CN110892914A - New application of lactobacillus paracasei K56 in improvement of asthma and related allergic symptoms - Google Patents
New application of lactobacillus paracasei K56 in improvement of asthma and related allergic symptoms Download PDFInfo
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Abstract
The invention provides a new application of lactobacillus paracasei K56 in improving asthma and related allergic symptoms. The Lactobacillus paracasei (Lactobacillus paracasei subsp. paracasei) K56 has a preservation number of CGMCC No.15139 or DSM 27447. The invention discovers that the single strain has the effect of remarkably improving asthma and related allergic diseases, can improve the over-contraction reaction of the respiratory tract, improve the activity of Th1 type cells, reduce the activity of Th2 type cells, reduce IgE secreted by B cells to improve allergy and improve the symptoms of respiratory inflammation caused by asthma.
Description
Technical Field
The invention relates to the technical field of microorganisms, in particular to a novel application of lactobacillus paracasei (Lactobacillus paracasei subsp. paracasei) K56 (with the preservation number of CGMCC No.15139 or DSM27447) in improving allergic asthma and related allergic symptoms.
Background
Asthma is a disease that is closely related to allergic constitution and causes chronic inflammation and stenosis of the bronchi due to environmental allergens, or non-allergic irritants, thereby causing chronic airway obstruction that may be recovered. Allergic asthma, also known as extrinsic asthma, usually occurs in young patients and atopic dermatitis may occur during infancy. Allergic rhinitis, urticaria and dermatitis may occur in families, allergic skin tests usually show positive reaction, increase of lgE in blood and positive allergen inhalation tests can induce asthma reaction.
CD4+T helper cells (Th) are a group of cells of considerable importance in the immune system, and this type of T helper cells can differentiate into at least four different T helper cells, including Th1 cells, Th2 cells, Th17 cells, and Regulatory T cells (tregs), depending on the situation. Th2 cell is key for promoting anaphylaxis, the cytokines (cytokines) mainly secreted by Th2 cell include IL-4, IL-5 and IL-13, IL-4 is the main cytokine for promoting IgE production; IL-5 promotes the differentiation and survival of erythroid leukocytes (eosinophil); IL-13 promotes mucus proliferation, increased Airway resistance (AHR), and allergic reactions. When CD4+Differentiation of T helper cells in the direction of Th1 cells negatively regulates proliferation and activation of Th2 cells. When the immune system is inclined to either a Th1 immune response or a Th2 immune response, different cytokines are secreted and different cell surface markers are expressed. And IL-12 secreted by macrophages and dendritic cells stimulates CD4+T helper cellDifferentiation toward Th1 cells, production of IFN-gamma, and inhibition of Th2 cell activation. Therefore, if the balance of Th1/Th2 can be achieved, the allergic reaction caused by Th2 can be improved
In recent years, probiotics are recognized by medical and scientific circles to be beneficial to human health and have no side effect. The Food Agriculture Organization (FAO) and the World Health Organization (WHO) of the United Nations define probiotics (probiotics) as: a live microorganism, when administered in an appropriate amount, can confer a health benefit to the host. The genus lactobacillus is a group of gram-positive facultative anaerobes, ubiquitous in the human gastrointestinal tract and vagina, capable of fermenting sugars and taking lactic acid as the major metabolite. There are many studies and clinical trials that have in turn demonstrated that there is an important correlation between lactobacilli and health. Currently, lactobacilli have found efficacy including: (1) improving the bacterial phase balance of the intestinal microbiota; (2) prevention of diarrhoea (diarrhea); (3) reducing the risk of colon cancer (colon); (4) stimulating normal development and function of the gastrointestinal epithelial mucosal system (gastroenteritic mucosal system); (5) production of various vitamins (vitamines) and nutrients (nutrients); and (6) prevention and treatment of vaginitis (vaginosis).
Disclosure of Invention
One object of the present invention is to provide a novel use of lactobacillus paracasei K56.
Lactobacillus paracasei (Lactobacillus paracasei subsp. paracasei) K56 strain has been deposited at the German Collection of microorganisms and Cell Cultures (German Collection of microorganisms and Cell Cultures) on day 27 6 month 2013 under accession number DSM 27447; in addition, Lactobacillus paracasei (Lactobacillus paracasei subsp. paracasei) K56 strain has also been stored in China General microbiological culture Collection Center (CGMCC) with the Collection number of CGMCC No.15139 in 29.12 months and 29.2017.
The invention discovers that a single strain of Lactobacillus paracasei (Lactobacillus paracasei subsp. paracasei) K56 has the effect of improving asthma and related allergic symptoms, can improve the excessive respiratory tract contraction reaction of allergic asthma mice, improves the activity of Th1 type cells of the allergic asthma mice, reduces the activity of Th2 type cells, reduces IgE secreted by B cells to improve allergy, and improves the respiratory tract inflammation symptoms of the allergic asthma mice caused by asthma.
Thus, the present invention provides the use of Lactobacillus paracasei (Lactobacillus paracasei) with the deposit number CGMCC No.15139 or DSM27447 for the preparation of a composition for improving asthma and related allergic symptoms.
According to a particular embodiment of the invention, the lactobacillus paracasei is used for preparing the composition in the form of a solid or liquid bacterial preparation.
According to a particular embodiment of the invention, the composition may comprise a food composition, a feed composition or a pharmaceutical composition.
According to a particular embodiment of the invention, the composition can be used in animals or humans. The composition may further comprise conventional ingredients in the art. For example, for pharmaceutical compositions, suitable excipients may be included, which may be excipients, diluents, fillers, absorption enhancers, and the like. For food compositions, the bifidobacterium lactis of the invention may be produced as in the prior art with bifidobacterium lactis-containing food products, the compositions being available in different forms depending on the needs of the recipient. Such as powders, lozenges, granules, microcapsules, liquid preparations, and the like.
According to a particular embodiment of the invention, the composition is for improving the hyper-systolic airway response in allergic asthmatic mice. In specific application, the application amount of the lactobacillus paracasei is 3.88 multiplied by 106CFU～3.88×1013CFU/day, preferably 3.88X 109CFU～3.88×1011CFU/day.
According to a specific embodiment of the present invention, the composition is for promoting the activity of Th1 type cells in allergic asthma mice. In specific application, the application amount of the lactobacillus paracasei is 3.88 multiplied by 106CFU～3.88×1013CFU/day, preferably 3.88109CFU～3.88×1011CFU/day.
According to a particular embodiment of the invention, the composition is for reducing the activity of a Th2 type cell. In specific application, the application amount of the lactobacillus paracasei is 3.88 multiplied by 106CFU～3.88×1013CFU/day, preferably 3.88X 109CFU～3.88×1011CFU/day.
According to a particular embodiment of the invention, the composition is for reducing IgE secretion by B-cells to ameliorate allergy. In specific application, the application amount of the lactobacillus paracasei is 3.88 multiplied by 106CFU～3.88×1013CFU/day, preferably 3.88X 109CFU～3.88×1011CFU/day.
According to a particular embodiment of the invention, the composition is for ameliorating the symptoms of airway inflammation associated with asthma in sensitized asthmatic mice. In specific application, the application amount of the lactobacillus paracasei is 3.88 multiplied by 106CFU～3.88×1013CFU/day, preferably 3.88X 109CFU～3.88×1011CFU/day.
In a specific embodiment of the present invention, the composition may also include a biocompatible excipient, in addition to lactobacillus paracasei K56, to prepare a dosage form of a solution, suspension, emulsion, powder, lozenge, pill, syrup, lozenge, tablet, chewing gum, or capsule for general or medicinal use.
In a specific embodiment of the invention, the composition is a food composition, and the food may be a fermented dairy product (e.g., fermented milk, flavored fermented milk, fermented milk beverage, etc.), cheese, milk-containing beverage, solid beverage, powdered milk, or the like.
In another specific embodiment of the invention, the composition is a feed composition. The other components of the feed composition may be selected with reference to conventional techniques in the field of probiotic feeds.
In another specific embodiment of the present invention, the composition is a pharmaceutical composition. The other components of the pharmaceutical composition may be selected with reference to conventional techniques in the field of probiotic medicine.
In conclusion, the invention provides a new application of lactobacillus paracasei K56, the lactobacillus paracasei K56 has the effects of remarkably improving asthma and related allergic symptoms, can improve the airway over-contraction reaction of allergic asthma mice, improve the activity of Th1 type cells of the allergic asthma mice, reduce the activity of Th2 type cells, reduce IgE secretion of B cells to improve allergy, and improve the respiratory inflammation symptoms of the allergic asthma mice caused by asthma, can be used for preparing foods, medicines, feeds and the like with the effects of improving asthma and related allergic symptoms, and has wide application prospects.
Drawings
FIG. 1 shows the effect of K56 on airway resistance in allergic asthma models in allergic asthma mice.
FIG. 2 shows the effect of K56 on Th1 type cytokines in lung washes of sensitized asthmatic mice in allergic asthma mode.
FIG. 3 shows the effect of K56 on Th2 type cytokines in lung washes of sensitized asthmatic mice in allergic asthma mode.
FIG. 4 shows the effect of K56 on proinflammatory cytokines in lung washes of sensitized asthmatic mice in allergic asthma mode.
FIG. 5 shows the effect of K56 on OVA-IgE production in serum of sensitized asthmatic mice in allergic asthma model.
Detailed Description
For a more clear understanding of the technical features, objects and advantages of the present invention, reference is now made to the following detailed description taken in conjunction with the accompanying specific embodiments, and the technical solutions of the present invention are described, it being understood that these examples are intended to illustrate the present invention and are not intended to limit the scope of the present invention.
In the examples, each raw reagent material is commercially available, and the experimental method not specifying the specific conditions is a conventional method and a conventional condition well known in the art, or a condition recommended by an instrument manufacturer.
Example 1: experiment on efficacy of lactobacillus paracasei K56 in improving asthma and related allergic symptoms
The lactic acid bacteria were administered to Balb/c mice in a tube-fed manner,the groups were divided into 3 groups, Control (CON), OVA, and K56, respectively. The feeding bacteria amount of the K56 bacteria is 109CFU/200μL H2The O, CON and OVA groups were all fed with 200 μ LH2And O, starting feeding from 0 day, feeding to 32 days, sacrificing the mice, and detecting the remission effect of the lactic acid bacteria on the anaphylactic reaction. On the 0 th day and the 14 th day of the experiment, OVA (20 mug OVA/2mg Alu/200 mug LPBS) was injected for sensitization, and on the 0 th day, the 15 th day and the 22 th day blood was collected to detect total IgE (total IgE) to confirm whether sensitization was successful, and after successful sensitization, 100 mug OVA/40 mug PBS was nasally dropped on the 28 th day, the 29 th day and the 30 th day to induce respiratory tract anaphylaxis of mice. Changes in Airway resistance (AHR) were detected on day 31 using a barometric wall-body pletype graph (Buxco Electronics, inc., Troy, NY). On day 32, the sacrifice mice were used to collect lung washing solution, serum, etc. for analysis of cytokines, OVA-IgE, etc.
Airway resistance (AHR): the respiratory tract was stimulated by nasal instillation of allergen OVA, and the change in respiratory tract resistance was measured by stimulating the contraction of mouse airway smooth muscle with aerosolized methacholine (methacholine) the day after three consecutive days of allergen stimulation. The group fed K56 showed that K56 was helpful in improving the airway hyper-constriction response in the mice with allergic asthma when the airway resistance was lower than that of the OVA group at different doses, as shown in FIGS. 1 to 5
The nasal cavity is used for dripping allergen OVA to stimulate respiratory tract, and atomized methacholine is used for stimulating the contraction of mouse airway smooth muscle after three consecutive days of allergen stimulation, so as to measure the change of respiratory tract resistance. FIG. 1 illustrates that the group fed K56 showed lower airway resistance than the OVA group at different doses, indicating that K56 was helpful in improving the airway hyper-constriction response in the allergic asthma mice.
BALF contains various cytokines, and the comparison of the concentration change of the cytokines before and after sensitization is helpful for clarifying the effect of lactic acid bacteria on improving asthma symptoms and regulating the immune balance of respiratory mucosa. FIG. 2 shows that the IFN-gamma expression level of K56 is higher than that of OVA group, and K56 is helpful for improving the Th1 cell activity of allergic asthma mice.
The expression level of Th2 type cytokines in lung washes is shown in FIG. 3, and the expression levels of IL-4, IL-5 and IL-13 are all lower than those of OVA group when K56 is fed, showing that K56 is helpful for reducing Th2 type cell activity of allergic asthma mice and further improving allergy.
The expression level of proinflammatory cytokines in lung flushing fluid is shown in figure 4, and the expression level of TNF- α fed with K56 is lower than that of OVA group, which shows that K56 is helpful for improving respiratory tract inflammation reaction of allergic asthma mice accompanied by asthma.
Mouse serum OVA-specific IgE: in the allergic asthma animal model, over-activation of Th2 is known to be an important key for pathogenesis, the trend of T cells also influences the situation that B cells secrete antibody in vivo, Th2 cells are activated, and B cells tend to secrete IgE. Fig. 5 shows that serum OVA-IgE fed with K56 is significantly lower than the OVA group, showing that K56 bacteria can reduce Th2 cell activation and consequently lower IgE secretion by B cells to improve allergy.
In conclusion, in the mouse model, the results of AHR test, BALF analysis and OVA-specific IgE expression all show that Lactobacillus paracasei subspecies K56 has excellent ability to improve allergic and asthmatic symptoms. Therefore, the lactobacillus paracasei subspecies paracasei K56 can be used as dairy products, drinks or additives of dairy products, and has the functions of improving and relieving asthma and even allergic symptoms. In addition, Lactobacillus paracasei subspecies Paracasei K56 may also be mixed with a biocompatible excipient to prepare a dosage form of solution, suspension, emulsion, powder, lozenge, pill, syrup, lozenge, tablet, chewing gum, or capsule for general or medicinal use.
Claims (10)
1. Use of Lactobacillus paracasei (deposited under the accession number cgmccno.15139 or DSM27447) for the preparation of a composition for ameliorating asthma and related allergic symptoms.
2. Use according to claim 1, wherein the lactobacillus paracasei is used for preparing the composition in the form of a solid or liquid bacterial preparation.
3. Use according to claim 1, wherein the composition comprises a food composition, a feed composition or a pharmaceutical composition.
4. The use according to claim 1, wherein the composition is for improving the hyper-systolic airway response in allergic asthmatic mice;
preferably, the application amount of the lactobacillus paracasei is 3.88 multiplied by 106CFU～3.88×1013CFU/day, more preferably 3.88X 109CFU～3.88×1011CFU/day.
5. The use according to claim 1, wherein the composition is for increasing the activity of Th1 type cells in allergic asthma mice;
preferably, the application amount of the lactobacillus paracasei is 3.88 multiplied by 106CFU～3.88×1013CFU/day, more preferably 3.88X 109CFU～3.88×1011CFU/day.
6. The use according to claim 1, wherein the composition is for reducing the activity of a Th 2-type cell;
preferably, the application amount of the lactobacillus paracasei is 3.88 multiplied by 106CFU～3.88×1013CFU/day, more preferably 3.88X 109CFU～3.88×1011CFU/day.
7. The use according to claim 1, wherein the composition is for reducing B-cell secretion of IgE to ameliorate allergy;
preferably, the application amount of the lactobacillus paracasei is 3.88 multiplied by 106CFU～3.88×1013CFU/day, more preferably 3.88X 109CFU～3.88×1011CFU/day.
8. The use according to claim 1, wherein the composition is for ameliorating the symptoms of airway inflammation associated with asthma in sensitized asthmatic mice;
preferably, the application amount of the lactobacillus paracasei is 3.88 multiplied by 106CFU～3.88×1013CFU/day, more preferably 3.88X 109CFU～3.88×1011CFU/day.
9. Use according to any one of claims 4 to 8, wherein the composition is a food composition.
10. Use according to claim 9, wherein the food product is a fermented dairy product, a cheese, a milk-containing drink, a solid drink or a powdered milk.
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