CN116889578A - Application of strain or microbial inoculum of leuconostoc mesenteroides intestinal subspecies with antiallergic effect - Google Patents
Application of strain or microbial inoculum of leuconostoc mesenteroides intestinal subspecies with antiallergic effect Download PDFInfo
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- CN116889578A CN116889578A CN202310635579.8A CN202310635579A CN116889578A CN 116889578 A CN116889578 A CN 116889578A CN 202310635579 A CN202310635579 A CN 202310635579A CN 116889578 A CN116889578 A CN 116889578A
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- leuconostoc mesenteroides
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Abstract
The invention relates to the technical field of microorganisms, and discloses application of a strain or a microbial agent of leuconostoc mesenteroides intestinal membrane subspecies with antiallergic effect. The application of the invention is the application of a strain or a microbial inoculum of leuconostoc mesenteroides subspecies intestinal membrane in preparing a product with any one of the following functions: 1) For up-regulating Th1 type cytokine levels; 2) For reducing the level of cytokines that enhance allergic reactions; 3) For reducing IgE content in a body; 4) For reducing histamine content; 5) For improving intestinal flora structure; 6) Is used for improving the content of short chain fatty acids in intestinal tracts; 7) For restoring intestinal barrier damage or enhancing intestinal mucosal barrier; 8) For alleviating or ameliorating weight loss; 9) For improving exercise capacity; 10 For antiallergic or allergy symptom relief. The leuconostoc mesenteroides intestinal subspecies have antiallergic effect, do not have toxic genes, and have no toxic or side effects in animal evaluation, and have advantages compared with the traditional medicine for treating allergy at present.
Description
Technical Field
The invention relates to the technical field of microorganisms, in particular to application of a strain or a microbial inoculum of leuconostoc mesenteroides intestinal membrane subspecies with antiallergic effect.
Background
Allergy belongs to type I allergy, is an inflammatory reaction caused by unbalance of Th1 and Th2 immune reactions mediated by IgE and is caused by external environmental factors acting on organisms, and is mainly characterized by infiltration of cells expressing Th2 cytokines in a large amount in tissues. Allergy is increasingly common in social groups, and causes allergic diseases such as allergic eczema, urticaria, recurrent allergic rhinitis, allergic asthma and the like. At present, drugs such as loratadine and cetirizine are often used for treating allergic diseases, but the drugs have more side effects, such as the loratadine often causes adverse reactions such as hypodynamia, headache, somnolence, dry mouth, gastrointestinal discomfort (including nausea and gastritis) and the like.
Existing studies indicate that probiotics can alleviate and prevent allergic reactions. For example, a strain of bifidobacterium infantis (with the preservation number of CGMCC 20868) is disclosed in Chinese patent application CN202110347183.4, and the strain has excellent antiallergic effect. Chinese patent application CN201910686679.7 discloses lactobacillus rhamnosus (preservation number is CGMCC 20868CGMCC No.16873), which can effectively regulate intestinal flora of organism, and prevent and control allergy.
Leuconostoc mesenteroides (Leuconostoc mesenteroides) is an important class of lactic acid bacteria and is widely used in foods due to its excellent fermentation properties and the ability to produce specific flavours. Leuconostoc mesenteroides is also an important starter in the dairy industry, for example, leuconostoc mesenteroides plays an important role in the cheese maturation process and contributes a special flavor to the cheese. In addition, the use of leuconostoc mesenteroides in other types of dairy products is also becoming increasingly mature. While leuconostoc mesenteroides subspecies (Leuconostoc mesenteroides subsp.mesencteroides) is a subspecies of leuconostoc mesenteroides, which is classified as one of the safe edible fungi by the ministry of health (bulletin of the health, no. 8 of 2012) and the united states food and drug administration (Food and Drug Administration, FDA). Leuconostoc mesenteroides has not been used in antiallergic applications at present.
Disclosure of Invention
The invention aims to provide a strain or a microbial inoculum of leuconostoc mesenteroides intestinal subspecies with antiallergic effect, and the leuconostoc mesenteroides intestinal subspecies have the antiallergic effect, can relieve anaphylaxis symptoms, reduce histamine content and IgE content, and regulate immune response balance of Th1/Th2, th2/Th17 and the like through verification.
The invention provides the following technical scheme:
use of a strain or agent of the leuconostoc mesenteroides subspecies of the intestinal membrana in the preparation of any one of the following products 1) or 2) or 3) or 4) followed by 5) or 6) or 7) or 8) or 9) or 10):
1) A product for up-regulating Th1 type cytokine levels;
2) A product for reducing the level of cytokines that enhance allergic reactions;
3) A product for reducing IgE content in a body;
4) Product for reducing histamine content
5) A product for improving the structure of the intestinal flora;
6) A product for increasing the content of short chain fatty acids in the intestinal tract;
7) Products for restoring intestinal barrier damage or enhancing intestinal mucosal barrier;
8) A product for alleviating or ameliorating weight loss;
9) Products for improving exercise capacity;
10 A product for antiallergic or allergy symptom relief;
the leuconostoc mesenteroides intestinal subspecies are preserved in the microorganism strain collection of Guangdong province at 10 months and 18 days of 2022, the preservation address is 100 # of Xian Lizhonglu in Guangzhou, and the preservation number is GDMCC No:62899 the microorganism classification is named Leuconostoc mesenteroides subspecies Leuconostoc mesenteroides subsp.
The strain of the leuconostoc mesenteroides intestinal subspecies used in the present invention was designated WHH1141. The in vitro cell test and the mouse animal test prove that the leuconostoc mesenteroides intestinal subspecies strain WH1141 has antiallergic capability, can increase the secretion of IFN-gamma and IL-10 in the spleen cells of the in vitro mice, and up-regulates Th1. Meanwhile, the content of IgE in serum of the OVA sensitized mice can be obviously reduced, the content of short chain fatty acid in feces of the sensitized mice is obviously increased, and histamine, IL-4, IL-5, IL-13 and IL-17 in serum and intestinal tracts of the sensitized mice are obviously reduced, so that the balance of Th1/Th2 immune response and the balance of Th2/Th17 immune response are regulated, the flora restoration is promoted, the allergic reaction symptoms of the mice are effectively relieved, the exercise capacity is improved, the weight reduction degree of the mice is relieved or improved, and the weight of the mice is recovered.
As a preferred feature of the present invention,
the microbial inoculum is a bacterial liquid containing the leuconostoc mesenteroides subspecies intestinal membrane strain,
or the microbial inoculum is a solid microbial inoculum prepared by adding a physiologically acceptable solid carrier and/or auxiliary agent to the leuconostoc mesenteroides subspecies intestinal membrane strain. The solid carrier or the auxiliary agent can be selected from common carriers or auxiliary agents such as cellulose acetate phthalate, hydroxypropyl methyl fiber phthalate, ethyl cellulose, polyethylene glycol, saccharides and the like.
As a preferred aspect of the invention, the up-regulating Th1 cytokine levels is increasing both IFN-gamma and IL-10 levels. It was found that strain WHH1141 was able to promote both IFN- γ and IL-10 production in mouse spleen cells in vitro relative to LGG, with secretion amounts of IFN- γ and IL-10 being 3.24 and 1.22 times that of LGG, respectively, compared to other leuconostoc mesenteroides strains or other lactic acid bacteria strains.
As a preferred aspect of the invention, the reduction in the level of a cytokine that enhances an allergic response is a reduction in the level of one or more of IL-4, IL-5, IL-13 and IL-17. Can effectively regulate the balance of Th2/Th17 immune response and relieve anaphylactic reaction of mice.
As a preferred aspect of the present invention, the improving the intestinal flora structure is increasing the relative abundance of musibaculum jejun_unclassified of bacteroidae or musibaculum spp. Muribaculaceae and Muribaculum spp. Have more positive effects on short chain fatty acid production and recovery of inflammatory response.
And/or, the improving the intestinal flora structure is to reduce the relative abundance of one or more of wart micro bacteria, desulfur vibrio and pseudomonas in the intestinal tract. The wart microzyme, the desulfur vibrio and the pseudomonas can produce toxin and increase organism inflammation.
Preferably, the method for improving the content of the short-chain fatty acid in the intestinal tract comprises the step of improving the content of one or more of butyric acid, acetic acid and propionic acid. Studies have demonstrated that SCFAs can act on type II helper T lymphocytes (Th 2), type II resident lymphocytes (ILC 2), eosinophils, mast cells and basophils to affect the development of allergic diseases. Besides affecting allergy, SCFAs can also enhance intestinal mucosa barrier, especially butyric acid can affect the function of the tight junction protein to maintain intestinal barrier function, and furthermore, SCFAs also have the effects of neutralizing inflammation, inhibiting the growth of conditional pathogenic bacteria, enhancing immunity and the like, so that the increase of short chain fatty acid content, especially the butyric acid content, is not only helpful for relieving allergic reaction, but also has health care effect.
Preferably, the method of the invention comprises the step of recovering the damage of the intestinal barrier or enhancing the intestinal mucosa barrier to improve the integrity of intestinal villi and/or improve the tightness of the intestinal mucosa lamina propria.
As a preferred feature of the present invention,
the product is a pharmaceutical product, a food product, a health product or a dietary nutritional supplement.
As a preferred feature of the present invention,
the food is fermented milk, cheese, milk beverage, milk powder or fermented fruits and vegetables.
A food, pharmaceutical, health product or dietary supplement contains the above strain or inoculant of Leuconostoc mesenteroides subspecies.
The beneficial effects of the invention are as follows:
(1) The leuconostoc mesenteroides intestinal subspecies strain WHH1141 provided by the invention can obviously increase the secretion of IFN-gamma and IL-10 in vitro mouse spleen cells, up-regulate Th1, and simultaneously can obviously reduce the serum IgE of sensitized mice and obviously reduce the histamine in serum and intestinal tracts;
(2) The leuconostoc mesenteroides intestinal subspecies strain WHH1141 provided by the invention obviously improves the expression of immune factors such as IL-4, IL-5, IL-13, IL-17 and the like caused by OVA sensitization, and effectively regulates the immune response balance of Th2/Th17, thereby relieving the anaphylactic reaction of mice;
(3) The leuconostoc mesenteroides intestinal subspecies strain WHH1141 provided by the invention can effectively promote the recovery of intestinal flora of allergic mice, remarkably increase the number of beneficial bacteria such as bacteroides and bacteroides in the intestinal tract, remarkably improve the levels of acetic acid, propionic acid and butyric acid in feces, and the levels of the acetic acid and the butyric acid are recovered to the normal level of the mice, so that the leuconostoc mesenteroides intestinal subspecies strain has a health care effect;
(4) The leuconostoc mesenteroides intestinal subspecies strain WHH1141 provided by the invention can effectively recover the intestinal barrier injury of the OVA sensitized mice, and increase the intestinal defenses and the protective effects;
(5) The leuconostoc mesenteroides intestinal subspecies strain WHH1141 provided by the invention can relieve and improve the weight reduction of mice, strengthen the weight of the mice, and simultaneously can also improve the exercise capacity of the mice and increase the exercise quantity of the mice;
(6) The leuconostoc mesenteroides intestinal subspecies strain WHH1141 provided by the invention can relieve the anaphylactic symptoms of mice, has an antiallergic effect, and shows that the strain has no toxic genes and no toxic or side effects in animal evaluation, and has certain advantages compared with the traditional medicines for treating allergy at present.
Drawings
FIG. 1 is an experimental plan for animals used in the present invention.
Fig. 2 is a graph showing the body weight of mice of leuconostoc mesenteroides intestinal subspecies WHH1141 intervention group, model group and blank group over time.
FIG. 3 is a graph comparing serum IgE levels of mice from Leuconostoc mesenteroides intestinal subspecies WH 1141 intervention, model and blank.
FIG. 4 is a graph comparing histamine levels in serum from mice of Leuconostoc mesenteroides intestinal subspecies WH 1141 intervention group, model group and blank group.
Fig. 5 is a graph comparing histamine levels in the colon of mice in the leuconostoc mesenteroides intestinal subspecies WHH1141 intervention group, model group, and blank group.
Fig. 6 is a graph comparing β diversity of intestinal flora of mice in the leuconostoc mesenteroides intestinal subspecies WHH1141 intervention group, model group and blank control group.
FIG. 7 is a graph comparing the relative abundance of different species of enterobacteria in mice of Leuconostoc mesenteroides WH 1141 intervention group, model group, and blank group: (a) bacteroides relative abundance, (b) desulphurisation mycoides relative abundance, (c) campylobacter relative abundance, (d) wart micro mycoides relative abundance, (e) bacteroidae relative abundance, (f) bacteroides relative abundance, (g) desulphurisation vibriaceae relative abundance, and (h) pseudomonas relative abundance.
FIG. 8 is a graph of the morphology of jejunal tissues of mice in the Leuconostoc mesenteroides intestinal subspecies WH 1141 intervention group, model group and blank group.
Detailed Description
The following is a further description of embodiments of the invention.
Unless otherwise indicated, all materials used in the present invention are commercially available or are commonly used in the art; unless otherwise indicated, the methods in the examples below are all conventional in the art.
First part, leuconostoc mesenteroides intestinal subspecies information for use in the present invention
The leuconostoc mesenteroides with antiallergic effect is obtained by screening fermented yogurt collected near the Ha-side of Hall in Xinjiang by the inventor, and is named as strain WHH1141, deposited in the microorganism strain collection in Guangdong province at 10 months and 18 days in 2022, and deposited at the deposition address of No. 100 in Xian Zhonglu in Guangzhou, and deposited with the number of GDMCC No:62899, identified as belonging to the leuconostoc mesenteroides subsp Leuconostoc mesenteroides subsp.
1. Biological Properties
Morphological features: the bacteria grows into milky white colonies in MRS agar culture medium, and the bacteria are round and smooth in surface. Gram staining is typically positive, and cells are observed under a microscope to be arranged in pairs or in short chains, without flagella, spores and movements;
culturing characteristics: the optimal growth temperature is 30 degrees, and the anaerobic growth is facultative.
2. Identification of strains
2.1 biological identification
For the sequence sequencing of the 16s rRNA gene, the obtained result is subjected to homology comparison analysis with NCBI GenBank database, and the result shows that the nucleic acid sequence similarity of the strain and the leuconostoc mesenteroides subspecies Leuconostoc mesenteroides subsp. The GenBank accession number of the 16s rRNA gene sequence is OQ832598.
API50 CHL system identification of 2.2WHH1141 strain:
WH1141 fermentation brothCentrifugation was performed at 1mL, and the bacterial sludge was resuspended in physiological saline. Adjusting OD 600 OD600 to 5mL of the resuspended bacteria liquid was about 0.5. The supernatant was removed by centrifugation and resuspended in 5mL of API50 medium. Add to API50 plate, 100 μl per well. Incubate at 37℃for 24h and 48h. The results show that the WH 1141 strain can utilize various carbon sources such as D-galactose, D-glucose, D-fructose, D-mannose and the like.
3. Safety of the strain
3.1 genome analysis of Leuconostoc mesenteroides intestinal subspecies WH1141
Extracting genome DNA of the leuconostoc mesenteroides intestinal membrane subspecies WH1141 strain, and completing genome sequencing by utilizing a BGI platform and a Nanopore platform. Assembling the Nanopore data by using Flye, and performing self-error correction on the assembled genome by using Medaka software and the Nanopore platform sequencing data; and further correcting the sequence by using the multipolish and Polca software and combining with high-quality BGI platform sequencing data to obtain a genome completion map. The results showed that the leuconostoc mesenteroides intestinal subspecies WHH1141 genome size was 2.04mb and gc ratio was 37.96%, with the genome containing 12 rRNA and 2026 genes. The setA which is verified by experiments with sequences in the VFDB database is an analysis data set, the sequence similarity is more than or equal to 80%, the coverage is more than or equal to 70% as a screening standard, and the result shows that no virulence related genes are detected. Using database CARD and software RGI, with Perfect and Strict algorithms as screening criteria, the results showed that no antibiotic resistance related genes were detected.
3.2 evaluation of safety of Leuconostoc mesenteroides intestinal subspecies animal experiments samples were administered to animals by both intraperitoneal injection and oral gavage, whereas Leuconostoc mesenteroides intestinal subspecies WH1141 were evaluated for pathogenicity of the different exposure routes to the animals. The results show that the stock solution of the leuconostoc mesenteroides intestinal membrane subspecies WH 1141 culture is injected into the mice in an intraperitoneal mode, the weight of the male mice and the weight of the female mice are not influenced, and the toxic reaction or death of the mice are not caused. The stock solution and the concentrated solution of the intestinal membrane subspecies WH 1141 culture of the leuconostoc mesenteroides are orally infused into the stomach of mice, and have no influence on the body weight of male mice and female mice, and no toxic reaction or death of the mice is observed. The strain has high safety, no toxicity and no pathogenicity.
First part, function of Leuconostoc mesenteroides intestinal subspecies used in the present invention
4. In vitro antiallergic function
Mice were intraperitoneally injected with 1mg/mL OVA, spleen cells from the mice were taken after 7 days, and spleen cell concentration was adjusted to 5X 10 after lysis 6 cells/mL (splenocyte final concentration 2.5X10) 6 cells/mL); after second-generation activation of probiotics, the OD600 was adjusted to 0.50±0.10. Inactivating at 95deg.C for 10min to obtain dead bacterial suspension (final concentration of bacterial liquid is 2.5X10) 7 CFU/mL); the final OVA concentration was 100. Mu.g/mL. After incubation of spleen cells, probiotics and OVA for 3 days, centrifugation was carried out at 2170rpm for 5min, the supernatant was aspirated and the cytokines were detected with ELISA kit. Lactobacillus rhamnosus LGG was used as positive control strain.
IFN-gamma is mainly produced by activated T cells and NK cells, belongs to Th1 type cytokines, and promotes Th cells to differentiate into Th1 when the organism generates anaphylactic reaction, thereby playing a role in inhibiting type I hypersensitivity reaction. IL-10 is an important cytokine in allergic reactions, has a strong immunosuppressive effect, can reduce IgE production, and has obvious changes in allergic diseases. The in vitro cell experiment of the invention detects the change of two cytokines, IFN-gamma and IL-10. Specific assay results are shown in Table 1 below, where IFN-gamma and IL-10 results are presented as ratios to IFN-gamma and IL-10 in LGG samples.
TABLE 1 detection of IFN-gamma and IL-10 secretion by spleen cells in vitro by different strains
As can be seen from the table, the different strains have a greater difference in the levels of cytokines produced by splenocytes in vitro, even though they are all leuconostoc mesenteroides subspecies, strain WHH3417 and strain WHH1559 have a lower ability to promote IFN-gamma and IL-10 production than LGG, strain WH 1098 and strain WH 3440 have a higher ability to promote IFN-gamma production than LGG, but have a lower ability to promote IL-10 production than LGG, and only strain WH 1141 has a higher ability to promote IFN-gamma and IL-10 production than LGG. By comprehensive comparison, the leuconostoc mesenteroides intestinal subspecies WH1141 has the most remarkable effect, and can obviously promote the production of IFN-gamma and IL-10 at the same time, which are 3.24 times and 1.22 times of LGG respectively.
5. Animal test of leuconostoc mesenteroides intestinal subspecies WHH1141
And (3) taking OVA as an allergen, establishing an allergic animal model by using a BALB/c mouse, then giving a sample to be tested to the animal, and detecting the influence of leuconostoc mesenteroides intestinal subspecies WH1141 on the allergic reaction of the mouse. The specific experimental scheme is shown in figure 1.
The invention selects BALB/c mice, and experimental mice are circulated for 12h/12h in a barrier environment at 25+/-1 ℃ and relative humidity of 50+/-5%. After the normal diet of 30 female BALB/c mice was adapted for 7 days, the mice were randomly divided into 3 groups of 10 mice each, which were the blank control group, model group, and Leuconostoc mesenteroides intestinal subspecies WHH1141 intervention group. The sensitization mode of the model group and the leuconostoc mesenteroides intestinal membrane subspecies WH 1141 intervention group is that the volume emulsion of 500 mug/mL OVA and Freund's adjuvant is injected into the abdominal cavity, and each sensitization is carried out 1 time in 0 day and 14 days; the excitation mode is that 50mg OVA is injected orally and the excitation is carried out for 1 time for 28 days, 30 days and 32 days, and 3 times in total. PBS was used for the control group. Weigh weekly. Faeces were collected on day 35, blood was collected from the orbit and the neck was broken and sacrificed. Dissecting, and taking the tissues of lung, colon, jejunum and the like to detect relevant indexes.
5.1 body weight
The results are weighed weekly during the course of the experiment and are shown in figure 2. The blank control group and the leuconostoc mesenteroides intestinal membrane subspecies WH1141 intervention group have no significant difference (p is more than 0.05), and have significant differences (p is less than 0.05) from the model group. This demonstrates that leuconostoc mesenteroides intestinal subspecies WHH1141 post-drying helped restore body weight in allergic mice. Thus, the leuconostoc mesenteroides intestinal subspecies WHH1141 strain has the ability to alleviate or improve weight loss.
5.2 serum IgE:
a significant feature of type I allergic reactions is the elevation of IgE levels in serum, which is generally thought to be caused by Thl and Th2 cell imbalance. After the animal test is finished, the orbit is subjected to blood sampling, the serum is collected by standing and centrifugation, and the change of IgE in the serum is detected, and the result is shown in figure 3.
Compared with the control group, the model group obviously improves the IgE content in the serum of the mice, namely, the model is successfully modeled. The serum IgE content of mice in the leuconostoc mesenteroides intestinal subspecies WHH1141 intervention group is significantly lower than that in the model group. This suggests that intervention by leuconostoc mesenteroides intestinal subspecies WHH1141 can alleviate allergic reactions by reducing IgE levels in the body.
5.3 histamine in serum and colon:
when allergic reaction occurs in the body, the concentration of histamine increases, which causes symptoms such as telangiectasia, increased vascular permeability, smooth muscle contraction, increased secretion of respiratory tract, and blood pressure drop, and is a main medium for causing allergic reaction. The ELISA kit is used for detecting the change of histamine in serum and colon, and the results are shown in figures 4 and 5.
The model group significantly increased the histamine content in the serum and colon of the mice compared to the control group. The histamine content in the serum and colon of mice from the leuconostoc mesenteroides intestinal subspecies WHH1141 intervention group was significantly lower than that of the model group.
5.4 changes in intestinal flora:
total DNA in the feces of each group of mice is extracted, amplified by using a 16S rDNA v3+v4 region primer, sequenced by an upper machine, and data analysis is collected, and the intestinal flora of the mice in a blank control group, a model group and an intestinal flora intervention group of leuconostoc mesenteroides WH1141 are compared. The results show that:
1) No significant differences in alpha diversity between groups; as shown in fig. 6, there was a significant difference in beta diversity between groups (P < 0.001);
2) At the gate level, as shown in fig. 7 (a) - (d), compared with the model group, the intestinal membrane subspecies WHH1141 of leuconostoc mesenteroides interfered with the group, the relative abundance of Bacteroidota of bacteroides, the relative abundance of desulfofobactota, campylobacter, verrucomicrobia, was significantly increased (P < 0.01);
3) Further in the comparison at the family or genus level, as shown in fig. 7 (e) to (h), it was found that the relative abundance of muribaculaceae_unclassified of the reclassified bacteroidae was significantly increased (P < 0.01) after the WHH1141 dry prognosis of leuconostoc mesenteroides subspecies compared to the model group; meanwhile, the relative abundance increases significantly (P < 0.01) and is close to the level of the blank group; the relative abundance of Desulfovibrio and Pseudomonas of the Vibrionaceae family was significantly reduced (P < 0.01) and was close to the level of the blank pair group. However, muribaculaceae and Muribaculum spp have more positive effects on short chain fatty acid production and recovery of inflammatory reaction; the wart microzyme, the desulfur vibrio and the pseudomonas can produce toxin and increase organism inflammation.
Therefore, the WH1141 strain can effectively improve the intestinal flora structure of the allergic mice, partial flora is close to that of blank control mice, the abundance of intestinal beneficial bacteria such as bacteroides is increased, the abundance of harmful bacteria such as wart micro bacteria, vibrio desulphurisation, pseudomonas and the like is obviously reduced, and the recovery of allergic symptoms of the mice is facilitated.
5.5 changes in short chain fatty acids in feces:
the SCFAs act on type II helper T lymphocytes (Th 2), type II resident lymphocytes (ILC 2), eosinophils, mast cells, basophils and other various allergic effector cells to affect the occurrence of allergic diseases, and the levels of the SCFAs in the allergic body are significantly reduced. SCFAs can also enhance intestinal mucosal barriers, particularly butyric acid, which can affect the function of tight junctions, including promoting the transfer of occludin and Zoneulin-1 from the cytoplasm to tight junctions, inhibiting the expression of permeabilized Claudin-2, and increasing transepithelial resistance in vitro.
The invention uses high performance liquid phase to detect short chain fatty acid in the feces, and the result is shown in Table 2.
TABLE 2 short chain fatty acid content in the faeces of mice of each group
From the above table, the dry prognosis of leuconostoc mesenteroides intestinal subspecies WHH1141 showed a very significant increase in the content of acetic acid, propionic acid and butyric acid compared to the model group. The dry prognosis of the leuconostoc mesenteroides intestinal membrane subspecies WH 1141 is illustrated, the content of the intestinal short-chain fatty acid can be obviously improved, the intestinal short-chain fatty acid (acetic acid and butyric acid) level of a normal mouse intestinal is approached, the intestinal mucosa barrier is enhanced, and the allergic symptoms are relieved.
5.6 cytokines in serum and colon:
IL-4 produced by Th2 cells is an important regulator in IgE synthesis, and can promote T, B cell proliferation and induce adhesion molecule expression. IL-5 can cooperate with IL-4 to stimulate B cells to synthesize IgE and induce eosinophil activation, proliferation, differentiation and migration; inhibit eosinophil apoptosis and prolong the survival period. IL-13 is a cytokine secreted by activated Th2 cells, and is mainly involved in T, B cell development, regulating Th1 and Th2 cell functions, promoting IgE synthesis, inducing EOS migration, and playing an important role in the pathogenesis of allergic diseases. Studies suggest that IL-13 may have a greater driving effect in allergic reactions than IL-4.IL-13 can down regulate secretion of IFN-gamma and other Thl cytokines, and participate in maintaining expression of Th2 cytokines, so that Th1/Th2 balance is inclined to Th2 response. IL-17 is produced primarily by T helper cells TH 17. IL-17 can directly or indirectly induce various cytokines, chemokines, inflammatory factors and antimicrobial proteins to identify target genes mediating autoimmune and chronic infections, and is involved in the induction of allergic diseases such as asthma, mediating the inflammatory processes thereof.
After the animal test is finished, the orbit is subjected to blood collection, the orbit is subjected to standing and centrifugation to collect serum, colon tissues are taken, and ELISA (enzyme-linked immunosorbent assay) kits are used for detecting the changes of various cytokines in the serum and the colon, and the results are shown in Table 3.
TABLE 3 serum and colon cytokine levels in mice of each group
Compared with the control group, the model group obviously improves the content of IL-4, IL-5, IL-13 and IL-17 in the serum and colon of the mice. The intestinal membrane leuconostoc mesenteroides WH 1141 dry prognosis shows that the content of IL-4, IL-5, IL-13 and IL-17 is extremely remarkably reduced (p < 0.001) compared with the model group. The leuconostoc mesenteroides intestinal membrane subspecies WH1141 dry prognosis can obviously reduce the level of cytokines for enhancing allergic reaction and effectively regulate the balance of Th2/Th17 immune response.
5.7 jejunal tissue morphology
As shown in fig. 8, the jejunal mucosa of the mice in the blank group is complete and smooth, the villi is complete, and the lamina propria is compact. Compared with the blank control group, the jejunum of the model group mice shows obvious villus shedding, loose lamina propria and reduced intestinal barrier protection capability. Compared with the model group, the jejunum villus of the mice with the leuconostoc mesenteroides intestinal subspecies WH1141 after dry prognosis is completed and the lamina propria is compact. It is demonstrated that leuconostoc mesenteroides intestinal membrane subspecies WH1141 can restore intestinal barrier damage of OVA sensitized mice and increase intestinal defenses and protective effects.
5.8 improvement of allergic symptoms
An allergic animal model was established for BALB/c mice using OVA as allergen. The mice were observed for allergic symptoms scoring during the course of the test for status, hair, vitality. The score is 0-5, 0 means that the mice are in good state, the hair is smooth, the vitality is normal, and the mice have no allergic symptoms. A score of 5 indicates that the mice were in a very poor state, unresponsive after stimulation, and messy hair. The specific allergy symptoms scores are shown in table 4.
Table 4 allergic symptoms score for each group of mice
From the above table, the hair of the model group mice was remarkably erected, the exercise amount was reduced, and the exercise ability was lowered. Compared with the model group, the leuconostoc mesenteroides intestinal membrane subspecies WH1141 dry prognosis mice have smoother hair, increased exercise and improved exercise capacity. It is demonstrated that the intervention of the leuconostoc mesenteroides intestinal subspecies WH1141 can significantly improve the allergic symptoms of mice.
It should be understood that the foregoing description is only of the preferred embodiments of the invention and is not intended to limit the invention in any way, and modifications and variations may be made in accordance with the above teachings, all of which are intended to fall within the purview of the appended claims.
Claims (10)
1. Use of a strain or agent of the leuconostoc mesenteroides subspecies intestinal membrane for the preparation of any one of the following products 1) or 2) or 3) or 4) or 5) or 6) or 7) or 8) or 9) or 10):
1) A product for up-regulating Th1 type cytokine levels;
2) A product for reducing the level of cytokines that enhance allergic reactions;
3) A product for reducing IgE content in a body;
4) Product for reducing histamine content
5) A product for improving the structure of the intestinal flora;
6) A product for increasing the content of short chain fatty acids in the intestinal tract;
7) Products for restoring damage to or enhancing the intestinal mucosal barrier;
8) A product for alleviating or ameliorating weight loss;
9) Products for improving exercise capacity;
10 A product for antiallergic or allergy symptom relief;
the leuconostoc mesenteroides intestinal subspecies are preserved in the microorganism strain collection of Guangdong province at 10 months and 18 days of 2022, the preservation address is 100 # of Xian Lizhonglu in Guangzhou, and the preservation number is GDMCC No:62899 the microorganism classification is named Leuconostoc mesenteroides subspecies intestinal membraneLeuconostoc mesenteroides subsp. Mesenteroides。
2. The use according to claim 1, wherein,
the microbial inoculum is a bacterial liquid containing the leuconostoc mesenteroides subspecies intestinal membrane strain,
or the microbial inoculum is a solid microbial inoculum prepared by adding a physiologically acceptable solid carrier and/or auxiliary agent to the leuconostoc mesenteroides subspecies intestinal membrane strain.
3. The use according to claim 1, wherein,
the up-regulation of Th1 type cytokine levels is simultaneous increase of IFN-gamma and IL-10 levels.
4. The use according to claim 1, wherein,
the reduction in the level of cytokines that enhance the allergic response is a reduction in the level of one or more of IL-4, IL-5, IL-13, and IL-17.
5. The use according to claim 1, wherein,
the intestinal flora improving structure is to increase the unclassified bacteroidae in intestinal tractsMuribaculaceaeUnclassified and/or BacteroidesMuribaculum The relative abundance of spp;
and/or, the improving the intestinal flora structure is to reduce the relative abundance of one or more of wart micro bacteria, desulfur vibrio and pseudomonas in the intestinal tract.
6. The use according to claim 1, wherein,
the content of the short-chain fatty acid in the intestinal tract is increased by one or more of butyric acid, acetic acid and propionic acid.
7. The use according to claim 1, wherein,
the recovery of damage to or enhancement of the intestinal mucosal barrier is to increase intestinal villus integrity and/or to increase the intimacy of the intestinal mucosal lamina propria.
8. The use according to claim 1, wherein,
the product is a pharmaceutical product, a food product, a health product or a dietary nutritional supplement.
9. The use according to claim 8, wherein,
the food is fermented milk, cheese, milk beverage, milk powder or fermented fruits and vegetables.
10. A food, pharmaceutical, nutraceutical or dietary nutritional supplement comprising the strain or inoculant of the leuconostoc mesenteroides subspecies intestinal membrane of claim 1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310635579.8A CN116889578A (en) | 2023-05-31 | 2023-05-31 | Application of strain or microbial inoculum of leuconostoc mesenteroides intestinal subspecies with antiallergic effect |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310635579.8A CN116889578A (en) | 2023-05-31 | 2023-05-31 | Application of strain or microbial inoculum of leuconostoc mesenteroides intestinal subspecies with antiallergic effect |
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| Publication Number | Publication Date |
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| CN116889578A true CN116889578A (en) | 2023-10-17 |
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| CN202310635579.8A Pending CN116889578A (en) | 2023-05-31 | 2023-05-31 | Application of strain or microbial inoculum of leuconostoc mesenteroides intestinal subspecies with antiallergic effect |
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