Degradation system promoters for enhanced recycling

Abstract

This invention provides novel formulations for promoting cell recycling and methods for increasing the responses of various genes related to cell recycling. It also includes degradation system promoters for promoting recycling, containing water-soluble nucleoproteins, particularly low molecular weight compounds (250 to 15000) obtained through enzymatic treatment of fish sclerotium; health foods containing these degradation system promoters; and methods for increasing the responses of various degradation system-related genes and promoting cell recycling through oral intake of these degradation system promoters.

Description

Technical Field

[0001] This invention relates to degradation systems that promote the recycling of cells, etc., and more specifically to degradation systems that promote the recycling of proteasomes, autophagy, apoptosis, etc., and degradation system promoters that promote the recycling of cells or organelles, etc.

[0002] Furthermore, the present invention also relates to a method for increasing the response of various degradation system-related genes (proteasome-related genes, autophagy-related genes, apoptosis-related genes) by orally ingesting the degradation system promoter. Background Technology

[0003] In recent years, with the increasing public concern for health, health foods have emerged that use deoxyribonucleic acid (DNA), ribonucleic acid (RNA), and nucleoproteins (also known as nucleoproteins or salmon bladder extracts), which are complexes of these nucleic acids and proteins, as raw materials or active ingredients. For example, nucleoproteins are known to have immunomodulatory, antioxidant, and vasodilatory effects. In addition, there are proposals for cancer cell proliferation inhibitory substances containing low-molecular-weight water-soluble nucleic acids, or such water-soluble nucleic acids and amino acids (Patent Document 1).

[0004] Existing technical documents

[0005] Patent documents

[0006] Patent Document 1: Japanese Patent Application Publication No. 2014-152144 Summary of the Invention

[0007] [The technical problem that the invention aims to solve]

[0008] To date, various effects and benefits, such as antioxidant activity, have been described through the ingestion of nucleic acids (DNA, RNA) or nuclear proteins.

[0009] However, to date, there have been no reports indicating that the intake of these substances affects the degradation of aging cells or oxidized unwanted proteins and defective mitochondria within cells, thereby affecting the recycling of the cells themselves and organelles.

[0010] As degradation systems used for recycling within cells, there exist the ubiquitin-proteasome system, which selectively degrades unwanted and damaged proteins within the cell, and autophagy, associated with vesicular autophagosomes, which participate in the self-degradation of similarly damaged or dysfunctional organelles within the cell. Furthermore, the selective removal of damaged mitochondria is specifically termed mitophagy. Autophagy's degradation targets are "non-selective," whereas the ubiquitin-proteasome system primarily achieves selective degradation by labeling the proteins to be degraded with ubiquitin proteins. Recent studies have reported associations between these phenomena and aging and disease. For example, it has been reported that in Drosophila, mutations in autophagy-related genes reduce average lifespan, while overexpression increases it. Additionally, there are reports of associations between autophagy and neurodegenerative diseases such as Alzheimer's disease and Parkinson's disease, where the loss of autophagy leads to the accumulation of amyloid proteins such as β-amyloid and α-synuclein.

[0011] However, there have been no reports to date on the relationship between these ubiquitin-proteasome systems and autophagy and antioxidant activity.

[0012] The present invention aims to provide novel formulations for promoting cell recycling and methods for increasing responses to various related genes related to promoting cell recycling.

[0013] The technical solution adopted to solve the technical problem.

[0014] This invention relates to degradation system promoters for enhancing recycling containing water-soluble nucleoproteins, and particularly to degradation system promoters comprising nucleoproteins obtained by enzymatic treatment of fish sclerotium and reduced to a molecular weight of 250 to 15,000. Furthermore, it particularly relates to degradation system promoters targeting proteasome degradation systems, autophagy degradation systems, or apoptosis.

[0015] Furthermore, the present invention also relates to health foods containing the degradation system promoter, for example, health foods in the form of tablets, beverages, capsules, granules, powders, pills or gels.

[0016] In addition, the present invention also relates to a method for increasing the response of proteasome-related genes, the response of autophagy-related genes, or the response of apoptosis-related genes by oral intake of the degradation system promoter, and also to a method for promoting the recycling of cells, organelles, or proteins by oral intake of the degradation system promoter.

[0017] Furthermore, the present invention also targets proteasome-related gene response enhancers containing water-soluble nucleoproteins, autophagy-related gene response enhancers, apoptosis-related gene response enhancers, and cell, organelle, or protein recycling enhancers.

[0018] [The effects of the invention]

[0019] The present invention comprises a degradation system promoter of water-soluble nucleoproteins, particularly low-molecular-weight nucleoproteins based on enzymatic treatment of fish sclerotium. When orally ingested, this promoter induces proteasome activation, autophagy activation, and apoptosis activation, thereby promoting the degradation and removal of oxidized proteins and unhealthy organelles such as mitochondria, and cell death itself, thus enhancing the recycling of these cells. Furthermore, it is anticipated that this enhanced recycling effect will have a positive impact on health maintenance, including anti-oxidation and disease prevention. Attached Figure Description

[0020] Figure 1 This is a graph showing the results of d-ROMs tests performed on the blood of subjects before and after ingestion in the test tablet group (which ingested test tablets containing water-soluble nucleoprotein (salmon leucocyanide extract) for 8 weeks) and the invalid control tablet group (which ingested invalid control tablets without the extract for 8 weeks); A) is the shift (measured value) of d-ROMs, and B) is the change in d-ROMs, which is the change based on the pre-ingestion baseline.

[0021] Figure 2 This is a graph showing the results of a comprehensive microarray-based gene expression analysis of total RNA extracted from human blood in human subjects after 8 weeks of ingestion of water-soluble nucleoprotein (salmon leucocyanide extract).

[0022] Figure 3 This figure shows the yeast surviving after 24 hours of treatment in a culture medium obtained by propagating yeast (Saccharomyces cerevisiae BY4741 strain (MATa; his3D1; leu2D0; met15D0; ura3D0)) using SD complete liquid medium, with 1% water-soluble nucleoprotein (salmon leucocyanide extract) added, and a control group using only SD complete liquid medium to propagate yeast (Saccharomyces cerevisiae BY4741 strain) culture medium. The culture medium was appropriately diluted and inoculated onto YPD plates containing 4 mM hydrogen peroxide. The yeast was then cultured at 30°C for 72 hours under hydrogen peroxide conditions.

[0023] Figure 4This graph shows the survival rate of yeast (Saccharomyces cerevisiae BY4741 strain) culture medium obtained by propagating yeast (Saccharomyces cerevisiae BY4741 strain) using SD complete liquid medium with 1% water-soluble nucleoprotein (salmon leucovorin extract) added, and the control group obtained by propagating yeast (Saccharomyces cerevisiae BY4741 strain) culture medium obtained by propagating yeast (Saccharomyces cerevisiae BY4741 strain) using only SD complete liquid medium. After 24 hours of treatment, the culture medium was appropriately diluted and inoculated onto YPD plates with 4 mM hydrogen peroxide and YPD plates without hydrogen peroxide. The survival rate was calculated by counting the number of colonies with and without hydrogen peroxide after 72 hours of incubation at 30°C.

[0024] Figure 5 This is a graph showing the results of a comprehensive microarray-based gene expression analysis of total RNA extracted from yeast after 24 hours of adding water-soluble nucleoprotein (salmon leucovorin extract) to yeast (Saccharomyces cerevisiae BY4741 strain). Detailed Implementation

[0025] The degradation system promoter of the present invention contains a water-soluble nucleoprotein. The inventors have newly discovered that this water-soluble nucleoprotein can activate the proteasome, autophagy, and apoptosis—which are intracellular components and the cell itself—in order to promote the degradation and removal of oxidized proteins, carbonylated proteins, undesirable mitochondria, damaged lysosomes, and other undesirable proteins and organelles, as well as cell death itself. Therefore, the degradation system promoter of the present invention is expected to remove the aforementioned undesirable proteins, cells, and organelles through oral ingestion, thus enabling these cells to be recycled.

[0026] The present invention will now be described in detail.

[0027] In the degradation system promoter of the present invention, the target degradation system is the proteasome degradation system, the autophagy degradation system, or apoptosis. These can also be described as different scales of so-called degradation systems. The proteasome is associated with the selective degradation of undesirable proteins, autophagy is associated with the degradation of undesirable proteins and organelles in the cytoplasm, and apoptosis is associated with the degradation of the cell itself (cell death). Promoting the degradation of these undesirable cells makes them easier to recycle into new cells, thereby improving cell recycling. In addition, as mentioned above, the proteasome and autophagy are involved in aging and various diseases. Variations and reductions in these degradation systems can lead to undesirable phenomena such as reduced lifespan. Therefore, it is believed that by taking up nucleic acids (DNA, RNA) or nuclear proteins, the autophagy and proteasome systems can be promoted. It is believed that in addition to the increased antioxidant effect caused by the simple intake of nucleic acids, anti-aging and health maintenance can also be expected.

[0028] The water-soluble nucleoproteins used in the degradation system promoters of the present invention, and the proteasome-related gene response enhancers, autophagy-related gene response enhancers, apoptosis-related gene response enhancers, and cell, organelle, or protein recycling promoters described later, can be exemplified by water-soluble nucleoproteins obtained by extracting nucleic acids from biological cells and reducing their molecular weight through enzymatic treatment. That is, in this specification, water-soluble nucleoproteins can refer to extracts of fish milt that have been reduced in molecular weight. As an example of the aforementioned water-soluble nucleoproteins, examples include water-soluble nucleoproteins made by reducing the molecular weight of fish milt through enzymatic treatment or other methods to make it water-soluble, such as salmon milt extract.

[0029] It should be noted that in this specification, water solubility refers to the property of being able to dissolve in water at a concentration of 0.1% by mass or more.

[0030] Examples of fish species mentioned above include salmon, trout, and herring. Among these, salmon milt, which contains a large amount of nucleic acid, is preferred as it has historically been largely discarded and unable to be effectively utilized as a resource.

[0031] Examples of enzymatic treatment of fish milt include treating the suspension obtained by crushing fish milt with protease, followed by treatment with nuclease, etc.

[0032] Typically, if DNA or similar substances are extracted from fish roe, the molecular weight can reach 1,000,000 or more, so they are reduced to low molecular weight through enzymatic treatment. Preferred water-soluble nucleoproteins include those containing at least 20% by mass, more preferably at least 30% by mass, of nucleosides / nucleotides / oligonucleotides / polynucleotides (hereinafter also referred to as low molecular weight compounds) with a molecular weight of 250–100,000, such as 250–20,000, 250–15,000, 500–13,200, or 250–10,000 or 250–6,600.

[0033] Even more preferably, the water-soluble nucleoprotein described in this invention, such as salmon milt extract, can be a water-soluble nucleoprotein containing more than 2% by mass of a low molecular weight compound with a molecular weight of 500 to 15,000 and more than 18% by mass of a low molecular weight compound with a molecular weight of 250 to 6,600.

[0034] Examples of such proteases include serine proteases, cysteine ​​proteases, metalloproteinases, and aspartic proteases. Fish milt contains proteins in addition to protamine, so proteases with low substrate specificity are particularly desirable. Examples of good proteases include those manufactured by Novozymes Japan Co., Ltd. Hydrolysis using these proteases can be carried out at temperatures ranging from 30 to 60°C, for example, from 40 to 50°C, and at a pH of 6 to 7, depending on the activation and inactivation of the protease.

[0035] The aforementioned nuclease is an enzyme that hydrolyzes the 3,5'-phosphodiester bonds of deoxyribonucleic acid (DNA) to generate 5'-nucleotides polymerized from oligomers. There are no particular limitations on the properties of this nuclease, but it is preferable to possess a certain degree of thermal stability. Such nucleases are commercially available from companies such as Amano Enzyme Co., Ltd. and Sigma-Aldrich. Considering the activation and inactivation of the nuclease, hydrolysis using the aforementioned nuclease can be carried out, for example, at a temperature range of 60–75°C and a pH range of 5–6.

[0036] The specific steps of the enzyme treatment are as follows: First, the fish milt, such as salmon milt, is treated with proteases to hydrolyze the proteins contained in the fish milt. After hydrolysis, nuclease treatment is performed to reduce the nucleic acids to low molecular weight.

[0037] This invention also includes health foods containing degradation system promoters as a target.

[0038] The health food of the present invention contains, as an active ingredient, the degradation system promoter, for example, a water-soluble nucleoprotein with a molecular weight of 250 to 15,000, or, for example, a low molecular weight compound of about 250 to 10,000, obtained by enzymatic treatment of fish milt. It may also contain other incorporated ingredients depending on the various product forms described below. The content of the recycling agent in the health food may be appropriately adjusted with reference to the clinical dosage described below.

[0039] The health food of the present invention can be mixed with various known ingredients such as sweeteners, acidulants, and vitamins to create products that meet the user's preferences. Furthermore, as product forms, it can be provided in the form of, for example, tablets, capsules, granules, powders, pills, syrups, liquids, emulsions, suspensions, etc., or in the form of beverages, gels, dairy products such as yogurt, seasonings, processed foods, desserts, snacks, etc.

[0040] For example, the oral tablets, capsules, granules, powders, pills, etc., can be formulated using conventional methods. They can be prepared using excipients such as white sugar, lactose, glucose, starch, mannitol; binders such as hydroxypropyl cellulose, syrup, gum arabic, gelatin, sorbitol, tragacanth gum, methylcellulose, polyvinylpyrrolidone; disintegrants such as starch, carboxymethyl cellulose or its calcium salt, microcrystalline cellulose, polyethylene glycol; lubricants such as talc, magnesium stearate or calcium stearate, silicon dioxide; and lubricants such as sodium laurate, glycerin, etc.

[0041] Furthermore, the aforementioned syrups, liquids, emulsions, suspensions, etc., can also be formulated using conventional methods. Solvents such as water, ethanol, isopropanol, propylene glycol, 1,3-butanediol, and polyethylene glycol can be used as active ingredients to promote the degradation system. Surfactants such as sorbitol fatty acid esters, polyoxyethylene sorbitol fatty acid esters, polyoxyethylene fatty acid esters, polyoxyethylene ethers of hydrogenated castor oil, and lecithin can be used. Suspension agents such as sodium carboxymethyl salt, cellulose derivatives such as methylcellulose, natural rubbers such as gum arabic and gum arabic, and preservatives such as esters of p-hydroxybenzoic acid, benzalkonium chloride, and sorbates can also be used.

[0042] In addition, the beverage preparations and gelling agents, as well as dairy products, seasonings, processed foods, desserts, snacks, etc., may contain oligoRNA, zinc, collagen, chondroitin, hyaluronic acid, vitamins (vitamin B1, vitamin B2, vitamin B6, vitamin B12 and other B vitamins, vitamin C, etc.) and glutathione (yeast extract containing glutathione, reduced glutathione, oxidized glutathione, etc.).

[0043] More specifically, for applications such as beverages, the use is not limited to this purpose and may also include pig collagen peptides, syrups containing trace amounts of sugar, edible yeast extracts containing RNA, edible yeast containing zinc, shark cartilage extracts containing chondroitin, rooster comb extracts / acidulants containing hyaluronic acid, vitamin C, arginine, magnesium carbonate, niacin, vitamin B6, vitamin B2, vitamin B1, folic acid, vitamin B12, fish extracts containing anserine, citrulline, salmon ovary membrane extracts, and pig placenta extracts. Furthermore, for applications such as tablets, the use is not limited to this purpose and may also include brewer's yeast, ginkgo leaf extract, and garlic extract containing S-allyl cysteine. Additionally, for applications such as gels, the use is not limited to this purpose and may also include fish collagen peptides.

[0044] In addition, the health food of the present invention may contain other compounds that are active in pharmaceuticals or veterinary medicines, besides the degradation system promoters mentioned above as effective ingredients.

[0045] The clinical dosage of the degradation system promoters, etc., described in this invention varies depending on age, weight, patient sensitivity, severity of symptoms, etc. Generally, the baseline effective dosage is approximately 100 mg to 500 mg per day for adults, for example, as the amount of water-soluble nucleoprotein (salmon leucocyanide extract). However, amounts exceeding the above range may be used as needed.

[0046] It should be noted that the above-mentioned effective dosage guidelines also apply to the health foods of the present invention. In the case of health foods, for example, the above-mentioned daily dosage guidelines can be used as a reference, and the amount of degradation system promoter incorporated into the health food can be appropriately adjusted based on the daily intake of the health food itself.

[0047] The degradation system promoter of the present invention increases the response genes associated with the above-mentioned degradation systems, thereby promoting these degradation systems.

[0048] That is, the present invention also includes methods for increasing the response of proteasome-related genes, the response of autophagy-related genes, and the response of apoptosis-related genes through oral intake of the degradation system promoter. Furthermore, since degradation of cells, etc., also promotes recycling, the present invention also includes methods for increasing the recycling of cells, organelles, or proteins. Among these methods for increasing gene responses and increasing recycling, the same clinical dose (standard dose) as the aforementioned degradation promoter is preferably directly applied.

[0049] Furthermore, as described above, the present invention also includes water-soluble nuclear proteins, such as proteasome-related gene response enhancers, autophagy-related gene response enhancers, apoptosis-related gene response enhancers, cell or organelle recycling enhancers, and cell, organelle, or protein recycling enhancers as objects.

[0050] These response enhancers and recycling promoters, as described above, contain water-soluble nucleoproteins, such as salmon milt extract, which can effectively utilize the water-soluble nucleoproteins, such as salmon milt extract, used in the aforementioned degradation promoters.

[0051] Furthermore, these response enhancers and recycling promoters can also be in the form of health foods containing them. These health foods may preferably use the forms and ingredients exemplified in the above-mentioned health foods containing degradation promoters.

[0052] Example

[0053] <Water-soluble nucleoprotein (salmon leucocyanide extract)>

[0054] The water-soluble nucleoprotein (salmon milt extract) obtained from salmon milt using hydrolytic enzymes such as proteases and nucleases contains 20-30% by mass nucleic acid (water-soluble nucleic acid with a molecular weight range of 250 to 15,000) and 50% by mass amino acids (including 15% by mass arginine).

[0055] <Manufacturing of Test Tablets>

[0056] According to Table 1, test tablets (tablet A) containing the water-soluble nucleoprotein (salmon leucocyanide extract) obtained by the above manufacturing example and ineffective control tablets (tablet B) not containing the extract were manufactured.

[0057] [Table 1]

[0058] [Table 1] Formulation of the test tablets

[0059]

[0060] ※Amount added: 1.5g per 6 tablets

[0061] <Experimental Example (1): Changes in Markers of Oxidative Stress>

[0062] A randomized, double-blind, controlled trial was conducted on 41 healthy individuals (male, without medication, underlying diseases, etc.) aged 40 to under 60 years.

[0063] Subjects were divided into two groups: an experimental tablet group (tablet A, 20 subjects) and a null control tablet group (tablet B, 21 subjects). Each experimental group received either tablet A or tablet B for 8 weeks. In the experimental tablet group, the dosage of tablet A was adjusted to 360 mg (6 tablets per day) of water-soluble nucleoprotein (salmon leukocyte extract). In the null control tablet group, the same dosage of tablet B (6 tablets per day) as tablet A was administered.

[0064] The d-ROMs (Reactive Oxygen Metabolites-derived compounds) test was conducted to comprehensively evaluate the state of oxidative stress in vivo by collecting blood samples from subjects before and after ingestion (week 0 and week 8) and measuring hydrogen peroxide, which is a metabolite produced by free radicals in the blood.

[0065] In addition, total RNA was extracted from the collected blood for gene expression analysis.

[0066] The results are shown in Figure 1 Results of d-ROMs tests ( Figure 1 (A) Measured value, (B) Change from pre-ingestion baseline).

[0067] like Figure 1 As shown, compared with the nucleoprotein (salmon leukocyte extract) group, the experimental tablet group that ingested tablet A containing water-soluble nucleoprotein for 8 weeks showed a significant decrease in the measured values ​​and changes in d-ROMs (reduction of oxidative stress markers).

[0068] <Experimental Example (2): Gene Expression Analysis>

[0069] Similar to Experiment (1), the subjects in the experimental tablet group (tablet A, 20 subjects) and the null control tablet group (tablet B, 21 subjects) were given tablets for 8 weeks. Total RNA was then extracted from the blood collected from the subjects for comprehensive gene expression analysis.

[0070] The conditions for comprehensive gene expression analysis are as follows:

[0071] <Comprehensive Gene Expression Analysis Based on Microarrays>

[0072] Microarray used: Applied Biosystems (registered trademark), Human, Thermo Fisher Scientific.

[0073] 1. Total RNA quality was assessed (RNA degradation evaluation) using an Agilent 2100 Bioanalyzer.

[0074] 2. Samples with RNA concentration of 100 ng / μl or higher, RNA amount of 300 ng or higher, A260 / 280 ratio of 1.8-2.1, and RIN value of 7.0 or higher should be used for testing.

[0075] 3. cRNA synthesis was performed using the GeneChip WT Plus Reagent Kit. Using 100 ng of total RNA as a template, first-strand cDNA was synthesized in the first cycle via reverse transcription, followed by second-strand cDNA synthesis. Subsequently, cRNA was synthesized via in vitro transcription. After purification, the cRNA yield was determined spectrophotometrically.

[0076] 4. Single-stranded cDNA derived from cRNA was synthesized using the GeneChip WT Plus Reagent Kit. The second-cycle single-stranded cDNA was synthesized from the first cRNA via reverse transcription. The synthesized single-stranded DNA was purified and the yield was determined spectrophotometrically.

[0077] 5. Single-stranded DNA fragmentation and labeling were performed using the GeneChip WT Terminal Labeling Kit (Thermo Fisher Scientific). For 5.5 μg of ssDNA, fragmentation was performed using uracil DNA glycosylase (UDG) and depurine / depyrimidine endonuclease 1 (APE1). Subsequently, the fragmented ssDNA was biotinylated using terminal deoxynucleotidyl transferase (TdT) and GeneChip DNA Labeling Reagent.

[0078] 6. Hybridization of Clariom S Array, Human was performed using the GeneChip Hybridization, Wash, and Stain Kit (Thermo Fisher Scientific). Fragmented and labeled ssDNA was added to hybridization buffer and hybridized on Clariom S Array, Human for 17 hours. After washing and staining, fluorescence was read using a scanner, and CEL files were created using GCC (GeneChip Command Console software) and then processed via the Expression Console. TM The software generates a CHP file.

[0079] 7. Pathway analysis was performed using the Transcriptome Analysis Console (Thermo Fisher Scientific) with a threshold of 1.3 or higher to evaluate the rate of change of transcripts.

[0080] The gene expression analysis results related to each degradation system (proteasome, autophagy, apoptosis) are shown in Table 2. Furthermore, the results of the combined gene expression analysis of the proteasome degradation system and autophagy are shown in Table 2. Figure 2 .

[0081] [Table 2]

[0082] [Table 2] Results of gene expression analysis

[0083]

[0084] As shown in Table 2, compared with the null control group, the experimental tablet group that ingested tablet A containing water-soluble nucleoprotein (salmon leucocyanide extract) for 8 weeks showed a significant increase in the expression of genes related to various degradation systems.

[0085] <Experimental Example (3): Evaluation of Oxidative Stress Resistance Using Yeast>

[0086] For the yeast (Saccharomyces cerevisiae), the BY4741 strain was isolated by plate culture at -80℃, and 2-3 independent colonies were picked and cultured in SD complete medium with shaking for 18 hours (160 rpm, 30℃) to increase the yeast count.

[0087] Yeast culture broth with 1 / 10 volume added to fresh SD complete liquid medium was prepared. An experimental group was prepared with 1% of the aforementioned water-soluble nucleoprotein (salmon leucanthus extract) relative to the medium, and a control group was prepared with only the aforementioned yeast culture in SD complete liquid medium (water-soluble nucleoprotein (salmon leucanthus extract)). All cultures were incubated for 24 hours. The yeast culture broth after 24 hours was washed twice with sterile distilled water to remove the water-soluble nucleoprotein (salmon leucanthus extract) from the medium. The resulting culture broth was used as a sample. The yeast culture broth was appropriately diluted to obtain independent colonies and inoculated onto YPD plates supplemented with hydrogen peroxide at 4 mM. After incubation at 30°C for 72 hours, the number of yeast colonies surviving in the presence of hydrogen peroxide was counted. The appearance of the experimental and control groups after 72 hours of incubation is shown in the figure. Figure 3 .

[0088] like Figure 3 As shown, compared with the control group without added water-soluble nucleoprotein (salmon leucocyanide extract), the experimental group with added water-soluble nucleoprotein (salmon leucocyanide extract) had more colonies identified.

[0089] In addition, for the same hydrogen peroxide-added system as the above-mentioned system, but without hydrogen peroxide (hydrogen peroxide not added) system [yeast + water-soluble nucleoprotein (salmon leucocyanide extract) group, yeast only (water-soluble nucleoprotein (salmon leucocyanide extract) not added) group], it was also incubated at 30°C for 72 hours.

[0090] The number of bacterial colonies in each culture medium was counted. Based on the presence or absence of water-soluble nucleoprotein (salmon leucocyanide extract), the colony count in the hydrogen peroxide-added medium was divided by the colony count in the hydrogen peroxide-free medium to calculate the yeast survival rate (%). The results are shown below. Figure 4 .

[0091] like Figure 4 As shown, the results indicate that the survival rate of yeast with added water-soluble nucleoprotein (salmon oolong extract) was significantly higher than that of yeast without added water-soluble nucleoprotein (salmon oolong extract).

[0092] <Experimental Example (4): Comprehensive Gene Expression Analysis Based on Microarrays Using Yeast>

[0093] Comprehensive microarray-based gene expression analysis was performed using total RNA extracted from yeast (Saccharomyces cerevisiae) (BY4741 strain (MATa; his3D1; leu2D0; met15D0; ura3D0)) supplemented with 1% of the above water-soluble nucleoproteins (salmon leucocyanide extract) after 24 hours.

[0094] Using a microarray: Thermo Fisher Scientific's Genechip Yeast Genome 2.0 Array

[0095] 1. Total RNA quality was tested (concentration determination) using an Agilent 2200 TapeStation.

[0096] 2. Using 250 ng of total RNA from each sample, a labeling reaction was performed using the GeneChip (registered trademark) 3'IVT PLIS Kit, and hybridization was carried out.

[0097] 3. Use Expression Console TM The signal values ​​are quantized using the RMA algorithm and then normalized using the quantile method.

[0098] 4. Pathway analysis was performed using the Transcriptome Analysis Console (Thermo Fisher Scientific) with a threshold of 1.3 or higher to evaluate the rate of change of transcripts.

[0099] The gene expression analysis results related to each degradation system (proteasome, autophagy) are shown in Table 3 and... Figure 5 .

[0100] [Table 3]

[0101] [Table 3] Results of gene expression analysis in yeast (after 24 hours)

[0102]

[0103]

[0104] As shown in Table 3, the group with water-soluble nucleoprotein (salmon leucocyanide extract) added to yeast for 24 hours showed a significant increase in the expression of relevant genes in various degradation systems compared to the null control group (without addition).

[0105] As mentioned above, water-soluble nucleoproteins (salmon leucovorin extract) help reduce oxidative stress. Furthermore, the results of increased expression of genes related to degradation systems such as proteasomes, autophagy, and apoptosis in both humans and yeast initially showed that nucleoproteins can help promote the degradation of defective or damaged proteins, organelles such as mitochondria, and the cells themselves. It also showed the possibility of recycling these degraded cells to generate new cells, and the potential to help reduce, inhibit, and increase oxidative stress and health.

Claims

1. A degradation system promoter for improving recycling, wherein, It contains water-soluble nucleoproteins.

2. The degradation system promoter according to claim 1, wherein, The water-soluble nucleoprotein is a low-molecular-weight nucleoprotein with a molecular weight of 250 to 15,000, obtained by enzymatic treatment of fish sclerotium.

3. The degradation system accelerator according to claim 1 or 2, wherein, The degradation system is a proteasome degradation system.

4. The degradation system accelerator according to claim 1 or 2, wherein, The degradation system is an autophagy degradation system.

5. The degradation system promoter according to claim 1 or 2, wherein, The degradation system is apoptosis.

6. A health food, wherein, The degradation system promoter includes any one of claims 1 to 5.

7. The health food according to claim 6, wherein, The health food products are available in the form of tablets, beverages, capsules, granules, powders, pills, or gels.

8. A method for increasing the response of proteasome-related genes, wherein, The degradation system promoter described in claim 1 or 2 is administered orally.

9. A method for increasing the response of autophagy-related genes, wherein, The degradation system promoter described in claim 1 or 2 is administered orally.

10. A method for increasing the response of apoptosis-related genes, wherein, The degradation system promoter described in claim 1 or 2 is administered orally.

11. A method for improving the recycling of cells, organelles, or proteins, wherein, The degradation system promoter described in claim 1 or 2 is administered orally.

12. A proteasome-associated gene response enhancer, wherein, It contains water-soluble nucleoproteins.

13. A response enhancer for autophagy-related genes, wherein, It contains water-soluble nucleoproteins.

14. A response enhancer for apoptosis-related genes, wherein, It contains water-soluble nucleoproteins.

15. A cell, organelle, or protein recycling enhancer, wherein, It contains water-soluble nucleoproteins.

Images