Composition for preventing, improving or treating muscle weakness-related diseases containing celery seed extract as an active ingredient

Abstract

The present invention relates to a composition for preventing, improving, or treating diseases associated with muscle weakness, comprising a celery seed extract. The celery seed extract according to the present invention is a natural product and can increase muscle mass and strength with few or no side effects. Therefore, it is expected that the celery seed extract according to the present invention can be useful for preventing, improving, or treating diseases associated with muscle weakness, metabolic diseases, liver diseases, etc., since it can increase muscle mass and strength with few or no side effects by using a natural product and can not only prevent muscle weakness, but also inhibit increases in body weight and body fat, and reduce blood lipid concentrations and liver toxicity indicators (GOT and GPT).

Description

[Technical Field]

[0001] The present invention relates to a composition for preventing, ameliorating or treating a muscle weakness-related disease, comprising a celery seed extract.

[0002] This application claims the benefit of priority based on Korean Patent Application No. 10-2021-0121287 filed on September 10, 2021, and Korean Patent Application No. 10-2022-0085416 filed on July 12, 2022, and all contents disclosed in the specifications and drawings of said applications are incorporated herein by reference. [Background technology]

[0003] Skeletal muscle is the largest organ in the human body, accounting for 40-50% of total body weight, and plays an important role in various metabolic functions, including energy homeostasis and heat generation. As the human body ages, changes in its composition occur, resulting in a redistribution of body fat and protein. At around age 50, the rate of protein synthesis in muscle cells slows down compared to the rate of protein breakdown, and muscle begins to rapidly degenerate.

[0004] Sarcopenia is a condition characterized by a loss of approximately 13-24% of one's normal body mass, and is characterized by a decrease in protein content, fiber diameter, muscle strength production, and fatigue resistance. Sarcopenia can occur for a variety of reasons, including sepsis, cancer, renal failure, glucocorticoid excess, denervation, muscle disuse, obesity, and the aging process. It is primarily caused by the gradual loss of skeletal muscle mass and quality that occurs as aging progresses.

[0005] Meanwhile, sarcopenic obesity, which occurs when muscle mass and strength decrease with aging while body fat mass increases, has become a problem. Obesity and sarcopenia in the elderly are thought to synergistically interact with each other, exacerbating the risk of functional impairment, metabolic disorders, and even death. From a pathological perspective, these conditions have been suggested to have a strong mutual interaction. Therefore, for overweight or obese individuals, or individuals with normal weight but high body fat, increasing muscle mass and reducing body fat can prevent and treat metabolic diseases such as diabetes and hypertension. Increasing muscle mass increases basal metabolic energy, enabling efficient weight loss without the yo-yo effect. Therefore, exercise, dietary therapy, and ergogenic aids are used to increase muscle mass. However, commercially available ergogenic aids contain chemical compounds that can cause fatal side effects.

[0006] Therefore, the present inventors have endeavored to develop a therapeutic agent for muscle weakness that has no side effects and is highly effective. As a result, they have confirmed that celery seed extract increases muscle mass and strength in obese or aging mice, thereby completing the present invention. Summary of the Invention [Problem to be solved by the invention]

[0007] The present inventors have endeavored to develop a naturally occurring active substance capable of preventing, ameliorating, or treating muscle weakness-related diseases. As a result, they have confirmed the excellent muscle mass-increasing effect and muscle metabolism-improving effect of celery seed extract in obese or aging mice, and have completed the present invention based on this finding.

[0008] Therefore, an object of the present invention is to provide a pharmaceutical composition for preventing or treating diseases associated with muscle weakness, which contains celery seed extract as an active ingredient.

[0009] Another object of the present invention is to provide a food composition for preventing or improving diseases associated with muscle weakness, which contains celery seed extract as an active ingredient.

[0010] Another object of the present invention is to provide a muscle-strengthening feed or feed additive containing celery seed extract as an active ingredient.

[0011] However, the technical problems that the present invention aims to achieve are not limited to those mentioned above, and other problems not mentioned will be clearly understood by those skilled in the art to which the present invention pertains from the following description. [Means for solving the problem]

[0012] To achieve the above-mentioned objects, the present invention provides a pharmaceutical composition for preventing or treating muscle weakness-related diseases, which comprises celery seed extract as an active ingredient.

[0013] The present invention also provides a food composition for preventing or ameliorating muscle weakness-related diseases, which comprises celery seed extract as an active ingredient.

[0014] The present invention also provides a muscle-strengthening composition containing celery seed extract as an active ingredient.

[0015] The present invention also provides a muscle-building feed or feed additive containing celery seed extract as an active ingredient.

[0016] In one embodiment of the present invention, the celery seed extract may be an extract using one or more solvents selected from the group consisting of water, alcohols having 1 to 6 carbon atoms, acetone, ether, benzene, chloroform, ethyl acetate, methylene chloride, hexane, cyclohexane, petroleum ether, subcritical fluids, and supercritical fluids, but is not limited thereto.

[0017] In another embodiment of the present invention, the muscle weakness-related disease may be, but is not limited to, one or more selected from the group consisting of sarcopenia, muscle atrophy, muscle dystrophy, and cardiac atrophy.

[0018] In yet another embodiment of the present invention, the sarcopenia may be, but is not limited to, aging sarcopenia or obesity sarcopenia.

[0019] In yet another embodiment of the present invention, the composition may have one or more of the following characteristics, but is not limited to these: (a) Increased muscle mass, (b) Suppression of muscle mass loss; (c) increased muscle strength, (d) inhibiting the increase in lipid content of muscle tissue; or (e) Inhibition of muscle tissue fibrosis.

[0020] In yet another embodiment of the present invention, the composition may further have one or more of the following characteristics, but is not limited thereto: (a) suppression of weight and body fat gain; (b) suppression of fibrosis of liver tissue or adipose tissue; (c) suppression of increases in plasma lipid concentrations; (d) suppression of an increase in lipid content in liver tissue; (e) suppression of increases in plasma hepatotoxicity indicators; (f) suppression of the increase in lipid peroxides and an increase in antioxidant capacity, or (g) Improving gut microbial imbalance.

[0021] The present invention also provides a method for preventing or treating a muscle weakness-related disorder; a metabolic disorder; or a liver disease, comprising the step of administering a composition comprising celery seed extract to an individual in need thereof.

[0022] The present invention also provides use of a composition containing celery seed extract for the prevention or treatment of muscle weakness-related diseases, metabolic diseases, or liver diseases.

[0023] The present invention also provides a use of a celery seed extract for the manufacture of a medicament for treating a muscle weakness-related disorder; a metabolic disorder; or a liver disorder.

[0024] The present invention also provides a composition for preventing, ameliorating, or treating metabolic diseases, which comprises celery seed extract as an active ingredient.

[0025] The present invention also provides a composition for preventing, ameliorating, or treating liver disease, which comprises celery seed extract as an active ingredient.

[0026] The present invention also provides a method for enhancing muscle strength, comprising administering a composition comprising celery seed extract to an individual in need thereof.

[0027] The present invention also provides a use of a composition containing celery seed extract for strengthening muscles.

[0028] The present invention also provides a use of celery seed extract for producing a drug for strengthening muscles. [Effects of the Invention]

[0029] The celery seed extract of the present invention is a natural product that can increase muscle mass and strength with few or no side effects, thereby preventing muscle weakness, suppressing weight and body fat gain, and reducing blood lipid levels and hepatotoxicity indicators (GOT and GPT). Therefore, it is expected to be useful in preventing, improving, or treating muscle weakness-related diseases, metabolic diseases, liver diseases, etc. [Brief explanation of the drawings]

[0030] [Figure 1] FIG. 1 is a simplified diagram showing the experimental design using an obesity-induced mouse model to evaluate the effect of celery seed extract on obesity-induced muscle loss. [Figure 2A]After 20 weeks of feeding, mice from the normal diet group (ND), high-fat diet group (HFD), and celery seed extract supplemented group (CS) were fed, and then the gastrocnemius, quadriceps, tibialis anterior, kidney, and liver were removed and weighed. The results are shown in weight per 100g of body weight (HFD vs. ND: *p<0.05, **p<0.01, ***p<0.001; HFD vs. CS: #p<0.05, ##p<0.01, ###p<0.001, and the same applies below). [Figure 2B] The weights of mice in the normal diet group (ND), high-fat diet group (HFD), and celery seed extract-supplemented group (CS) were measured at weekly intervals over a 20-week period, and the graph compares the changes in weight (left) and the efficacy of suppressing weight gain (right) over the feeding period. [Figure 2C] This figure compares the results of measurements of food intake, energy intake, and food efficiency (FER) in mice from a normal diet group (ND), a high-fat diet group (HFD), and a celery seed extract-supplemented group (CS). [Figure 2D] Mice were fed for 20 weeks in the normal diet group (ND), high-fat diet group (HFD), and celery seed extract-supplemented group (CS), after which white adipose tissue (WAT) was excised from each area and weighed. The weights were plotted as weights per 100 g of body weight (Epididymal: epididymis, Perirenal: perirenal, Retroperitoneum: retroperitoneal cavity, Mesenteric: mesenteric, Visceral: visceral, Subcutaneous: subcutaneous, interscapular WAT: interscapular white fat, interscapular BAT: interscapular brown fat). [Figure 3A] These figures show the morphological changes in muscle tissue at 20 weeks after 20 weeks of feeding in mice from the normal diet group (ND), high-fat diet group (HFD), and celery seed extract supplemented group (CS) (top: H&E staining; bottom: Sirius red staining). Scale bar = 100 μm. [Figure 3B]These figures show morphological changes in the epididymal white adipose tissue at 20 weeks after 20 weeks of feeding in mice from the normal diet group (ND), high-fat diet group (HFD), and celery seed extract supplemented group (CS) (top: H&E staining; bottom: Masson's trichrome staining). Scale bar = 100 μm. [Figure 3C] These figures show the morphological changes in liver tissue at 20 weeks after 20 weeks of feeding in mice from the normal diet group (ND), high-fat diet group (HFD), and celery seed extract supplemented group (CS) (top: H&E staining; bottom: Masson's trichrome staining). Scale bar = 100 μm. [Figure 4] This figure shows the results of measuring thigh thickness at 4 and 20 weeks (top row) and tensile strength at 19 weeks (bottom row) for mice in the normal diet group (ND), high-fat diet group (HFD), and celery seed extract supplemented group (CS). [Figure 5A] FIG. 1 shows the results of measuring the lipid content in muscle tissue of mice in the normal diet group (ND), high-fat diet group (HFD), and celery seed extract supplemented group (CS) (FA: free fatty acids, TG: triglycerides, CHOL: cholesterol). [Figure 5B] This figure shows the results of measuring the lipid content and lipid metabolic enzyme (PAP) activity in liver tissue of mice in the normal diet group (ND), high-fat diet group (HFD), and celery seed extract supplemented group (CS) (FA: free fatty acid, TG: triglyceride, CHOL: cholesterol). [Figure 6] FIG. 1 is a graph showing a comparison of changes in the expression of muscle cell growth-related proteins in mice from a normal diet group (ND), a high-fat diet group (HFD), and a celery seed extract supplemented group (CS). [Figure 7] This figure compares the weekly measurement results of plasma triglycerides (left) and plasma total cholesterol (right) in mice from a normal diet group (ND), a high-fat diet group (HFD), and a celery seed extract-supplemented group (CS). [Figure 8]FIG. 1 shows a comparison of the measurement results of plasma GOT (glutamic oxaloacetic transaminase) and GPT (glutamic pyruvic transaminase) in mice from a normal diet group (ND), a high-fat diet group (HFD), and a celery seed extract-supplemented group (CS). [Figure 9A] FIG. 1 shows a comparison of the results of measuring paraoxonase (PON) in plasma and liver tissue of mice in a normal diet group (ND), a high-fat diet group (HFD), and a celery seed extract-supplemented group (CS). [Figure 9B] FIG. 1 shows a comparison of the measurement results of lipid peroxides (TBARS) in erythrocytes and liver tissue of mice in a normal diet group (ND), a high-fat diet group (HFD), and a celery seed extract supplemented group (CS). [Figure 9C] FIG. 1 shows a comparison of the measurement results of glutathione (GSH) in red blood cells and liver tissues of mice in a normal diet group (ND), a high-fat diet group (HFD), and a celery seed extract-supplemented group (CS). [Figure 10] FIG. 1 is a simplified diagram showing the experimental design using an aging mouse model to evaluate the effect of celery seed extract on age-related muscle loss. [Figure 11] The body weights of young mice (YC), aged mice (NC), and celery seed extract-treated mice (CS) were measured at weekly intervals over a 12-week period, and the results are shown in the graph below to confirm changes in body weight due to the rearing period (YC vs. NC: *p<0.05, **p<0.01, ***p<0.001; NC vs. CS: &p<0.05, &&p<0.01, &&&p<0.001, same below). [Figure 12] This figure shows a comparison of the food intake and food efficiency (FER) measurements for young mice (YC), aged mice (NC), and celery seed extract-supplemented mice (CS). [Figure 13]Young mice (YC), aged mice (NC), and celery seed extract-supplemented mice (CS) were fed the experimental diet for 12 weeks, after which white adipose tissue (WAT) from each region was excised and weighed. The weights are shown in the graph as weight per 100g of body weight (Epididymal: epididymis, Perirenal: perirenal, Retroperitoneum: retroperitoneal, Mesenteric: mesenteric, Visceral: visceral, Subcutaneous: subcutaneous, interscapular WAT: interscapular white fat, interscapular BAT: interscapular brown fat). [Figure 14A] After feeding the experimental diet to young mice (YC), aged mice (NC), and celery seed extract-supplemented mice (CS) for 12 weeks, the gastrocnemius, quadriceps, and tibialis anterior muscles were removed and weighed, and the results are shown in weight per 100g of body weight. [Figure 14B] FIG. 1 shows the results of measuring the tensile strength of the young mouse group (YC), aged mouse group (NC), and celery seed extract-supplemented group (CS) at 12 weeks. [Figure 14C] FIG. 1 shows the results of measuring thigh thickness at 4 and 12 weeks in the young mouse group (YC), aged mouse group (NC), and celery seed extract-supplemented group (CS). [Figure 15] FIG. 1 shows the results of measuring the lipid content in muscle tissue of young mice (YC), aged mice (NC), and celery seed extract-treated mice (CS) (FA: free fatty acids, TG: triglycerides, CHOL: cholesterol). [Figure 16] These figures show the morphological changes in muscle tissue at week 12 after feeding the experimental diet to young mice (YC), aged mice (NC), and celery seed extract-supplemented mice (CS) for 12 weeks (top: H&E staining; bottom: Sirius red staining). Scale bar = 100 μm. [Figure 17]Immunochemical analysis of IGF-1 and myostatin expression was performed in young mice (YC), aged mice (NC), and celery seed extract-supplemented mice (CS) after feeding the experimental diet for 12 weeks. Scale bar = 100 μm. [Figure 18] This figure shows the results of examining the expression of muscle atrophy-related genes (FoxO1, FoxO3, Atrogin, and MuRF1) in young mice (YC), aged mice (NC), and celery seed extract-supplemented mice (CS) after feeding the experimental diet for 12 weeks. [Figure 19] This figure shows the results of confirming the expression of myofibrillar proteins (Igf-1R, FoxO1, and sirt3) in young mice (YC), aged mice (NC), and celery seed extract-supplemented mice (CS) after feeding the experimental diet for 12 weeks. [Figure 20] This figure shows the results of glucose tolerance testing in young mice (YC), aged mice (NC), and celery seed extract-supplemented mice (CS) after feeding the experimental diet for 12 weeks. [Figure 21A] This figure shows the liver weights measured in young mice (YC), aged mice (NC), and celery seed extract-supplemented mice (CS) after feeding them the experimental diet for 12 weeks, and shows the weight per 100 g of body weight. [Figure 21B] This figure shows the results of measuring plasma GOT and GPT, which are indicators of liver damage, in young mice (YC), aged mice (NC), and celery seed extract-supplemented mice (CS) after feeding the experimental diet for 12 weeks. [Figure 21C] FIG. 1 shows the results of measuring the lipid content in liver tissue of young mice (YC), aged mice (NC), and celery seed extract-supplemented mice (CS) after feeding the experimental diet for 12 weeks. [Figure 22A] This figure shows the results of measuring the glutathione (GSH) content in liver tissue, red blood cells, and plasma of young mice (YC), aged mice (NC), and celery seed extract-supplemented mice (CS) after feeding the experimental diet for 12 weeks. [Figure 22B]This figure shows the results of measuring superoxide dismutase (SOD) activity in liver tissue and erythrocytes in young mice (YC), aged mice (NC), and celery seed extract-supplemented mice (CS) after feeding the experimental diet for 12 weeks. [Figure 22C] This figure shows the results of measuring the H2O2 content in mitochondria, cytosol, and red blood cells in young mice (YC), aged mice (NC), and celery seed extract-supplemented mice (CS) after feeding the experimental diet for 12 weeks. [Figure 23] This figure shows the results of measuring the contents of short-chain fatty acids, acetic acid, propionate, and butyrate, in young mice (YC), aged mice (NC), and celery seed extract-supplemented mice (CS), after feeding the experimental diet for 12 weeks. [Figure 24A] This figure shows the results of analyzing the intestinal microorganisms of young mice (YC), aged mice (NC), and celery seed extract-supplemented mice (CS) after feeding them the experimental diet for 12 weeks, and shows the results of analyzing the intestinal microorganisms involved in colitis and inflammatory responses. [Figure 24B] This figure shows the results of analyzing the intestinal microorganisms of young mice (YC), aged mice (NC), and celery seed extract-supplemented mice (CS) after feeding them the experimental diet for 12 weeks, and shows the results of analyzing the intestinal microorganisms involved in obesity and the production of short-chain fatty acids. BEST MODE FOR CARRYING OUT THE INVENTION

[0031] The present inventors have endeavored to develop a naturally occurring active substance capable of preventing, ameliorating, or treating muscle weakness-related diseases. As a result, they have confirmed the excellent muscle mass-increasing effect and muscle metabolism-improving effect of celery seed extract in obese or aging mice, and have completed the present invention based on this finding.

[0032] Accordingly, the present invention provides a pharmaceutical composition for preventing or treating muscle weakness-related diseases, which comprises celery seed extract as an active ingredient.

[0033] The present invention also provides a muscle-strengthening composition containing celery seed extract as an active ingredient.

[0034] The "celery seeds" of the present invention are seeds of the celery plant of the Umbelliferae family, which are millet-sized, yellowish-brown in color, and have a celery-like aroma and are used as a spice. Celery seeds are characterized by a grassy smell and bitter taste, and are known to have anti-inflammatory, diuretic, sedative, aphrodisiac, anti-rheumatic, and arthritis effects.

[0035] In the present invention, the term "extract" includes the extract itself and all formulations that can be produced using the extract, such as the extract obtained by the extraction process of the celery seeds, diluted or concentrated solutions of the extract, dried products obtained by drying the extract, crude or purified products of the extract, or mixtures thereof.

[0036] The method for extracting celery seeds according to the present invention is not particularly limited and may be any method commonly used in the art. Non-limiting examples of the extraction method include hot water extraction, ultrasonic extraction, filtration, reflux extraction, etc., which may be performed alone or in combination of two or more methods.

[0037] In the present invention, the type of extraction solvent used to extract the celery seeds is not particularly limited and can be extracted using a conventional method known in the art for extracting extracts from natural products, i.e., using a conventional solvent under conventional temperature and pressure conditions. For example, in the present invention, the celery seed extract may be extracted with one or more solvents selected from the group consisting of water, C1-C6 organic solvents, and subcritical or supercritical fluids. The C1-C6 organic solvent may be one or more selected from the group consisting of C1-C6 alcohols, acetone, ether, benzene, chloroform, ethyl acetate, methylene chloride, hexane, cyclohexane, and petroleum ether, but is not limited thereto. In the present invention, the celery seed extract may preferably be extracted with ethanol.

[0038] In the present invention, the celery seed extract is added to 70 to 130 g, 70 to 110 g, 70 to 100 g, 70 to 80 g, 80 to 130 g, 100 to 130 g, 110 to 130 g, 90 to 110 g, 95 to 105 g, 70 g, 80 g, 90 g, or 100 g of celery seed powder at a concentration of 40 to 95%, 40 to 85%, 40 to 75%, 40 to 65%, 40 to 55%, 40 to 50%, 40 to 47%, 40 to 45%, 40 to 43%, 40 to 41%. , 50-55%, 55-65%, 65-75%, 68-75%, 70-75%, 73-75%, 65-73%, 65-70%, 65-68%, 75-85%, 85-95%, 88-95%, 90-95%, 90-92%, 90-91%, 40%, 50%, 60%, 65%, 68%, 69%, 70%, 71%, 72%, 75%, 78%, 80%, or 90% ethanol concentration, 700mL-1.3L, 700mL-1.1L Add 700mL to 1L, 700mL to 900mL, 900mL to 1L, 1L to 1.1L, 1L to 1.3L, 1.2L to 1.3L, 700mL, 800mL, 900mL, or 1L to adjust the temperature to 30 to 90°C, 30 to 70°C, 30 to 50°C, 40 to 50°C, 50 to 70°C, 55 to 75°C, 60 to 70°C, 60 to 65°C, 60 to 63°C, 70 to 90°C, 80 to 90°C, 85 to 90°C, 30°C, 40°C, 50°C, 55°C, or 60°C. Extraction may be performed 1 to 5 times, 1 to 3 times, 1 to 2 times, 2 to 4 times, 4 to 5 times, 1 time, 2 times, or 3 times at 2°C for 1 hour 30 minutes to 3 hours 30 minutes, 1 hour 30 minutes to 3 hours, 1 hour 30 minutes to 2 hours, 2 hours to 2 hours 30 minutes, 2 hours 30 minutes to 3 hours, 3 hours to 3 hours 30 minutes, 2 hours, 2 hours 30 minutes, 2 hours 50 minutes, or 3 hours, and the extracted liquid may be filtered, concentrated under reduced pressure, and freeze-dried, but is not limited to this.

[0039] The prepared extract may then be filtered, concentrated, or dried to remove the solvent, or all of the filtration, concentration, and drying processes may be performed. For example, filtration may be performed using filter paper or a vacuum filter, concentration may be performed using a vacuum concentrator, and drying may be performed using a freeze-drying method, but is not limited thereto.

[0040] In the present invention, the term "active ingredient" refers to an ingredient that exhibits a desired activity alone or that can exhibit a desired activity together with a carrier or the like that is inactive by itself.

[0041] In the present invention, the term "muscle weakness-related disorder" refers to any disorder in which muscle tissue or muscle cells are reduced or lost. The muscle weakness may be limited to a single muscle, one side of the body, the upper or lower limbs, or may be generalized and may be congenital or acquired. Furthermore, subjective symptoms of muscle weakness, including muscle fatigue and muscle pain, can be objectively quantified through physical examination.

[0042] The muscle weakness-related disease refers to any disease that can occur due to muscle weakness, including, but not limited to, sarcopenia, muscle atrophy, muscle dystrophy, or cardiac atrophy.

[0043] In the present invention, "sarcopenia" refers to a condition in which the body's muscles (muscle mass, muscle strength) are abnormally reduced or weakened due to various reasons, such as aging and obesity, making physical activity difficult. If symptoms worsen, this can lead to disability and an increased risk of death. Sarcopenia has a wide-ranging effect on the entire body, not just the muscles themselves, but also on bones, blood vessels, nerves, liver, heart, and pancreas. In particular, because bones maintain their density through stress (stimulation) from muscles, sarcopenia and osteoporosis are closely related. Furthermore, muscle loss is known to hinder the development of new blood vessels and nerves, increasing the risk of cognitive decline, fatty liver, and diabetes.

[0044] In the present invention, the sarcopenia may be, but is not limited to, aging sarcopenia or obesity sarcopenia.

[0045] The term "senile sarcopenia" refers to a gradual decrease in muscle mass or a gradual weakening of muscle density and function due to aging, which directly induces a decrease in muscle strength and may result in a decrease and impairment of various bodily functions.

[0046] The term "obesity-related sarcopenia" refers to the phenomenon in which obesity causes fat deposition within muscles, resulting in a decrease in muscle mass, an increase in inflammatory factors derived from mast cells, and a decline in mitochondrial function within the muscles, resulting in weakened muscle function. When obesity causes abnormalities in insulin secretion, energy cannot be adequately supplied to cells, which can cause impairments in muscle development.

[0047] "Muscular atrophy" is a condition in which the muscles of the limbs gradually atrophy, and can induce progressive degeneration of motor nerve fibers and cells in the spinal cord, leading to amyotrophic lateral sclerosis (ALS) and spinal progressive muscular atrophy (SPMA).

[0048] The term "muscle dystrophy" refers to a degenerative muscle disease that is a disease in which gradual muscle atrophy and weakness appear, and is pathologically characterized by necrosis of muscle fibers.

[0049] The "cardiac atrophy" is a condition in which the heart shrinks due to external or internal factors, and manifests as a symptom in which myocardial fibers become thin due to starvation, wasting disease, or senility, leading to a decrease in adipose tissue.

[0050] The term "muscle strengthening" as used herein refers to the effects of enhancing exercise performance, enhancing maximal endurance, increasing muscle mass, enhancing muscle recovery, reducing muscle fatigue, improving energy balance, or a combination thereof.

[0051] In one example of the present invention, in a high-fat diet-induced muscle-losing obese mouse model, it was confirmed that celery seed extract not only increased muscle mass and strength, inhibited muscle tissue fibrosis, inhibited an increase in the lipid content of muscle tissue, increased the expression of the muscle atrophy inhibitor-related protein PGC1α and proteins MyD88 and Traf6 involved in protein accumulation, and activated Akt and Pi3k proteins involved in the inhibition of muscle cell differentiation and death, but also inhibited an increase in body weight and body fat, inhibited fibrosis of liver tissue and adipose tissue, inhibited an increase in lipid content of liver tissue, inhibited an increase in plasma lipid concentrations, inhibited an increase in plasma hepatotoxicity index, inhibited an increase in lipid peroxides, and increased antioxidant capacity (see Example I).

[0052] In another example of the present invention, in an aging mouse model, it was confirmed that celery seed extract not only increased muscle mass and strength, inhibited muscle tissue fibrosis, inhibited an increase in lipid content in muscle tissue, increased the expression of IGF-1, which is involved in muscle tissue synthesis, decreased the expression of myostatin, which is involved in muscle tissue degradation, decreased the expression of muscle atrophy-related genes, increased the expression of IGF-1R, which is involved in the synthesis of muscle fiber proteins, and decreased the expression of FoxO1, which is involved in muscle atrophy, but also inhibited an increase in body fat, lowered blood glucose, inhibited an increase in plasma lipid concentrations, inhibited an increase in plasma hepatotoxicity index, inhibited an increase in lipid content in liver tissue, increased antioxidant capacity, increased short-chain fatty acids, and improved intestinal microbial imbalance (see Example II).

[0053] Therefore, the celery seed extract can be applied not only to the prevention, improvement or treatment of muscle weakness-related diseases, but also to the prevention, improvement or treatment of metabolic diseases or liver diseases.

[0054] Therefore, in the present invention, the composition of the present invention can prevent, improve, or treat muscle weakness-related diseases by satisfying one or more of the following characteristics: (a) Increased muscle mass, (b) Suppression of muscle mass loss; (c) increased muscle strength, (d) inhibiting the increase in lipid content of muscle tissue; or (e) Inhibition of muscle tissue fibrosis.

[0055] In addition, in the present invention, the composition of the present invention can prevent, improve, or treat muscle weakness-related diseases, metabolic diseases, or liver diseases by further satisfying one or more of the following characteristics: (a) suppression of weight and body fat gain; (b) suppression of fibrosis of liver tissue or adipose tissue; (c) suppression of increases in plasma lipid concentrations; (d) suppression of an increase in lipid content in liver tissue; (e) suppression of increases in plasma hepatotoxicity indicators; (f) suppression of the increase in lipid peroxides and an increase in antioxidant capacity, or (g) Improving gut microbial imbalance.

[0056] In the present invention, "improving imbalance of intestinal microorganisms" means promoting the proliferation or growth of beneficial intestinal bacteria and suppressing the proliferation or growth of harmful intestinal bacteria, while maintaining a balance between beneficial and harmful intestinal bacteria. Microorganisms begin to live in the intestinal organs, and when their population reaches equilibrium, they form intestinal microorganisms. The individual microorganisms that make up the intestinal microorganisms are involved in vitamin supply, infection prevention, intestinal function, etc., and therefore the composition of intestinal microorganisms is known to be closely related to the occurrence of constipation, intestinal-related diseases, etc.

[0057] The term "intestinal beneficial bacteria" refers collectively to microorganisms that reside in the intestine and have beneficial effects on the human body. For example, the intestinal beneficial bacteria may include probiotics. For example, the intestinal beneficial bacteria may include, but are not limited to, the genus Bifidobacterium, Lactobacillus, Lactococcus, Streptococcus, Akkermansia, Faecalibacterium, Roseburia, Lachnospiraceae, or Enterococcus. However, while some Enterococcus strains are classified as beneficial intestinal bacteria and have antibacterial activity along with Lactobacillus, some enterococci are classified as harmful bacteria that cause diseases such as inflammation due to vancomycin-resistant enterococci (VRE), urinary tract infections, and infective endocarditis due to their high antibiotic resistance. Therefore, further research is needed on the potential and stability of probiotics from Enterococcus.

[0058] The term "harmful intestinal bacteria" refers collectively to microorganisms that reside in the intestines and have harmful effects on the human body, such as enteritis, etc. For example, harmful intestinal bacteria may include, but are not limited to, Escherichia coli, Fusobacterium, Clostridium, Staphylococcus, Desulfovibrio, Desulfovibrionaceae, Erysipelatoclostridium, Enterorhabdus, or Porphyromonas.

[0059] In the present invention, "short-chain fatty acids" refer to fatty acids having six or fewer carbon atoms, which are major metabolic products of intestinal microorganisms. Increased levels of short-chain fatty acids can acidify the intestinal environment, stimulate bowel movements, inhibit fat absorption, reduce body fat, and reduce blood triglycerides, thereby preventing, ameliorating, or treating metabolic diseases such as obesity. According to one embodiment of the present invention, the short-chain fatty acids are acetic acid, propionic acid, and butyric acid.

[0060] In the present invention, "obesity" refers to a state in which an imbalance between energy intake and consumption results in excessive energy accumulation in the body and an abnormal increase in adipose tissue. Even if a person appears to be of normal weight, if the proportion of body fat is high, the person is considered obese. Obesity is caused by a combination of various factors rather than a single cause, and possible causes include incorrect eating habits, including Westernized eating habits, decreased activity, emotional factors, and genetic factors. In the present invention, the obesity may be, but is not limited to, sarcopenic obesity.

[0061] In the present invention, "sarcopenic obesity" refers to a state in which muscle mass and strength are reduced and body fat mass is increased due to aging or obesity, i.e., a combination of obesity and sarcopenia caused by the conversion of muscle to fat. For example, among people with similar body mass indexes, those with increased body fat mass and decreased muscle mass are known to be at higher risk of developing functional limitations and metabolic diseases than those with a balanced body fat and muscle mass.

[0062] In the present invention, "metabolic disease" refers to a condition or disease closely related to or caused by obesity, and may be, but is not limited to, dyslipidemia; hepatotoxic diseases including drug-induced liver injury, viral liver injury, hepatitis, cirrhosis, liver cancer, or hepatic coma; or fatty liver.

[0063] In the present invention, the "liver disease" may be any one or more selected from the group consisting of non-alcoholic fatty liver disease and liver cancer.

[0064] In the present invention, "nonalcoholic fatty liver disease (NAFLD)" refers to a type of fatty liver that occurs when fat accumulates in the liver in patients who do not consume excessive amounts of alcohol. It refers to a wide range of diseases, including simple fatty liver without an inflammatory response, and the progression of this disease to hepatic inflammatory responses, liver fibrosis, and cirrhosis. If nonalcoholic fatty liver disease is detected in its early stages, good outcomes can be achieved, but if not, it can progress to nonalcoholic steatohepatitis (NASH) for various reasons, which can further lead to cirrhosis and liver cancer.

[0065] NAFLD is divided into primary and secondary depending on the cause, with primary NAFLD being caused by hyperlipidemia, diabetes, or obesity, which are characteristics of metabolic syndrome, and secondary NAFLD being caused by nutritional causes (rapid weight loss, starvation, intestinal bypass surgery), various drugs, toxic substances (poisonous mushrooms, bacterial toxins), metabolic causes, and other factors. The incidence of NAFLD, which is associated with diabetes and obesity, which are key characteristics of metabolic syndrome and is a primary cause, is approximately 50% of diabetic patients and approximately 76% of obese patients, and it is known that NAFLD occurs in most obese diabetic patients.

[0066] The non-alcoholic fatty liver disease may be any one or more selected from the group consisting of simple fatty liver disease, nutritional fatty liver disease, starvation fatty liver disease, obesity fatty liver disease, diabetic fatty liver disease, steatohepatitis, liver fibrosis, and liver cirrhosis, but is not limited thereto.

[0067] The "pharmaceutical composition" according to the present invention may further comprise suitable carriers, excipients, and diluents commonly used in the manufacture of pharmaceutical compositions, such as one or more selected from the group consisting of diluents, binders, disintegrants, lubricants, adsorbents, moisturizers, film coating materials, and controlled-release additives.

[0068] The pharmaceutical composition according to the present invention may be formulated by a conventional method into the form of a powder, granules, sustained-release granules, enteric-coated granules, liquid, eye drops, elixir, emulsion, suspension, spirit, troche, perfume, lemonade, tablet, sustained-release tablet, enteric-coated tablet, sublingual tablet, hard capsule, soft capsule, sustained-release capsule, enteric-coated capsule, pill, tincture, soft extract, dry extract, fluid extract, injection, capsule, perfusion solution, plaster, lotion, paste, spray, inhalant, patch, sterile injection solution, or external preparation such as aerosol, and the external preparation may have the form of a cream, gel, patch, spray, ointment, plaster, lotion, liniment, paste, or cataplasm.

[0069] Carriers, excipients and diluents that may be included in pharmaceutical compositions according to the invention include lactose, dextrose, sucrose, oligosaccharides, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate and mineral oil.

[0070] When the formulation is made, it is prepared using diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, surfactants, etc.

[0071] Additives for the tablets, powders, granules, capsules, pills, and lozenges according to the present invention include excipients such as corn starch, potato starch, wheat starch, lactose, sucrose, glucose, fructose, D-mannitol, precipitated calcium carbonate, synthetic aluminum silicate, calcium hydrogen phosphate, calcium sulfate, sodium chloride, sodium bicarbonate, purified lanolin, microcrystalline cellulose, dextrin, sodium alginate, methylcellulose, sodium carboxymethylcellulose, kaolin, urea, colloidal silica gel, hydroxypropyl starch, hydroxypropylmethylcellulose (HPMC) 1928, HPMC 2208, HPMC 2906, HPMC 2910, propylene glycol, casein, calcium lactate, and Primogel; gelatin, gum arabic, ethanol, agar powder, cellulose acetate phthalate, carboxymethylcellulose, calcium carboxymethylcellulose, glucose, purified water, and casein sodium. Binders such as cellulose, glycerin, stearic acid, sodium carboxymethylcellulose, sodium methylcellulose, methylcellulose, microcrystalline cellulose, dextrin, hydroxycellulose, hydroxypropyl starch, hydroxymethylcellulose, purified shellac, starch paste, hydroxypropyl cellulose, hydroxypropylmethylcellulose, polyvinyl alcohol, and polyvinylpyrrolidone may be used; disintegrants such as hydroxypropylmethylcellulose, corn starch, agar powder, methylcellulose, bentonite, hydroxypropyl starch, sodium carboxymethylcellulose, sodium alginate, calcium carboxymethylcellulose, calcium citrate, sodium lauryl sulfate, anhydrous silicic acid, 1-hydroxypropylcellulose, dextran, ion exchange resins, polyvinyl acetate, formaldehyde-treated casein and gelatin, alginic acid, amylose, guar gum, baking soda, polyvinylpyrrolidone, calcium phosphate, gelling starch, gum arabic, amylopectin, pectin, sodium polyphosphate, ethylcellulose, white sugar, magnesium aluminum silicate, D-sorbitol liquid, and hard anhydrous silicic acid;Lubricants such as calcium stearate, magnesium stearate, stearic acid, hydrogenated vegetable oil, talc, lycopodium, kaolin, petrolatum, sodium stearate, cocoa butter, sodium salicylate, magnesium salicylate, polyethylene glycol (PEG) 4000, PEG 6000, liquid paraffin, hydrogenated soybean oil (Lubri wax), aluminum stearate, zinc stearate, sodium lauryl sulfate, magnesium oxide, macrogol, synthetic aluminum silicate, silicic anhydride, higher fatty acids, higher alcohols, silicone oil, paraffin oil, polyethylene glycol fatty acid ether, starch, sodium chloride, sodium acetate, sodium oleate, dl-leucine, and hard silicic anhydride may also be used.

[0072] Examples of additives that may be used in the liquid preparation according to the present invention include water, dilute hydrochloric acid, dilute sulfuric acid, sodium citrate, sucrose monostearates, polyoxyethylene sorbitol fatty acid esters (twin esters), polyoxyethylene monoalkyl ethers, lanolin ethers, lanolin esters, acetic acid, hydrochloric acid, aqueous ammonia, ammonium carbonate, potassium hydroxide, sodium hydroxide, prolamine, polyvinylpyrrolidone, ethyl cellulose, and sodium carboxymethyl cellulose.

[0073] The syrup according to the present invention may contain a solution of sucrose, other sugars or sweeteners, and may contain, as required, flavoring agents, coloring agents, preservatives, stabilizers, suspending agents, emulsifiers, thickeners, etc.

[0074] The emulsion of the present invention may contain purified water, and may contain emulsifiers, preservatives, stabilizers, fragrances, etc., as needed.

[0075] As the suspending agent according to the present invention, a suspending agent such as acacia, tragacanth, methylcellulose, carboxymethylcellulose, sodium carboxymethylcellulose, microcrystalline cellulose, sodium alginate, hydroxypropylmethylcellulose (HPMC), HPMC1828, HPMC2906, or HPMC2910 may be used, and if necessary, a surfactant, preservative, stabilizer, coloring agent, or fragrance may also be used.

[0076] Injections according to the present invention include solvents such as distilled water for injection, 0.9% sodium chloride injection, Ringer's injection, dextrose injection, dextrose + sodium chloride injection, PEG, lactated Ringer's injection, ethanol, propylene glycol, non-volatile oils - sesame oil, cottonseed oil, peanut oil, soybean oil, corn oil, ethyl oleate, isopropyl myristate, and benzene benzoate; solubilizers such as sodium benzoate, sodium salicylate, sodium acetate, urea, urethane, monoethylacetamide, butazolidine, propylene glycol, Tweens, nicotinamide, hexamine, and dimethylacetamide; weak acids and their salts (acetic acid and sodium acetate), weak bases and their salts (ammonia and ammonium acetate), and organic compounds. It may contain buffers such as proteins, albumin, peptone, and gums; isotonicity agents such as sodium chloride; stabilizers such as sodium bisulfite (NaHSO3), carbon dioxide gas, sodium metabisulfite (Na2S2O5), sodium sulfite (Na2SO3), nitrogen gas (N2), and ethylenediaminetetraacetic acid; sulfating agents such as sodium bisulfite 0.1%, sodium formaldehyde sulfoxylate, thiourea, disodium ethylenediaminetetraacetic acid, and acetone sodium bisulfite; soothing agents such as benzyl alcohol, chlorobutanol, procaine hydrochloride, glucose, and calcium gluconate; and suspending agents such as sodium CMC, sodium alginate, Tween 80, and aluminum monostearate.

[0077] Suppositories according to the present invention may contain any of the following: cocoa butter, lanolin, Witepsol, polyethylene glycol, glycerogelatin, methylcellulose, carboxymethylcellulose, a mixture of stearic and oleic acids, Subanal, cottonseed oil, peanut oil, coconut oil, cocoa butter plus cholesterol, lecithin, lanet wax, glycerol monostearate, Tween or Span, Imhausen, Monolene (propylene glycol monostearate), glycerin, Adeps solidus, Buytyrum Tego-G, Cebes Pharma 16, Pharma 16), Hexalide Base 95, Cotomar, Hydrokote SP, S-70-XXA, S-70-XX75 (S-70-XX95), Hydrokote 25, Hydrokote 711, Idropostal, Massa Estralium Bases such as estrarium, A, AS, B, C, D, E, I, T), Massa-MF, Maspol, Maspol-15, Neospostal-N, Paramound-B, Sposhiro (OSI, OSIX, A, B, C, D, H, L), suppository base IV type (AB, B, A, BC, BBG, E, BGF, C, D, 299), Spostal (N, Es), Wecoby (W, R, S, M, Fs), and Tezestar triglyceride base (TG-95, MA, 57) may also be used.

[0078] Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and such solid preparations are prepared by mixing the extract with at least one or more excipients, such as starch, calcium carbonate, sucrose or lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used.

[0079] Liquid preparations for oral administration include suspensions, oral liquids, emulsions, syrups, etc., and may contain various excipients such as wetting agents, sweeteners, flavoring agents, preservatives, etc. in addition to commonly used simple diluents such as water and liquid paraffin.

[0080] Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried preparations, and suppositories. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate.

[0081] The pharmaceutical composition according to the present invention is administered in a pharmaceutically effective amount. In the present invention, the term "pharmaceutically effective amount" refers to an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to any medical treatment, and the effective dose level can be determined by factors including the type and severity of the patient's disease, the activity of the drug, sensitivity to the drug, the time of administration, the route of administration and excretion rate, the duration of treatment, concurrently used drugs, and other factors well known in the medical field.

[0082] The pharmaceutical composition according to the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered singly or in multiple doses. Taking all of the above factors into consideration, it is important to administer an amount that can achieve the maximum effect with the minimum amount without side effects, and this can be easily determined by a person of ordinary skill in the art to which the present invention pertains.

[0083] The pharmaceutical compositions of the present invention may be administered to an individual by a variety of routes, including, but not limited to, oral ingestion, subcutaneous injection, intraperitoneal administration, intravenous injection, intramuscular injection, paraspinal space (intrathecal) injection, sublingual administration, buccal administration, rectal insertion, vaginal insertion, ocular administration, otic administration, nasal administration, inhalation, spraying through the mouth or nose, dermal administration, transdermal administration, and the like, all of which are foreseeable.

[0084] The pharmaceutical composition of the present invention will be determined by the type of drug as the active ingredient, as well as various related factors such as the disease to be treated, the administration route, the patient's age, sex, weight, and severity of the disease.

[0085] The present invention also provides a method for preventing or treating a muscle weakness-related disorder, comprising administering a composition comprising a celery seed extract to an individual in need thereof.

[0086] The present invention also provides a use of a composition containing celery seed extract for the prevention or treatment of muscle weakness-related diseases.

[0087] The present invention also provides a use of celery seed extract for the manufacture of a medicament for treating a disease associated with muscle weakness.

[0088] In the present invention, the term "individual" refers to a subject in need of treatment for a disease, and more specifically refers to mammals such as human or non-human primates, mice, rats, dogs, cats, horses, and cows.

[0089] In the present invention, "administration" means providing a given composition of the present invention to an individual by any suitable method.

[0090] In the present invention, "prevention" means any action that suppresses or delays the onset of the target disease, and "treatment" means any action that improves or beneficially alters the target disease and its associated metabolic disorder symptoms by administering the pharmaceutical composition according to the present invention.

[0091] In another aspect, the present invention provides a food composition for preventing or ameliorating muscle weakness-related diseases, which contains celery seed extract as an active ingredient.

[0092] In the present invention, "improvement" refers to any action of reducing a parameter related to a target disease, for example, the severity of symptoms, by administering the composition according to the present invention. In this case, the functional health food composition may be used simultaneously with or separately from a therapeutic drug for the prevention or improvement of muscle weakness-related diseases, metabolic diseases, or liver diseases, before or after the onset of the disease.

[0093] When the celery seed extract of the present invention is used as a food additive, it can be added directly or in combination with other foods or food ingredients, and can be used appropriately by conventional methods. The amount of the active ingredient to be added can be appropriately determined depending on the intended use (prevention, health, or therapeutic treatment). Generally, when producing a food or beverage, the celery seed extract of the present invention may be added in an amount of 15% by weight or less, or 10% by weight or less, based on the raw materials. However, in the case of long-term intake for health and hygiene purposes or health regulation purposes, the amount may be less than the above range, and since there is no safety issue, the active ingredient may be used in an amount greater than the above range.

[0094] The type of food is not particularly limited. Examples of foods to which the substance can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea, energy drinks, alcoholic beverages, and vitamin complexes, and include all health functional foods in the usual sense.

[0095] The health drink composition of the present invention may contain various flavorings or natural carbohydrates as additional ingredients, as in conventional beverages. The natural carbohydrates mentioned above include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. Examples of sweeteners that can be used include natural sweeteners such as thaumatin and stevia extract, and synthetic sweeteners such as saccharin and aspartame. The content of the natural carbohydrates is about 0.01 to 0.20 g or about 0.04 to 0.10 g per 100 mL of the composition of the present invention.

[0096] In addition to the above, the compositions of the present invention may contain various nutrients, vitamins, electrolytes, flavors, colorants, pectinic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonation agents used in carbonated beverages, etc. The compositions of the present invention may also contain fruit pulp for the production of natural fruit juices, fruit juice beverages, and vegetable beverages. These ingredients may be used independently or in combination. The proportion of these additives is not critical, but is typically selected in the range of 0.01 to 0.20 parts by weight per 100 parts by weight of the composition of the present invention.

[0097] In the present invention, the food composition may be a functional health food composition, but is not limited thereto.

[0098] In the present invention, the term "health functional food" is the same as "food for special health use (FoSHU)" and refers to a food with high medical and therapeutic effects that is processed to efficiently exhibit bioregulatory functions in addition to providing nutrients. The food may be manufactured in various forms such as tablets, capsules, powder, granules, liquid, pills, etc. to obtain useful effects in preventing or improving muscle weakness-related diseases, metabolic diseases, or liver diseases.

[0099] In the present invention, the "health functional food composition" is characterized by comprising one or more of a carrier, a diluent, an excipient, and an additive, and being formulated into one selected from the group consisting of tablets, pills, powders, granules, powders, capsules, and liquids.

[0100] The health functional food of the present invention can be manufactured by methods commonly used in the art, and can be manufactured by adding raw materials and ingredients commonly used in the art. Moreover, unlike general medicines, since it is made from food as raw materials, it has the advantage of being free from side effects that can occur with long-term use of medicines, and is highly portable.

[0101] In another aspect of the present invention, there is provided a muscle-strengthening feed or feed additive containing celery seed extract as an active ingredient.

[0102] In the present invention, the term "feed" refers to a substance that supplies organic or inorganic nutrients necessary for sustaining the life of an animal. The feed contains nutrients such as energy, protein, lipids, vitamins, and minerals required by animals such as livestock, and may be, but is not limited to, plant-based feed such as grains, roots and fruits, food processing by-products, algae, fibers, oils and fats, starches, gourds, and grain by-products, or animal-based feed such as proteins, minerals, oils and fats, minerals, and single-cell proteins.

[0103] In the present invention, the term "feed additive" refers to a substance added to feed to improve the productivity and health of animals, and may further include, but is not limited to, amino acid supplements, vitamin supplements, enzyme preparations, flavoring agents, silicate preparations, buffering agents, extractants, oligosaccharides, etc. for growth promotion, disease prevention, etc.

[0104] The content of celery seed extract contained in the muscle-strengthening feed or feed additive is not particularly limited, but may be 0.001 to 1% (w / w), preferably 0.005 to 0.9% (w / w), and most preferably 0.01 to 0.5% (w / w). DETAILED DESCRIPTION OF THE INVENTION

[0105] In order to facilitate understanding of the present invention, preferred examples are presented below. However, the following examples are provided merely to facilitate understanding of the present invention, and the contents of the present invention are not limited to the following examples.

[0106] [Example] Example I. Effect of celery seed on obesity-related muscle loss <Experimental Materials and Methods> 1. Preparation of celery seed extract The celery seed powder used in the experiment was purchased from ES Food Ingredients Co., Ltd. 100g of celery seed powder was added to 1L of 70% ethanol and extracted at 60℃ for 3 hours, three times. The extracted liquid was filtered, concentrated under reduced pressure, and freeze-dried. The celery seed extract powder was obtained, stored frozen, and then used to manufacture diet. The extraction yield was 10.02%.

[0107] 2. Experimental Animal Models 2-1. Laboratory animals To confirm the effects of celery seed extract supplementation, an experimental design was conducted using a high-fat diet-induced muscle-losing obesity mouse model, as shown in Figure 1. Specifically, 4-week-old male C57BL / 6J mice (JA BIO, Korea) were purchased from JOONGAH BIO. After 1 week of pelleted diet adaptation, the mice were divided into a normal diet group (ND), a high-fat diet group (HFD; 20% fat, 1% cholesterol), and a celery seed extract-supplemented group (CS, HFD + 0.1% (w / w) celery seed ethanol extract) in a randomized complete block design and fed the experimental diets for 20 weeks. The animal room was maintained under constant conditions of temperature (24±2°C), humidity (50±5%), and a 12-hour photoperiod (6:00 AM to 6:00 PM). Experimental food and water were provided ad libitum to each animal in an individual cage.

[0108] 2-2. Composition of experimental diet The compositions of the experimental diets for the ND, HFD, and CS groups are shown in Table 1. The ND group was a normal diet group and was fed an AIN-76 semisynthetic diet. The HFD group was fed a high-fat diet containing 20% ​​fat and 1% cholesterol by adding lard and cholesterol to the AIN-76 diet. The CS group was fed a high-fat diet supplemented with celery seed extract at a dose of 0.1% of the diet.

[0109] [Table 1]

[0110] 3. Sample Collection 3-1. Collection of blood samples During the breeding period, the animals were fasted for 12 hours every four weeks, and then blood samples were taken from the tail without anesthesia to measure blood lipid concentrations. At the end of the breeding period, the animals were fasted for 12 hours, and then anesthetized with isoflurane (5 mg / kg body weight, Baxter, USA), and blood was collected from the inferior vena cava. The collected blood was treated with heparin and centrifuged at 3,000 rpm at 4°C for 15 minutes. Plasma was then collected and stored at -70°C until analysis.

[0111] 3-2. Collection of tissue samples After blood was collected from the abdominal inferior vena cava in the above 3-1, the mice were sacrificed and the liver, kidney, epididymal white fat, perirenal white fat, interscapular white fat, brown fat, and muscle tissue were harvested. The tissues were rinsed several times with 0.9% saline solution, and then the surface water was removed. They were weighed, classified according to experimental purpose, rapidly cooled in liquid nitrogen, and stored at -70°C until analysis.

[0112] 4. Measurement of thigh thickness and tensile strength (whole-limb grip strength) During the feeding period, the thickness of the thighs of the mice was measured using calipers at 4-week intervals.

[0113] To measure the tensile strength of mice at 19 weeks after feeding the experimental diet, they were allowed to adapt to the measurement device for three days, and then measurements were taken for five consecutive days. The mouse was placed on the top edge of the grid of the measurement device, and while keeping its body parallel to the grid, it was grabbed by the tail and gently and slowly pulled backwards. The maximum muscle strength with which the mouse could grasp the grid was recorded.

[0114] 5. Analysis of Plasma Lipid Concentrations Plasma triglyceride (TG), total cholesterol, and HDL cholesterol concentrations were measured using enzymatic kits from Asan Pharmaceutical Co., Seoul, Korea. Free fatty acid (FA) content was measured using a non-esterified fatty acid (NEFA) kit (Shinyang Chemical) based on an enzymatic colorimetric method. Plasma apolipoprotein A-1 (Apo A-1) and apolipoprotein B (Apo B) concentrations were measured using Apo A-1 and Apo B measurement kits (Nitto Boseki Co., Tokyo, Japan).

[0115] 6. Measurement of Tissue Lipid Content To measure lipid content, lipids from liver and muscle tissues were extracted by the method of Folch et al. (1957). The extract was evaporated with nitrogen gas at 37°C and diluted with isopropanol. For quantification, the enzyme reagent was mixed with 3 mM cholic acid as an emulsifier and 0.5% Triton® X-100 to remove turbidity that occurs during color development. Lipid components were extracted and quantified in the same manner as for plasma triglycerides, cholesterol, and free fatty acids.

[0116] 7. Measurement of Enzyme Activity 7-1. Separation of erythrocyte zymogen According to the method of McCord and Fridovich (1969), heparinized blood was centrifuged at 3,000 rpm at 4°C for 15 minutes to completely remove the plasma and buffy coat, and then washed three times with 0.9% saline. The washed red blood cells were hemolyzed with an equal volume of distilled water and used to measure antioxidant enzyme activity.

[0117] 7-2. Isolation of zymogen from liver tissue To isolate zymogens from liver tissue, we used a modified version of the isolation method by Hulcher et al. (1973). 0.5 g of liver tissue was added to a buffer solution containing 0.1 M triethanolamine, 0.02 M ethylenediamine tetracetate (EDTA, pH 7.4), and 0.002 M dithiothreitol (DTT). The mixture was homogenized in an ice-cold glass Teflon homogenizer (Glascol, 099C K44, USA). The homogenized tissue was then centrifuged at 3,000 rpm for 15 minutes at 4°C. The supernatant was then centrifuged again at 13,000 rpm for 15 minutes at 4°C. The supernatant and the separated sediment were used as the mitochondrial fraction, and the supernatant was ultracentrifuged (Beckman, Optima TLX-120, USA) at 32,500 rpm and 4°C for 1 hour to obtain the cytosol fraction. The microsome fraction was obtained by adding the same buffer solution to the pellet separated from the supernatant cytosol fraction and ultracentrifuging it again at 33,000 rpm and 4°C for 40 minutes. The pellet was then dissolved in 1 ml of buffer solution and stored at -70°C before being used for analysis and protein quantification.

[0118] 7-3. Measurement of lipid metabolism enzyme activity (Phosphatidate phosphohydrolase) PAP (Phosphatidate phosphohydrolase) activity was measured according to the method of Walton et al. (1985). 50 μl of the reaction mixture with or without 0.05 M Tris-HCl (pH 7.0), 1.25 mM Na2-EDTA, and 1 mM MgCl2 was dissolved in 0.9% NaCl solution, and 50 μl of substrate containing 1 mM phosphatidate and phosphatidylcholine was added. Then, 0.1 mL of microsome fraction was added to initiate the reaction. After 15 minutes of incubation at 37°C, 0.1 mL of 1.8 M H2SO4 was added to stop the reaction. 0.1 mL of 0.13% sodium dodecyl sulfate (SDS) solution, 0.25 mL each of 1.25% ascorbic acid and 0.32% ammonium molybdate were then added, and the mixture was incubated at 45°C for 20 minutes for color development, after which the absorbance was measured at 820 nm.

[0119] 7-4. Measurement of antioxidant enzyme activity 7-4-a. Measurement of paraoxonase activity Paraoxonase (PON) activity was measured by a modified and supplemented method of Mackness et al. (1991). The reaction mixture consisted of 940 μl of 0.1 M Tris-HCl buffer (pH 8.0) containing 2 mM CaCl2, to which 30 μl of plasma and liver tissue microsomes (enzyme source) was added, and 30 μl of 100 mM paraoxon (O,O-diethyl-Op-nitrophenylphosphate, Sigma Chemical Co.) (substrate) was added. The activity of p-nitrophenol (extinction coefficient: 17,000 M) was measured for 90 seconds at 25°C and 405 nm. -1 cm -1 The increase in absorbance of the solution was measured.

[0120] 7-4-b. Measurement of glutathione (GSH) content Glutathione content, including both oxidized and reduced GSH, was measured by a modified and supplemented method of Fiala et al. (1976). 0.5 g of liver tissue was added to a buffer solution containing 0.1 M triethanolamine, 0.02 M EDTA (pH 7.4), and 0.002 M DTT, and homogenized using a glass-Teflon homogenizer (Glascol, 099C K44, USA) under ice-cold conditions. 0.3 mL of distilled water and 0.5 mL of 4% sulfosalicylic acid were added to 0.2 mL of the homogenate, and the mixture was centrifuged at 25,000 rpm at 4°C for 10 minutes to obtain the supernatant. To 0.3 mL of the supernatant, 2.7 mL of 0.1 M disulfide reagent (5.5'-dithiobis + 0.1 M sodium phosphate buffer, pH 8.0) was added, and the mixture was allowed to react at room temperature for 20 minutes, after which the absorbance was measured at 412 nm.

[0121] 8. Measurement of lipid peroxide (TBARS) content in red blood cells and liver tissue 8-1. Measurement of lipid peroxide content in red blood cells The lipid peroxide content of erythrocytes was measured using the method of Tarladgis et al. (1964). 3 mL of 5% trichloroacetic acid (TCA) and 1 mL of 0.06 M thiobarbituric acid (TBA) were added to 50 μl of erythrocytes and incubated at 80°C for 90 minutes. The mixture was cooled to room temperature and centrifuged at 2,000 rpm and 25°C for 15 minutes. The supernatant was then collected and its absorbance measured at 535 nm. Lipid peroxidation (MDA) standard solution was prepared by hydrolyzing tetramethoxypropane (TMP). The amount of TBA reactant released at 267 nm was calculated using the MDA extinction coefficient. Specifically, 1 mmol of TMP was dissolved in 100 mL of 0.01 N HCl solution and incubated at 50°C for 60 minutes. The mixture was then cooled to room temperature, and TMP was hydrolyzed with MDA. 1 mL of the hydrolyzed TMP solution was diluted in 100 mL of 0.01 M Na3PO4 (pH 7.0) buffer to prepare an MDA standard solution (1 × 10 -4 The absorption spectrum of the MDA standard solution was obtained at 267 nm, and the exact concentration was calculated and corrected from the extinction coefficient. A TBA-MDA chromophore standard curve was then obtained, and the amount of TBA reactant was calculated from this curve using the MDA extinction coefficient.

[0122] 8-2. Measurement of lipid peroxide content in liver tissue The lipid peroxide content of liver tissue was determined using the method of Ohkawa et al. (1979). 0.5 g of liver tissue was homogenized in a buffer solution containing 0.1 M triethanolamine, 0.02 M EDTA (pH 7.4), and 0.002 M DTT under ice-cold conditions using a glass-Teflon homogenizer (Glascol, 099C K44, USA). 0.2 mL of the homogenate, 0.2 mL of 8.1% sodium dodecyl sulfate (SDS) solution, and 0.6 mL of distilled water were mixed and left at room temperature for 5 minutes. Then, 1.5 mL of 20% acetic acid buffer (pH 3.5) and 1.5 mL of 0.8% TBA were added and incubated at 95°C for 1 hour. After the reaction, the sample was cooled to room temperature, and 1 mL of distilled water and 5 mL of n-butanol:pyridine (15:1) solution were added. After centrifugation at 3,000 rpm and 20°C for 15 minutes, the absorbance of the supernatant was measured at 532 nm. The MDA standard solution was prepared by hydrolyzing TMP, and the amount of TBA reactant released at 267 nm was calculated using the MDA extinction coefficient.

[0123] 9. Tissue Morphological Analysis For tissue morphological observation, liver, epididymal white adipose tissue, and muscle tissue samples were fixed in 10% formaldehyde for 24 hours, then rehydrated in two changes of the same solution, dehydrated in two volumes of ethanol, and embedded in paraffin. Poly-L-lysine-treated sections were prepared and 5 μm thick. They were then stained with hematoxylin and eosin (H&E) and observed under a light microscope at 200x magnification. To stain collagen and muscle fibers, liver and adipose tissue were stained with Masson's trichrome (connective tissue stained blue, nuclei stained dark red, and cytoplasm stained pink), and muscle tissue was stained with Sirius red (muscle fibers stained yellow and collagen stained red), and observed under a light microscope at 200x magnification.

[0124] 10. Western blot measurement 0.1 g of muscle tissue was homogenized with 3 mm beads and 1 ml of lysis buffer (T-PER buffer, Thermo Scientific, Rockford, IL, USA), then centrifuged at 14,000 rpm for 15 minutes to collect the supernatant, which was then transferred to a new tube and used for protein extraction. Protein quantification was performed using the Quick Start Kit. TMBradford Reagent (Bio-Rad, Hercules, CA, USA) was used. Equal amounts of protein were electrophoresed on SDS-polyacrylamide gels (SDS-PAGE). Proteins separated by electrophoresis were transferred to polyvinylidene fluoride (PVDF) membranes (Merck Millipore, New Jersey, USA) and blocked with 5% skim milk / Tris-buffered saline with Tween 20 (TBST; 20 mM Tris-HCl, pH 7.4, 150 mM NaCl, 0.1% Tween 20) at room temperature for 1 hour, followed by incubation with primary antibodies at 4°C overnight. The primary antibodies were anti-PGC1α(1:1000,abcam,ab191838), anti-Akt(1:1000,cell signaling,#9272), anti-phospho Akt(1:1000,cell signaling,#4058), anti-Pi3k(1:1000,cell signaling,#4292), anti-phospho Pi3k(1:1000,cell signaling,#4228), anti-mTOR(1:1000,cell signaling,#2983), anti-MyD88(1:1000,cell signaling,#4283), anti-Sirt1(1:1000,cell Antibodies were used: anti-Traf6 (1:1000, Cell Signaling, #3931) and anti-Traf6 (1:1000, Abcam, ab33915) diluted in 5% BSA; alpha tubulin (1:1000, Cell Signaling, #2125) was used as a loading control. The secondary antibody was horseradish peroxidase (HRP)-conjugated anti-rabbit IgG (1:5000; Cell Signaling, #7074S) diluted in 5% skim milk and incubated at room temperature for 1 hour. After each incubation, the cells were washed three times for 10 minutes with TBST buffer before proceeding.The washed membranes were visualized using an Enhanced Chemiluminescent (ECL) kit (Super-Signal West Pico Plus, 34580, Thermo Scientific, Rockford, IL, USA) to develop bands, which were then quantified and analyzed using a G-box (50S; BI System Co.).

[0125] 11.Statistical analysis The experimental results were calculated using the SPSS package program, version 25.0 (Statistical Package for the Social Sciences, SPSS Inc., Chicago), a computer statistical program. Student's t-tests were used to test for significance between the ND and HFD groups, and between the HFD and CS groups. All results are expressed as mean ± SE (standard error).

[0126] <Experimental Results> 1. Analysis of changes in tissue weight and body weight due to celery seed extract 1-1. Changes in muscle and organ weight To analyze changes in muscle and organ weights, the gastrocnemius, femoral, tibialis anterior, kidney, and liver were removed and weighed, and then expressed as weight per 100 g of body weight for comparison.

[0127] As shown in Figure 2A, muscle tissue weights at all muscle sites in the HFD group were significantly decreased compared to the ND group, while gastrocnemius and thigh muscle weights in the CS group were significantly increased compared to the HFD group. Kidney weights in the HFD group were significantly decreased compared to the ND group, while kidney weights in the CS group were significantly increased compared to the HFD group. Liver weights in the HFD group were significantly increased compared to the ND group, while liver weights in the CS group were significantly decreased compared to the HFD group. These results confirmed that celery seed extract has the effects of increasing muscle mass, inhibiting liver weight gain, and inhibiting kidney weight loss.

[0128] 1-2. Weight change To examine the effect of celery seed extract intake on body weight gain, body weight and food intake were measured once a week during the feeding period.

[0129] As a result, as shown in Figure 2B, the HFD group showed a rapid increase in body weight compared to the ND group, whereas the CS group showed a significant decrease in body weight compared to the HFD group from the 12th week of feeding the experimental diet.

[0130] Furthermore, when the body weight gain over 20 weeks was converted to daily weight gain (g / day), it was confirmed that weight gain in the CS group was significantly suppressed compared to the HFD group.

[0131] On the other hand, as shown in Figure 2C, there were no significant differences in average daily food intake and energy intake between the CS and HFD groups. However, the food efficiency ratio (FER), which indicates weight gain relative to energy intake, was significantly higher in the high-energy HFD group compared to the low-energy ND group, whereas it was significantly lower in the CS group compared to the HFD group. These results demonstrate that celery seed extract significantly reduces body weight under conditions of equal energy intake.

[0132] 1-3. Changes in adipose tissue weight To analyze changes in adipose tissue weight, the adipose tissue was excised from the mice, weighed, and then expressed as weight per 100 g of body weight for comparison.

[0133] As shown in Figure 2D, the adipose tissue weights at all sites in the HFD group were significantly higher than those in the ND group, confirming that obesity was induced by a high-fat diet. In contrast, the CS group supplemented with celery seed extract significantly reduced the weights of perirenal and mesenteric white adipose tissue, which correspond to visceral fat, and visceral fat and total white adipose tissue. These results suggest that celery seed extract significantly reduces the weight of white adipose tissue, which stores energy, thereby reducing body fat.

[0134] 2. Analysis of the effects of celery seed extract on morphological changes in liver, adipose, and muscle tissues 2-1.Muscle tissue H&E staining of the gastrocnemius muscles of mice revealed that the morphology and structure of muscle cell bundles were irregular in the HFD group compared to the ND group, as shown in Figure 3A. However, in the CS group, muscle cell morphology was confirmed to have improved to a normal level (upper panel of Figure 3A).

[0135] Furthermore, Sirius red staining was performed for histological visualization of collagen fibers in muscle tissue. Results showed that more collagen staining was observed in muscle tissue in the HFD group than in the ND group, but that muscle tissue fibrosis induced by a high-fat diet was suppressed in the CS group (Fig. 3A, bottom panel).

[0136] 2-2.Adipose tissue As shown in Figure 3B, H&E staining of epididymal white adipose tissue (EPI) revealed that the adipocytes in the HFD group were larger than those in the ND group, and that the adipocytes in the CS group were smaller than those in the HFD group (upper row of Figure 3B).

[0137] Furthermore, to examine the effect of celery seed extract on adipose tissue fibrosis, Masson's trichrome staining was performed. Results showed that fibrotic connective tissue was significantly more prominent in the adipocytes of the HFD group than in the ND group, and that adipose tissue fibrosis induced by a high-fat diet was suppressed in the CS group (Figure 3B, bottom).

[0138] 2-3.Liver tissue H&E staining of the liver tissue revealed that, as shown in Figure 3C, the HFD group showed the greatest accumulation of lipid droplets, mainly in the portal vein of the liver tissue, whereas the number of lipid droplets in the CS group supplemented with celery seed extract was reduced compared to the HFD group.

[0139] In addition, to examine the effect of celery seed extract on liver tissue fibrosis, Masson's trichrome staining was performed. The results showed that in the HFD group, a large amount of connective tissue had accumulated around the hepatic portal vein, while in the CS group, liver tissue fibrosis caused by a high-fat diet was suppressed.

[0140] 3. Analysis of the effects of celery seed extract on thigh thickness and muscle strength As shown in Figure 4, the thigh thickness of the hind limbs measured after 4 weeks of feeding the experimental diet was significantly higher in the HFD group than in the ND group, but no significant difference was observed between the CS and HFD groups. However, after 20 weeks of feeding the experimental diet, the thigh thickness of both the right and left hind limbs of the CS group was significantly increased compared to the HFD group.

[0141] Furthermore, the results of measuring tensile strength after 19 weeks of feeding the experimental diet showed no significant difference between the ND and HFD groups, but the CS group showed a significantly higher tensile strength than the HFD group. Therefore, these results confirmed that celery seed extract has the effect of improving thigh thickness, which had been reduced by a high-fat diet, and increasing muscle strength.

[0142] 4. Analysis of the effects of celery seed extract on tissue lipid content To analyze the effect of celery seed extract on the lipid content of tissues, the contents of free fatty acids, triglycerides, and cholesterol per unit weight of tissue were compared, and the results are shown in FIG.

[0143] 4-1. Lipid content of muscle tissue As shown in Figure 5A, the contents of triglycerides, free fatty acids, and cholesterol in muscle tissue were significantly increased in the HFD group, whereas the contents of free fatty acids and cholesterol in the CS group were significantly decreased compared to the HFD group. These results indicate that the increase in muscle weight and thickness in 1-1 and 3 above is not due to lipid accumulation, since celery seed extract inhibits the increase in lipid content in muscle tissue.

[0144] 4-2. Lipid content and lipid metabolic enzyme activity in liver tissue The activity of phosphatidate phosphohydrolase (PAP), an enzyme involved in the synthesis of triglycerides in liver tissue, was measured. As shown in Figure 5B, the HFD group showed significantly higher activity than the ND group. It was confirmed that the PAP activity, which was increased by the high-fat diet, was significantly reduced by supplementation with celery seed extract.

[0145] Furthermore, the lipid content of liver tissue in the HFD group, which consumed a high-fat diet, was significantly increased compared to the ND group, which consumed a normal diet, while the free fatty acid and total cholesterol content of liver tissue in the CS group, which was supplemented with celery seed extract, was significantly decreased compared to the HFD group. These results confirmed that celery seed extract reduces the lipid content of liver tissue.

[0146] 5. Analysis of the effects of celery seed extract on the expression of proteins related to muscle cell growth To confirm the effect of celery seed extract on the growth mechanism of muscle tissue cells, we measured the expression levels of proteins in the Pi3k / Akt / mTOR pathway, which promotes muscle cell differentiation and is involved in the accumulation of myofibrillar proteins.

[0147] As a result, as shown in Figure 6, the expression of PGC1α (peroxisome proliferator-activated receptor gamma coactivator 1-alpha), which is involved in suppressing inflammation and muscle atrophy in muscle tissue, and MyD88 (Myeloid differentiation primary response 88) and Traf6 (Tumor necrosis factor receptor associated factor 6), which are upstream factors of the Pi3k / Akt / mTOR pathway, was significantly increased in the CS group compared to the HFD group.

[0148] Meanwhile, in the case of Akt (Protein kinase B) and Pi3k (phosphoinositide 3-kinase), which are involved in the differentiation and suppression of myocyte death, it was confirmed that the expression of pAkt and pPi3k, which are activated forms of Akt and Pi3k, increased. These results confirmed that celery seed extract has the effect of improving muscle metabolism.

[0149] 6. Analysis of the effects of celery seed extract on plasma lipid concentrations 6-1. Plasma triglyceride and total cholesterol concentrations Plasma triglyceride and total cholesterol concentrations were measured once a week during the experimental period. As shown in Figure 7, the total cholesterol concentration in the HFD group increased significantly compared to the ND group from 4 weeks after feeding the experimental diet. In the CS group, plasma total cholesterol concentration decreased significantly compared to the HFD group after 16 weeks of feeding the experimental diet.

[0150] On the other hand, there was no significant difference in triglyceride concentration between the groups.

[0151] 6-2.Plasma lipid concentration Plasma free fatty acids, triglycerides, total cholesterol, and non-HDL cholesterol concentrations were significantly decreased in the CS group compared to the HFD group, whereas the HDL cholesterol to total cholesterol ratio (HDL-C / Total C ratio; HTR) was significantly increased in the CS group compared to the HFD group. Furthermore, the atherogenic index (AI) and plasma apolipoprotein B (Apo B) levels were significantly lower in the CS group compared to the HFD group. These results demonstrate that celery seed extract reduces plasma lipid concentrations.

[0152] [Table 2]

[0153] 7. Analysis of the effects of celery seed extract on plasma hepatotoxicity indices To confirm the effect of celery seed extract on hepatotoxicity, the activities of glutamic oxaloacetic transaminase (GOT) and glutamic pyruvic transaminase (GPT), which are closely related to liver damage, were measured using enzyme kits from Asan Pharm Co., Seoul, Korea.

[0154] As a result, as shown in Figure 8, plasma GOT and GPT levels were significantly increased in the HFD group compared to the ND group, and plasma GOT and GPT levels were significantly decreased in the CS group compared to the HFP group. These results confirmed that celery seed extract has the effect of suppressing hepatotoxicity.

[0155] 8. Analysis of the effects of celery seed extract on antioxidant-related biomarkers 8-1. Paraoxonase (PON) activity As shown in Figure 9A, plasma PON activity, which inhibits the oxidation of plasma lipoprotein particles, tended to decrease in the HFD group compared to the ND group, whereas the CS group showed a significant increase compared to the HFD group. Furthermore, liver PON activity was lowest in the HFD group, whereas the CS group showed a significant increase compared to the HFD group.

[0156] 8-2. Lipid peroxides (thiobarbituric acid reactive substances; TBARS) Comparison of TBARS in erythrocytes and liver tissues showed that, as shown in Figure 9B, TBARS in erythrocytes was significantly increased in the HFD group compared to the ND group, but significantly decreased in the CS group compared to the HFD group. Furthermore, TBARS in liver tissues was significantly increased in the HFD group compared to the ND group, but tended to decrease in the CS group.

[0157] 8-3. Glutathione (GSH) The results of comparing the GSH content in red blood cells and liver tissue, an antioxidant biomarker, showed that the high-fat diet reduced GSH content in liver tissue compared to the ND group, whereas the CS group increased GSH content compared to the HFD group, as shown in Figure 9C. These results confirmed that celery seed extract has the effect of suppressing the increase in lipid peroxides and increasing antioxidant capacity.

[0158] Example II. Effect of celery seeds on age-related muscle loss <Experimental Materials and Methods> 1. Preparation of celery seed extract Celery seed powder was purchased from ES Food Ingredients Co., Ltd. 100g of celery seed powder was added with 1L of 70% ethanol and extracted three times at 60℃ for 3 hours. The extracted liquid was filtered, concentrated under reduced pressure, and freeze-dried. The celery seed alcohol extract powder was obtained, stored frozen, and used to prepare diet. The extract yield was 10.02%.

[0159] 2. Experimental Animal Models 2-1. Laboratory animals Experimental animals were 8-week-old male C57BL / 6J mice and 50-week-old male C57BL / 6J mice (JA BIO, Korea) purchased from JOONGAH BIO. As shown in Figure 10, after 2 weeks of adaptation to the pelleted diet, the animals were divided into three groups using a randomized block design: young mice (YC, Young control, n = 8), aged mice (NC, Negative control, n = 7), and aged mice receiving a diet supplemented with celery seed extract (CS, 0.1% (w / w) celery seed ethanol extract, n = 7). They were housed on the experimental diet for 12 weeks. The animal room was maintained under constant temperature (24 ± 2°C), humidity (50 ± 5%), and a 12-hour photoperiod (6:00 AM - 6:00 PM). Each animal was housed in an individual cage and provided with experimental diet and water ad libitum. During the 12-week experimental period, food intake and body weight were measured once a week, and the food efficiency ratio (FER) was calculated by dividing the daily weight gain by the daily energy intake.

[0160] 2-2. Composition of experimental diet The composition of the experimental diet is shown in Table 3. All groups were fed the prepared AIN-93G diet, and the CS group was fed a normal diet supplemented with celery seed extract at a dose of 0.1% of the diet.

[0161] [Table 3]

[0162] 3. Sample Collection After fasting for 12 hours, the animals were anesthetized with isoflurane (5 mg / kg body weight, Baxter, USA), and blood was collected from the inferior vena cava. The collected blood was treated with heparin and centrifuged at 3,000 rpm at 4°C for 15 minutes. Plasma was then collected and stored at -70°C until analysis.

[0163] The liver, kidney, epididymal white fat, perirenal white fat, interscapular white and brown fat, and muscle tissue were removed from the experimental animals and rinsed several times with 0.9% saline solution. After removing the surface moisture, the tissue was weighed, classified according to experimental purpose, rapidly cooled in liquid nitrogen, and stored at -70°C until analysis.

[0164] 4. Measurement of thigh thickness and tensile strength During the experimental period, the thickness of the thighs of the mice was measured using a caliper at 4-week intervals.

[0165] To measure the tensile strength of the experimental animals at 12 weeks after feeding the experimental diet, the animals were allowed to adapt to the measuring equipment for 3 days, and then the tensile strength was measured for 5 consecutive days.

[0166] Specifically, the experimental animal was placed on the top edge of the grid of the measuring device, and was allowed to grasp the grid with all four paws. While keeping the mouse's body parallel to the grid, the mouse's tail was grasped and gently and slowly pulled backwards, and the maximum muscle strength with which the mouse grasped the grid was recorded.

[0167] 5.Biochemical analysis 5-1. Analysis of plasma lipid concentrations Plasma triglyceride, total cholesterol, and HDL cholesterol concentrations were measured using an enzyme kit from Asan Pharmaceutical Co., Seoul, Korea. Free fatty acid content was measured using a non-esterified fatty acid (NEFA) kit from Shinyang Chemical, which uses an enzymatic colorimetric method.

[0168] 5-2. Measurement of lipid content in tissues The lipid content of liver and muscle tissues was extracted by the method of Folch et al. (1957), and the extract was evaporated with nitrogen gas at 37°C and diluted with isopropanol. For quantification, the enzyme reagent was mixed with 3 mM cholic acid as an emulsifier and 0.5% Triton® X-100 to remove turbidity that occurs during color development. Lipid components were extracted and quantified in the same manner as for plasma triglycerides, cholesterol, and free fatty acids.

[0169] 5-3. Separation of zymogen 5-3-a. Isolation of erythrocyte zymogen Erythrocytes were prepared according to the method of McCord and Fridovich (1969) by centrifuging heparinized blood at 3,000 rpm at 4°C for 15 minutes to completely remove plasma and buffy coat, followed by washing three times with 0.9% saline. The washed erythrocytes were hemolyzed with an equal volume of distilled water and used to measure antioxidant enzyme activity.

[0170] 5-3-b. Isolation of zymogen from liver tissue To isolate zymogens from liver tissue, we used a modified version of the method described by Hulcher et al. (1973). A buffer solution containing 0.1 M triethanolamine, 0.02 M ethylenediaminetetraacetate (EDTA, pH 7.4), and 0.002 M dithiothreitol (DTT) was added to 0.5 g of liver tissue and homogenized using a glass-Teflon homogenizer (Glascol, 099C K44, USA) under ice-cold conditions. The homogenized tissue was then centrifuged at 3,000 rpm for 15 minutes at 4°C. The supernatant was then centrifuged again at 13,000 rpm for 15 minutes at 4°C. The resulting sediment was used as the mitochondrial fraction, and the supernatant was then centrifuged at 32,500 rpm for 1 hour at 4°C in an ultracentrifuge (Beckman, Optima TLX-120, USA) to obtain the cytosol fraction. The microsomal fraction was prepared by adding the same buffer solution to the pellet separated from the cytosolic fraction of the supernatant, followed by another ultracentrifugation at 33,000 rpm at 4°C for 40 minutes. The pellet was then dissolved in 1 ml of buffer solution and stored at -70°C before being used for analysis and protein quantification.

[0171] 5-4. Measurement of antioxidant enzyme activity 5-4-a. Measurement of glutathione content Glutathione (GSH) content, including both oxidized and reduced GSH, was measured by a modified and supplemented method of Fiala et al. (1976). 0.5 g of liver tissue was added to a buffer solution containing 0.1 M triethanolamine, 0.02 M EDTA (pH 7.4), and 0.002 M DTT, and homogenized using a glass-Teflon homogenizer (Glascol, 099C K44, USA) under ice-cold conditions. 0.3 mL of distilled water and 0.5 mL of 4% sulfosalicylic acid were added to 0.2 mL of the homogenate, and the mixture was centrifuged at 25,000 rpm at 4°C for 10 minutes to obtain the supernatant. To 0.3 mL of the supernatant, 2.7 mL of 0.1 M disulfide reagent (5.5'-dithiobis + 0.1 M sodium phosphate buffer, pH 8.0) was added, and the mixture was allowed to react at room temperature for 20 minutes, after which the absorbance was measured at 412 nm.

[0172] 5-4-b. Measurement of superoxide dismutase (SOD) activity SOD is a superoxide anion radical (O 2- It is an enzyme that catalyzes the decomposition of HCl into H2O2 and O2, and its activity was measured by modifying and supplementing the method of Marklund et al. (1974) using the degree of color development due to the autoxidation of pyrogallol under alkaline conditions. 0.1 mL of liver tissue and erythrocyte zymogen and 0.1 mL of 7.2 mM pyrogallol solution were added to 1.5 mL of 50 mM Tris-hydroxymethyl-aminomethane buffer (pH 8.5) containing 10 mM EDTA, and the mixture was incubated at 25°C for 10 minutes. 50 μL of 1 N HCl solution was added to terminate the reaction. The absorbance change was measured at 420 nm (Beckman 650 spectrophotometer, USA) and calculated as the SOD units required to prevent the autoxidation of pyrogallol relative to the cytosolic protein and erythrocyte hemoglobin (SOD unit / mg cytosolic protein, SOD unit / g hemoglobin).

[0173] 6. Intraperitoneal glucose tolerance test After fasting for 12 hours, the mice were intraperitoneally administered 0.5 g of glucose solution per kg of body weight. Blood was collected via the tail vein at 0, 30, 60, and 120 minutes, and blood glucose was measured using a blood glucose meter.

[0174] 7. Analysis of liver tissue toxicity through measurement of plasma GOT and GPT activity The activities of GOT and GPT, which are closely related to hepatocellular injury, were measured using enzyme kits from Asan Pharm Co., Seoul, Korea.

[0175] 8. Morphological Analysis of Tissue Cells For tissue morphological observation, a portion of muscle tissue removed at the time of animal sacrifice was fixed in 10% formaldehyde solution for 24 hours, then rehydrated in the same solution twice, dehydrated in 2 volumes of ethanol, and embedded in paraffin. 5 μm-thick tissue sections were treated with poly-L-lysine and then stained with H&E and observed under a light microscope at 200x magnification. To stain muscle fibers, Sirius Red staining was performed and observed under a light microscope at 200x magnification. For immunochemical analysis, Igf-1R and myostatin were stained using immunohistochemistry (IHC) and observed under a light microscope at 400x magnification.

[0176] 9. Real-time PCR RNA was isolated from muscle tissue and cDNA was synthesized using a PrimeScript Real-time Reagent Kit (Takara, Japan). The cDNA was diluted in RNAse-free water, stored at -20°C, and used for real-time RT-PCR. A SYBR Green PCR kit (Takara, Japan) was used for real-time RT-PCR gene expression analysis. Primers for analyzing gene expression were synthesized by Geno Tech Co., Ltd. (Daejeon, Korea). The fluorescent signal was monitored at each cycle, and the threshold cycle (Ct) was analyzed. mRNA expression between experimental groups was quantified using a CFX96 Real-Time System (Bio-Rad, USA). The primers used are listed in Table 4.

[0177] [Table 4]

[0178] 10. Western Blot Measurement 0.1 g of frozen muscle tissue was homogenized with 3 mm beads and 1 ml of lysis buffer (T-PER buffer, Thermo Scientific, Rockford, IL, USA), then centrifuged at 14,000 rpm for 15 minutes. The supernatant was collected and transferred to a new tube, and the protein was extracted and used in the experiment. Protein quantification was performed using Quick Start. TM Bradford Reagent (Bio-Rad, Hercules, CA, USA) was used. Equal amounts of protein were electrophoresed on SDS-polyacrylamide gels (SDS-PAGE). Proteins separated by electrophoresis were transferred to polyvinylidene fluoride (PVDF) membranes (Merck Millipore, New Jersey, USA) and blocked with 5% skim milk / Tris-buffered saline with Tween 20 (TBST; 20 mM Tris-HCl, pH 7.4, 150 mM NaCl, 0.1% Tween 20) at room temperature for 1 hour, followed by incubation with primary antibodies at 4°C overnight. The secondary antibody, horseradish peroxidase (HRP)-conjugated anti-rabbit IgG (1:5000; Cell Signaling, #7074S), was diluted in 5% skim milk and incubated at room temperature for 1 hour. After each incubation, the membrane was washed three times for 10 minutes with TBST buffer before proceeding. The washed membrane was visualized using an enhanced chemiluminescent (ECL) kit (Super-Signal West Pico Plus, 34580, Thermo Scientific, Rockford, IL, USA) to develop bands, and quantified and analyzed using a G-box (50S; BI System Co.).

[0179] 11. Analysis of short-chain fatty acids A certain amount of the collected raw feces was weighed, diluted with triple distilled water according to the ratio, thoroughly stirred, and then centrifuged at 13,000 rpm for 3 minutes at room temperature (RT).

[0180] 150 μL of the supernatant was transferred to a short-chain fatty acid analysis vial, and 150 μL of GC buffer was added. The vial was sealed with a short-chain fatty acid analysis cap and analyzed for 10 minutes using an Agilent Headspace 7697A equipped with an HP-Innowax column and an Agilent GC7890B flame ionization detector. The short-chain fatty acids to be analyzed were prepared at their optimal analytical concentrations, mixed, and serially diluted with triple-distilled water. Starting with the lowest concentration, each concentration was analyzed five times. Results were obtained by repeating the analysis five times.

[0181] 12. Methods for analyzing intestinal microorganisms Raw fecal bacterial DNA was extracted using the Mag-Bind® Universal Pathogen Kit (Omega Bio-tek). Following the manufacturer's protocol, fecal samples were suspended in SLX-Mlus buffer, then crushed, separated, and washed. The extracted fecal microbial DNA was amplified with a 16S amplicon PCR forward primer (5'-TCGTCGGCAGCGTCAGATGTATAAGAGACAGCCTACGGGNGGCWGCAG-3') and a 16S amplicon PCR reverse primer (5'-GTCTCGTGGGCTCGGAGATGTGTATAAGACCAGAGTATCACTACH). PCR was performed at 95°C for 3 minutes, followed by eight cycles of 95°C for 30 seconds, 55°C for 30 seconds, 72°C for 30 seconds, and 72°C for 5 minutes, followed by a 4°C hold for the PCR reaction. After the cleanup step, library concentrations were confirmed using Qubit 4.0 (ThermoFisher Scientific) with 1x dsDNA HS Analysis Solution (ThermoFisher Scientific) and then sequenced using an Illumina Miseq system. Reads were aligned using unique barcodes for each PCR product. Sequencing results were analyzed using the Qiime2 bioinformatics pipeline, and taxonomic assignments were made using the Silva reference database.

[0182] 13.Statistical analysis The experimental results were calculated using the SPSS package program, version 25.0 (Statistical Package for the Social Sciences, SPSS Inc., Chicago), a computer statistical program. Student's t-tests were performed to test for significance between each YC group and the NC group, and between the NC group and the CS group. All results are expressed as mean ± SE (standard error).

[0183] <Experimental Results> 1. Weight loss with celery seed extract The average changes in body weight over the 12 weeks of feeding the experimental diets are shown in Figure 11. There was a significant difference in body weight between the YC and NC groups from before the start of the experiment to the end, but there was no significant difference in body weight or weight gain between the NC and CS groups.

[0184] 2. Effect of celery seed extract on food intake and food efficiency As shown in Figure 12, the average daily food intake (g / day) was higher in the NC group than in the YC group, but there was no significant difference between the CS and NC groups. The food efficiency ratio (FER) was significantly higher in the YC group, whose body weight increased due to growth, compared with the NC group, whose body weight remained almost unchanged, and there was no significant difference between the NC and CS groups.

[0185] 3. Effect of celery seed extract on adipose tissue weight As a result of measuring the weight of adipose tissue per unit body weight, as shown in Figure 13, the weights of epididymal white adipose tissue, perirenal white adipose tissue, mesenteric white adipose tissue, visceral white adipose tissue, interscapular white adipose tissue, and total white adipose tissue in the NC group were significantly increased compared to the YC group, and it was confirmed that supplementation with celery seed extract significantly decreased the weights of epididymal white adipose tissue, perirenal white adipose tissue, mesenteric white adipose tissue, visceral white adipose tissue, interscapular white adipose tissue, and total white adipose tissue.

[0186] 4. Effect of celery seed extract on muscle tissue weight, tensile strength and thigh thickness of hind limbs The changes in muscle tissue weight and muscle strength per 100g of body weight are shown in Figures 14A and 14B. After the experiment, the gastrocnemius, femoral muscles, and tibialis anterior muscles of the experimental animals were removed and measured. As shown in Figure 14A, the weights of the gastrocnemius, tibialis anterior, and total muscle tissue in the NC group were significantly reduced compared to the YC group, while in the CS group, the gastrocnemius and total muscle mass, which had decreased due to aging, were significantly increased compared to the NC group.

[0187] The results of the tensile force measurements conducted on the 12th week of the experiment, as shown in Figure 14B, showed that the tensile force of the NC group was significantly reduced compared to the YC group, whereas the reduced tensile force of the CS group was normalized to the level of the YC group.

[0188] During the experimental diet feeding period, the thickness of the hind leg thighs was measured. As shown in Figure 14C, no significant difference was observed in any group at 4 weeks after feeding the experimental diet. However, at 12 weeks after feeding the experimental diet, the thickness of both the right and left hind leg thighs in the CS group was significantly increased compared to the NC group.

[0189] 5. Effect of celery seed extract on the lipid content of muscle tissue The content of free fatty acids (FA), triglycerides (TG), and cholesterol (CHOL) per unit weight of muscle tissue was compared. As shown in Figure 15, the content of triglycerides, free fatty acids, and cholesterol in muscle tissue significantly increased in the NC group, and by supplementation with celery seed extract, they significantly decreased in the CS group compared to the NC group.

[0190] 6. Morphological Analysis of Muscle Tissue H&E staining was performed to analyze the morphology of the gastrocnemius muscle. As shown in Figure 16, the morphology and structure of muscle cell bundles were irregular in the NC group compared to the YC group, but supplementation with celery seed extract improved this to normal levels. Sirius red staining was performed to visualize collagen fibers in the muscle tissue histologically. Collagen staining in the muscle tissue was significantly higher in the NC group than in the YC group, confirming that supplementation with celery seed extract suppressed muscle tissue fibrosis in the CS group.

[0191] 7. Immunochemical Analysis of Muscle Tissue The effects of celery seed extract on the expression of IGF-1 and myostatin, which are involved in the synthesis and breakdown of muscle tissue, were examined. As shown in Figure 17, IGF-1 was expressed at a lower level between muscle tissue cells in the NC group compared to the YC group, but supplementation with celery seed extract increased the decreased IGF-1 expression.

[0192] Furthermore, it was confirmed that myostatin expression was reduced to a level similar to that of the YC group by supplementation with celery seed extract.

[0193] 8. Effect of celery seed extract on the expression of genes related to muscle function The expression of muscle atrophy-related genes (FoxO1, FoxO3, Atrogin, and MuRF1) in the gastrocnemius muscle was compared. As shown in Figure 18, the expression of all muscle atrophy-related genes was significantly increased in the NC group compared to the YC group, and was significantly decreased by supplementation with celery seed extract.

[0194] 9. Effect of celery seed extract on myofibrillar protein expression To confirm the mechanism by which celery seed extract supplementation promotes muscle cell growth, we measured the expression levels of proteins in the Igf-1R pathway, which promotes muscle cell differentiation and is involved in the accumulation of myofibrillar proteins. As shown in Figure 19, the expression of Igf-1R (Igf-1 Receptor) protein, which is involved in the synthesis of myofibrillar proteins, was highest in the CS group, whereas expression levels were reduced in the NC group. FoxO1, which is involved in inflammatory metabolism and muscle atrophy in muscle tissue, was most highly expressed in the NC group compared to the YC group, whereas expression was reduced in the CS group.

[0195] In addition, it was confirmed that the expression of Sirt3 protein, which regulates antioxidant metabolism in mitochondria, was increased in the CS group compared to the NC group.

[0196] 10. Intraperitoneal glucose tolerance test As a result of the glucose tolerance test conducted 12 weeks after feeding the experimental diet, as shown in Figure 20, 30 minutes after intraperitoneal injection of glucose, blood glucose levels increased in all groups, and after 120 minutes, blood glucose levels in the YC and CS groups had significantly decreased compared to the NC group, and were confirmed to be normal values.

[0197] 11. Effect of celery seed extract on plasma lipid concentrations After 12 weeks of feeding the experimental diets, the mice were sacrificed and plasma samples were collected. The concentrations of plasma total cholesterol and other lipids measured are shown in Table 5. As shown in Table 5, the total plasma cholesterol and non-HDL cholesterol concentrations in the CS group were significantly decreased compared to the NC group. In contrast, the HDL cholesterol concentration and the ratio of HDL cholesterol to total cholesterol (HDL-C / Total C ratio; HTR) were significantly increased in the CS group compared to the NC group.

[0198] [Table 5]

[0199] 12. Effect of celery seed extract on liver tissue weight, hepatotoxicity index, and lipid content As shown in FIG. 21A, the weight of liver tissue per 100 g of body weight was found to be enlarged in the NC group compared to the YC group.

[0200] GOT and GPT, which are indicators of hepatotoxicity, are enzymes present in hepatocytes, and generally, high plasma GOT and GPT levels indicate liver damage. As shown in Figure 21B, plasma GOT and GPT levels were significantly increased in the NC group compared to the YC group, and were significantly reduced by celery seed extract supplementation.

[0201] Furthermore, as shown in Figure 21C, the lipid content of the liver tissue of the NC group was significantly increased compared to the YC group, whereas the lipid content of the liver tissue of the CS group was significantly decreased compared to the NC group.

[0202] 13. Antioxidant-related biomarkers Comparison of the GSH content, an antioxidant-related biomarker, revealed that, as shown in Figure 22A, the GSH content in liver tissue, red blood cells, and plasma was decreased in the NC group, but increased to the level of the YC group by supplementation with celery seed extract.

[0203] Furthermore, as shown in Figure 22B, the metabolic activity of SOD in liver tissue and erythrocytes was significantly decreased in the NC group compared to the YC group, but was significantly increased by supplementation with celery seed extract.

[0204] The H2O2 contents in liver tissue (mitochondria and cytosol) and erythrocytes were significantly increased in the NC group compared with the YC group, as shown in Figure 22C, and were significantly decreased by supplementation with celery seed extract.

[0205] 14. Analysis of intestinal microorganisms The results of a comparative analysis of short-chain fatty acid content and microbial flora using raw feces collected from mice at 11-12 weeks of feeding are shown in Figure 23. As shown in Figure 23, the short-chain fatty acid analysis results showed higher values ​​in the YC group compared to the NC group, and a significant difference was observed in acetate in particular. Furthermore, it was found that supplementation with celery seed extract normalized the short-chain fatty acid content, which had decreased due to aging, to the level of the YC group.

[0206] Analysis of the gut microbial flora was performed at the genus level. Analysis of microorganisms associated with the development of inflammatory bowel disease and colitis confirmed that aging-related increases in levels were normalized to the YC group level by celery seed extract supplementation, as shown in Figure 24A. Furthermore, as shown in Figure 24B, microorganisms associated with gut microbial imbalance (Desulfovibrionaceae, Erysipelatoclostridium) were significantly increased in the NC group compared to the YC group, and significantly decreased in the CS group compared to the NC group. Microorganisms associated with obesity / metabolic disease (Lactobacillus) were significantly increased in the CS group compared to the NC group. Analysis of microorganisms involved in the synthesis of short-chain fatty acids showed that the Lachnospiraceae, which is involved in the production of acetate, tended to decrease in the NC group compared to the YC group, while the Roseburia genus tended to increase with the supplementation of celery seed extract.

[0207] The above description of the present invention is for illustrative purposes only, and those skilled in the art will understand that the present invention can be easily modified into other specific forms without changing the technical spirit or essential features of the present invention. Therefore, it should be understood that the above-described embodiments are illustrative in all respects and are not limiting. [Industrial Applicability]

[0208] The celery seed extract according to the present invention is a natural product and can increase muscle mass and strength with little or no side effects. This not only prevents muscle weakness, but also inhibits weight and body fat gain, and reduces blood lipid levels and hepatotoxicity indicators (GOT and GPT). Therefore, it can be useful in preventing, improving, or treating muscle weakness-related diseases, metabolic diseases, liver diseases, etc., and has industrial applicability.

Claims

1. A pharmaceutical composition for preventing or treating a muscle weakness-related disease, comprising: Contains ethanol extract of celery seeds as an active ingredient. The pharmaceutical composition for preventing or treating a muscle weakness-associated disease, wherein the muscle weakness-associated disease is one or more selected from the group consisting of sarcopenia, muscle atrophy, muscle dystrophy, and cardiac atrophy.

2. The pharmaceutical composition according to claim 1, wherein the sarcopenia is aging-related sarcopenia or obesity-related sarcopenia.

3. 10. The pharmaceutical composition of claim 1, wherein the composition has one or more of the following characteristics: (a) increased muscle mass, (b) suppression of muscle mass loss; (c) increased muscle strength; (d) inhibiting an increase in lipid content in muscle tissue; or (e) Inhibition of muscle tissue fibrosis.

4. The pharmaceutical composition of claim 3, wherein the composition further has one or more of the following effects: (a) suppression of weight and body fat gain; (b) suppression of fibrosis of liver tissue or adipose tissue; (c) suppressing an increase in plasma lipid levels; (d) suppression of an increase in lipid content in liver tissue; (e) suppression of increases in plasma hepatotoxicity indicators; (f) suppressing the increase of lipid peroxides and increasing antioxidant capacity, or (g) Improvement of gut microbial imbalance.

5. A food composition for preventing or improving a muscle weakening-related disease, Contains ethanol extract of celery seeds as an active ingredient. The food composition for preventing or improving a muscle weakening-associated disease, wherein the muscle weakening-associated disease is one or more selected from the group consisting of sarcopenia, muscle atrophy, muscle dystrophy, and cardiac atrophy.

6. The food composition according to claim 5, wherein the sarcopenia is aging-related sarcopenia or obesity-related sarcopenia.

7. The food composition according to claim 5, characterized in that the composition has one or more of the following characteristics: (a) increased muscle mass, (b) suppression of muscle mass loss; (c) increased muscle strength; (d) inhibiting an increase in lipid content in muscle tissue; or (e) Inhibition of muscle tissue fibrosis.

8. The food composition according to claim 7, characterized in that the composition further has one or more of the following characteristics: (a) suppression of weight and body fat gain; (b) suppression of fibrosis of liver tissue or adipose tissue; (c) suppressing an increase in plasma lipid levels; (d) suppression of an increase in lipid content in liver tissue; (e) suppression of increases in plasma hepatotoxicity indicators; (f) suppressing the increase of lipid peroxides and increasing antioxidant capacity, or (g) Improvement of gut microbial imbalance.

9. A muscle-strengthening feed or feed additive containing an ethanol extract of celery seeds as an active ingredient.

10. 1. Use of an ethanol extract of celery seeds for the manufacture of a medicament for the treatment of a muscle weakness-related disease, comprising: The muscle weakness-related disease is one or more selected from the group consisting of sarcopenia, muscle atrophy, muscle dystrophy, and cardiac atrophy.

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