Medicinal and edible composition for improving senile muscle attenuation

Abstract

The invention discloses a medicinal and edible composition for improving muscle attenuation of old people, the composition comprises fructus piperis longi and radix asparagi extracts, and the composition is combined with highland barley, common turnip, maca, D-psicose and other active substances to develop a series of medicinal and edible compositions for improving muscle attenuation. The composition disclosed by the invention has a good protective effect in improvement of old-age muscle attenuation, and can be applied to muscle attenuation and exercise ability decline accompanied by aging, so that the activity of telomerase is enhanced; meanwhile, the traditional Chinese medicine composition is helpful for adjusting the proportion of immune cell subtypes, improving functional activity decline of an immune system and reducing chronic low-level inflammation states, and has both safety and effectiveness. Molecular mechanism research shows that the compound activates glutathione metabolism and inhibits ferroptosis signals to reverse skeletal muscle cell senescence. The composition disclosed by the invention can be developed into foods and health-care products for improving muscle mass, volume, strength and functions, and has a very good application prospect.

Description

Technical Field

[0001] This invention relates to the field of food and health care, and more particularly to a food-medicine homology composition for improving muscle atrophy in the elderly. Background Technology

[0002] Sarcopenia, also known as muscle wasting syndrome, is a progressive, systemic skeletal muscle disease characterized by a decline in muscle mass and motor function. It is a major age-related phenotypic feature in older adults. Sarcopenia is classified into primary and secondary types, and is more prevalent in two groups: first, older adults without underlying diseases, which may be termed geriatric sarcopenia; and second, patients with multiple underlying chronic or wasting diseases, which may induce secondary sarcopenia.

[0003] With the accelerating aging process in modern society, muscle atrophy, decreased mobility, and metabolic disorders in the elderly have gradually become an increasingly prominent public health problem. Extensive surveys and research data show that muscle atrophy and motor dysfunction are prevalent among people aged 65 and above, with nearly half suffering from varying degrees of distress, placing enormous pressure and burden on society and families. Faced with this serious health challenge, existing intervention methods have significant limitations. Common anti-aging drugs, such as the well-known combination therapy of dasatinib and quercetin, can target and regulate the skeletal muscle aging-related gene p16. Ink4a and p21 Cip1 While it can significantly inhibit apoptosis, this treatment may cause side effects such as anemia, thrombocytopenia, and neutropenia, and it also has certain cytotoxicity, which poses potential health risks to the elderly.

[0004] In the non-pharmaceutical sector, some functional foods or dietary supplements on the market already exist, primarily based on single or a few natural active ingredients, such as products with caffeine, taurine, and protein bars as core components. However, these products generally suffer from limited efficacy and single target, and are ill-suited to the slower metabolism and multiple coexisting conditions of the elderly. Geriatric sarcopenia is a complex physiological and pathological process involving multiple pathways and targets. Single ingredients often only target one or a few aspects, failing to comprehensively improve the multi-system damage associated with aging. This results in unstable or insignificant effects, failing to meet the urgent needs of the elderly market. Therefore, there is an urgent need in this field for a safe, effective, and comprehensive solution that can improve the symptoms of geriatric sarcopenia while ensuring safety and efficacy. Summary of the Invention

[0005] To address at least one of the aforementioned problems, the present invention provides a food-medicine composition for improving age-related muscle atrophy, and the application of a composition containing Piper longum and Asparagus in improving age-related muscle atrophy syndrome, thereby enhancing the efficacy of food-medicine materials in improving age-related muscle atrophy.

[0006] To achieve the above-mentioned invention, the first objective of this invention is to provide a medicinal and edible composition for improving muscle atrophy in the elderly, comprising, by weight percentage: 30%-70% Piper longum extract and 30%-70% Asparagus extract.

[0007] Furthermore, the preparation method of the composition includes the following specific steps:

[0008] (1) The preparation method of Piper longum extract is as follows: Piper longum is used as raw material, and it is obtained by water extraction, filtration, concentration and freeze drying;

[0009] (2) The preparation method of Asparagus extract is as follows: Asparagus is used as raw material, and it is obtained by water extraction, filtration, concentration and freeze drying;

[0010] (3) By mass percentage, 30%-70% of Piper longum extract and 30%-70% of Asparagus extract are mixed and compounded to form a Piper longum and Asparagus composition;

[0011] Further, in step (1), 5 to 15 times the weight of the raw material of purified water is added to Piper longum, and the mixture is refluxed for 1 to 1.5 hours. The residue is filtered through a 10 to 100 μm membrane, and the residue is extracted repeatedly. The two extracts are combined, and the supernatant is concentrated under reduced pressure at 50 to 80°C and 0.1 to 1 MPa to 10% to 30% of the original volume. Then, the mixture is freeze-dried to obtain Piper longum extract.

[0012] Further, in step (2), 5 to 15 times the weight of the raw material of purified water is added to the asparagus, and the mixture is refluxed for 1 to 1.5 hours. The residue is filtered through a 10 to 100 μm membrane, and the residue is extracted repeatedly. The two extracts are combined, and the supernatant is concentrated under reduced pressure at 50 to 80°C and 0.1 to 1 MPa to 10% to 30% of the original volume. Then, the mixture is freeze-dried to obtain the asparagus extract.

[0013] In this invention, the medicinal and edible composition for improving muscle atrophy in the elderly further comprises, by weight percentage: 50% to 80% of the above-mentioned composition of Piper longum and Asparagus extracts, and any or all of the following components:

[0014] (1) 20%-50% highland barley;

[0015] (2) 20%-50% coriander root;

[0016] (3) 20%-50% Maca;

[0017] (4) 20%-50% D-alokulose.

[0018] Furthermore, the barley undergoes a pre-treatment of crushing, being crushed to a mesh size of 50-200.

[0019] Furthermore, the coriander root is a coriander root extract obtained through water extraction, filtration, concentration, and freeze-drying.

[0020] Furthermore, the maca is a maca extract obtained through water extraction, filtration, concentration, and freeze-drying.

[0021] Furthermore, the chemical formula of the D-allulose is C6H. 12 O6.

[0022] A second object of the present invention is to provide the use of the composition in the preparation of a medicine or food for improving muscle loss in the elderly.

[0023] Furthermore, the application includes its use in the preparation of medicinal food products for the prevention and / or improvement of muscle atrophy or muscle movement capacity in the elderly.

[0024] Furthermore, the medicinal food products are functional foods, dietary supplements, traditional Chinese medicine preparations, or drugs.

[0025] Furthermore, the traditional Chinese medicine preparations or drugs mentioned include tablets, capsules, oral liquids, lozenges, granules, pills, powders, ointments, elixirs, suspensions, powders, suppositories, ointments, plasters, creams, sprays, drops, patches, and drop pills.

[0026] Furthermore, by adding excipients to the food-medicine homology composition of the present invention, a traditional Chinese medicine preparation or drug is prepared; wherein the excipients include mannitol, sorbitol, sodium metabisulfite, sodium bisulfite, sodium thiosulfate, cysteine ​​hydrochloride, thioglycolic acid, methionine, vitamin C, disodium EDTA, sodium calcium EDTA, carbonates of monovalent alkali metals, acetates, phosphates or aqueous solutions, hydrochloric acid, acetic acid, sulfuric acid, phosphoric acid, amino acids, sodium chloride, potassium chloride, sodium lactate, xylitol, maltose, glucose, fructose, dextran, starch, sucrose, mannitol, silicon derivatives, cellulose and its derivatives, alginate, gelatin, polyvinylpyrrolidone, glycerol, Tween 80, agar, calcium carbonate, calcium bicarbonate, surfactants, polyethylene glycol, cyclodextrin, phospholipids, kaolin, talc, calcium stearate, and magnesium stearate.

[0027] Furthermore, the application in the prevention and / or improvement of muscle loss includes its use in primary and secondary sarcopenic syndromes such as asthenia, myasthenia, muscle stiffness, muscle dysfunction, or muscle atrophy.

[0028] Furthermore, the application in preventing and / or improving muscle loss also includes applications in enhancing and / or protecting muscle mass, volume, strength, and function.

[0029] The beneficial effects of this invention are:

[0030] (1) This invention develops a series of compound compositions for improving muscle loss in the elderly by combining extracts of Piper longum and Asparagus with active substances such as barley, coriander root, maca and D-alokulose. Foods, health products and drugs prepared using this compound composition are both safe and effective in improving muscle mass and exercise capacity in the elderly sarcopenia syndrome, and have very good application prospects.

[0031] (2) Based on the combination of Piper longum and Asparagus, this invention combines one or more food materials that are "dual-purpose medicine and food" or "new resource food". Compared with current anti-aging drugs, it not only improves muscle loss in the elderly, but also ensures the safety of raw materials. At the same time, this invention also helps to regulate the proportion of immune cell subtypes such as lymphocytes, neutrophils, monocytes, eosinophils and basophils in elderly individuals, improve the decline of immune system function and activity, reduce chronic low-level inflammation, and improve the state of innate immunosuppression.

[0032] (3) This invention, through behavioral experiments and physiological and biochemical index detection in aging mice, confirmed that the combination of Piper longum and Asparagus has a good protective effect against muscle atrophy in the elderly, and demonstrated the application of this combination in improving age-related muscle mass and motor ability from a molecular mechanism perspective. RNA-seq sequencing of mouse skeletal muscle tissue identified key regulatory genes, revealing that it mainly reverses skeletal muscle cell aging by activating glutathione-glutathione peroxidase 4 (GSH-GPX4) and inhibiting key targets in ferroptosis signals such as prostaglandin intraperoxidase 2 (Ptgs2) and transferrin receptor 1 (TfR1). This invention provides a theoretical basis and practical case for studying the molecular mechanism of food-medicine homologous combinations such as Piper longum and Asparagus extracts in improving muscle atrophy in the elderly. Attached Figure Description

[0033] Figure 1 The images are H&E staining images of mouse hind limb skeletal muscle tissue from Example 1 and Comparative Examples 1-3 of this invention.

[0034] Figure 2 The content of immune cells in whole blood of mice in Example 1 and Comparative Examples 1-3 of this invention.

[0035] Figure 3 The images show RNA-seq sequencing analysis of mouse hindlimb skeletal muscle tissue from Examples 1 and 1-2 of this invention.

[0036] Figure 4 The levels of GSH and ferroptosis-related genes in mouse skeletal muscle tissue of Examples 1 and 1-2 of this invention are shown.

[0037] Figure 5 The images are H&E staining images of mouse hind limb skeletal muscle tissue from Examples 2-6 and Comparative Examples 4-7 of this invention.

[0038] in, Figure 1 The images above are partial images of the examples below, magnified 200 times and 400 times respectively, with corresponding scale bars of 200μm and 100μm.

[0039] Figure 2 middle, Figure 2 a is the total blood cell count of mice in Example 1 and Comparative Examples 1-3; Figure 2 b shows neutrophil images from mice in Example 1 and Comparative Examples 1-3; Figure 2 c shows the lymphocytes of mice in Example 1 and Comparative Examples 1-3; Figure 2 d shows the mononuclear cells of mice in Example 1 and Comparative Examples 1-3; Figure 2 e shows eosinophils from mice in Example 1 and Comparative Examples 1-3; Figure 2 f shows basophils from mice in Example 1 and Comparative Examples 1-3.

[0040] Figure 3 middle, Figure 3 a is a statistical graph showing the number of differentially expressed genes (DEGs) in the skeletal muscle of mice in Example 1 and Comparative Example 2 groups; Figure 3 b is the volcano diagram of DEGs; Figure 3 c represents the heatmap analysis of DEGs; Figure 3 d is a graph showing the enrichment of the KEGG signal pathway in DEGs.

[0041] Figure 4 middle, Figure 4 a represents the level of glutathione in the serum of mice in Example 1 and Comparative Examples 1-2; Figure 4 b represents the serum glutathione peroxidase level; Figure 4 c is a graph showing the GPX4 gene expression level in mouse skeletal muscle; Figure 4 d shows the expression level of the Ptgs2 gene in mouse skeletal muscle; Figure 4 e is a graph showing the expression level of the TfR1 gene in mouse skeletal muscle.

[0042] Figure 5 The images above are partial images of the examples below, magnified 200 times and 400 times respectively, with corresponding scale bars of 200μm and 100μm. Detailed Implementation

[0043] To illustrate the invention in detail, the following embodiments, in conjunction with the accompanying drawings, describe the technical solutions of the invention, which will help those skilled in the art to further understand the invention, but do not limit the invention in any way. It should be noted that the embodiments are preferred solutions of the invention and are intended to illustrate the implementation conditions that can be used to implement the invention, rather than limiting the experimental conditions. All other embodiments obtained by those skilled in the art based on the embodiments of the invention without creative effort are within the scope of protection of the invention.

[0044] Example 1:

[0045] A medicinal and edible composition for improving muscle loss in the elderly: by weight percentage, it comprises: 30%-70% Piper longum extract and 30%-70% Asparagus extract.

[0046] 1. Preparation method of 50% Piper longum and 50% Asparagus extract and their composition:

[0047] Step 1: After drying and pulverizing Piper longum, add 10 times the weight of the herb in purified water and reflux at 95°C for 1.5 hours. Filter through a 100μm membrane and collect the supernatant. Repeat the above process on the residue and collect the supernatant again. Combine the two supernatants, concentrate under reduced pressure, concentrate 10 times, collect the supernatant, and freeze-dry to obtain Piper longum extract.

[0048] Step 2: After drying and pulverizing the asparagus, add 10 times the weight of the medicinal material in purified water, reflux at 95℃ for 1.5 hours, filter through a 100μm membrane, and collect the supernatant. Repeat the above process for the residue and collect the supernatant again. Combine the two supernatants, concentrate under reduced pressure, concentrate 10 times, collect the supernatant, freeze dry, and obtain the asparagus extract.

[0049] Step 3: Mix 50% of the Piper longum extract from Step 1 and 50% of the Asparagus extract from Step 2 in a 1:1 ratio to form a Piper longum and Asparagus combined extract.

[0050] 2. Feeding and intervention of experimental mice

[0051] The experiment of this invention was conducted on C57BL / 6 mice (8 weeks old and 18 months old) one week after they were adapted to the experimental environment. Ten mice were randomly divided into groups, and the intervention lasted for 2 months. The 8-week-old mice were considered young mice, and the 18-month-old mice were considered old mice. After 2 months of intervention, various indicators were tested and analyzed, at which time the mice were 4 months and 20 months old, respectively.

[0052] Mouse housing conditions: The animal experimental facility continuously maintains the barrier environment standard. The main environmental indicators are controlled within the following ranges: temperature 23.1±0.9℃, relative humidity 52.8±3.0%, air exchange rate 10-20 times / hour, light:dark ratio 12h:12h. Husbandry and management are carried out by qualified personnel. Bedding and cages are changed and disinfected twice a week to keep the environment clean and dry. Feed and water are provided daily, allowing animals free access to eat and move around.

[0053] Intervention plan: Daily gavage administration for 2 months.

[0054] Intervention dose: 200 mg / 10 g mouse body weight.

[0055] 3. Testing the motor abilities of mice

[0056] (1) Forelimb grip strength measurement

[0057] The forelimb gripping force of mice was assessed using a gripping force meter (BIO-GS3). The mouse's forelimbs were placed on the metal mesh of the gripping force meter, allowing its paws to grip the wire. Simultaneously, the mouse's tail was lifted, making its body parallel to the table, and it was gently pulled backward until the mouse's paws lost their grip on the wire. The gripping force (N) was recorded for each test, with three trials per mouse, and the average gripping force was calculated.

[0058] (2) Fatigue rotating bar test

[0059] Before the formal test, the mice were trained by placing them on a fatigue rotundus (ZB-200) and having them maintain balance for 1 minute. Then, the rotundus speed was set to 5 rpm, and the mice were instructed to move in the opposite direction of the rotundus for 5 minutes without falling. After all mice had completed training, the formal experiment was conducted. Within 1 minute, the rotundus speed was gradually increased to 15 rpm, and mice were placed on the rotundus in sequence according to the training order. When a mouse became fatigued and fell off the rotundus, the instrument automatically recorded the time (in seconds) the mouse spent on the rotundus.

[0060] (3) Climbing ability test

[0061] To determine whether mice had motor dysfunction, each mouse was tested for climbing ability. During the test, the mouse was placed head-up at the top of a vertical bar (1 cm in diameter and 60 cm high), and the mouse would descend the bar from top to bottom. The total time (s) required for the mouse to descend was recorded. Each mouse was tested three times, and the average value was calculated.

[0062] 4. Mouse immune cell analysis

[0063] Fresh anticoagulated whole blood samples were collected for complete blood cell analysis. A fully automated hematology analyzer (BC-5000Vet, Mindray, China) was used to perform a whole blood cell (WBC) count, including lymphocytes (Lym), neutrophils (Neu), monocytes (Mon), eosinophils (Eos), and basophils (Bas).

[0064] 5. Tests for muscle loss-related indicators

[0065] (1) Muscle mass measurement

[0066] After behavioral testing, the mice were immediately euthanized by cervical dislocation. The skeletal muscle tissue of the mice was then dissected on ice, and the mass (mg) was precisely weighed on an analytical balance. The tissue was then flash-frozen in liquid nitrogen and transferred to -80°C.

[0067] (2) Telomerase assay

[0068] Approximately 20 mg of mouse skeletal muscle tissue was collected, and the level of telomerase (TE) in the skeletal muscle tissue was measured using an ELISA kit.

[0069] (3) Determination of GSH content and GSH-Px activity

[0070] Approximately 20 mg of mouse skeletal muscle tissue was collected, and the content of reduced glutathione (GSH) and the activity of glutathione peroxidase (GSH-Px) in the skeletal muscle tissue were determined using a biochemical reagent kit (microplate method).

[0071] (4) Histopathological observation

[0072] Mouse skeletal muscle tissue was fixed by immersion in 4% paraformaldehyde solution for 24 hours for histological and pathological observation. The tissues were sequentially dehydrated using 70%, 80%, 90%, and 100% ethanol, embedded in paraffin, and then sectioned to a thickness of 4-5 μm using ultrasectioning. The tissue sections were then stained with hematoxylin and eosin (H&E) staining. Colon tissue sections were scanned and imaged using a digital section scanner, and the scanned images were analyzed using Case Viewer v2.3 software.

[0073] 6. Skeletal muscle transcriptomic analysis

[0074] (1) Sample processing

[0075] Take an appropriate amount of skeletal muscle tissue and extract total RNA from the sample using the TRIzol method. Weigh 20 mg of skeletal muscle tissue and add 500 μL of FreeZol Reagent. After lysis at room temperature for 5 min, add Dilution Buffer and let stand for 5 min. Centrifuge at 11200 rpm for 15 min. Collect the supernatant and add an equal volume of isopropanol. Mix well and let stand for 10 min. Centrifuge at 11200 rpm for 10 min. Add 75% mL of ethanol to the white precipitate, resuspend, centrifuge at 9100 rpm for 3 min, discard the supernatant, air dry, and dissolve the precipitate in enzyme-free water.

[0076] (2) Quality control and sequencing

[0077] The concentration and purity of the extracted RNA were determined. Agarose gel electrophoresis was used to check the integrity of the RNA in each sample, and the RQN value (>6.5) was measured using an Agilent 5300. For total RNA that passed quality control, AT base pairing was performed using Oligo(dT) magnetic beads to separate and enrich mRNA. Fragmentation buffer was added to fragment the mRNA (300bp), and cDNA was synthesized using random primers via reverse chromatography, followed by double-strand synthesis. End Repair Mix was added to blunt the sticky ends of the double-stranded cDNA, and an A base was added to the 3' end. Finally, the ligation adapter product was purified, fragmented, amplified by PCR, and enriched into a library. Quantification was performed using a second-generation high-throughput sequencing platform (NovaSeq X Plus) using Qubit 4.0, and the data were then sequenced. Bridge PCR amplification was performed on a cBot to generate clusters, which were then sequenced.

[0078] (3) Data Analysis

[0079] The raw sequencing reads were filtered using FASTP software to remove adapter sequences, trim low-quality bases at the ends, remove reads with N% > 10%, and discard sequences < 20 bq in length, resulting in high-quality sequencing data (clean data). This data was then used for quality assessment and subsequent analysis. HiSat2 software was used to align the clean data with a reference genome, obtaining mapped data for transcript assembly and expression level calculation, and the results were quality controlled. By locating genomic regions, RSEM software was used to calculate gene and transcript expression levels, obtaining gene read counts. Differential expression analysis and Kyoto Genome Encyclopedia (KEGG) enrichment analysis were performed on genes from different groups using DEGseq, Goatools, and Python scipy packages.

[0080] 7. Real-time quantitative polymerase chain reaction

[0081] (1) Extraction of tissue RNA

[0082] RNA was extracted from mouse hind limb skeletal muscle tissue according to the instructions of the RNA kit. The extracted RNA was measured using a micro spectrophotometer to ensure that the OD260 / 280 of the RNA was within 1.8-2.2, and the integrity of the RNA was checked by an RNA agarose gel assay before further processing.

[0083] (2) cDNA synthesis

[0084] According to the kit instructions, the total amount of RNA in each sample was adjusted to be consistent. Then, the mixture shown in Table 1 was prepared in RNase-free centrifuge tubes. After mixing, the reaction was performed according to the following procedure: 50℃, 15min; 85℃, 5s.

[0085] Table 1 Reverse transcription reaction system

[0086]

[0087] (3) RT-qPCR

[0088] Dilute the obtained cDNA reaction solution to an appropriate concentration. Prepare the RT-qPCR reaction system according to the kit instructions; the specific amplification system is shown in Table 2.

[0089] Table 2 RT-qPCR amplification system

[0090]

[0091] After preparing the RT-qPCR reaction system, place it in a quantitative PCR instrument. The reaction program should be set according to the instruction manual; the primers used are shown in Table 3. Perform at least three replicates for each sample, using β-actin as a reference gene, and apply 2^(- ) qPCR. ΔΔCt () indicates the relative mRNA expression of each target gene.

[0092] Table 3. Gene primer sequences for RT-qPCR

[0093]

[0094] Comparative Example 1 of this invention uses 8-week-old young mice, which are subjected to no treatment and are directly used in animal experiments as the blank control group of this invention.

[0095] Comparative Example 2 of this invention uses 18-month-old senescent mice. Comparative Example 2 of this invention uses the same amount of distilled water as the extract in Example 1 for animal experiments and serves as the experimental control group.

[0096] Comparative Example 3 of this invention uses 18-month-old senescent mice. Comparative Example 3 of this invention uses resveratrol as the positive control, with the same amount of extract as in Example 1. Animal experiments are conducted in Comparative Example 3 as a positive control group.

[0097] The mice in Example 1 and Comparative Examples 1-3 were subjected to functional tests, and the test results are shown in Table 4:

[0098] Table 4 Test Results of Example 1

[0099]

[0100] As shown in Table 4, compared with the young mice in the blank control group of Comparative Example 1, the aging mice in the experimental control group of Comparative Example 2 had significantly reduced motor ability, muscle mass and telomerase level. In contrast, the homologous composition in Example 1, which used 50% Piper longum and 50% Asparagus extract, had a more significant improvement effect compared with the positive control group of Comparative Example 3. Specifically, it significantly improved the forelimb grip strength of aging mice, reduced the exercise time on the fatigue rotarod test and reduced the climbing time, improved skeletal muscle tissue quality and reduced TE level.

[0101] according to Figure 1 H&E staining images of mouse hind limb skeletal muscle showed that, compared with the young mice in the blank control group of Comparative Example 1, the skeletal muscle cells of the aged mice in the experimental control group of Comparative Example 2 were loosely arranged, the cross-sectional area of ​​muscle fibers was reduced, and they showed slight swelling. Local cell nuclei were pyknoid or ruptured, and immune cells infiltrated and accumulated locally. In contrast, Example 1, which used 50% Piper longum and 50% Asparagus extract in combination, showed that the skeletal muscle cells of the mice were tightly arranged, the cross-sectional area of ​​muscle fibers was significantly increased, and fewer cracks were observed. The effect was significantly better than that of the mice in Comparative Example 3.

[0102] from Figure 2 Analysis of the content of immune cells in whole blood showed that, compared with the young mice in the blank control group (Comparative Example 1), the number of total blood cells in the peripheral blood of the aged mice in the experimental control group (Comparative Example 2) was significantly reduced. Figure 2 a). In Example 1, the combined use of Piper longum and Asparagus extract significantly increased the level of immune cell populations and improved the innate immunosuppressive state compared to the experimental control group in Comparative Example 2. Furthermore, it was found that the levels of the major immune cell populations—neutrophils, lymphocytes, eosinophils, and basophils—were significantly increased. Figure 2 b~ Figure 2 f), while the number of monocytes was significantly reduced (f). Figure 2 d). Since monocytes and macrophages are key factors in promoting low-grade chronic inflammation, extracts of Piper longum and Asparagus cochinchinensis may help regulate the proportions of various immune cell subtypes, improve the decline in immune system function, and reduce chronic low-level inflammation in older individuals.

[0103] This invention, based on RNA-seq sequencing, further explored the potential transcriptional regulatory mechanism of the combined use of Piper longum and Asparagus extracts on skeletal muscle aging in aging mice. According to... Figure 3 As shown, compared with the young mice in the blank control group of Comparative Example 1, a total of 696 differentially expressed genes (DEGs) were screened from the skeletal muscle of the aged mice in the experimental control group of Comparative Example 2. Among them, 307 differentially expressed genes were significantly upregulated and 389 genes were significantly downregulated. Figure 3 a, Figure 3 b). According to the heatmap of differential gene expression, the use of Piper longum and Asparagus extracts significantly reversed the skeletal gene expression profile in aging mice. Figure 3 c).

[0104] To further clarify the regulatory mechanism of Piper longum and Asparagus extracts on skeletal muscle in aging mice, this invention performs KEGG bio-enrichment analysis on differentially expressed genes in mouse skeletal muscle. Figure 3 d). The results showed that DEGs were mainly enriched in ferroptosis, ABC transporter signaling, and glutathione metabolism.

[0105] from Figure 4 It can be seen that, compared with the young mice in the blank control group (Comparative Example 1), the levels of GSH and GSH-Px in the skeletal muscle of the aged mice in the experimental control group (Comparative Example 2) were significantly reduced. Figure 4 a, Figure 4 b). In Example 1, the combined use of Piper longum and Asparagus extract significantly increased the levels of both compared to the control group in Comparative Example 2. RT-qPCR analysis of key genes in ferroptosis signaling revealed that the ferroptosis repressor gene GPX4 was significantly activated by Piper longum and Asparagus extract. Figure 4 c), and significantly inhibited key genes that induce ferroptosis, such as Ptgs2 and TfR1. Figure 4 d, Figure 4 e).

[0106] Example 2

[0107] This embodiment provides a composition for improving sarcopenia in the elderly, comprising 35% Piper longum extract, 35% Asparagus root extract, and 30% barley, resulting in a food-medicine homology composition. Other testing conditions are consistent with those in Example 1. The barley used is barley flour.

[0108] The method for preparing the barley flour is as follows: dry the barley and then pulverize it to a powder of 100 mesh.

[0109] Example 3

[0110] This embodiment provides a composition for improving sarcopenia in the elderly, comprising 35% Piper longum extract, 35% Asparagus root extract, and 30% Daphne genkwa extract, resulting in a food-medicine homology composition. Other testing conditions are consistent with those in Example 1. The Daphne genkwa extract is used in this embodiment.

[0111] The preparation method of the *Corydalis yanhusuo* root extract is as follows: after drying and pulverizing the *Corydalis yanhusuo* root, add 10 times the weight of the medicinal material in purified water, reflux extract at 95℃ for 1.5 h, filter through a 100 μm membrane, collect the supernatant, repeat the above process on the residue once, and collect the supernatant again; combine the two supernatants, concentrate under reduced pressure, concentrate 10 times, collect the supernatant, freeze dry, and obtain the *Corydalis yanhusuo* root extract.

[0112] Example 4

[0113] This embodiment provides a composition for improving sarcopenia in the elderly, comprising 35% Piper longum extract, 35% Asparagus root extract, and 30% Maca, resulting in a food-medicine homology composition. Other testing conditions are consistent with those in Example 1. The Maca used is a Maca extract.

[0114] The preparation method of the maca extract is as follows: the root of the plant is dried and pulverized, and 10 times the weight of the medicinal material in purified water is added. The mixture is refluxed at 95°C for 1.5 hours, filtered through a 100μm membrane, and the supernatant is collected. The residue is extracted once more using the above process, and the supernatant is collected again. The two supernatants are combined, concentrated under reduced pressure, and concentrated 10 times. The supernatant is collected and freeze-dried to obtain the maca extract.

[0115] Example 5

[0116] This embodiment provides a composition for improving sarcopenia in the elderly, comprising 40% Piper longum extract, 40% Asparagus extract and 20% D-allulose, to obtain a food-medicine homology composition, with other test conditions consistent with Example 1.

[0117] Example 6

[0118] This embodiment provides a composition for improving sarcopenia in the elderly, comprising 30% Piper longum extract, 30% Asparagus root extract, 10% highland barley flour, 10% coriander root, 10% maca and 10% D-aloxone, to obtain a food-medicine homology composition, with other test conditions consistent with Example 1.

[0119] Comparative Example 4

[0120] The specific implementation method is the same as in Example 1, except that the Piper longum extract is omitted.

[0121] Comparative Example 5

[0122] The specific implementation method is the same as in Example 1, except that the asparagus extract is omitted.

[0123] Comparative Example 6

[0124] The specific implementation method is the same as in Example 1, except that the Piper longum extract in the composition is replaced with resveratrol.

[0125] Comparative Example 7

[0126] The specific implementation method is the same as in Example 1, except that the asparagus extract in the composition is replaced with resveratrol.

[0127] Table 5 Test results of Examples 2-6

[0128]

[0129]

[0130] As shown in Table 5, compared to the combined use of Piper longum and Asparagus extract in Example 1, the combination of Piper longum and Asparagus extract with active substances such as highland barley, coriander root, maca extract, and D-allulose exhibits a significant synergistic effect, greatly improving the forelimb grip strength and movement time on the fatigue rotundus in aging mice; increasing pole climbing speed and reducing pole climbing time; delaying skeletal muscle mass decline; and increasing telomerase levels. Compared to the positive group mice in Comparative Example 3 that were fed resveratrol alone and the mice treated with the combined intervention of Piper longum and Asparagus extract in Example 1, the combination of Piper longum and Asparagus extract with multiple active substances such as highland barley, coriander root, maca, and D-allulose demonstrates a greater effect in improving senile sarcopenia, while significantly improving muscle mass, strength, and motor ability in aging mice, and increasing telomerase levels.

[0131] As can be seen from Table 5, through comparative examples 4 to 7, in the combination of Piper longum extract and Asparagus extract, omitting or replacing any of the components affects the effect of the composition on improving the decline in motor ability and muscle loss in aging mice.

[0132] according to Figure 5 H&E staining images of skeletal muscle showed that in Examples 2-6, the skeletal muscle cells in mice were tightly packed, the cross-sectional area of ​​muscle fibers was significantly increased, and fewer cracks were observed. These results improved the attenuation of mouse skeletal muscle tissue to varying degrees, and the effects were significantly better than those in Comparative Examples 4-7.

[0133] The above are merely optional embodiments of the present invention and do not limit the patent scope of the present invention. All equivalent structural transformations made using the contents of the present invention specification and drawings, or direct / indirect applications in other related technical fields, are included within the patent protection scope of the present invention, as will be understood by those skilled in the art under the technical concept of the present invention.

Claims

1. A medicinal and edible composition for improving muscle atrophy in the elderly, characterized in that, The composition comprises, by weight percentage: 30%-70% Piper longum extract and 30%-70% Asparagus extract.

2. A medicinal and edible composition for improving muscle atrophy in the elderly, characterized in that, The composition, by weight percentage, comprises: 50% to 80% of the composition as described in claim 1, and any or all of the following components: (1) 20%-50% highland barley; (2) 20%-50% coriander root; (3) 20%-50% Maca; (4) 20%-50% D-alokulose.

3. The medicinal and edible composition for improving muscle atrophy in the elderly as described in claim 1, characterized in that, The preparation method of the composition includes the following steps: (1) Piper longum extract was prepared by water extraction, filtration, concentration and freeze drying. (2) Asparagus extract was prepared by water extraction, filtration, concentration and freeze drying. (3) By mass percentage, 30%-70% of the Piper longum extract from step (1) and 30%-70% of the Asparagus extract from step (2) are mixed and compounded to form a Piper longum and Asparagus composition.

4. The medicinal and edible composition for improving muscle atrophy in the elderly as described in claim 3, characterized in that, The preparation method of Piper longum extract in step (1) is as follows: add 5 to 15 times the weight of the raw material of purified water to Piper longum, reflux extract for 1 to 1.5 hours, filter with a 10 to 100 μm membrane, extract the residue repeatedly, combine the two extracts, concentrate under reduced pressure at 50 to 80°C and 0.1 to 1 MPa to 10% to 30% of the original volume, freeze dry to obtain Piper longum extract; The preparation method of the asparagus extract in step (2) is as follows: add 5 to 15 times the weight of the raw material of purified water to the asparagus, reflux for 1 to 1.5 hours, filter with a 10 to 100 μm membrane, extract the residue repeatedly, combine the two extracts, concentrate under reduced pressure at 50 to 80°C and 0.1 to 1 MPa to 10% to 30% of the original volume, freeze dry, and obtain the asparagus extract.

5. The use of the composition according to any one of claims 1-4 in the preparation of a medicinal food product for improving muscle loss in the elderly.

6. The application as described in claim 5, characterized in that, The applications include use in food and medicinal products for the prevention and / or improvement of age-related muscle loss or muscle mobility, including functional foods, dietary supplements, or medicines.

7. The application as described in claim 6, characterized in that, The applications in preventing and / or improving muscle loss also include applications that enhance and / or protect muscle mass, volume, strength, and function.

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