CN109966321A - The composition being used for from particulate matter toxicity protection cell and tissue comprising lactobacillus casei bacterial strain - Google Patents

The composition being used for from particulate matter toxicity protection cell and tissue comprising lactobacillus casei bacterial strain Download PDF

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CN109966321A
CN109966321A CN201811557788.0A CN201811557788A CN109966321A CN 109966321 A CN109966321 A CN 109966321A CN 201811557788 A CN201811557788 A CN 201811557788A CN 109966321 A CN109966321 A CN 109966321A
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particulate matter
lactobacillus casei
bacterial strain
toxicity
composition
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CN109966321B (en
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姜京秀
判哲镐
车光贤
洪性哲
金周妍
李昭暎
崔一东
李明熙
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Korea Advanced Institute of Science and Technology KAIST
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
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    • A61K35/74—Bacteria
    • A61K35/741—Probiotics
    • A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
    • A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K10/00—Animal feeding-stuffs
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    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K50/00—Feeding-stuffs specially adapted for particular animals
    • A23K50/40—Feeding-stuffs specially adapted for particular animals for carnivorous animals, e.g. cats or dogs
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/99—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from microorganisms other than algae or fungi, e.g. protozoa or bacteria
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P39/00—General protective or antinoxious agents
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q17/00—Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
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    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2200/00—Function of food ingredients
    • A23V2200/30—Foods, ingredients or supplements having a functional effect on health

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Abstract

The present invention discloses the composition comprising lactobacillus casei bacterial strain as effective component and to particulate matter toxicity with protective action.The composition includes the extract of lactobacillus casei bacterial strain, its fragment, its culture or the bacterial strain, fragment or culture as effective component, to inhibit the cellular damage as caused by particulate matter toxicity, and improves the repellence to particulate matter toxicity.

Description

It is used for comprising lactobacillus casei bacterial strain from particulate matter toxicity protection cell and tissue Composition
Technical field
The present invention is disclosed comprising lactobacillus casei bacterial strain as effective component and to particulate matter toxicity with protective action Composition.
Background technique
Recently, the yellow sand occurred by NORTH CHINA, the desertification of Mongolian dry ground band contains harmful weight such as cadmium, copper, lead Metal and various harmful bacterias and mould are identified as serious healthy adverse factor.In addition, recently, due to using fossil Fuel is also being sharply increased by the compound of the carbon compound of combustion generation, sulfate, nitrate, harmful heavy metal.According to It knows, this micro combustions product (particulate matter, fine particle) generates many side effects to human body.
Particulate matter (particulate matter, PM) refer in air for a long time floating dust, 10 microns of partial size Hereinafter referred to as PM10,5 microns of hereinafter referred to as PM5 of partial size, 2.5 microns of hereinafter referred to as PM2.5 of partial size, 1 micron of partial size with It is referred to as PM1 down.In these particulate matters, by 2.5 microns of partial size referred to as fine particles below.
These particulate matters directly penetrate into inside of human body by respiratory system or skin, directly injury human body, moreover, dropping When rain, mixes in rainwater, have an effect on crops and domestic animal.In particular, 10 microns of particulate matters below of partial size pass through breathing infiltration To bronchus, the various respiratory diseases such as asthma, chronic closure tuberculosis are induced, it is aerial in the exposure such as skin and eyeball Physical feeling causes various inflammation.In addition, and occur cerebral hemorrhage, migraine, the cranial vascular diseases such as dementia, cardiovascular disease, The inducement of the other diseases such as depression, intestinal dysfunction, reproductive dysfunction, locomitivity decline, fetal growth obstacle, once It is absorbed into vivo, would not be discharged easily, but accumulate, induce cancer or the inducement as genetic disease, it is therefore desirable to the greatest extent It games fastly.
Citation
Patent document
Patent document 1:KR 10-2017-0025450 A
Summary of the invention
Technical problem
It is an advantage of the invention to provide one kind comprising lactobacillus casei bacterial strain as effective component and for Grain object toxicity has the composition of protection effect.
Another object of the present invention is to provide one kind comprising lactobacillus casei bacterial strain as effective component and inhibition by The composition of cellular damage caused by particulate matter toxicity.
It is a further object of the present invention to provide one kind comprising lactobacillus casei bacterial strain be effective component and enhance to The composition of the repellence of grain object toxicity.
Technical solution
On the one hand, the present invention provides a kind of comprising lactobacillus casei bacterial strain, its fragment, its culture or the bacterium The extract of strain, fragment or culture is as effective component for preventing or treating the disease as caused by particulate matter toxicity Pharmaceutical composition.
In embodiment, the bacterial strain can be Lactobacillus casei HY2782 (Lactobacillus casei HY2782), Lactobacillus casei KCTC3109 (Lactobacillus casei KCTC3109) or Lactobacillus casei KCTC13086 (Lactobacillus casei KCTC13086)。
In embodiment, the growth inhibition as caused by particulate matter toxicity can be prevented or be treated to described pharmaceutical composition.
In embodiment, the barrier of the reproductive function as caused by particulate matter toxicity can be prevented or be treated to described pharmaceutical composition Hinder.
In embodiment, the Motor execution ability as caused by particulate matter toxicity can be prevented or be treated to described pharmaceutical composition Decline.
On the other hand, the present invention provides a kind of comprising lactobacillus casei bacterial strain, its fragment, its culture or the bacterium The extract of strain, fragment or culture is as effective component for inhibiting the food of the cellular damage as caused by particulate matter toxicity Product composition.
In embodiment, the bacterial strain can be Lactobacillus casei HY2782 (Lactobacillus casei HY2782), Lactobacillus casei KCTC3109 (Lactobacillus casei KCTC3109) or Lactobacillus casei KCTC13086 (Lactobacillus casei KCTC13086)。
In embodiment, the food compositions can improve the growth inhibition as caused by particulate matter toxicity.
In embodiment, the food compositions can inhibit the Damage of Germ Cells as caused by particulate matter toxicity.
In embodiment, the food compositions can improve the reproductive dysfunction as caused by particulate matter toxicity.
In embodiment, the food compositions can improve the decline of the Motor execution ability as caused by particulate matter toxicity.
On the other hand, the present invention provides a kind of comprising lactobacillus casei bacterial strain, its fragment, its culture or the bacterium The extract of strain, fragment or culture is as effective component for inhibiting the change of the cellular damage as caused by particulate matter toxicity Cosmetic compositions.
In embodiment, the bacterial strain can be Lactobacillus casei HY2782 (Lactobacillus casei HY2782), Lactobacillus casei KCTC3109 (Lactobacillus casei KCTC3109) or Lactobacillus casei KCTC13086 (Lactobacillus casei KCTC13086)。
On the other hand, the present invention provides a kind of comprising lactobacillus casei bacterial strain, its fragment, its culture or the bacterium The extract of strain, fragment or culture is as effective component for inhibiting the feeding of the cytotoxicity as caused by particulate matter toxicity Feed composition.
In embodiment, the bacterial strain can be Lactobacillus casei HY2782 (Lactobacillus casei HY2782), Lactobacillus casei KCTC3109 (Lactobacillus casei KCTC3109) or Lactobacillus casei KCTC13086 (Lactobacillus casei KCTC13086)。
In embodiment, the fodder compound can improve the growth inhibition as caused by particulate matter toxicity.
In embodiment, the fodder compound can inhibit the Damage of Germ Cells as caused by particulate matter toxicity.
In embodiment, the fodder compound can improve the reproductive dysfunction as caused by particulate matter toxicity.
In embodiment, the fodder compound can improve the decline of the Motor execution ability as caused by particulate matter toxicity.
In embodiment, the fodder compound can have selected from ox, pig, sheep, horse, rabbit, dog, cat, chicken, turkey, duck, The raising purposes of one or more of tortoise, snake, fish, earthworm, spider, insect and nematode.
In embodiment, the nematode can be Caenorhabditis elegans.
Invention effect
The present invention provides the new applications of lactobacillus casei bacterial strain.
On the one hand, as effective component and malicious for particulate matter comprising lactobacillus casei bacterial strain the present invention provides one kind Property have protection or relaxation effect composition.
On the other hand, the present invention provides one kind as effective component and inhibits by particle comprising lactobacillus casei bacterial strain The composition of cellular damage caused by object toxicity.
On the other hand, the present invention provides one kind as effective component and enhances to particle comprising lactobacillus casei bacterial strain The composition of the repellence of object toxicity.
Detailed description of the invention
Fig. 1 is to confirmed Lactobacillus casei HY2782 bacterial strain used in the present invention for by hydrocarbon containing Ppolynuclear aromatic CCD-18Co human body cell caused by the particulate matter 1 of compound (polycyclic aromatic hydrocarbons, PAH) withers The chart of the protection effect died.
For CCD-18Co cell, individually put into particulate matter 1 (500 μ g/mL) or Lactobacillus casei HY2782 bacterial strain (1 × 106CFU/mL particulate matter 1 and Lactobacillus casei HY2782 bacterial strain), or are simultaneously put into, after culture 72 hours, measurement cell is raw Deposit rate.Statistical significance is shown together, compared with the individually group of investment particulate matter, statistical significance is P < 0.001 * * *.From In the chart, can clearly it confirm, lactobacillus casei bacterial strain imitates the protection of the human body cell apoptosis as caused by particulate matter 1 Fruit.
Fig. 2 is to confirmed that Lactobacillus casei HY2782 bacterial strain is for by the particulate matter containing heavy metal used in the present invention The chart of protection effect of CCD-18Co human body cell caused by 2.
For CCD-18Co cell, individually put into particulate matter 2 (500 μ g/mL) or Lactobacillus casei HY2782 bacterial strain (1 × 106CFU/mL particulate matter 2 and Lactobacillus casei HY2782 bacterial strain), or are simultaneously put into, after culture 72 hours, measurement cell is raw Deposit rate.Statistical significance is shown together, with individually investment particulate matter group compared with, statistical significance be * P < 0.05, * * * P < 0.001.From the chart, can clearly it confirm, lactobacillus casei bacterial strain is for the human body cell apoptosis as caused by particulate matter 2 Protect effect.
Fig. 3 is the Lactobacillus casei HY2782 bacterial strain used in the present invention for by containing Ppolynuclear aromatic hydrocarbonization Close CCD-18Co human body cell apoptosis caused by the particulate matter 1 of object (polycyclic aromatic hydrocarbons, PAH) Protection effect, it is aobvious by fluorescence after being dyed simultaneously using Hoechst33342 and propidium iodide (propidium iodide) The photo of micro mirror observation.
For CCD-18Co cell, individually put into particulate matter 1 (500 μ g/mL) or Lactobacillus casei HY2782 bacterial strain (1 × 106CFU/mL particulate matter 1 and Lactobacillus casei HY2782 bacterial strain), or is simultaneously put into utilize cell after culture 24 hours After Hoechst33342 (blue) and the dyeing of propidium iodide (red) fluorescent reagent, using fluorescence microscope from CCD- The fluorescence detected in 18Co.Hoechst33342 has the characteristic dyed for all cells, and propidium iodide has The characteristic that only cell that cell membrane is destroyed is dyed.Therefore, when being dyed simultaneously with two kinds of fluorescent reagents, survival Normal cell only by Hoechst33342 reagent dyeing, it is blue, and the cell that cell membrane is destroyed simultaneously quilt Hoechst33342 reagent and propidium iodide reagent dyeing show pink colour fluorescence.Experimental result, when individually investment particulate matter 1, It is very high by the cell (PI-positive cells) of propidium iodide stain, on the contrary, putting into Lactobacillus casei HY2782 bacterial strain simultaneously When with particulate matter 1, significantly reduced by the cell of propidium iodide stain.It is possible thereby to clearly confirm, Lactobacillus casei for by Protection effect of human body cell apoptosis caused by grain object.
Fig. 4 is to utilize the fluorescence microscope result of Fig. 3 to be dyed by propidium iodide (propidium iodide, PI) Ratio of the cell in the CCD-18Co cell being all colored carry out the result of quantificational expression.Statistical significance is shown together Out, compared with the control group, statistical significance is * P < 0.05, P < 0.001 * * *, by the group of independent investment particulate matter 1 with throw simultaneously When entering the group of particulate matter 1 and Lactobacillus casei HY2782 bacterial strain and making comparisons,###P<0.001.From the chart, it can define true Recognize, the protection effect of lactobacillus casei bacterial strain for the human body cell apoptosis as caused by particulate matter.
Fig. 5 is to confirmed that Lactobacillus casei HY2782 bacterial strain is for particulate matter 1 used in the present invention by zoopery Use Caenorhabditis elegans (Caenorhabditis elegans) as mould for the experiment with the chart of 2 protection effect Type animal.
For Caenorhabditis elegans, Escherichia coli OP50 (E.coli OP50) or the present invention of the feeding as conventional bait Lactobacillus casei HY2782 bacterial strain four days.At this point, the toxicity in order to confirm particulate matter 1 and 2, the further feeding particle of difference Object.In order to confirm influence that particulate matter generates the growth of animal pattern, the height of Caenorhabditis elegans is measured after four days, And measure every group 50 heights.Statistical significance is shown together, compared with the only control group of feeding Escherichia coli, system Meter conspicuousness is P < 0.001 * * *.From the chart, can clearly it confirm, lactobacillus casei bacterial strain by particulate matter 1 and 2 for being drawn The growth inhibiting protection effect risen.
Fig. 6 is to confirmed that Lactobacillus casei HY2782 bacterial strain is for particulate matter 1 used in the present invention by zoopery The Caenorhabditis elegans as animal pattern has been investigated also, for the experiment with the chart of 2 protection effect The fecundity of (Caenorhabditis elegans).
For Caenorhabditis elegans, Escherichia coli OP50 (E.coli OP50) or the present invention of the feeding as conventional bait Lactobacillus casei HY2782 bacterial strain four days.At this point, the toxicity in order to confirm particulate matter 1 and 2, the further feeding particle of difference Object.In order to verify the influence that particulate matter generates the reproductive function of animal pattern, every group of oviposition sum is observed.Experimental result, really Particulate matter toxicity of accepting has an impact reproductive function, and the particulate matter toxicity for influencing reproductive function, Lactobacillus casei Bacterial strain has protection effect.Using every group of 30 Caenorhabditis elegans as object, oviposition sum is measured.Together by statistical significance It shows, compared with the only control group of feeding Escherichia coli, compared with the only control group of feeding Escherichia coli, statistical significance is * * P<0.01.From the chart, can clearly it confirm, lactobacillus casei bacterial strain presses down the reproductive function as caused by particulate matter 1 and 2 The protection effect of system.
It is that confirmed that Lactobacillus casei HY2782 bacterial strain is for particulate matter used in the present invention by zoopery in Fig. 7 The chart of 1 protection effect, also, in order to investigate the Caenorhabditis elegans as animal pattern in the experiment The locomitivity of (Caenorhabditis elegans).
For Caenorhabditis elegans, Escherichia coli OP50 (E.coli OP50) or the present invention of the feeding as common bait Lactobacillus casei HY2782 bacterial strain.At this point, the toxicity in order to confirm particulate matter 1, the further feeding particulate matter 1 of difference.In order to The influence that confirmation particulate matter generates the locomitivity of animal pattern measures it using every group of 15 Caenorhabditis elegans as object Body number of bends in 20 seconds.In group of the feeding as the Escherichia coli OP50 of conventional bait, particulate matter drops significantly The low locomitivity of Caenorhabditis elegans, and in the group of feeding Lactobacillus casei HY2782 bacterial strain, do not occur this movement The case where ability reduces.Statistical significance is shown together, compared with the only control group of feeding Escherichia coli, statistical significance For P < 0.001 * * *.From the chart, can clearly it confirm, lactobacillus casei bacterial strain is for the locomitivity as caused by particulate matter Protection effect of inhibition.
Be in Fig. 8 confirmed according to the present invention used in lactobacillus casei bacterial strain type and for by particulate matter toxicity Protection effect that caused reproductive function weakens, and be by the chart of the data drawing list of following table 4.
In the Caenorhabditis elegans of feeding Escherichia coli OP50 (E.coli OP50), with independent investment Escherichia coli The group of OP50 is compared, while the oviposition sum put into the group of Escherichia coli OP50 and particulate matter 2 substantially reduces, show by Genotoxicity caused by grain object 2.Equally, it in the Caenorhabditis elegans of feeding Lactobacillus gasseri KCTC3163 bacterial strain, also shows The genotoxicity as caused by particulate matter 2 out.On the contrary, feeding Lactobacillus casei HY2782 bacterial strain and Lactobacillus casei at the same time In the Caenorhabditis elegans of KCTC3109 bacterial strain, there is not the genotoxicity as caused by particulate matter 2.Together by statistical significance It shows, compared with the only control group of feeding Escherichia coli OP50, statistical significance is P < 0.01 * * or P < 0.001 * * *.Work as investment When various microorganisms, the difference of the group for individually putting into microorganism and the group for putting into microorganism and particulate matter simultaneously for #P < 0.05 or##P< 0.01, also, when there is no statistical significance, it is indicated with n.s..It from the chart, can clearly confirm, be used in the present invention Lactobacillus casei bacterial strain effect is protected with weakening for the reproductive function as caused by particulate matter toxicity, and confirm this effect Difference is shown according to the type of bacterial strain.
Specific embodiment
Hereinafter, will be explained in the present invention.
The present invention is provided to prepare the lactobacillus casei bacterial strain of particulate matter toxicity alleviation composition, its fragment, its Culture, the extract of the bacterial strain, the extract of the fragment or the culture extract purposes.
In embodiment, the particulate matter may include Ppolynuclear aromatic hydrocarbon (polycyclic ) or heavy metal hydrocarbons.
In embodiment, the particulate matter can be PM10.The PM10 refers to 10 microns of particulate matters below of partial size.
Lactobacillus casei bacterial strain is the representative lactic acid bacteria for producing fermented dairy product, so far, it is known that cheese cream Bacillus improves intestinal flora, increases intestinal beneficial bacterium, reduce harmful intestinal tract bacteria, improve bowel movement function, strengthen immunity, reduced Quick and asthma symptoms etc., but alleviate effect without the particulate matter toxicity of open Lactobacillus casei.
Activity needed for " effective component " refers to independent show in this specification can be with itself inactive load Body etc. shows required active ingredient together.
On the one hand, there is the composition of this disclosure particulate matter toxicity remission effect, particulate matter toxicity to protect The repellence of effect, the effect for inhibiting the cellular damage as caused by particulate matter toxicity and enhancing to particulate matter toxicity.
In embodiment, the composition can prevent or treat the growth inhibition as caused by particulate matter toxicity.
In embodiment, the composition can inhibit the Damage of Germ Cells as caused by particulate matter toxicity, or prevention Or treatment reproductive function as caused by harmful substance toxicity weakens.
In embodiment, the composition can prevent or to treat the Motor execution ability as caused by particulate matter toxicity low Under.Motor execution ability refers to the ability moved using muscle.
Bacterial strain in this specification be through the invention belonging to technical field in well known conventional physics, chemistry it is prominent Change method etc. improves or is modified to the activity being identical with this or than its better activity.
In embodiment, the lactobacillus casei bacterial strain can be Lactobacillus casei HY2782 (Lactobacillus Casei HY2782) bacterial strain.
In embodiment, the Lactobacillus casei HY2782 can be according in Korean Patent No spy 1997-0009934B1 Method disclosed in (day for announcing: on June 19th, 1997, application number: KR 10-1993-0026829) obtains.Above-mentioned document is as ginseng Document is examined, full text is contained in the application.
In another embodiment, the Lactobacillus casei HY2782 is as the bacterium being widely used in fermented dairy product production Strain, can be easily separated and use from the commercially available fermented dairy product beverage containing Lactobacillus casei HY2782.The institute used It states fermented dairy product beverage and is free of other bacterial strains other than Lactobacillus casei HY2782.One bottle of chlorella yakult is bought on the market (name of product: chlorella yakult, manufacturer: chlorella yakult Co., Ltd, South Korea).In above-mentioned one bottle of chlorella yakult (Yakult) containing 2 × 108The Lactobacillus casei HY2782 bacterial strain of CFU or more.Using the salt water by sterilizing, chlorella yakult beverage is diluted to 105~106 After times, 1mL dilution is applied on MRS agar (agar) plating medium.Then, it is cultivated under 37 DEG C, aerobic condition At 48 hours, the bacterium colony of Lactobacillus casei HY2782 bacterial strain is formed.By a colony inoculation being formed by bacterium colony in MRS liquid It after on body culture medium, is cultivated 24 hours at 37 DEG C, obtains the culture solution containing Lactobacillus casei HY2782.In general, manufacturing When beverage, by sterilization treatment process, so that it is free of other bacterial strains, it is therefore, aobvious and easy to those skilled in the art See, the bacterium colony that the fermented dairy product beverage culture containing Lactobacillus casei HY2782 is formed is exactly cheese cream bar Bacterium HY2782.Following table 1 is MRS agar, indicates that the composition of the MRS solid medium of 1L, table 2 are MRS meat soup, indicates 1L's The composition of MRS fluid nutrient medium.
Table 1
Peptone (Peptone) 10.0g
Beef extract (Beef extract) 10.0g
Yeast extract (Yeast extract) 5.0g
Glucose (Dextrose) 20.0g
Polysorbate80 (Polysorbate 80) 1.0g
Ammonium citrate (Ammonium citrate) 2.0g
Sodium acetate (Sodium acetate) 5.0g
Magnesium sulfate (Magnesium sulfate) 0.1g
Manganese sulfate (Manganese sulfate) 0.05g
Dipotassium hydrogen phosphate (Dipotassium phosphate) 2.0g
Agar (Agar) 15.0g
Table 2
Peptone (Peptone) 10.0g
Beef extract (Beef extract) 10.0g
Yeast extract (Yeast extract) 5.0g
Glucose (Dextrose) 20.0g
Polysorbate80 (Polysorbate 80) 1.0g
Ammonium citrate (Ammonium citrate) 2.0g
Sodium acetate (Sodium acetate) 5.0g
Magnesium sulfate (Magnesium sulfate) 0.1g
Manganese sulfate (Manganese sulfate) 0.05g
Dipotassium hydrogen phosphate (Dipotassium phosphate) 2.0g
In embodiment, the lactobacillus casei bacterial strain can be selected from Lactobacillus casei HY2782, Lactobacillus casei One or more of KCTC3109 and Lactobacillus casei KCTC13086.
In embodiment, the bacterial strain is by strain culturing and after culture solution is centrifuged, with by sterilizing Physiological saline washed after, be scattered in solvent, such as sterile milk, be lyophilized, in the form of freeze-dried powder manufacture and use.
The fragment can refer to bacterial strain self by chemically or physically strength it is broken obtained by product.
The culture can refer to that will cultivate some or all substances contained in the culture medium of bacterial strain is included Substance, but regardless of being liquid or solid form, for example, can refer to comprising as strain culturing product metabolin or secretion The substance of object or its fragment, bacterial strain itself also may include in culture.In addition, the culture can be culture solution.
The extract can refer to the fragment of bacterial strain itself, bacterial strain, the culture of bacterial strain or their mixture Product obtained from extraction is included in but regardless of extracting method, the form of Extraction solvent, the ingredient being extracted or extract After extraction can the substance as obtained from other methods processing or the processing such as being fractionated, be concentrated extensive concept.
In embodiment, the composition can be pharmaceutical compositions.
The pharmaceutical compositions can also further contain preservative, stabilizer, wettable powder other than effective component The pharmacies adjuvant such as agent or emulsification promoter, osmotic pressure adjusting salt and/or buffer and other be beneficial to treatment object Various oral preparations or non-oral formulation is made according to conventional methods in matter.
The oral preparation includes such as tablet, pill, hard capsule, soft capsule, liquor, suspending agent, emulsifier, syrup Agent, pulvis, powder, granula subtilis, granule, micropill preparation etc., these dosage forms, can also be living containing surface other than effective component Property agent, diluent (example: lactose, glucose, sucrose, mannitol, D-sorbite, cellulose, glycine), lubricant (example: dioxy SiClx, talcum, stearic acid, magnesium stearate, calcium stearate and polyethylene glycol).Tablet can also contain such as aluminium-magnesium silicate, starch The adhesive of paste, gel, bassora gum, methylcellulose, sodium carboxymethylcellulose and polyvinyl chloride pyrrolidones etc., according to feelings Condition can contain starch, agar, alginic acid or disintegrating agent, absorbent, colorant, flavouring agent and the sweetener of its sodium salt etc. Pharmacy additive.This tablet can be made up of conventional mixing, granulation or coating method etc..
In addition, the form of the non-oral formulation can be Percutaneously administrable preparation, for example, it may be injection, drops, soft Cream, lotion, gel, creams (cream), spray, suspending agent, emulsion, suppository, patch etc., but not limited to this.
The determination of the dosage of the effective component is in the level of those skilled in the art, although the daily dosage meeting of drug Change with many factors such as the occurring degree of administration object, disease time, age, health status, complication, but at It is artificial quasi-, on the one hand can be administered with 1 μ g/kg to 2000mg/kg, and on the other hand can with 50 μ g/kg to 500mg/kg, often Bu 1 to 3 time administration, the dosage do not limit the scope of the invention in any way.
The pharmaceutical compositions can be a kind of skin preparations for extenal use, and the skin preparations for extenal use refers to may include any smearing The general name of product outside skin, wherein containing the pharmaceuticals of various dosage forms.
In embodiment, the composition can be food compositions.
The food compositions can be liquid or solid dosage form, such as, it may include various foodstuffs, beverage, mouth are fragrant Sugar, tea, compound vitamin, healthy accesary foods class etc., can be with powder, particle, tablet, capsule, acidified milk or beverage etc. Form uses.The food compositions of various dosage forms, can also be by those skilled in the art according to dosage form other than effective component Or purpose is used, without difficulty it is suitable for selection and the ingredient that mixed phase answers field usually used, makes simultaneously when with other raw materials Used time, it may occur that synergistic effect.
For the liquid component that may contain other than the effective component of this disclosure, it is not particularly limited, As common beverage, multiple scents or natural carbohydrate etc. may include as supplementary element.As the day The example of right carbohydrate has the polysaccharide, all of disaccharides, maltose, sucrose of monosaccharide, glucose, fructose etc. etc. Common sugar alcohols such as carbohydrate and xylitol, D-sorbite, erythritol of such as dextrin, cyclodextrin etc..As the fragrance Agent, it may be advantageous to which (thaumatin (Thaumatin), stevia rebaudianum (stevia) extract are (for example, sweet tea using natural flavours Chrysanthemum disaccharide glucoside A, glycyrrhizin etc.) and synthesis flavouring agent (such as saccharin, Aspartame etc.).The ratio of the natural carbohydrate Example, which can be, is usually about 1 to 20 gram in the composition of every 100 milliliters of this disclosure, and on the other hand about 5 to 12 grams.
On the one hand, the food compositions may include a variety of nutritional agents, vitamin, mineral (electrolyte), such as resultant wind Flavouring agent, colorant and the flavoring agent (cheese, chocolate etc.) of taste agent and natural flavour mountaineous dose etc., pectic acid and its salt, alginic acid and Its salt, organic acid, protective colloid thickener, pH adjusting agent, stabilizer, preservative, glycerol, alcohol, the carbon for soda Acidulant etc..On the other hand, it may include for making fruit juice and vegetable beverage pulp.It can mentioned component is independent or group It closes and uses.The ratio of above-mentioned additive can be various, but is generally selected from the group of this disclosure relative to every 100 parts by weight In the range of 0.001 to about 20 parts by weight for closing object.
In embodiment, the composition can be cosmetic composition.
The cosmetic composition can also further include functional additive and general other than effective component Ingredient contained in cosmetic composition.The functional additive may include selected from water soluble vitamin, fat-soluble dimension Ingredient in raw element, macromolecule peptide, macromolecule polysaccharide, sphingolipid and marine algae extract.Match synthesis as in addition to this included Point, can there are lubricant component, moisturizer, emollient, surfactant, organic/inorganic pigment, organic powder, sun-screening agent, anti-corrosion Agent, antioxidant, plant extracts, pH adjusting agent, alcohol, pigment, fragrance, blood circulation accelerant, coolant, stops fungicide Sweat agent, Purified Water etc..
The dosage form of the cosmetic composition is not particularly limited, can suitably be selected by purpose.For example, can be made into choosing From powder, surfactant, toner, lotion, lotion, milky lotion, moisturizing emulsion, nutritional emulsions, massage cream, nourishing cream, protect Wet frost, hand lotion, foundation emulsion, essence dew, nutrition essence, facial mask, shampoo, perfumed soap, cleansing cream, facial cleanser, wash one's face frost, body One or more of body cream, bath oil dosage form, but not limited to this.
When dosage form of the invention is cream, frost or when gel, as carrier components, can be used animal origin, plant fiber, Wax, paraffin, starch, bassora gum, cellulose derivative, polyvinyl alcohol, organosilicon (silicone), bentonite, silica, cunning Stone or zinc oxide etc..
When dosage form of the invention is pulvis or spray, as carrier components, lactose, talcum, silica, hydrogen can be used Aluminium oxide, calcium silicates or polyamide powder especially when for spray, can further include such as chlorofluorocarbons, propane/fourth The promotor of alkane or dimethyl ether.
When dosage form of the invention is solution or emulsion, as carrier components, solvent, resolvating agent or emulsifying are used Agent, it may for example comprise water, ethyl alcohol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, Ergol, propylene glycol, 1,3- fourth Base glycol oily (1,3-butylglycoloil), glycerol aliphatic (acid) ester, polyethylene glycol or D-sorbite aliphatic ester.
When dosage form of the invention is suspension, as carrier components, such as water, ethyl alcohol or the liquid of propylene glycol can be used State diluent, such as ethoxylated isostearyl alcohol (Ethoxylated isostearyl alcohol), polyoxyethylene sorbitol Ester (polyoxyethylene sorbitol ester) and polyoxyethylene sorbitan ester (Polyoxyethylene Sorbitan ester) suspending agent, microcrystalline cellulose, inclined aluminium hydroxide (Aluminum methahydroxide), swelling Soil, agar or bassora gum etc..
It may include fatty alcohol as carrier components when dosage form of the invention is the detergent containing surfactant Sulfate, fatty alcohol sulphuric acid ether, sulfosuccinic acid monoesters, isethionate, imidazolidine derivatives, methyl tauride, flesh Propylhomoserin salt, fatty acid amide ether sulfate, alkyl amido betaine, aliphatic alcohol, fatty glyceride, fatty acid diethanol acyl Amine, vegetable oil, lanolin derivative or oxygroup fatty acid glyceride etc..
When dosage form of the invention is soap, as carrier components, alkali metal salt, the fatty acid half of fatty acid can be used Ester, fatty acid protein matter hydrolysate, isethionate, lanolin derivative, aliphatic alcohol, vegetable oil, glycerol, sugar etc..
In embodiment, the composition can be fodder compound.
In embodiment, the composition can be livestock feed composition.
In embodiment, the composition can have selected from ox, pig, sheep, horse, rabbit, dog, cat, chicken, turkey, duck, tortoise, The raising purposes of one or more of the fish such as snake, flatfish, sea eel, earthworm, spider, insect and nematode.
In embodiment, the nematode can be Caenorhabditis elegans.
Hereinafter, by present invention will be further described in detail through examples.It will be apparent to those skilled in the art that , these embodiments are merely to illustrate the present invention, and the scope of the present invention is not limited to this this embodiment.
Embodiment 1: the acquisition of particulate matter
There are two types of the particulate matters used in the present specification, buys and uses from SIGMA-ALDRICH.Two kinds of particles object is equal For PM10 (particulate matter 10, partial size are the particulate matter of 10 microns), particulate matter 1 is characterized in largely containing There is polycyclic aromatic hydrocarbons (polycyclic aromatic hydrocarbons, PAHs), particulate matter 2 is characterized in A large amount of heavy metals for containing such as arsenic, cadmium, nickel, lead.
Embodiment 2: the acquisition of lactobacillus casei bacterial strain
The bacterial strain used in the present embodiment is well known Lactobacillus casei HY2782 (Lactobacillus casei HY2782) bacterial strain, the bacterial strain powder are obtained and are used from chlorella yakult Ping Ze probiotics factory, manufacturer (strain) South Korea.It is described Lactobacillus casei HY2782 lactic acid bacteria powder (credit number: 20140017107, product manufacturing number: 201400171077, approval Date: on July 19th, 2016), be a kind of free from extraneous odour, odorless with solid dulcet pale yellow powder character.It is described dry Lactobacillus paracasei HY2782 bacterial strain in preservation on December 19 in 2017 to South Korea, preservation mechanism Culture Collection (KCTC), and Obtain deposit number KCTC 13438BP.
In addition, as the known bacterial strain that those skilled in the art are easy to get, from South Korea's Culture Collection (KCTC) buy commercially available Lactobacillus casei KCTC3109 (is with the Lactobacillus casei ATCC393 being easy to get from U.S. ATCC Identical bacterial strain), Lactobacillus casei KCTC13086, lactobacillus paracasei KCTC3189, Lactobacillus gasseri KCTC3163 (with from The Lactobacillus gasseri ATCC33323 that U.S. ATCC is easy to get is identical bacterial strain) bacterial strain and be used for following experiments.
In following experimental examples, by evaluation Lactobacillus casei in goldbeater's skin cell (in vitro) and Caenorhabditis elegans (in body) mould Particulate matter in type protects effect.Based on this, intend providing have effects that the composition alleviated for particulate matter toxicity.
Experimental example 1: the human body cell apoptosis effect as caused by particulate matter, and the Apoptosis based on lactobacterium casei strains Protection effect
In this experimental example, it is thus identified that the human body cell apoptosis effect as caused by particulate matter, and it is based on Lactobacillus casei The Apoptosis protection effect of strain.For this purpose, having used two kinds of particles object, particulate matter 1 is largely to contain Ppolynuclear aromatic hydrocarbonization The particulate matter of object is closed, particulate matter 2 is particulate matter largely containing heavy metal.
CCD-18Co cell as human normal colorectal cell strain is bought and is made from bank, Korea Cell system (Soul) With.CCD-18Co cell utilize added with 10% fetal calf serum (heat-inactivated fetal bovine serum, FBS), RPMI culture medium (the Roswell Park Memorial Institute of 10U/mL penicillin, 100 μ g/mL streptomysins Medium), cultivate and keep under the conditions of 5% carbon dioxide and 37 DEG C.
CCD-18Co cell is assigned in 48 orifice plates, so that every hole reaches 2 × 104A cell is cultivated 24 hours, and is made It adheres to.Then, particulate matter 1 (500 μ g/mL), particulate matter 2 (500 μ g/mL) or Lactobacillus casei HY2782 bacterial strain are individually put into (1×106CFU/mL particulate matter and Lactobacillus casei HY2782 bacterial strain), or are simultaneously put into, is cultivated 72 hours.Culture terminates Afterwards, cells survival rate is measured using Ez-Cytox (Daeil Lab Science Co., Ltd., South Korea) kit (Kit). After Ez-Cytox solution is added in the culture medium of 10 times of dilution, cultivate 1 hour.Then, it is gone completely by centrifuge separation After particulate matter, the absorbance under 450nm is only measured with supernatant.Cells survival rate is directly proportional to absorbance, and sets The cells survival rate of control group is 100%, the cells survival rate of experiment with computing group.
Experimental result, as the individually investment largely particulate matter 1 containing polycyclic aromatic hydrocarbons (500 μ g/mL), The survival rate of CCD-18Co cell significantly reduces.When put into simultaneously particulate matter 1 and Lactobacillus casei HY2782 bacterial strain (1 × 106When CFU/mL), it is able to confirm that with only compared with the experimental group that particulate matter 1 is individually handled, cells survival rate significantly restores (figure 1).Similarly, largely the particulate matter 2 containing heavy metal also reduces cells survival rate, by putting into particulate matter 2 and cheese cream simultaneously Bacillus HY2782 bacterial strain inhibits Apoptosis (Fig. 2).In conclusion lactobacterium casei strains are for by containing Ppolynuclear aromatic carbon Human body cell apoptosis caused by the particulate matter of hydrogen compound or heavy metal has remission effect.
Experimental example 2: reaffirmed by fluorescence microscope the human body cell apoptosis effect as caused by particulate matter with And the Apoptosis protection effect based on lactobacterium casei strains
In this experimental example, the degree of injury that cell membrane is observed by fluorescence microscope, has been reaffirmed by particulate matter Caused human body cell apoptosis effect and Apoptosis protection effect based on lactobacterium casei strains.For this purpose, being contaminated with DNA fluorescence Toner Hoechst33342 (blue-fluorescence) and propidium iodide (red fluorescence) while staining cell, and utilize fluorescence microscope It is observed.Hoechst33342 reagent has a permeability of cell membrane, and propidium iodide is without membrane permeability, therefore living cells quilt Hoechst33342 reagent dyeing, blue, in contrast, the cell of damaged membrane is then simultaneously by Hoechst33342 and iodine Change the third pyridine reagent dyeing, shows pink colour fluorescence.
CCD-18Co cell as human normal colorectal cell strain is bought and is made from bank, Korea Cell system (Soul) With.CCD-18Co cell utilize added with 10% fetal calf serum (heat-inactivated fetal bovine serum, FBS), RPMI culture medium (the Roswell Park Memorial Institute of 10U/mL penicillin, 100 μ g/mL streptomysins Medium), cultivate and keep under the conditions of 5% carbon dioxide and 37 DEG C.
CCD-18Co cell is assigned in 6 orifice plates, so that every hole reaches 3 × 105After a cell, cultivate 24 hours, And make its attachment.Then, particulate matter 1 (500 μ g/mL) or Lactobacillus casei HY2782 bacterial strain (1 × 10 are individually put into6CFU/ ML particulate matter 1 and Lactobacillus casei HY2782 bacterial strain), or are simultaneously put into, is cultivated 24 hours.Then, Dulbecco phosphoric acid is utilized Salt buffer salt water (Dulbecco's phosphate-buffered saline) solution wash cell twice, be added to containing After in the culture medium of Hoechst33342 (2ng/mL) and propidium iodide (20 μ g/mL) reagent, contaminated under 37 DEG C of dark conditions Color 15 minutes.After recycling dulbecco phosphate buffered saline solution to wash 1 time in the cell after dyeing, in room temperature dark Under the conditions of dry two hours.Then, fluorescent microscopy images are shot using fluorescence microscope (Nikon, TE-2000).In order to Quantitative comparison cell membrane damage degree, each experimental group arbitrarily shoot three sentence after, for the cell dyed by propidium iodide (PI) Ratio (%) in the cell being all colored is calculated and compared.
Experimental result, in control group (control) and only with Lactobacillus casei HY2782 bacterial strain (1 × 106CFU/mL) single Whole fluorescence only blue in only processed cell, it follows that absolutely not there is cell membrane damage.When independent investment is big When measuring particulate matter 1 (the 500 μ g/mL) containing polycyclic aromatic hydrocarbons, it increased significantly in the cell of pink colour fluorescence, thus The cell membrane of confirmation CCD-18Co cell is seriously destroyed, and damaged membrane (Fig. 3, Fig. 4) occurs due to particulate matter 1. Particulate matter 1 and Lactobacillus casei HY2782 bacterial strain (1 × 10 are put into when simultaneously6When CFU/mL), the ratio of pink colour cell compares particle The independent processing group of object 1 substantially reduces.Therefore, reaffirmed lactobacillus casei bacterial strain for the human body as caused by particulate matter toxicity The protection effect of Apoptosis is extremely significant.
Experimental example 3: the animal pattern Caenorhabditis elegans growth inhibitory effect as caused by particulate matter and Lactobacillus casei bacterium Recovery efficiency of the strain to particulate matter toxicity
In order to confirm the toxicity of particulate matter and based on the recovery effects of lactobacillus casei bacterial strain, Caenorhabditis elegans is used as Animal model.Caenorhabditis elegans used from U.S. nematode heredity center (Caenorhabditis Genetics Center, CGC) the wild type N2 bacterial strain (strain) bought.As bait, feeding Escherichia coli OP50 is kept for 20 DEG C, and is tested. Escherichia coli OP50 is also to buy and use from U.S. nematode heredity center.
It is dynamic that Caenorhabditis elegans (Caenorhabditis elegans) is commonly used for model in biology, chemical research Object.Caenorhabditis elegans is tested instead of rodent, advantageous in terms of zooscopy ethics.Pharmacy and functional food are combined Object, the active in body (in vivo) of individual level can be assessed by being further developed into from existing isolated experiment (in vitro) And effect.In addition, recently, in more famous research papers, this Caenorhabditis elegans to be widely used in the function of lactic acid bacteria In the excavation of energy property and the research of mechanism of action.
The ovum of Caenorhabditis elegans is only collected, and is tested for toxicity assessment.By Escherichia coli OP50 and cheese cream bar Bacterium HY2782 bacterial strain is coated in respectively on NGM (nematode growth medium) agar medium, in order to evaluate toxicity, with The concentration of 1mg/mL is added to particulate matter 1 and particulate matter 2.In the particulate object and Escherichia coli or cheese cream bar so prepared The ovum of Caenorhabditis elegans is spread on the culture medium of bacterium HY2782 bacterial strain, is cultivated 3-4 days.Then, stereoscopic microscope observing show is utilized The form of beautiful hidden rhabditida, and microscope photo is shot, and utilize Image J program (National Health Institute, the U.S.) measure the length of Caenorhabditis elegans.According to each experimental group, the length of 25-50 nematode is measured respectively Degree.
Experimental result, compared with as the Escherichia coli OP50 of the conventional bait of Caenorhabditis elegans, Lactobacillus casei bacterium Strain does not impact the growth of the Caenorhabditis elegans as animal pattern.That is, it is thus identified that as the dry of bait Lactobacillus paracasei bacterial strain has no too many differences compared with as the Escherichia coli of conventional bait.But it is big when being put into Escherichia coli When measuring the particulate matter 1 containing polycyclic aromatic hydrocarbons or the particulate matter 2 largely containing heavy metal, as animal pattern The growth of Caenorhabditis elegans substantially reduces (Fig. 5).Thus, it is possible to confirm that particulate matter induces toxicity in individual level.On the contrary, When putting into particulate matter and Lactobacillus casei HY2782 bacterial strain simultaneously, the growth absolutely not toxicity to Caenorhabditis elegans is confirmed It influences, thereby confirms that lactobacillus casei bacterial strain can alleviate the embryotoxicity (Fig. 5) of the individual level induced by particulate matter.It is this Effect is unrelated with the type of particulate matter, can all be confirmed in particulate matter 1 and particulate matter 2.
Experimental example 4: the animal pattern Caenorhabditis elegans reproductive function inhibitory effect as caused by particulate matter and cheese cream bar Recovery effects of the bacteria strain to particulate matter toxicity
In this experimental example, in order to verify the toxicity by particulate matter and recovery effect based on Lactobacillus casei again, no The size of Caenorhabditis elegans is only observed, and observes oviposition sum, thus the influence that observation generates fecundity.
The ovum of Caenorhabditis elegans is only collected, and is tested for toxicity assessment.By Escherichia coli OP50 and cheese cream bar Bacterium HY2782 bacterial strain is coated in respectively on NGM (nematode growth medium) agar medium, in order to evaluate toxicity, with The concentration of 1mg/mL is added to particulate matter 1 and particulate matter 2.In the particulate object and Escherichia coli or cheese cream bar so prepared The ovum of Caenorhabditis elegans is spread on the culture medium of bacterium HY2782 bacterial strain, is cultivated 64 hours.Then, daily by the beautiful hidden bar line of each group Worm moves on to the culture medium for being coated with Escherichia coli OP50, and observes oviposition quantity daily.Oviposition quantity is calculated divided by 30 daily 5 days in total oviposition quantity is all added the oviposition sum as every animal pattern by every oviposition quantity.
Experimental result, compared with as the Escherichia coli OP50 of Caenorhabditis elegans routine bait, lactobacillus casei bacterial strain Significant impact is not caused to the reproduction of the Caenorhabditis elegans as animal pattern.That is, it is thus identified that as bait When feeding lactobacillus casei bacterial strain, compared with as the Escherichia coli of conventional bait, too many differences are had no in terms of fecundity. But when to Escherichia coli investment largely particulate matter 1 (1mg/mL) containing polycyclic aromatic hydrocarbons, as model The fecundity of the Caenorhabditis elegans of animal declines, also, works as the investment largely particulate matter 2 (1mg/mL) containing heavy metal When, the fecundity of the Caenorhabditis elegans as animal pattern is remarkably decreased (Fig. 6).Thus, it is possible to confirm particulate matter in individual Toxicity is induced in level.And, it is thus identified that when putting into particulate matter and Lactobacillus casei HY2782 bacterial strain simultaneously, to beautiful hidden bar The fecundity of nematode absolutely not toxic effect thereby confirms that lactobacillus casei bacterial strain can alleviate induced by particulate matter The genotoxicity (Fig. 6) of body level.This effect is unrelated with the type of particulate matter, can be in particulate matter 1 and particulate matter 2 all It is confirmed.
Experimental example 5: the animal pattern Caenorhabditis elegans locomitivity inhibitory effect as caused by particulate matter and cheese cream bar The recovery effect of bacteria strain to particulate matter toxicity
In this experimental example, in order to verify the toxicity as caused by particulate matter again and the recovery function based on Lactobacillus casei Effect, not only observes the size and oviposition sum of Caenorhabditis elegans, but also observes the movement of Caenorhabditis elegans, demonstrates The influence that locomitivity is generated.
The ovum of Caenorhabditis elegans is only collected, and is tested for toxicity assessment.By Escherichia coli OP50 and cheese cream bar Bacterium HY2782 bacterial strain is coated in respectively on NGM (nematode growth medium) agar medium, in order to evaluate toxicity, with The concentration of 1mg/mL is added to the largely particulate matter 1 containing polycyclic aromatic hydrocarbons.In the particulate so prepared After spreading the ovum of Caenorhabditis elegans on the culture medium of object and Escherichia coli or Lactobacillus casei HY2782 bacterial strain, cultivate 96 hours. Then, each group Caenorhabditis elegans is transplanted to respectively on the glass slide of physiological buffered saline.Then with stereoscope and Microscope camera (Jenopitk, Progress Gryphax, Germany) has taken video.Caenorhabditis elegans has been measured 20 The number of body bending (body bending) in second, and every group of 15 nematodes are investigated.
Experimental result, under conditions of putting into the Escherichia coli OP50 as Caenorhabditis elegans routine bait, particulate matter 1 Considerably reduce the body number of bends of Caenorhabditis elegans.In contrast, when independent investment Lactobacillus casei HY2782 bacterial strain Or when putting into Lactobacillus casei HY2782 bacterial strain and particulate matter 1 simultaneously, compared with the control group, body number of bends absolutely not subtracts Few (Fig. 7).It thereby confirms that by absorbing Lactobacillus casei, the decline of the locomitivity as caused by particulate matter can be restored significantly.
Experimental example 6: the type of lactobacillus casei bacterial strain compares recovery effect of particulate matter toxicity
In this experimental example, in order to which more different types of Lactobacillus casei alleviates effect to particulate matter toxicity, according to institute Method documented by experimental example 3 and 4 is stated, various microorganisms is tested respectively and the growth of Caenorhabditis elegans and fecundity is produced Raw influence.
Experimental result, as shown in table 3, when putting into Escherichia coli, the growth inhibition ratio as caused by particulate matter is 90.5%, And the growth inhibition ratio of Lactobacillus casei HY2782, KCTC13086 are respectively 100.3% and 96.7%, confirm Lactobacillus casei The growth inhibition toxicity as caused by particulate matter is effectively relieved in bacterial strain.On the contrary, lactobacillus paracasei KCTC3189 and Lactobacillus gasseri The growth inhibition ratio of KCTC3163 is respectively 82.3% and 89.1%, it is thus identified that the bacterial strain does not have particulate matter toxicity to alleviate function Effect.Therefore, even being able to confirm that the bacterial strain for belonging to identical lactobacillus (genus), when kind of (species) is different, root According to the type of bacterial strain, it is also different that particulate matter toxicity alleviates effect.In conclusion being able to confirm that lactobacillus casei bacterial strain effectively delays The toxicity as caused by particulate matter is solved.
In addition, when putting into Escherichia coli, the reproduction inhibiting rate as caused by particulate matter is as shown in table 4 and Fig. 8 61.3%, and the reproduction inhibiting rate of Lactobacillus casei HY2782, Lactobacillus casei KCTC3109 bacterial strain are respectively 82.2% and 82.7%, confirmation Lactobacillus casei is effectively relieved the reproduction as caused by particulate matter and inhibits toxicity, on the contrary, Lactobacillus gasseri The reproduction inhibiting rate of KCTC3163 is 66.7%, confirms that no particulate matter toxicity alleviates effect.Therefore, even being able to confirm that category In the bacterial strain of identical lactobacillus (genus), when kind of (species) is different, according to the type of bacterial strain, particulate matter toxicity is slow It is also different to solve effect.In conclusion being able to confirm that lactobacillus casei bacterial strain effectively alleviates the toxicity as caused by particulate matter.
Table 3
Table 4
The preparation example of the composition related in one aspect to of this specification is illustrated below, it is various but it is also suitable for other Preparation, these preparation examples are not intended to limit the present invention, are only used for illustrating.
" preparation example 1 " tablet
Mix the starch of the Lactobacillus casei culture of 150mg, the glucose of 100mg, the red ginseng extract of 50mg, 96mg And the magnesium stearate of 4mg, and 30% ethyl alcohol of 40mg is added, it is formed after particle, in 60 DEG C of dryings, is pressed into tablet press machine Tablet.
" preparation example 2 " granule
Mix the Lactobacillus casei culture of 150mg, the glucose of 100mg, the red ginseng extract of 50mg and 600mg Starch adds 30% ethyl alcohol of 100mg, is formed after particle, after 60 DEG C of dryings form granule, is filled in packet.It will The final weight of content is adjusted to 1g.
" preparation example 3 " potus
By the citric acid of the Lactobacillus casei culture of 150mg, the glucose of 10mg, the red ginseng extract of 50mg, 2g with And the Purified Water mixing of 187.8g, and be fitted into bottle.The termination capacity of content is adjusted to 200mL.
The production of " preparation example 4 " health food
Lactobacillus casei culture ... ... ..1000mg
Vitamin mixtures
Vitamine A acetate ... ... ..70 μ g
Vitamin E ... ... ... ..1.0mg
Vitamin B1 ... ... ... .0.13mg
Vitamin B2 ... ... ... .0.15mg
Vitamin B6 ... ... ... .0.5mg
0.2 μ g of vitamin B12 ... ... ...
Vitamin C ... ... ... ..10mg
10 μ g of biotin ... ... ... ...
Niacinamide ... ... ... ... 1.7mg
Folic acid ... ... ... ... ..50 μ g
Calcium pantothenate ... ... ... ... 0.5mg
Mixture of inorganic substance
Ferrous sulfate ... ... ... .1.75mg
Zinc oxide ... ... ... ... 0.82mg
Magnesium carbonate ... ... ... ... 25.3mg
Potassium dihydrogen phosphate ... ... ... ..15mg
Calcium monohydrogen phosphate ... ... ... .55mg
Potassium citrate ... ... ... .90mg
Calcium carbonate ... ... ... ... 100mg
Magnesium chloride ... ... ... ... 24.8mg
The ratio of components of said vitamin and mineral mixture is relatively to be suitble to the food of health food according to preferred Embodiment mixing composition, but can arbitrarily change its mixing ratio, and according to conventional health food preparation method, can will be upper After stating ingredient mixing, particle is made, and according to the conventional method, be used in the preparation of health food.
The production of " preparation example 5 " health drink
Lactobacillus casei culture ... ... ..1000mg
Citric acid ... ... ... ... 1000mg
Oligosaccharide ... ... ... ... 100g
Plum concentrate ... ... ... .2g
Taurine ... ... ... ... 1g
The .900mL in addition Purified Water is total ... ...
According to conventional method for preparing health drink, after mentioned component is mixed, about 1 hour of agitating and heating at 85 DEG C After, the solution of preparation is filtered, 2 liters of containers by sterilizing are packed into, after sealing sterilizing, refrigeration keeping.Later, it is used in this In the preparation of the health beverage composition of invention.
The ratio of components is will to be relatively suitble to the ingredient of hobby beverage to mix composition in accordance with the preferred embodiment, but can be with The areas such as stratum, demand country, usage, national preference degree according to demand, arbitrarily change its mixing ratio.Art technology Personnel can carry out within the scope of the present invention a variety of applications and deformation according to above content.
More than, the specific part of the content of present invention is described in detail.To those skilled in the art, this The specific technology of kind is a preferred embodiment, and it's not limited to that for the scope of the present invention.Therefore, reality of the invention Range is limited by appended claims and its equivalent.

Claims (8)

1. the extract of the extract of lactobacillus casei bacterial strain, its fragment, its culture, the bacterial strain, the fragment or The extract of the culture is preparing the purposes in particulate matter toxicity alleviation composition.
2. purposes according to claim 1, wherein
The lactobacillus casei bacterial strain is selected from Lactobacillus casei HY2782, Lactobacillus casei KCTC3109 and Lactobacillus casei One or more of KCTC13086.
3. purposes according to claim 1, wherein
The particulate matter includes polycyclic aromatic hydrocarbons.
4. purposes according to claim 1, wherein
The particulate matter contains heavy metal.
5. purposes according to claim 1, wherein
The particulate matter is PM10.
6. according to claim 1 or purposes described in any one of 5, wherein
The composition is food compositions.
7. according to claim 1 or purposes described in any one of 5, wherein
The composition is cosmetic composition.
8. according to claim 1 or purposes described in any one of 5, wherein
The composition is fodder compound.
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