester (Jinan Yiyan Technology Development Co., Ltd., Jinan, China) at weeks 0, 6 and 12 of the experiment.

[0048] The specific procedure is as follows: Place the rat's limbs on the metal grid of the force gauge, allowing it to grasp naturally. The experimenter holds the rat's tail and pulls it horizontally backward at a constant speed, recording the maximum gripping force (peak force) during the pulling process. Each rat is measured three times, and the average value is taken as the raw gripping force data. The final gripping force value is standardized by body weight and calculated using the formula: Final gripping force = Average raw gripping force / Rat body weight (g / g).

[0049] The results are attached. Figure 2As shown, compared with the NC group, the MC group showed significantly lower absolute grip strength, body weight-normalized grip strength, gastrocnemius muscle mass index, and quadriceps femoris muscle mass index. After dietary intervention, compared with the MC group, the low, medium, and high dose groups all significantly improved the absolute grip strength and body weight-normalized grip strength of rats; the medium dose group significantly increased the gastrocnemius muscle mass index of rats, while the low dose group significantly increased the quadriceps femoris muscle mass index of rats.

[0050] (2) Blood biochemistry test

[0051] After the experiment, the animals were anesthetized and blood was collected. The four serum lipid parameters were detected using a Mindray BS-800 fully automated blood biochemistry analyzer. Commercial test kits provided by Nanjing Jiancheng Bioengineering Research Institute were used, and the operation was strictly carried out in accordance with the instructions of the kits. The concentrations of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), malondialdehyde (MDA, a lipid peroxidation marker) and superoxide dismutase (SOD, an antioxidant enzyme) were detected.

[0052] The results are attached. Figure 3 As shown, compared to the NC group, the MC group had significantly increased levels of IL-6, TNF-α, and MDA, while significantly decreased levels of SOD. After dietary intervention, compared to the MC group, all three dose groups (low, medium, and high doses) significantly reduced serum IL-6 and MDA levels; the medium and high dose groups significantly reduced TNF-α levels, but only the high dose group significantly increased SOD levels. There were no significant changes in the four lipid profiles.

[0053] (3) Fecal short-chain fatty acid detection

[0054] Preparation of mixed standard solutions: Accurately weigh 10 mg of each of the eight standards, add 50% acetonitrile solution, vortex to mix, and dilute to 1 mL to prepare stock solutions of each standard. Mix 100 μL of each stock solution and dilute to 1 mL to obtain mixed standard stock solution A. Dilute mixed standard stock solution A 50 times to obtain mixed standard stock solution B with a concentration of 20 μg / mL. Take 40 μL of mixed standard stock solution B and add the following in sequence: 20 μL of 200 mM 3-nitrophenylhydrazine hydrochloride (3NPH·HCl) and 20 μL of 120 mM 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC·HCl, containing 6% pyridine). The above mixture is derivatized at 40℃ for 30 min, and then diluted to 200 μL with 50% acetonitrile. The solutions are then serially diluted to prepare the L1-L14 series of working solutions (dispensed into 1.5 mL EP tubes).

[0055] Sample pretreatment: Accurately weigh 20 mg of fecal sample and add 500 μL of extraction buffer (methanol:water = 4:1, v / v). Grind using a cryogenic grinder (-10℃, 50 Hz) for 6 min, then sonicate in an ice-water bath (5℃, 40 kHz) for 30 min. Let the sample stand for 30 min, centrifuge at 13000 rcf for 15 min at 4℃, take 20 μL of supernatant, add: 20 μL of 200 mM 3NPH·HCl, 20 μL of 120 mM EDC·HCl (containing 6% pyridine), derivatize at 40℃ for 30 min, and then bring the volume to 1000 μL with 50% acetonitrile for analysis.

[0056] UHPLC-MS / MS analytical conditions:

[0057] Chromatographic system: ExionLC AD ultra-high performance liquid chromatography (equipped with QTRAP® 6500+ mass spectrometer detector, AB Sciex, USA); column: Waters BEH C18 column (150 × 2.1 mm, 1.7 μm); column temperature: 40℃. Mobile phase: Phase A: 0.01% formic acid aqueous solution; Phase B: 0.01% formic acid acetonitrile solution.

[0058] Gradient elution procedure:

[0059] | Time period (min) | Phase B ratio (%) | Flow rate (mL / min) |

[0060] |---------------|--------------|----------------|

[0061] | 0-2 | 10 | 0.35 |

[0062] | 2-11 | 10→55 | 0.35 |

[0063] | 11-12 | 55→95 | 0.35 |

[0064] | 12-13 | 95 | 0.35 |

[0065] | 13-13.1 | 95→10 | 0.35 |

[0066] | 13.1-16 | 10 | 0.35 |

[0067] Mass spectrometry conditions:

[0068] Ion source: Electrospray ionization (ESI), negative ion mode; source parameters: temperature 450℃, curtain gas 35 psi, collision gas medium, half of the ion source gas 40 psi, spray voltage -4500 V

[0069] Quality control measures:

[0070] For every 5-10 samples analyzed, insert quality control samples (QC), using mixed samples or medium-concentration standard solutions, and calculate the relative standard deviation (RSD) of each target analyte. The RSD should be <15% to ensure system stability.

[0071] Data processing:

[0072] The AB Sciex OS quantitative software was used for automatic integration, combined with manual verification. A standard curve was plotted with the analyte peak area as the ordinate and the concentration as the abscissa. The sample peak area was substituted into the linear equation to calculate the actual concentration, and the internal standard method was used for quantification.

[0073] The results are attached. Figure 4 As shown, compared to the NC group, the levels of acetic acid, propionic acid, isobutyric acid, valeric acid, isovaleric acid, and hexanoic acid were significantly decreased in the MC group. Compared with the model control group, the low, medium, and high dose groups all significantly increased serum acetic acid levels; the high dose group significantly increased propionic acid, butyric acid, valeric acid, and hexanoic acid levels.

[0074] (4) 16S high-throughput sequencing

[0075] DNA Extraction and Illumina Sequencing: After collecting fecal samples under sterile conditions, all samples were immediately flash-frozen in liquid nitrogen and stored at -80°C. Total microbial genomic DNA was extracted from feces using a rapid pure fecal DNA isolation kit (Shanghai Meiji Biomedical Technology Co., Ltd.) according to the manufacturer's instructions. DNA quality and concentration were determined by 1.0% agarose gel electrophoresis and a NanoDrop® ND-2000 spectrophotometer (Thermo Scientific, USA). The V3-V4 hypervariable region of the bacterial 16S rRNA gene was amplified using primer pairs 338F (5'-ACTCCTACGGGAGGCAGCAG-3') and 806R (5'-GGACTACHVGGGTWTCTAAT-3') on a BIO-RAD T100 gradient PCR instrument (USA). The PCR reaction system consisted of: 4 μL 5×FastPfu buffer, 2 μL 2.5 mM dNTPs, 0.8 μL each of primers (5 μM), 0.4 μL Fast Pfu polymerase, 10 ng template DNA, and ddH2O to a final volume of 20 μL. All samples were amplified three times. The PCR products were purified by 2% agarose gel electrophoresis and quantified using a Synergy HTX multi-mode microplate reader (Biotek, USA). The purified amplicons were mixed in equimolar amounts and sequenced at both ends using a standard procedure on an Illumina NextSeq 2000 PE300 sequencing platform (Illumina, San Diego, USA).

[0076] Sequence Processing and Data Analysis: After sample splitting, the original sequences were quality filtered using fastp (v0.19.6) and assembled using FLASH (v1.2.11). High-quality sequences were denoised using the DADA2 plugin in the Qiime2 (version 2020.2) workflow (using recommended parameters). This algorithm achieves single nucleotide resolution based on in-sample error features, generating amplicon sequence variants (ASVs). Based on ASV information, α-diversity indices such as the ACE index and Shannon index were calculated using Mothur v1.30.2. Principal coordinate analysis (PCoA) was performed using the Bray-curtis dissimilarity matrix from the Vegan v2.4.3 package to assess the similarity of microbial communities among different samples. The linear discriminant analysis effect size (LEfSe) method (LDA score > 2.5, P < 0.05) was used to screen bacterial groups with significant differences between groups.

[0077] The results are attached. Figure 5-9As shown, compared with the MC group, the high-dose group significantly increased the ACE and GMHI indices of the gut microbiota. At the β-diversity level, the microbial composition of the intervention group showed significant separation from the MC group with increasing intervention dose, and was similar to that of the NC group. At the genus level, the relative abundance of Lactobacillus, norank_f_Muribaculaceae, and Lachnospiraceae_NK4A136_group increased in the high-dose group, while the relative abundance of Romboutsia decreased. Compared to the NC group, the MC group exhibited a significantly disordered microbial composition, specifically with a significant increase in the abundance of Romboutsia, Blautia, Turicibacter, Bifidobacterium, Faecalibaculum, and Acetitomaculum, while the abundance of Lachnospiraceae_NK4A136_group, Eubacterium_siraeum_group, norank_f_Ruminococcaceae, Lachnospiraceae_UCG-006, and Roseburia was significantly decreased. In contrast, compared to aged rats, the high-dose group showed a significant decrease in the relative abundance of some harmful bacteria, such as Romboutsia, Turicibacter, and Eubacterium_ruminatium_group, while the abundance of other bacteria, such as norank_f_Muribaculaceae, Phascolarctobacterium, Alistipes, and Lachnospiraceae_UCG-006, was significantly increased. The results of linear discriminant analysis (LDA) also supported these significant changes, showing that the abundance of 6, 6 and 16 genera were significantly enriched in the NC group, MC group and the three intervention groups, respectively.

[0078] (5) Detection of serum amino acid profile

[0079] Sample Preparation: Transfer 15 μL of sample to a 1.5 mL Eppendorf centrifuge tube. Add 35 μL of ultrapure water and 200 μL of extraction solvent (acetonitrile:methanol = 1:1 volume ratio, containing an isotopic internal standard mixture, pre-cooled to -40°C). Vortex for 30 seconds until completely mixed. Sonicate the mixture in an ice-water bath for 15 minutes. Then, incubate the sample solution at -40°C for 1 hour. Centrifuge at 12,000 rpm (corresponding to a centrifugal force of 13,800 × g and a rotor radius of 8.6 cm) for 15 minutes at 4°C. Take 100 μL of the supernatant and evaporate it to dryness using a rotary evaporator. Redissolve the residue in 100 μL of an aqueous solution containing 50% methanol. Add 100 μL of derivatization reagent and 50 μL of 1M NaHCO3 solution, and vortex to mix. Place the mixture in a 40°C water bath for derivatization reaction for 1 hour. After removal and cooling to room temperature, add 50 μL of 2M HCl solution and evaporate again to dryness. Finally, redissolve the residue in 200 μL of methanol for instrumental analysis.

[0080] Preparation of standard solutions: Weigh each standard substance separately, dissolve or dilute to a final concentration of 10 mmol / L, and prepare single-standard stock solutions. Transfer an appropriate amount of each stock solution to a 10 mL volumetric flask to prepare a mixed working standard solution. By serially diluting this mixed standard solution (containing an internal standard mixture of isotopes with the same concentration as the sample), a series of calibration standard solutions with concentration gradients are obtained.

[0081] UHPLC-MRM-MS analysis:

[0082] Chromatographic conditions: A Thermo Vanquish ultra-high performance liquid chromatography system (Thermo Fisher) was used, equipped with a Waters ACQUITY UPLC BEH C18 column (100 × 2.1 mm, 1.7 μm). Mobile phase A was 5 mM ammonium acetate aqueous solution, and mobile phase B was acetonitrile. The column temperature was set to 45 °C, the autosampler temperature to 4 °C, and the injection volume to 2 μL. Mass spectrometry conditions: A Thermo Altis TSQ Plus mass spectrometer (Thermo Fisher, USA) equipped with an electrospray ionization source was used. Typical ionization parameters: spray voltage -3300 V, sheath gas 40 Arb, auxiliary gas 10 Arb, purge gas 1 Arb, ion transfer tube temperature 325 °C, and evaporator temperature 350 °C. Method optimization: Multiple reaction monitoring (MRM) parameters for each target analyte were optimized by flow injection analysis. Standard solutions of individual analytes were directly injected into the API ion source of the mass spectrometer. In MRM scan mode, the most sensitive precursor / daughter ion pairs (Q1 / Q3) were selected, and collision energies were optimized. For each analyte, the Q1 / Q3 pair with the highest response and best selectivity was selected as the quantifier, while the remaining ion pairs were used as qualifiers to identify the target analyte. MRM data acquisition was performed using Xcalibur software (version 4.4.16.14, Thermo Fisher), and data processing was performed using Skyline software.

[0083] The results are attached. Figure 10 As shown, compared with the MC group, the intervention group showed a significant increase in the levels of nine amino acids. Of particular note was the decrease in the levels of total branched-chain amino acids (BCAA), L-valine, L-isoleucine, and L-leucine in the MC group, while the high-dose intervention group significantly increased the levels of these amino acids. Furthermore, the levels of other amino acids such as L-2-aminobutyric acid, L-aspartic acid, D-glutamic acid, L-phenylalanine, L-citrulline, and L-kynurenine were increased in the medium- and high-dose intervention groups.

[0084] (6) Western blot detection

[0085] Protein extraction and immunoblotting analysis:

[0086] Protein extraction from muscle tissue (30–50 mg): Tissue was lysed using RIPA lysis buffer (Shanghai Beyotime Biotechnology Co., Ltd.) containing a 1% phosphatase inhibitor mixture and a 1% protease inhibitor mixture (RPP), under ice bath conditions. The tissue was centrifuged at 15,000 × g for 15 minutes at 4°C, and the supernatant was collected as the clarified lysis buffer.

[0087] Protein quantification: Protein content was determined using the BCA protein concentration assay kit (Shanghai Beyotime Biotechnology Co., Ltd.).

[0088] Protein separation and transfer: Proteins were separated by 12% SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and then transferred to polyvinylidene fluoride (PVDF) membranes (Millibert & Co., Inc., USA).

[0089] Immune testing:

[0090] Blocking: PVDF membranes were blocked with blocking buffer containing 5% skim milk powder at room temperature for 1 hour; Primary antibody incubation: Incubation with primary antibody overnight at 4°C; Secondary antibody incubation: Incubation was then performed with horseradish peroxidase (HRP)-labeled secondary antibody; Color development and imaging: Protein bands were detected using a Tanon 5500 chemiluminescence imaging system (Shanghai Tianneng Technology Co., Ltd.). Band grayscale values ​​were calculated using ImageJ software (version 1.5, USA).

[0091] Antibody information: p70 S6K protein (Abcam, catalog number ab2571), AMPK-α1 protein (Abcam, catalog number ab32047), GAPDH internal reference protein (Abcam, catalog number ab8245).

[0092] The results are attached. Figure 11 As shown, compared with the NC group, the expression levels of AMPK-α1 and p70 S6K in the muscle of the MC group were significantly decreased. However, compared with the MC group, the protein expression level of AMPK-α1 in the muscle of the high-dose group was significantly increased. Similarly, compared with the MC group, the protein expression level of p70 S6K in the medium-dose group was also significantly increased.

[0093] Data from this invention are expressed as mean ± standard deviation (mean ± SD) for normally distributed data and as median and interquartile range (IQR) for non-normally distributed data. One-way ANOVA was used to analyze grip strength, muscle mass, and related biochemical parameters, with Tukey's multiple comparisons method used for post-hoc tests. All analyses were performed using SPSS 18.0 software (SPSS Inc., USA), and a p-value < 0.05 was considered statistically significant.

[0094] The various embodiments in this specification are described in a progressive manner, with each embodiment focusing on the differences from other embodiments. The same or similar parts between the various embodiments can be referred to each other.

[0095] The above description of the disclosed embodiments enables those skilled in the art to make or use the invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the invention. Therefore, the invention is not to be limited to the embodiments shown herein, but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A dietary composition for the prevention and treatment of sarcopenia, characterized by, Combinations of proportions comprising Lactobacillus paracasei LC86, Lactobacillus rhamnosus LRa05, Lactobacillus plantarum Lp05 and plant protein, according to the ratio of (8.3~50)×10 9 CFU Lactobacillus paracasei LC86, (8.3~50)×10 9 CFU Lactobacillus rhamnosus LRa05 and (8.3~50)×10 9 CFU Lactobacillus plantarum Lp05 per 5 grams of plant protein.

2. The dietary composition for preventing and treating sarcopenia according to claim 1, characterized by, According to the proportion of 50 x 10 9 CFU Lactobacillus paracasei LC86, 50 x 10 9 CFU Lactobacillus rhamnosus LRa05 and 50 x 10 9 CFU Lactobacillus plantarum Lp05 in the ratio combination.

3. Use of the dietary composition for the prevention and treatment of sarcopenia according to claim 1 or 2 for the manufacture of a product for the prevention and treatment of sarcopenia.

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