KR102260925B1 - Composition for protecting cells and tissues against toxicity induced by particulate matter comprising lactobacillus casei - Google Patents

Abstract

Disclosed herein is a composition comprising a Lactobacillus casei strain as an active ingredient and having a protective effect against fine dust toxicity. The composition is Lactobacillus casei strain; lysate thereof; its culture; Alternatively, by including an extract of the strain, lysate or culture as an active ingredient, there is an effect of suppressing cell damage due to fine dust toxicity and enhancing resistance to fine dust toxicity.

Description

Composition for cell and tissue protection against fine dust toxicity, including Lactobacillus casei strain {COMPOSITION FOR PROTECTING CELLS AND TISSUES AGAINST TOXICITY INDUCED BY PARTICULATE MATTER COMPRISING LACTOBACILLUS CASEI}

Disclosed herein is a composition comprising a Lactobacillus casei strain as an active ingredient and having a protective effect against fine dust toxicity.

Recently, yellow dust caused by desertification of dry areas in northern China and Mongolia has been identified as a serious health hazard because it contains harmful heavy metals such as cadmium, copper, and lead, and various harmful bacteria and fungi. In addition, in recent years, carbon compounds, sulfates, nitrates, and harmful heavy metal compounds generated by combustion due to the use of fossil fuels are also rapidly increasing. These fine combustion products (fine dust, ultrafine dust) are known to exhibit various adverse effects on the human body.

Particulate Matter (PM) refers to fine dust that floats in the air for a long period of time. PM10 for particle diameter of 10 μm or less, PM5 for particle diameter of 5 μm or less, PM2.5 for particle diameter of 2.5 μm or less, PM1 for particle diameter of 1 μm or less. is called Among these fine dust, particles with a particle diameter of 2.5 μm or less are called ultra-fine dust.

They can directly penetrate into the human body through the respiratory tract or through the skin, causing direct harm to the human body, as well as being mixed with rainwater during rainfall to affect crops and livestock. In particular, fine dust with a particle diameter of 10 μm or less penetrates into the bronchial tubes through respiration and causes various respiratory diseases such as asthma and chronic obstructive pulmonary disease, and causes various inflammations in parts of the body exposed to the air, such as the skin and eyes. In addition, it causes the occurrence of other diseases such as cerebrovascular diseases such as stroke, migraine, dementia, cardiovascular disease, depression, intestinal dysfunction, reproductive function disorders, and fetal growth disorders, and once absorbed into the body, it is not easily excreted but accumulates It is known to cause cancer or cause genetic diseases, so urgent countermeasures are required.

Korean Patent Publication No. 10-2017-0025450

In one aspect, an object of the present specification is to provide a composition comprising the Lactobacillus casei strain as an active ingredient and having a protective effect against the toxicity of fine dust.

In another aspect, an object of the present specification is to provide a composition comprising the Lactobacillus casei strain as an active ingredient and inhibiting cell damage caused by the toxicity of fine dust.

In another aspect, an object of the present specification is to provide a composition comprising the Lactobacillus casei strain as an active ingredient and enhancing resistance to the toxicity of fine dust.

In one aspect, the technology disclosed herein is a Lactobacillus casei strain; lysate thereof; its culture; Or it provides a pharmaceutical composition for the prevention or treatment of diseases caused by fine dust toxicity comprising the extract of the strain, lysate or culture as an active ingredient.

In an exemplary embodiment, the strain may be a Lactobacillus casei HY2782 (Lactobacillus casei HY2782) strain having an accession number of KCTC 13438BP.

In an exemplary embodiment, the pharmaceutical composition may prevent or treat growth inhibition caused by fine dust toxicity.

In an exemplary embodiment, the pharmaceutical composition may prevent or treat reproductive dysfunction caused by fine dust toxicity.

In another aspect, the technology disclosed herein is a Lactobacillus casei strain; lysate thereof; its culture; Or it provides a food composition for inhibiting cell damage due to fine dust toxicity comprising the extract of the strain, lysate or culture as an active ingredient.

In an exemplary embodiment, the strain may be a Lactobacillus casei HY2782 (Lactobacillus casei HY2782) strain having an accession number of KCTC 13438BP.

In an exemplary embodiment, the food composition may be to improve growth inhibition by fine dust toxicity.

In an exemplary embodiment, the food composition may inhibit germ cell damage caused by fine dust toxicity.

In an exemplary embodiment, the food composition may be to improve reproductive dysfunction caused by fine dust toxicity.

In another aspect, the technology disclosed herein is a Lactobacillus casei strain; lysate thereof; its culture; Or it provides a cosmetic composition for inhibiting cell damage caused by fine dust toxicity comprising the extract of the strain, lysate or culture as an active ingredient.

In an exemplary embodiment, the strain may be a Lactobacillus casei HY2782 (Lactobacillus casei HY2782) strain having an accession number of KCTC 13438BP.

In another aspect, the technology disclosed herein is a Lactobacillus casei strain; lysate thereof; its culture; Or it provides a feed composition for inhibiting cytotoxicity due to fine dust toxicity comprising the extract of the strain, lysate or culture as an active ingredient.

In an exemplary embodiment, the strain may be a Lactobacillus casei HY2782 (Lactobacillus casei HY2782) strain having an accession number of KCTC 13438BP.

In an exemplary embodiment, the feed composition may be to improve growth inhibition by fine dust toxicity.

In an exemplary embodiment, the feed composition may inhibit germ cell damage caused by fine dust toxicity.

In an exemplary embodiment, the feed composition may be to improve reproductive dysfunction caused by fine dust toxicity.

In an exemplary embodiment, the feed composition is selected from the group consisting of cattle, pigs, sheep, horses, rabbits, dogs, cats, chickens, turkeys, ducks, turtles, snakes, fish, earthworms, spiders, insects and nematodes. It may have one or more breeding uses.

In an exemplary embodiment, the nematode may be a C. elegans.

The technology disclosed herein has the effect of providing a new use of the Lactobacillus casei strain, more specifically the Lactobacillus casei HY2782 (Lactobacillus casei HY2782) strain.

In one aspect, the technology disclosed herein has an effect of providing a composition comprising a Lactobacillus casei strain as an active ingredient and having a protective effect against the toxicity of fine dust.

In another aspect, the technology disclosed herein has an effect of providing a composition comprising the Lactobacillus casei strain as an active ingredient and inhibiting cell damage caused by the toxicity of fine dust.

In another aspect, the technology disclosed herein has an effect of providing a composition comprising the Lactobacillus casei strain as an active ingredient and enhancing resistance to the toxicity of fine dust.

1 is a graph confirming the protective efficacy against CCD-18Co human cell death by fine dust 1 containing polycyclic aromatic hydrocarbons (PAH) of the Lactobacillus casei strain used in the present specification.
Fine dust 1 (500 μg/mL) or Lactobacillus casei strain (10 ⁶ CFU/mL) was administered alone to CCD-18Co cells, or fine dust 1 and Lactobacillus casei strain were administered simultaneously and cultured for 72 hours. Survival rates were determined. The statistical significance was also shown, and the statistical significance was *** P < 0.001 when compared with the fine dust alone group. In this graph, it can be clearly seen that the Lactobacillus casei strain has a protective effect against human cell death by fine dust 1 .
2 is a graph confirming the protective efficacy against CCD-18Co human cell death by fine dust 2 containing a heavy metal of the Lactobacillus casei strain used in the present specification.
Fine dust 2 (500 μg/mL) or Lactobacillus casei strain (10 ⁶ CFU/mL) was administered alone to CCD-18Co cells, or fine dust 2 and Lactobacillus casei strain were administered simultaneously and cultured for 72 hours. Survival rates were determined. Statistical significance was also shown, and the statistical significance was * P < 0.05, *** P < 0.001 when compared with the fine dust alone group. In this graph, the protective efficacy against human cell death by fine dust 2 of the Lactobacillus casei strain can be clearly confirmed.
3 is a graph confirming the protective efficacy against fine dust of the Lactobacillus casei strain used in the present specification in an animal experiment, and for this experiment, a model animal, Caenorhabditis elegans ) was used.
E. coli OP50 or Lactobacillus casei of the present invention was fed for 4 days as food to C. elegans. At this time, in order to check the toxicity caused by fine dust 1 and 2, fine dust was added and fed respectively. To check the effect of fine dust on the growth of model animals, the body length of C. elegans was measured after 4 days, and the body length of 50 animals per group was measured. Statistical significance was also shown, and the statistical significance was *** P < 0.001 when compared with the control group fed only E. coli. In this graph, it can be clearly confirmed that the Lactobacillus casei strain has a protective effect on growth inhibition by fine dust 1 and 2.
4 is a graph confirming the protective efficacy against fine dust of the Lactobacillus casei strain used in the present specification through an animal experiment, and for this experiment, the fertility of a model animal, Caenorhabditis elegans , was investigated.
E. coli OP50 or Lactobacillus casei of the present invention was fed to the C. elegans as food. At this time, in order to check the toxicity caused by fine dust 1 and 2, fine dust was added and fed respectively. To confirm the effect of fine dust on reproduction of model animals, the effect of fine dust on reproductive function was verified by observing the total number of eggs laid by each group. The total number of eggs was measured for 30 C. elegans per group. Statistical significance was also shown, and the statistical significance was ** P < 0.01 when compared with the control group fed only E. coli. In this graph, it can be clearly seen that the Lactobacillus casei strain has a protective effect on the inhibition of reproductive function by fine dust 1 and 2.

Hereinafter, the present invention will be described in detail.

The techniques disclosed herein include Lactobacillus casei strains; lysate thereof; its culture; Or comprising the extract of the strain, lysate or culture as an active ingredient, and provides a composition having a protective effect against fine dust toxicity.

In an exemplary embodiment, the fine dust may include polycyclic aromatic hydrocarbons or heavy metals.

Lactobacillus casei strain is a representative lactic acid bacteria used in the production of fermented milk products. Until now, Lactobacillus casei strain has been known to improve intestinal flora, increase intestinal beneficial bacteria, reduce intestinal harmful bacteria, improve bowel function, improve immunity, and reduce allergy or asthma symptoms. However, there is no known use of Lactobacillus casei for protection against cell damage, growth inhibition, or reproductive toxicity caused by fine dust.

As used herein, the term "active ingredient" refers to a component capable of exhibiting the desired activity alone or in combination with a carrier having no activity by itself.

In one aspect, the composition disclosed herein has a protective effect against the toxicity of fine dust, an effect of inhibiting cell damage due to the toxicity of fine dust, and an effect of enhancing resistance to the toxicity of fine dust.

In an exemplary embodiment, the composition may improve growth inhibition or growth inhibition caused by the toxicity of fine dust.

In an exemplary embodiment, the composition may inhibit reproductive cell damage due to the toxicity of fine dust or improve reproductive function degradation due to the toxicity of harmful substances.

The strain according to the present specification may be improved or improved to have an activity equivalent to or superior to this by a conventional physicochemical mutation method known in the art to which the present specification belongs.

In an exemplary embodiment, the strain may be a Lactobacillus casei HY2782 (Lactobacillus casei HY2782) strain having an accession number of KCTC 13438BP.

In an exemplary embodiment, the strain is cultured and the culture solution is centrifuged, washed with sterile physiological saline, dispersed in a solvent, such as sterile oil, and lyophilized to prepare and use in the form of a freeze-dried powder.

The lysate may refer to a product obtained by crushing the strain itself by chemical or physical force.

The culture may refer to a material including some or all materials contained in the culture medium regardless of the form of the culture, for example, a material containing a metabolite or secretion resulting from the strain culture, or a lysate thereof may mean, and the strain itself may also be included in the culture. In addition, the culture may mean including a fermented product.

The extract may refer to a product obtained by extracting the strain itself, the lysate of the strain, the culture of the strain, or a mixture thereof regardless of the extraction method, the extraction solvent, the extracted component or the form of the extract, and fractionation, concentration after extraction It is a broad concept that includes all substances that can be obtained by processing or processing by other methods, such as.

According to an exemplary embodiment, the composition may be a pharmaceutical composition.

In addition to the active ingredient, the pharmaceutical composition may further contain pharmaceutical adjuvants such as preservatives, stabilizers, wetting agents or emulsification accelerators, salts and/or buffers for osmotic pressure control, and other therapeutically useful substances, in a conventional method. It can be formulated in the form of various oral administration or parenteral administration according to the needs.

The oral administration agents include, for example, tablets, pills, hard and soft capsules, solutions, suspensions, emulsifiers, syrups, powders, powders, fine granules, granules, pellets, etc., and these formulations contain surfactants in addition to active ingredients. , diluents (e.g. lactose, dextrose, sucrose, mannitol, sorbitol, cellulose and glycine), lubricants (e.g. silica, talc, stearic acid and its magnesium or calcium salts and polyethylene glycol) . Tablets may also contain binders such as magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose and polyvinylpyrrolidine, optionally starch, agar, alginic acid or its sodium salt It may contain pharmaceutical additives such as disintegrants, absorbents, colorants, flavoring agents, and sweetening agents. The tablet may be prepared by a conventional mixing, granulating or coating method.

In addition, the parenteral dosage form may be a transdermal dosage form, for example, injections, drops, ointments, lotions, gels, creams, sprays, suspensions, emulsions, suppositories, patches, etc. may be, but is not limited thereto.

Determination of the dosage of the active ingredient is within the level of those skilled in the art, and the daily dosage of the drug varies depending on various factors such as the progress of the subject to be administered, the onset time, age, health status, complications, etc. Based on 1 μg/kg to 2000 mg/kg of the composition in one aspect, and 50 μg/kg to 500 mg/kg in another aspect, it can be administered in divided doses 1 to 3 times a day, and the dosage does not limit the scope of the present invention in any way.

The pharmaceutical composition may be an external preparation for the skin, and the external preparation for skin is a generic term that may include anything applied outside the skin, and pharmaceuticals of various formulations may be included here.

According to an exemplary embodiment, the composition may be a food composition.

The food composition may be in a liquid or solid form, for example, various foods, beverages, gum, tea, vitamin complexes, health supplements, etc., to be used in the form of powders, granules, tablets, capsules or beverages. can In addition to the active ingredient, the food composition of each formulation can be appropriately selected and formulated by those skilled in the art without difficulty depending on the formulation or purpose of use in addition to the active ingredient, and a synergistic effect may occur when applied simultaneously with other raw materials.

There is no particular limitation on the liquid ingredients that may be contained in addition to the active ingredients disclosed herein, and may include various flavoring agents or natural carbohydrates as additional ingredients, like conventional beverages. Examples of the natural carbohydrate include monosaccharides, disaccharides such as glucose and fructose, polysaccharides such as maltose and sucrose, conventional sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol etc. As the flavoring agent, natural flavoring agents (taumatin, stevia extract (eg, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (eg, saccharin, aspartame, etc.) can be advantageously used. The proportion of the natural carbohydrate may be generally about 1 to 20 g, in one aspect, about 5 to 12 g per 100 ml of the composition disclosed herein.

In one aspect, the food composition includes various nutrients, vitamins, minerals (electrolytes), synthetic flavoring agents and flavoring agents such as natural flavoring agents, coloring agents and thickening agents (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its salts. salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. In another aspect, it may include pulp for the production of natural fruit juices and vegetable beverages. The above components may be used independently or in combination. The proportion of the additive may vary, but is generally selected in the range of 0.001 to about 20 parts by weight per 100 parts by weight of the composition disclosed herein.

In an exemplary embodiment, the composition may be a cosmetic composition.

In addition to the active ingredient, the cosmetic composition may further include functional additives and ingredients included in general cosmetic compositions. The functional additive may include a component selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, high-molecular peptides, high-molecular polysaccharides, sphingolipids, and seaweed extract. Other ingredients included include oils and fats, moisturizers, emollients, surfactants, organic and inorganic pigments, organic powders, UV absorbers, preservatives, fungicides, antioxidants, plant extracts, pH adjusters, alcohols, pigments, fragrances, blood circulation An accelerator, a cooling agent, a limiting agent, purified water, etc. are mentioned.

The formulation of the cosmetic composition is not particularly limited, and may be appropriately selected according to the purpose. For example, powder, skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nourishing lotion, massage cream, nourishing cream, moisture cream, hand cream, foundation, essence, nourishing essence, pack, shampoo , soap, cleansing foam, cleansing lotion, cleansing cream, body lotion and body cleanser may be prepared in any one or more formulations selected from the group consisting of, but is not limited thereto.

When the formulation of the present invention is a paste, cream or gel, animal fiber, vegetable fiber, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as a carrier component. can

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component, and in particular, in the case of a spray, additional chlorofluorohydrocarbon, propane /may contain propellants such as butane or dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a solvent, solvating agent or emulsifying agent is used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 ,3-butylglycol oil, glycerol fatty esters, fatty acid esters of polyethylene glycol or sorbitan.

When the formulation of the present invention is a suspension, as a carrier component, water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol esters and polyoxyethylene sorbitan esters, microcrystalline Cellulose, aluminum metahydroxide, bentonite, agar or tracanth may be used.

When the formulation of the present invention is a surfactant-containing cleansing agent, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate, fatty acid amide as carrier components Ether sulfate, alkylamidobetaine, aliphatic alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, linolin derivative or ethoxylated glycerol fatty acid ester and the like can be used.

When the formulation of the present invention is soap, alkali metal salts of fatty acids, fatty acid hemiester salts, fatty acid protein hydrolysates, isethionate, lanolin derivatives, aliphatic alcohols, vegetable oils, glycerol, sugar, etc. may be used as carrier components. have.

In an exemplary embodiment, the composition may be a feed composition.

In an exemplary embodiment, the composition may be a feed composition for livestock.

In an exemplary embodiment, the composition is composed of cattle, pigs, sheep, horses, rabbits, dogs, cats, chickens, turkeys, ducks, turtles, snakes, flounder, fish such as eels, earthworms, spiders, insects and nematodes. It may have one or more breeding uses selected from the group.

In an exemplary embodiment, the nematode may be a C. elegans.

Hereinafter, the present invention will be described in more detail through examples. These examples are only for illustrating the present invention, and it will be apparent to those of ordinary skill in the art that the scope of the present invention is not to be construed as being limited by these examples.

Example 1. Securing fine dust

There were two types of fine dust used in this specification, and it was purchased from Sigma-Aldrich and used. Both types of fine dust were PM10 (particulate matter 10, fine dust characterized by a particle size of about 10 μm), and fine dust 1 was characterized by containing a large amount of polycyclic aromatic hydrocarbons (PAHs). Fine dust 2 is characterized by containing a large amount of heavy metals such as arsenic, cadmium, nickel, and lead.

Example 2. Lactobacillus Casey acquisition of strains

The strain used in this example is a known Lactobacillus casei HY2782 strain, and the strain powder was obtained from the manufacturer Korea Yakult Co., Ltd. Pyeongtaek Probiotics Plant and used (http://www.foodb2b.kr/kfda) /product/view/1187872/). The Lactobacillus casei HY2782 lactic acid bacteria powder (licensing number 20140017107, item manufacturing number 201400171077, license date July 19, 2016) has already had the appearance of a light yellow powder with no odor and unique flavor, and the Lactobacillus casei HY2782 The strain was deposited at the Korea Research Institute of Bioscience and Biotechnology Biological Resources Center as of December 19, 2017, and was given deposit number KCTC 13438BP.

In the following experimental example, the protective efficacy against harmful substances of the Lactobacillus casei strain is evaluated in serous cell (in vitro ) and C. elegans nematode ( in vivo ) models, and based on this, a composition having a protective effect against fine dust toxicity is provided want to

Experimental example 1. Human cell death effect by fine dust and Lactobacillus Casey Apoptosis protective effect by the strain

In this experimental example, the human apoptosis effect of fine dust and the apoptosis protective effect by the Lactobacillus casei strain were confirmed. For this purpose, two types of fine dust were used. Fine dust 1 was fine dust containing a large amount of polycyclic aromatic hydrocarbons, and fine dust 2 was fine dust containing a large amount of heavy metals.

CCD-18Co cells, a normal human colon cell line, were purchased from the Korea Cell Line Bank (Seoul) and used. CCD-18Co cells were 5% using RPMI medium (Roswell Park Memorial Institute medium) supplemented with 10% heat-inactivated fetal bovine serum (FBS), 10 U/mL penicillin, and 100 μg/mL streptomycin. It was cultured and maintained under conditions of carbon dioxide and 37°C.

CCD-18Co cells were aliquoted in a 48-well plate to 2×10 ⁴ cells per well, and then cultured for 24 hours to attach. After that, fine dust 1 (500 μg/mL), fine dust 2 (500 μg/mL) or Lactobacillus casei strain (10 ⁶ CFU/mL) were administered alone, or fine dust and Lactobacillus casei strain were administered simultaneously 72 incubated for hours. After the end of the culture, the cell viability was measured using the Ez-Cytox (Dail Lab Science, Korea) kit. Ez-Cytox solution was added to the medium diluted 10 times to the medium, and then cultured for 1 hour. After centrifugation was performed to completely remove fine dust, the absorbance was measured at 450 nm using only the supernatant. The cell viability is proportional to the absorbance, and the cell viability of the control group was 100%, and the cell viability of the rest of the experimental group was calculated.

As a result of the experiment, the viability of CCD-18Co cells was significantly reduced when fine dust 1 (500 μg/mL) containing a large amount of polycyclic aromatic hydrocarbons was administered. At this time, when fine dust 1 and Lactobacillus casei strain (10 ⁶ CFU/mL) were administered simultaneously, it was confirmed that the cell viability was significantly recovered compared to the experimental group treated with fine dust 1 alone ( FIG. 1 ). Similarly, fine dust 2 containing a large amount of heavy metal also reduced the cell viability, and it was confirmed that cell death was inhibited by simultaneous administration of the Lactobacillus casei strain (FIG. 2). In conclusion, it was confirmed that the Lactobacillus casei strain has a protective effect against human cell death caused by fine dust containing polycyclic aromatic hydrocarbons or heavy metals.

Experimental Example 2. Effect of inhibiting the growth of C. elegans, a model animal, caused by fine dust and recovery effect of fine dust toxicity by Lactobacillus casei strain

To confirm the toxicity of fine dust and the recovery ability by the Lactobacillus casei strain, C. elegans was used as an animal model. C. elegans was used after receiving a wild-type N2 strain from the Caenorhabditis Genetics Center (CGC, USA). E. coli OP50 was given as food, and the experiment was carried out while maintaining at 20 °C. Escherichia coli OP50 was also purchased from the Caenorhabditis Genetics Center (CGC, USA) and used.

Caenorhabditis elegans is a model animal commonly used as a model animal in biological and chemical research. C. elegans is advantageous in terms of animal research ethics as a substitute for rodent experiments, and it takes pharmaceutical and functional food compositions one step further from the existing in vitro-level experiments (in vitro ) to improve the individual-level in vivo activity and efficacy. There are merits that can be evaluated. In addition, recently, several prominent research papers have widely used these C. elegans nematodes for functional excavation of lactic acid bacteria and research on the mechanism of action.

Only eggs of C. elegans were collected and used for toxicity evaluation experiments. E. coli OP50 and Lactobacillus casei strains were each coated on NGM (nematode growth medium) agar medium, and fine dust 1 and fine dust 2 were added at a concentration of 1 mg/mL for toxicity evaluation. C. elegans eggs were sprinkled on the medium coated with fine dust and E. coli or Lactobacillus casei strain prepared in this way, and then cultured for 4 days. Afterwards, the shape of C. elegans was observed using a stereoscopic microscope and micrographs were taken, and the size of C. elegans was measured using the Image J program (National Health Institute, USA). The size of 50 nematodes in each experimental group was measured.

As a result of the experiment, the Lactobacillus casei strain of the present invention did not affect the growth of the model animal, C. elegans, compared with E. coli OP50, which is a general food for C. elegans. That is, it was confirmed that there was no significant difference between the Lactobacillus cassei strain as food and E. coli, which is a general food. However, when fine dust 1 (500 μg/mL) containing a large amount of polycyclic aromatic hydrocarbons or fine dust 2 containing a large amount of heavy metals was administered to E. coli, the growth of C. elegans, a model animal, was significantly reduced (Fig. 3). ). Through this, it was confirmed that fine dust causes toxicity at the individual level. In addition, it was confirmed that there was no toxic effect on the growth of C. elegans when fine dust and Lactobacillus casei strain were administered at the same time, and through this, Lactobacillus casei strain was able to protect individual-level growth toxicity caused by fine dust. It was confirmed that it is possible (FIG. 3). These effects were confirmed in both fine dust 1 and fine dust 2 regardless of the type of fine dust.

Experimental example 3. Model animals caused by fine dust Pretty Little elegans reproductive suppression effect and Lactobacillus Casey Efficacy of recovery of fine dust toxicity caused by strains

In this experimental example, the effect on fertility was also observed by observing the total number of eggs laid as well as the size of C. elegans in order to re-verify the recovery efficacy by the toxicity and Lactobacillus casei.

Only eggs of C. elegans were collected and used for toxicity evaluation experiments. E. coli OP50 and Lactobacillus casei strains were each coated on NGM (nematode growth medium) agar medium, and fine dust 1 and fine dust 2 were added at a concentration of 1 mg/mL for toxicity evaluation. C. elegans eggs were sprinkled on the medium coated with fine dust and E. coli or Lactobacillus casei strain prepared in this way, and then cultured for 64 hours. Afterwards, each group of C. elegans was transferred to a medium coated with E. coli OP50, respectively, and the number of eggs laid each day was observed. The number of eggs laid by 30 eggs in each group was observed, and the number of eggs laid per day was calculated by dividing the number of eggs laid each day by 30. was made in number.

As a result of the experiment, the Lactobacillus casei strain of the present invention did not significantly affect the reproduction of the model animal, C. elegans, compared with E. coli OP50, which is a general C. elegans food. That is, when the Lactobacillus casei strain was fed as food, it was confirmed that there was no significant difference in fertility compared to the general food E. coli OP50 strain. However, when fine dust 1 (500 μg/mL) containing a large amount of polycyclic aromatic hydrocarbons was administered to E. coli, the fertility of the model animal, C. elegans, was reduced, and fine dust 2 (500 μg/mL) containing a large amount of heavy metals ), the fertility of the model animal, C. elegans, was significantly reduced ( FIG. 4 ). Through this, it was confirmed that fine dust causes toxicity at the individual level. At this time, it was confirmed that when fine dust and Lactobacillus casei strain were administered at the same time, there was no toxic effect on the reproduction of C. elegans, and through this, Lactobacillus casei strain could protect individual level toxicity caused by fine dust. was confirmed (FIG. 4). These effects were confirmed in both fine dust 1 and fine dust 2 regardless of the type of fine dust.

Formulation examples of the composition according to one aspect of the present specification will be described below, but other various formulations are also applicable, which is not intended to limit the present invention, but merely to describe it in detail.

[Formulation Example 1] Tablet

After mixing 150 mg of Lactobacillus casei culture, 100 mg of glucose, 50 mg of red ginseng extract, 96 mg of starch and 4 mg of magnesium stearate, 40 mg of 30% ethanol was added to form granules, dried at 60° C. was used for tableting.

[ Formulation example 2] granules

After mixing 150 mg of Lactobacillus casei culture, 100 mg of glucose, 50 mg of red ginseng extract and 600 mg of starch, 100 mg of 30% ethanol was added to form granules, dried at 60° C. did. The final weight of the contents was 1 g.

[Formulation Example 3] Drinks

150 mg of Lactobacillus casei culture, 10 g of glucose, 50 mg of red ginseng extract, 2 g of citric acid and 187.8 g of purified water were mixed and filled in a bottle. The final volume of the contents was 200 ml.

[ Formulation example 4] Manufacturing of health food

Lactobacillus casei culture.... 1000 mg

vitamin mixture

Vitamin A Acetate ..............70 μg

Vitamin E .....................1.0 mg

Vitamin B1 ....................0.13 mg

Vitamin B2 .... 0.15 mg

Vitamin B6 ............... 0.5 mg

Vitamin B12 ............... 0.2 μg

Vitamin C.............................. 10 mg

Biotin .............................. 10 μg

Nicotinic acid amide . . . . . . . . . . . 1.7 mg

Folic acid .................... 50 μg

Calcium pantothenate............... 0.5 mg

mineral mixture

Ferrous sulfate ............... 1.75 mg

Zinc Oxide ............... 0.82 mg

Magnesium carbonate ............... 25.3 mg

Potassium phosphate monobasic .................... 15 mg

Dibasic Calcium Phosphate ................... 55 mg

Potassium citrate ............... 90 mg

Calcium carbonate .............. 100 mg

Magnesium Chloride................... 24.8 mg

The composition ratio of the vitamin and mineral mixture is a composition that is relatively suitable for health food in a preferred embodiment, but the mixing ratio may be arbitrarily modified, and the ingredients are mixed according to a conventional health food manufacturing method. , to prepare granules, and can be used for preparing health food compositions according to a conventional method.

[ Formulation example 5] Manufacture of health drinks

Lactobacillus casei culture.... 1000 mg

Citric acid ....................1000 mg

Oligosaccharide ............... 100 g

Plum Concentrate ............... 2 g

Taurine ............................... 1 g

Purified water was added to the total ....... 900 ㎖

After mixing the above ingredients according to a conventional health drink manufacturing method, stirring and heating at 85° C. for about 1 hour, the resulting solution was filtered and obtained in a sterilized 2L container, sealed and sterilized, and then stored in a refrigerator. Then, it was used to prepare the health drink composition of the present invention.

Although the composition ratio is prepared by mixing ingredients suitable for relatively favorite beverages in a preferred embodiment, the mixing ratio may be arbitrarily modified according to regional and ethnic preferences such as demand class, demanding country, and use purpose. Those of ordinary skill in the art to which the present invention pertains will be able to make various applications and modifications within the scope of the present invention based on the above contents.

Above, a specific part of the present invention has been described in detail, for those of ordinary skill in the art, this specific description is only a preferred embodiment, and the scope of the present invention is not limited thereby. It will be obvious. Accordingly, the substantial scope of the present invention will be defined by the appended claims and their equivalents.

Korea Institute of Bioscience and Biotechnology Biological Resources Center KCTC13438BP 20171219

Claims (7)

As a composition for alleviating the toxicity of fine dust,
The composition comprises Lactobacillus casei HY2782 (Lactobacillus casei HY2782) strain, its lysate, its culture, the extract of the strain, the extract of the lysate or the extract of the culture as an active ingredient,
The fine dust is a food composition for alleviating toxicity of fine dust, including polycyclic aromatic hydrocarbons or heavy metals.
The method of claim 1,
The fine dust is PM10, a food composition for alleviating toxicity of fine dust.
As a composition for alleviating the toxicity of fine dust,
The composition comprises Lactobacillus casei HY2782 (Lactobacillus casei HY2782) strain, its lysate, its culture, the extract of the strain, the extract of the lysate or the extract of the culture as an active ingredient,
The fine dust is a cosmetic composition for alleviating toxicity of fine dust, including polycyclic aromatic hydrocarbons or heavy metals.
4. The method of claim 3,
The fine dust is PM10, a cosmetic composition for alleviating toxicity of fine dust.
As a composition for alleviating the toxicity of fine dust,
The composition comprises Lactobacillus casei HY2782 (Lactobacillus casei HY2782) strain, its lysate, its culture, the extract of the strain, the extract of the lysate or the extract of the culture as an active ingredient,
The fine dust is a feed composition for alleviating the toxicity of fine dust, including polycyclic aromatic hydrocarbons or heavy metals.
6. The method of claim 5,
The fine dust is PM10, a feed composition for alleviating the toxicity of fine dust. delete

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