CN114958674A - Probiotics for barrier repair and skin anti-inflammation and application - Google Patents
Probiotics for barrier repair and skin anti-inflammation and application Download PDFInfo
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- CN114958674A CN114958674A CN202210663811.4A CN202210663811A CN114958674A CN 114958674 A CN114958674 A CN 114958674A CN 202210663811 A CN202210663811 A CN 202210663811A CN 114958674 A CN114958674 A CN 114958674A
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- lactobacillus rhamnosus
- gforu
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- lactobacillus
- inflammation
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- C—CHEMISTRY; METALLURGY
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- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
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- 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
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- A61K35/66—Microorganisms or materials therefrom
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- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
- A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
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- 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
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- A—HUMAN NECESSITIES
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- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P11/00—Drugs for disorders of the respiratory system
- A61P11/02—Nasal agents, e.g. decongestants
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61P17/00—Drugs for dermatological disorders
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- A—HUMAN NECESSITIES
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- A—HUMAN NECESSITIES
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- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
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- A—HUMAN NECESSITIES
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- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
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- 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
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- 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
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/175—Rhamnosus
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- C12R2001/00—Microorganisms ; Processes using microorganisms
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- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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Abstract
The invention mainly relates to the technical field of microorganisms, and particularly relates to probiotics for barrier repair and skin anti-inflammation and application thereof. The strain GforU-8 is preserved in China center for type culture Collection with the preservation number of CCTCC NO: M20211532. The strain GforU-8 has the functions of maintaining and repairing skin barrier, resisting inflammation and treating acne and nasosinusitis, and can be used for foods, medicines, cosmetics and the like.
Description
Technical Field
The invention mainly relates to the technical field of microorganisms, and particularly relates to probiotics for barrier repair and skin anti-inflammation and application thereof.
Background
The skin is the largest organ in the human body, the total weight accounts for about 16% of the body weight of an individual, and the skin is the first defense line for maintaining the stability of the body and resisting the invasion of external adverse factors. Studies have shown that skin diseases are induced if the external environment causes abnormalities in the relevant genes in the skin barrier.
The skin barrier is a structural barrier formed by the epidermal keratinocytes of the stratum corneum and the lipids between the cutin. The skin barrier prevents the release of excess water from the human body and prevents harmful substances such as chemicals or microorganisms from entering our body. The corneocyte cortex, which constitutes the surface of dead keratinocytes, plays an important role in the stability of intercellular lipids. Skin barrier damage can cause skin dryness, skin aging, atopic dermatitis, eczema, psoriasis, ichthyosis, and daily rotatory optical dermatitis, skin sensitivity, irritant dermatitis, and hormone dependent dermatitis, and seborrheic diseases such as acne, rosacea, and seborrheic dermatitis.
The probiotic is used in cosmetics, and can balance skin epidermal flora and repair skin barrier. As reported in the literature, the topical application of probiotics can treat atopic dermatitis, promoting the repair of the skin barrier. Meanwhile, researches find that the probiotics can prevent and treat various inflammatory diseases by regulating the phagocytic capacity of macrophages and enhance the immunity of the skin.
Acne is a common chronic inflammatory skin disease, and is closely related to factors such as excessive sebum secretion, blockage of pilosebaceous ducts, bacterial infection and inflammatory reaction. Research has shown that Propionibacterium acnes (Propionibacterium acnes), considered as the main pathogenic bacterium causing acne, can induce and activate the initial loop of acne inflammation and convert glycerol into fatty acids, leading to an inflammatory response; simultaneously, protease, hyaluronidase and chemotactic factor are produced, so that the hair follicle is hyperkeratotic and acne is formed. On the other hand, the skin resident staphylococcus epidermidis and propionibacterium acnes can compete with each other antagonistically, and increasing the amount of staphylococcus epidermidis can inhibit the proliferation of propionibacterium acnes. Therefore, reducing the flora ratio of propionibacterium acnes/staphylococcus epidermidis can effectively relieve acne inflammation, thereby maintaining skin health by regulating the skin micro-ecological balance.
Nasosinusitis is a common disease in otorhinolaryngology, and is mainly caused by respiratory tract allergy, nasal-paranasal sinus mucosa cleaning dysfunction, respiratory tract microbial infection, abnormal local anatomical structure of nose and the like. Research shows that Haemophilus influenzae (Haemophilus influenzae) is an important pathogenic bacterium of nasosinusitis, and Lactobacillus sake (Lactobacillus sakei) has a positive effect on maintaining normal functions of nasal cavities. Therefore, the proportion of the flora of the haemophilus influenzae/the sake lactobacillus is reduced, and the treatment of the nasosinusitis is facilitated.
At present, probiotic related products developed by utilizing a microecology technology can effectively solve the problems.
Disclosure of Invention
To achieve the technical objects, the problems of the prior art are solved. The lactobacillus rhamnosus GforU-8 separated and identified from the feces of healthy boys has the functions of repairing skin barriers, resisting inflammation and treating acne and nasosinusitis, and has great market potential in the fields of foods, medicines, maintenance products or cosmetics and the like.
The technical scheme of the invention is as follows:
firstly, the invention provides a strain of Lactobacillus rhamnosus (Lactobacillus rhamnosus) which is preserved in China center for type culture Collection with the preservation number of CCTCC NO: M20211532.
The Lactobacillus rhamnosus is derived from feces of a healthy 8-year-old boy, is numbered as GforU-8 by the inventor, and is identified as Lactobacillus rhamnosus (Lactobacillus rhamnous) through 16S rRNA. The strain is gram-positive and rod-shaped under a microscope; the bacterial colony grows on an MRS plate, can form a round bacterial colony with a smooth and opaque surface, is white and has a regular edge; the strain grows uniformly and turbulently in MRS liquid culture medium, and the strain is white precipitate after long-term storage, and the optimal growth temperature is 37 ℃.
Through research and test, the lactobacillus rhamnosus GforU-8 disclosed by the invention has the effects of repairing skin barriers, resisting inflammation, treating acne and nasosinusitis and the like. Therefore, the present application has heard of the use of the lactobacillus rhamnosus, which can be used for preparing foods, medicines, cosmetics, etc., according to its characteristics.
Furthermore, when the lactobacillus rhamnosus GforU-8 is used for preparing food, medicines and cosmetics, the lactobacillus rhamnosus GforU-8 can be thalli of the lactobacillus rhamnosus GforU-8 or/and derivatives or/and metabolites thereof.
The thallus of the lactobacillus rhamnosus GforU-8 can be live thallus or/and inactivated thallus;
the derivative is lysate or/and extract of lactobacillus rhamnosus GforU-8, including but not limited to cell wall, intracellular protein, intracellular polysaccharide and the like;
the metabolite is understood to be a primary metabolite or a secondary metabolite of lactobacillus rhamnosus GforU-8, such as a prebiotic, a compound containing an immunogenic component, an exopolysaccharide, etc., and may also be directly the supernatant of the fermentation broth.
Has the advantages that:
in vitro cell experiments show that the lactobacillus rhamnosus GforU-8 has the effect of promoting epidermal cell repair, and the survival rate of HaCaT cells damaged by Sodium Dodecyl Sulfate (SDS) improved by the GforU-8 reaches 103.24% -106.73%; (ii) a The gene has the effect of up-regulating the expression of skin barrier repair related factors including a filaggrin gene FLG, an involucrin gene IVL, an OVO-like transcription factor 1 gene OVOL1 and a lorin gene LOR, and the gene expression level is up-regulated by 1.27-2.17 times; has the function of reducing the release of inflammatory factors, and can reduce the generation amount of Nitric Oxide (NO) of mouse macrophage Raw264.7 induced by Lipopolysaccharide (LPS) from 44.91 to 71.07 percent; has the function of down regulating the expression of HaCaT cell inflammation related factors IL-6, IL-8, IL-22 and TRPV1 genes induced by staphylococcus aureus, and the expression quantity of the genes is down regulated by 15 to 32 percent.
In addition, in vitro experiments show that the lactobacillus rhamnosus GforU-8 has the function of reducing the ratio of propionibacterium acnes to staphylococcus epidermidis so as to treat acne; also has the function of reducing the ratio of haemophilus influenzae to lactobacillus sake, thereby treating nasosinusitis.
Preservation information
Preservation time: 12 months 03 days 2021;
the name of the depository: china center for type culture Collection;
the preservation number is: CCTCC NO: M20211532;
the address of the depository: eight paths of Wuhan university No. 299 in Wuhan district, Wuhan city, Hubei province;
and (3) classification and naming: lactobacillus rhamnosus (Lactobacillus rhamnosus).
Detailed Description
The present invention will be described in further detail with reference to the following examples, but it should not be construed that the scope of the above subject matter is limited to the following examples. All the technologies realized based on the above contents of the present invention belong to the scope of the present invention, and the following embodiments are all completed by adopting the conventional prior art except for the specific description. EXAMPLE 1 isolation of Lactobacillus rhamnosus GforU-8
Feces were collected from healthy 8-year-old boys. Properly treating a sample, then uniformly shaking the sample in normal saline, taking the supernatant, streaking the supernatant on an MRS solid plate, culturing the MRS solid plate at a constant temperature of 37 ℃ for 24-48 hours, and then selecting a white colony to repeatedly inoculate and screen until a uniform single colony is obtained, wherein the uniform single colony is named GforU-8.
Gram staining microscopy: the strain GforU-8 is gram-positive and rod-shaped under a microscope; growing on an MRS plate to form white round microcolonies with smooth, mellow and opaque surfaces and regular edges; the strain grows in MRS liquid culture medium in a uniform turbid way, and the strain is white and precipitated after being placed for a long time.
Example 2 nucleic acid identification of Lactobacillus rhamnosus GforU-8
1. 16s rRNA Gene sequence analysis:
picking single colony in MRS liquid culture medium, culturing overnight at 37 ℃, centrifuging and collecting thallus, and operating according to the steps of DNA extraction kit. The primers are bacterial universal primers 27F and 1492R, the PCR amplification system is a 50 mu L system, and the pre-denaturation is carried out for 5min at 95 ℃; 15s at 94 ℃, 15s at 57 ℃, 40s at 72 ℃ and 35 cycles; extension at 72 ℃ for 10 min.
2. Results
The homology comparison (BLASTN) of the PCR product sequencing result with the published standard sequence in GenBank indicates that the strain GforU-8 is Lactobacillus rhamnosus (Lactobacillus rhamnosus).
Example 3 Lactobacillus rhamnosus GforU-8 promotion of SDS-induced HaCaT cell injury repair experiment
1. Preparing GforU-8 bacterial liquid:
culturing activated lactobacillus rhamnosus GforU-8 bacterial liquid in MRS liquid culture medium in an incubator at 37 ℃ for standing culture for 16-18 h, detecting, and diluting the bacterial liquid to adjust OD (optical density) by using PBS (phosphate buffer solution) 600 After centrifugation, the supernatant was filtered through a 0.22 μm filter, and the cells were washed twice with PBS and resuspended in PBS to OD 600 The supernatant and the cell samples were finally autoclaved at 121 ℃ for 30min for inactivation, 0.2.
2. Experiment for promoting HaCaT cell repair
Inoculation of HaCaT cells (5X 10) 4 cell/well) to 96-well plates and cultured overnight until cells adhere. Preparing SDS of 50 mu g/ml, adding 100 mu l of SDS into each well, incubating for 8h, adding 5% supernatant (V/V, corresponding to the control group 1 being PBS with the same volume) into the experimental group 1, adding 10% inactivated thallus (V/V, corresponding to the control group 2 being PBS with the same volume) into the experimental group 2, and incubating for 24 h. 10 mul of CCK-8 solution is added respectively, incubated for 4h, and the absorbance at 450nm is detected.
Survival rate of cells ═ a Experiment of /A Control ×100％；
Wherein, A in the formula Experiment of For detection of the absorbance at 450nm in the experimental group, A Control The absorbance was measured at 450nm for the control.
As a result:
GforU-8 promotes HaCaT cell repair
According to the data in the table, the supernatant and the inactivated thallus of the lactobacillus rhamnosus GforU-8 have a repairing effect on SDS (sodium dodecyl sulfate) damage of HaCaT cells.
Example 4 Lactobacillus rhamnosus GforU-8 promotion HaCaT Barrier repair related Gene expression experiment
1. Preparation of GforU-8 bacterial solution (same as example 3)
2. Inoculation of human immortalized keratinocytes HaCaT (5X 10) 5 cell/well) to 6-well plates and cultured overnight until cells adhere. Adding 10 percent (V/V) of inactivated strain thallus, adding PBS with the same volume as the control group, respectively culturing for 24h, adding lysate, extracting total RNA of cells, detecting the concentration and purity of the RNA, adjusting all samples to 1 mu g, carrying out reverse transcription to obtain cDNA, taking GAPDH as an internal reference gene, and carrying out qPCR (quantitative polymerase chain reaction) to detect the expression of FLG (FLG) and OVOL (OVOL 1) genes. Expression fold changes were calculated according to the formula.
The formula: f ═ 2 -ΔΔCT ；
Wherein:
△CT experiment of ＝CT Experiment of -CT Internal reference (experiment) ；
△CT Control of ＝CT Control -CT Internal reference (contrast) ；
△△CT＝△CT Experiment of -△CT Control 。
As a result:
GforU-8 inactivated thallus up-regulated barrier repair related gene expression
The results in the table show that lactobacillus rhamnosus GforU-8 has the function of up-regulating the expression of cell repair related factors, namely a silk polymerization protein gene FLG, an involucrin gene IVL, an OVO-like transcription factor 1 gene OVOL1 and a lorir of a loricrin gene, and the effect of promoting skin barrier repair of the strain is fully demonstrated.
Example 5 Lactobacillus rhamnosus GforU-8 decreases the amount of NO produced by Raw264.7 cells
1. Preparation of GforU-8 bacterial liquid
GforU-8 was cultured overnight in MRS medium and OD was detected 600 And adjusting the concentration of the bacterial liquid to OD with PBS buffer 600 After centrifugation, the supernatant was filtered through a 0.22 μm filter, and the cells were washed twice with PBS and resuspended in PBS to OD 600 The supernatant and the cell samples were finally autoclaved at 121 ℃ for 30min for inactivation, 0.2.
2. Preparation of Raw264.7 cells
Raw264.7 cells were digested and then digested at 2X 10 5 cells/well were inoculated into 24-well plates and incubated overnight at 37 ℃ in a 5% carbon dioxide incubator.
3. GforU-8 addition and LPS stimulation
Culturing Raw264.7 cells overnight, adding 5 percent (V/V, equal volume of PBS is added to a corresponding control group) of the supernatant into an experimental group 1, and adding 10 percent (V/V, equal volume of PBS is added to a corresponding control group) of inactivated thallus into an experimental group 2; and after 2h, adding 0.5ml of LPS solution with the concentration of 0.2 mu g/ml to induce Raw264.7 cells to be inflamed, taking cell culture supernatant after 20h, drawing a standard curve according to the method of the Biyunyan NO detection kit, and calculating the concentration and inhibition rate of NO in the sample.
As a result:
GforU-8 reduces NO production of Raw264.7 cells
The results show that the lactobacillus rhamnosus GforU-8 and the fermentation liquor thereof have anti-inflammatory effect, can reduce the NO production amount of Raw264.7 cells induced by LPS, and compared with an LPS control group, the supernate is reduced by 71.07 percent, and the inactivated thallus is reduced by 44.91 percent.
Example 6 Lactobacillus rhamnosus GforU-8 downregulates expression of HaCaT cell inflammatory factors
1. GforU-8 sample preparation
GforU-8 was cultured overnight with MRS and OD was detected 600 And adjusting the concentration of the bacterial liquid to OD with PBS buffer 600 After centrifugation, the cells were removed and the supernatant was filtered through a 0.22 μm filter and finally inactivated by autoclaving at 121 ℃ for 30 min.
2. HaCaT cell preparation
HaCaT cells were digested and then treated at 2X 10 5 cells/well were inoculated into 24-well plates and incubated overnight at 37 ℃ in a 5% carbon dioxide incubator.
3. Preparation and addition of staphylococcus aureus
Inoculating Staphylococcus aureus to nutrient broth, shaking culturing at 37 deg.C overnight, and culturing with MEMAdjusting the concentration of the bacterial liquid to OD in a clear culture medium 600 To 6, 100 μ l per well was added overnight cultured HaCaT cells to stimulate production of inflammatory factors, 3h later the cell culture medium was discarded, PBS washed 5 times and 1ml of MEM serum-free medium was added to each well.
4. GforU-8 sample addition
GforU-8 supernatant was added at 5% to staphylococcus aureus-stimulated HaCaT cells, 3 replicates per group, and incubated overnight.
5. qPCR method for detecting relative expression multiple of cell inflammatory factor mRNA
And (3) after removing a culture medium from the cells, extracting RNA by using an RNA extraction kit, detecting the concentration and purity of the RNA, adjusting all samples to 1 mu g, performing RT-PCR (reverse transcription-polymerase chain reaction) and qPCR by using a reverse transcription kit and a SYBRGreen qPCR kit, and calculating the relative expression multiple F of the related genes of the inflammatory factors.
The formula: f is 2 -ΔΔCT
As a result:
GforU-8 down-regulation of expression of inflammatory factor related genes
As shown in the results, GforU-8 can down-regulate the expression level of HaCaT inflammatory factors IL-6, IL-8, IL-22 and TRPV1 genes induced by staphylococcus aureus, and the expression level is reduced by 15-32%. Therefore, GforU-8 has an anti-inflammatory effect.
EXAMPLE 7 Effect of Lactobacillus rhamnosus GforU-8 on altering the flora ratio for the treatment of acne
1. Preparing a lactobacillus rhamnosus GforU-8 bacterial liquid:
culturing activated Lactobacillus rhamnosus GforU-8 bacterial liquid in an MRS liquid culture medium in an incubator at 37 ℃ for static culture for 16-18 h, detecting, and adjusting OD (origin-destination) by using an MRS blank culture medium 600 Inactivating at 121 deg.C for 30min, centrifuging to obtain supernatant, and filtering with 0.22 μm filter membrane to obtain sample to be tested.
2. Preparing a skin flora bacterial liquid:
mixing acne propionic acidBacillus CGMCC 1.5003 and Staphylococcus epidermidis CGMCC 1.4260 are cultured in BHI culture medium at 37 deg.C for 18 hr, detected and OD regulated with BHI blank culture medium 600 ＝0.2。
3. Experiment for influencing growth of skin flora by adding supernatant
The experimental group was cultured at 37 ℃ for 16h with 10% (V/V) inactivated GforU-8 supernatant added to two kinds of skin flora bacterial solutions (control group added with the same amount of MRS medium), and the bacterial concentrations (OD) of the control group and the experimental group were used 600 ) The effect on skin flora growth was evaluated in relative ratio.
4. Results
Effect of GforU-8 on growth of acne-related flora
The results show that GforU-8 has significant inhibitory effect on Propionibacterium acnes, but much lower inhibitory effect on Staphylococcus epidermidis. The GforU-8 can obviously change the proportion of skin flora, thereby achieving the purpose of treating acne.
EXAMPLE 8 Effect of Lactobacillus rhamnosus GforU-8 on altering the flora ratio for the treatment of sinusitis
1. Preparing a lactobacillus rhamnosus GforU-8 bacterial liquid:
culturing activated lactobacillus rhamnosus GforU-8 bacterial liquid in an MRS liquid culture medium in an incubator at 37 ℃ for standing culture for 16-18 h, detecting and adjusting to OD by using the MRS liquid culture medium 600 Inactivating at 121 deg.C for 30min, centrifuging to obtain supernatant, and filtering with 0.22 μm filter membrane to obtain sample to be tested.
2. Preparing nasal cavity flora bacterial liquid:
culturing Haemophilus influenzae ATCC 49766 in BHI culture medium and Lactobacillus sake ATCC 15521 in MRS culture medium at 37 deg.C for 18h, detecting, and adjusting to OD with original culture medium 600 ＝0.2。
3. Experiment for influencing growth of nasal flora by adding supernatant
The experimental group was cultured with 10% (V/V) inactivated GforU-8 supernatant added to two bacterial solutions (control group added with the same amount of MRS medium) at 37 deg.C for 3h, and relative concentration (OD) of Haemophilus influenzae/Lactobacillus sake 600 ) The ratio was evaluated for the effect on nasal flora growth.
4. Results
Effect of GforU-8 on the flora associated with sinusitis
The result shows that GforU-8 has obvious inhibition effect on haemophilus influenzae and certain proliferation promoting effect on lactobacillus sake. The GforU-8 can obviously change the proportion of nasal flora, thereby effectively treating the nasosinusitis.
In addition, the fermentation supernatant or the inactivated bacteria referred to in all the above examples have the corresponding effects, such as the derivatives (lysate or extract) of GforU-8 obtained by further processing, extraction, etc., or the primary metabolites or secondary metabolites of GforU-8 in the fermentation supernatant.
Claims (10)
1. A probiotic bacterium for barrier repair and skin inflammation resistance is provided, the strain is Lactobacillus rhamnosus (Lactobacillus rhamnosus) which is preserved in China center for type culture Collection with the preservation number of CCTCC NO: M20211532.
2. The application of the lactobacillus rhamnosus is characterized in that the lactobacillus rhamnosus is used for preparing food, medicines and cosmetics, and the lactobacillus rhamnosus is preserved in China center for type culture Collection with the preservation number of CCTCC NO: M20211532.
3. The use of lactobacillus rhamnosus according to claim 2 wherein the lactobacillus rhamnosus has the function of repairing the skin barrier.
4. The use of lactobacillus rhamnosus according to claim 2, wherein the lactobacillus rhamnosus has an anti-inflammatory action.
5. The use of lactobacillus rhamnosus according to claim 2, wherein the lactobacillus rhamnosus has the function of treating acne.
6. The use of lactobacillus rhamnosus according to claim 2, wherein the lactobacillus rhamnosus has the function of treating sinusitis.
7. The use of lactobacillus rhamnosus according to claim 2, wherein the lactobacillus rhamnosus is its thallus or/and its derivatives or/and its metabolites.
8. The use of lactobacillus rhamnosus according to claim 7, wherein the bacteria of lactobacillus rhamnosus are live bacteria or/and inactivated bacteria.
9. Use of lactobacillus rhamnosus according to claim 7 wherein the derivative is a lysate or/and extract of lactobacillus rhamnosus.
10. Use of lactobacillus rhamnosus according to claim 7 wherein the metabolites are primary or secondary metabolites of lactobacillus rhamnosus.
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