CN115154487B

CN115154487B - Polysaccharide salt fog mixture containing probiotics fermentation liquor and used for protecting respiratory tract and application thereof

Info

Publication number: CN115154487B
Application number: CN202211015334.7A
Authority: CN
Inventors: 王静; 王晓娟
Original assignee: Jiangxi Salt Selection Technology Co ltd
Current assignee: Jiangxi Salt Selection Technology Co ltd
Priority date: 2022-08-24
Filing date: 2022-08-24
Publication date: 2023-04-21
Anticipated expiration: 2042-08-24
Also published as: CN115154487A

Abstract

The invention discloses a polysaccharide salt fog mixture containing probiotics fermentation liquor and used for protecting respiratory tract and application thereof. The main components of the polysaccharide in the invention are methyl hesperidin, modified lentinan and linalool oxide. The invention discovers that the methyl modified hesperidin is more beneficial to the absorption of human bodies and has better protective effect on respiratory tracts. The modified lentinan has better anti-tumor and antibacterial effects. The probiotics fermentation liquor and methyl hesperidin, the sulfated modified lentinan and linalool oxide have a certain synergistic effect, so that the effects of antivirus, antivirus protection and respiratory tract protection are more effective and durable.

Description

Polysaccharide salt fog mixture containing probiotics fermentation liquor and used for protecting respiratory tract and application thereof

Technical Field

The invention belongs to the technical field of biology, and relates to a polysaccharide mixture for protecting respiratory tract, in particular to a polysaccharide mixture for protecting respiratory tract and a salt fog mixture thereof.

Background

In recent years, the prevalence rate of lung diseases such as slow lung obstruction, asthma and the like is high due to bad living habits, environmental pollution and the like; large-scale outbreaks of respiratory infections such as human avian influenza and influenza a also present new challenges for the prevention and treatment of respiratory diseases. The mucosal immune system, which is the first line of defense of the body, plays an important role in resisting invasion of pathogenic microorganisms, and thus, related research on respiratory tract mucosal immunity is becoming one of the hot spots in clinical and scientific research. The research on the relationship between traditional Chinese medicine and respiratory tract mucosa immunity is also increasing, and scholars often study the relationship between traditional Chinese medicine and respiratory tract mucosa immunity from the approaches of opening and bending the lung to the nose, and the exterior and interior of the lung and large intestine.

As a common means for treating respiratory diseases, aerosol inhalation therapy is a technique for obtaining therapeutic effects by atomizing bittern water by special equipment and then inhaling the bittern water by physiotherapy people, and has ideal therapeutic effects. The basic principle of aerosol inhalation therapy is: the medical atomizer deagglomerates and atomizes the powder into powder aerosol, which is inhaled by the patient to reach the focus position. Compared with the administration modes such as oral administration, injection and the like, the aerosol inhalation therapy has the advantages of rapid drug effect, small toxic and side effects and wide applicable population.

Methyl hesperidin is a natural flavonoid compound derived from fruits and vegetables, but the content of methyl hesperidin in plant seeds is very small, and most of the methyl hesperidin is modified. The prior literature proves that the methyl hesperidin has excellent antibacterial, antiviral and antifungal effects, but no literature currently proves the effect on respiratory tract infection. The lentinan has the effects of improving immunity, resisting tumor and the like, the biological activity of the lentinan and the derivatives thereof is closely related to the molecular structure of the lentinan, and the sulfated lentinan has high water solubility and obvious anti-tumor effect.

A large number of experiments prove that the methyl hesperidin and the modified lentinan are matched, so that the compound has obvious effect of resisting respiratory tract infection. The salt fog mitigant is prepared by combining the advantages of sulfated lentinan and methyl hesperidin and linalool oxide easy to absorb. The salt fog mixture can not only influence the secretion of bronchus, but also reduce the formation of condensed secretion in the tube, regulate the surface active substances of alveoli and improve the surface tension of alveoli. The probiotics fermentation liquor has certain antibacterial and bactericidal capacity. The salt particles can dehydrate the bacterial protein structure, and have the functions of sterilizing, resisting inflammation, dissolving phlegm, improving the ventilation condition of the trachea, and the like.

The invention aims to provide a mixture of salt fog and probiotics fermentation supernatant for protecting respiratory tract, which has the effect of antivirus and protecting respiratory tract.

Disclosure of Invention

In order to achieve the above purpose, the present invention adopts the following technical scheme:

a polysaccharide mixture for protecting respiratory tract and a salt mist mixture thereof, which are characterized in that: according to the mass percentage, the microbial fermentation liquid comprises 4-5% of methyl hesperidin, 5-10% of modified lentinan, 0.5-2% of linalool oxide, 10-20% of salt solution and 20-30% of probiotics fermentation supernatant.

Preferably, the modified lentinan is sulfated lentinan.

The salt according to the invention may be pure sodium chloride or sea salt containing other minerals.

The probiotics used in the present invention are streptococcus thermophilus, which is considered a "generally accepted safety (GRAS)" component, widely used in the production of some important fermented dairy products, including yogurt and cheese. Streptococcus thermophilus also has some functional activities such as the production of extracellular polysaccharides, bacteriocins and vitamins. In addition, streptococcus thermophilus can also be used as potential beneficial bacteria, and experiments prove that the streptococcus thermophilus has transport activity and certain gastrointestinal adhesiveness.

The invention discovers that the streptococcus thermophilus fermentation liquor can have synergistic effect with methyl hesperidin, modified lentinan and linalool oxide, so that the streptococcus thermophilus fermentation liquor has certain effects of protecting respiratory tract, resisting virus and resisting bacteria. Presumably, the reason is that sulfated lentinan has an effect of enhancing immunity, whereas methyl hesperidin is capable of maintaining normal permeability of blood vessels, so that the immunity of modified lentinan is enhanced. In addition, the fermentation liquor of streptococcus thermophilus contains a large amount of substances such as vitamins, and the methyl hesperidin can enhance the vitamin capacity, so that the effect of enhancing the immunity can be achieved. The experiment also carries out experiments of lentinan, methyl hesperidin and probiotics fermentation liquor, and discovers that the water solubility of the non-modified lentinan is poor, and the lentinan is difficult to be matched with the probiotics fermentation liquor and the methyl hesperidin to play a better role.

The invention prepares the mixture of the probiotics, the methyl hesperidin and the sulfated lentinan into salt mist, mixed solution and the like, and discovers that the salt mist mixture can better act on the respiratory tract and better protect the respiratory tract.

The salt fog mixture prepared by the invention can be atomized by special atomizing equipment for physiotherapy of people, or concentrated and soaked into a porous medium such as sponge to prepare a chip which is placed in a mask for physiotherapy.

The preparation method of the polysaccharide and the salt fog mixture thereof comprises the following steps:

s1, weighing 30-50g of hesperidin, adding 200-400mL of water into a three-neck flask, stirring to form a suspension, then adding 10-20g of calcium hydroxide, slowly dropwise adding 10-15mL of dimethyl carbonate, reacting at the reaction temperature of 30-60 ℃ for 6 hours, extracting with n-butanol for 2 times, combining organic phases, washing with water for 1 time, washing with saturated salt for 1 time, recovering n-butanol under reduced pressure to obtain a crude methyl hesperidin product, adding 240mL of ethanol with the mass fraction of 95%, heating to 60 ℃ for dissolution, filtering while the crude methyl hesperidin product is hot, cooling and crystallizing the filtrate at room temperature for 2 days, filtering, and drying at 80 ℃ to obtain orange yellow crystal powder, namely methyl hesperidin.

S2, placing 10mL of anhydrous pyridine into a 500mL round bottom flask, cooling to 0 ℃ in an ice-water bath, slowly adding 20mL of chlorosulfonic acid under magnetic stirring, continuously stirring at room temperature for 30min, sealing, and refrigerating in a refrigerator.

S3, dissolving a certain amount of lentinan in 40mLN, N-dimethylformamide, stirring for 30-60min at room temperature, adding an esterification reagent, and stirring for 2-3h in a water bath at 60 ℃. After the reaction is finished, adding precooled redistilled water to dilute the solution, adding sodium hydroxide to neutralize the solution, centrifuging, dialyzing with running water for 3 days, dialyzing with distilled water for 1 day, concentrating by rotary evaporation, adding absolute ethyl alcohol, placing at the temperature of 4 ℃ for alcohol precipitation for 24 hours, centrifuging, and freeze-drying to obtain white sulfated lentinan.

S4, placing linalool and formic acid in a water bath, controlling the reaction temperature in the water bath to be 60-65 ℃, then dropwise adding 30% aqueous hydrogen peroxide solution for 30min, controlling the temperature in the dropwise adding process to be 65-70 ℃, preserving heat and stirring for 2h after dropwise adding, cooling, adding saturated aqueous sodium chloride solution, extracting with benzene, combining extract liquid, washing the extract liquid with 10% aqueous sodium carbonate solution and water to be neutral, and drying with anhydrous sodium sulfate; removing benzene residues by evaporation after drying, then distilling under reduced pressure, and collecting fractions with the following boiling ranges at 799.8Pa, wherein the temperature ranges are 70-74 ℃ (I); 75-79 ℃ (II); 80-84 ℃ (III); 85-102 ℃ (IV); and (3) taking fraction (II), namely linalool oxide prepared by the method.

S5, taking an environment-friendly hidden streptococcus thermophilus, inoculating the streptococcus thermophilus to a TPY culture medium, performing anaerobic culture for 24 hours at 40 ℃, selecting single bacterial colony with good growth vigor, inoculating the single bacterial colony to the TPY culture medium for activation, and repeating the steps twice.

S6, inoculating the activated strain into a culture medium according to the volume ratio of 4%, standing and culturing for 8 hours in an anaerobic incubator at 40 ℃, filtering, centrifuging for 20 minutes at 3000r/min, and taking supernatant for ultraviolet sterilization to obtain the probiotic fermentation liquid.

S7, adding the prepared mixture of the methyl hesperidin and the modified lentinan into the probiotics fermentation liquor according to a certain mass ratio, and then adding a salt solution to fully mix the probiotics fermentation liquor with the salt solution to obtain the prepared salt fog mixture.

S8, atomizing the prepared salt mist mixture through professional atomizing equipment, or concentrating the salt mist mixture, soaking a mask filter element by using the salt mist mixture, and placing the mask filter element into a mask for physiotherapy.

Compared with the prior art, the invention has the beneficial effects that:

1. the salt fog and the probiotics fermentation liquor have a certain synergistic effect, so that the effect of protecting the respiratory tract is more obvious. The streptococcus thermophilus fermentation liquor can have synergistic effect with methyl hesperidin and modified lentinan, so that the streptococcus thermophilus fermentation liquor has certain effects of protecting respiratory tract, resisting virus and resisting bacteria. Presumably, the reason is that sulfated lentinan has an effect of enhancing immunity, whereas methyl hesperidin is capable of maintaining normal permeability of blood vessels, so that the immunity of modified lentinan is enhanced. In addition, the fermentation liquor of streptococcus thermophilus contains a large amount of substances such as vitamins, and the methyl hesperidin can enhance the vitamin capacity, so that the effect of enhancing the immunity can be achieved.

2. The salt fog mixture prepared by the invention has good water solubility and is easier to absorb.

3. The salt fog mixture prepared by the method is not applicable to toxic and harmful components in the preparation process, and is safer.

4. As the fermentation liquor of streptococcus thermophilus contains a large amount of vitamin substances, and linalool oxide can have synergistic effect with vitamin E and other components, the antioxidation effect is enhanced.

Drawings

FIG. 1 is a graph showing cell viability of examples 1-4, comparative examples 1-6 and negative control groups prepared according to the present invention.

FIG. 2 shows the secretion amount of TNF-alpha pro-inflammatory factor by examples 1-4, comparative examples 1-6 and blank control, lipopolysaccharide control group prepared according to the present invention

FIG. 3 shows the secretion amount of IL-6 pro-inflammatory factor by examples 1-4, comparative examples 1-6, and blank and lipopolysaccharide control groups prepared according to the present invention.

FIG. 4 shows the secretion amount of IL-1β pro-inflammatory factor by examples 1-4, comparative examples 1-6, and blank and lipopolysaccharide control groups prepared according to the present invention.

FIG. 5 shows the molecular structure of methyl hesperidin prepared in example 3 of the present invention.

FIG. 6 is an infrared spectrum of the modified lentinan prepared in example 4 of the present invention.

FIG. 7 shows linalool oxide prepared according to the present invention.

FIG. 8 is a hydrogen spectrum of linalool oxide prepared according to the present invention.

FIG. 9 is a graph of the carbon spectrum of linalool oxide prepared according to the present invention.

Detailed Description

The present invention will be described in further detail with reference to specific embodiments in order to make the objects, technical solutions and advantages of the present invention more apparent. It should be understood that the specific embodiments described herein are for purposes of illustration only and are not intended to limit the scope of the invention. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.

Example 1

S1, weighing 30g of hesperidin, adding 200mL of water into a three-neck flask, stirring, adding 10g of calcium hydroxide, slowly dropwise adding 10mL of dimethyl carbonate, reacting at the reaction temperature of 30 ℃ for 6 hours, extracting 2 times with n-butanol, merging organic phases, washing with water for 1 time, washing with saturated salt for 1 time, recovering n-butanol under reduced pressure to obtain a crude product of methyl hesperidin, adding 240mL of ethanol with the mass fraction of 95%, heating to 60 ℃ for dissolution, filtering while the crude product is hot, cooling and crystallizing the filtrate at room temperature for 2 days, filtering, and drying at 80 ℃ to obtain orange yellow crystal powder, namely methyl hesperidin.

S3, dissolving a certain amount of lentinan in 40mL of N, N-dimethylformamide, stirring for 30min at room temperature, adding an esterification reagent, and stirring for 2h in a water bath at 60 ℃. After the reaction is finished, adding precooled redistilled water to dilute the solution, adding sodium hydroxide to neutralize the solution, centrifuging, dialyzing with running water for 3 days, dialyzing with distilled water for 1 day, concentrating by rotary evaporation, adding absolute ethyl alcohol, placing at the temperature of 4 ℃ for alcohol precipitation for 24 hours, centrifuging, and freeze-drying to obtain white sulfated lentinan.

S4, placing linalool and formic acid in a water bath, controlling the reaction temperature in the water bath to be 60 ℃, then dropwise adding 30% aqueous hydrogen peroxide solution for 30min, controlling the temperature in the dropwise adding process to be 65 ℃, keeping the temperature and stirring for 2h after the dropwise adding is finished, adding saturated aqueous sodium chloride solution after cooling, extracting with benzene, combining extract liquid, washing the extract liquid with 10% aqueous sodium carbonate solution and water to be neutral, and drying with anhydrous sodium sulfate; removing benzene residues by evaporation after drying, then distilling under reduced pressure, and collecting fractions with the following boiling ranges at 799.8Pa, wherein the temperature ranges are 70-74 ℃ (I); 75-79 ℃ (II); 80-84 ℃ (III); 85-102 ℃ (IV); and (3) taking fraction (II), namely linalool oxide prepared by the method.

S7, mixing the prepared methyl hesperidin, the modified lentinan, linalool oxide, a salt solution, a probiotics fermentation supernatant and water according to the mass ratio: 4%, 5%, 0.6%, 10%, 20%, 60.4% to obtain salt fog mixture.

Example 2

The preparation procedure of example 2 is substantially the same as that of example 1, except that the mass ratio of the salt spray mixture in step S7 is: 5% of methyl hesperidin, 10% of modified lentinan, 1.0% of linalool oxide, 20% of salt solution, 30% of probiotics fermentation supernatant and 34% of water are mixed.

Example 3

The preparation procedure of example 3 is substantially the same as that of example 1, except that the mass ratio of the salt spray mixture in step S7 is: mixing methyl hesperidin 4%, modified lentinan 10%, linalool oxide 1.0%, salt solution 20%, probiotic fermented supernatant 20%, and water 45%.

Example 4

The preparation procedure of example 4 was substantially the same as that of example 1, except that the mass ratio of the salt spray mixture in step S7 was: 5% of methyl hesperidin, 8% of modified lentinan, 0.6% of linalool oxide, 15% of salt solution, 13% of probiotics fermentation supernatant and 58.4% of water.

Comparative example 1

Comparative example 1 was prepared in substantially the same manner as in example 1, except that no sea salt and no probiotic fermented liquid were added in step S7.

Comparative example 2

Comparative example 2 was prepared in substantially the same manner as in example 1, except that methyl hesperidin and the probiotic fermented liquid were not added in step S7.

Comparative example 3

Comparative example 3 was substantially the same as the preparation procedure of example 1, except that methyl hesperidin and sea salt were not added in step S7.

Comparative example 4

Comparative example 4 was substantially the same as the preparation procedure of example 1, except that no sea salt was added in step S7.

Comparative example 5

Comparative example 5 was substantially the same as the preparation procedure of example 1, except that methyl hesperidin was not added in step S7.

Comparative example 6

Comparative example 6 was substantially identical to the preparation procedure of example 1, except that no probiotic fermented liquid was added in step S7.

1. Cytotoxicity assays

Observing the growth state of the cells, scraping the cells when the cells grow logarithmically, centrifuging, counting the cells, and regulating the concentration to a final concentration of 4×10 ⁵ Cells/ml and were plated at a volume of 100 μl per well into 96 well plates overnight. The samples prepared in the experiment are dissolved by a proper amount of dimethyl sulfoxide with the concentration less than or equal to 0.1%, the DMEM is used for preparing a sample stock solution with the concentration of 20 mug/ml, a negative control group and a sample group are arranged, and 3 parallel holes are formed in each group. After overnight incubation, 50 μl (5 mg/mL) of MTT was added, the supernatant was spun off after 4 hours, and dimethyl sulfoxide was added to the wells, and the absorbance was measured at 570nm at 150 μl per well with no statistical difference between the groups.

As can be seen from FIG. 1, the examples prepared according to the present invention and the comparative examples are nontoxic to cells. From the data, it can be seen that the cell viability of the examples is higher than that of the comparative examples, which is in close and inseparable relationship with methyl hesperidin, modified lentinan, and the like.

2. Influence of sample induced IL-1 beta, TNF-alpha, IL-6 cytokine secretion

Observing the growth state of the cells until the cells grow to a concentration of about 4×10 per milliliter ⁵ Cells were plated onto 24-well plates at 500 μl per well overnight. The experiments were performed in groups, i.e. blank, lipopolysaccharide and sample. The sample set was added with the sample stock prepared according to the present invention and supplemented to 250. Mu.L with DMEM to a final concentration of 2. Mu.g/ml. The other groups were added with an equal amount of DMEM, while each experimental group was given three parallel wells. One hour after administration, 250. Mu.l of lipopolysaccharide (1. Mu.g/ml) was added to the blank, lipopolysaccharide and sample groups, and after 12 hours of incubation, the supernatant was split-packed and stored at-80℃for each cytokine measurement without statistical differences between groups.

The reaction strips were removed from the sealed bags, 20-fold dilutions of primary antibodies were added to the corresponding wells with coating solution, 100 μl per well, the plates sealed with a sealing plate membrane, and overnight at 4 ℃. The liquid in the plate is thrown off, and the plate is washed four times by washing liquid. 200. Mu.L of the sample diluent was added, the plate was sealed, and the plate was allowed to stand at room temperature and shaken for 1 hour. The liquid in the plate is thrown off, and the plate is washed four times by washing liquid. 100 μl of diluted IL-1β, TNF- α, IL-6 standard and test sample were added, respectively, and the mixture was shaken at room temperature for two hours. The liquid in the plate is thrown off, and the plate is washed four times by washing liquid. 200-fold dilutions of IL-1. Beta., TNF-. Alpha., IL-6 were added, respectively, and 100. Mu.l per well was shaken at room temperature for one hour. The liquid in the plate is thrown off, and the plate is washed four times by washing liquid. Adding 100 μl of horseradish peroxidase, shaking for 30min at normal temperature to remove liquid in the plate, and washing the plate with washing solution for five times. 3,3', 5' -tetramethyl benzidine substrate chromogenic solution was added, and developed for 15 minutes in a dark place at 100ml per well. 100. Mu.L of stop solution was added. OD (450 nm) of each reaction well was measured by an enzyme-labeled instrument. And (5) making a standard curve according to the standard substance of each concentration and the corresponding OD value. And calculating the concentration of the corresponding cytokine in each sample to be tested according to the standard curve.

The test results are shown in the following table:

TABLE 1

TABLE 2

Lipopolysaccharide can enter the body, and induce the expression of related protein genes in inflammatory signaling pathways of the body. TNF-alpha, IL-6, IL-1 beta are pro-inflammatory factors which in turn induce secretion of a number of inflammatory mediators, such as prostaglandins, nitric oxide, leukotrienes, etc., which induce a "waterfall effect".

FIG. 1 is a graph of cell viability for the negative control group and the sample group. From the figures, it can be seen that the survival rates of the example group prepared by the invention and the blank negative control group are not obviously different, so that the samples prepared by the invention are nontoxic and harmless to cells. In addition, it can be seen that the samples prepared by the invention have a certain promotion effect on the growth of cells.

FIGS. 2, 3 and 4 show the amounts of three pro-inflammatory factors TNF-alpha, IL-6 and IL-1β secreted by the samples prepared according to the present invention and the blank and lipopolysaccharide control groups, respectively. When lipopolysaccharide enters the body, it can induce the release of many active substances such as cytokine, proinflammatory factor, etc., such as IL-1β, TNF- α, IL-6, etc. IL-6 is an important and long-acting cytokine that stimulates effector cells to release inflammatory mediators and acute phase proteins, which play an important role in the homeostasis of the hematopoietic system and in neuroendocrine and bone metabolism, and is secreted in increased amounts in the body under conditions of infection, injury, and other stresses. Such deregulated high concentrations of IL-6 may trigger a series of inflammatory processes. TNF is associated with heat production and promotes the expression of other inflammatory mediators, which play a critical role in the inflammatory process, and TNF- α products are also believed to be associated with the induction of other pro-inflammatory mediators and can damage blood vessels at the site of inflammation, causing microvascular hemorrhagic responses. IL-1β is highly expressed in alveolar lavage fluid of patients with acute lung injury animal models and respiratory distress syndrome, and can stimulate effector cells to produce a large amount of cytokines such as IL-8, monocyte chemotactic protein and macrophage inflammatory protein, and cause aggregation of inflammatory cells such as leucocytes, mononuclear macrophages and the like. IL-1β can also cause epithelial cell damage and has the potential to cause pulmonary fibrosis, and can also increase vascular permeability, cause alveolar surfactant abnormalities, and exacerbate pulmonary edema.

As can be seen from FIGS. 2, 3 and 4, the content of IL-1 beta, TNF-alpha and IL-6 in the lipopolysaccharide group reached 18.12ng/ml, 139.52. Mu.g/ml and 12.32. Mu.g/ml, but the content of IL-1 beta, TNF-alpha and IL-6 in examples 1-4 was greatly reduced, which indicates that the anti-inflammatory effect of the example group was remarkable. The reason is probably that the methyl hesperidin and the modified lentinan in the example group have a synergistic effect and can inhibit the expression of some protein genes, thereby inhibiting the secretion of some pro-inflammatory factors in cells. Although the content of IL-1 beta, TNF-alpha and IL-6 in comparative examples 1-6 was decreased, the decrease was small, and it was difficult to exert the effects of anti-inflammatory and protecting the respiratory tract. Meanwhile, by comparing the examples with the comparative examples, it can be seen that the substances in the examples have good synergistic effect.

As can be seen from the figure, the sample prepared by the invention has good anti-inflammatory effect. It is demonstrated that the samples prepared according to the present invention can attenuate the occurrence of inflammation by inhibiting the secretion of cytokines. In addition, the comparative sample prepared by the invention has weak resistance to inflammation, but the sample of the example has good resistance to inflammation, which shows that the interaction between the methyl hesperidin and the modified lentinan, the probiotic fermentation liquor and the salt fog exists in the example prepared by the invention, so that the samples have good effects of resisting inflammation and protecting respiratory tract.

Finally, it should be noted that: the foregoing description is only illustrative of the preferred embodiments of the present invention, and although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that modifications may be made to the embodiments described, or equivalents may be substituted for elements thereof, and any modifications, equivalents, improvements or changes may be made without departing from the spirit and principles of the present invention.

Claims

1. A polysaccharide salt fog mixture containing probiotics fermentation liquor and used for protecting respiratory tract, which is characterized by comprising the following components in percentage by mass: 4-5% of methyl hesperidin, 5-10% of modified lentinan, 0.5-2% of linalool oxide, 10-20% of salt solution, 20-30% of probiotics fermentation supernatant and the balance of water; the modified lentinan is obtained by performing sulfuric acid esterification treatment on lentinan; the probiotics are streptococcus thermophilus;

the salt is pure sodium chloride or sea salt containing other minerals;

the preparation method of the fermentation broth comprises the following steps: s1, taking an environment-friendly hidden streptococcus thermophilus, inoculating the streptococcus thermophilus on a TPY culture medium, performing anaerobic culture, selecting a single colony with good growth vigor, inoculating the single colony into the TPY culture medium for activation, and repeating the steps twice; s2, inoculating the activated strain into a culture medium according to the volume ratio of 4%, standing and culturing in an anaerobic incubator, filtering, centrifuging, and taking supernatant for ultraviolet sterilization to obtain the probiotic fermentation liquid.

2. The polysaccharide salt mist mixture for protecting respiratory tract comprising probiotic fermented liquid according to claim 1, wherein the preparation method of methyl hesperidin is as follows: weighing hesperidin, adding 200-400mL of water into a three-neck flask, stirring to form a suspension, then adding calcium hydroxide, slowly dropwise adding dimethyl carbonate, reacting at 30-60 ℃, extracting 2 times with n-butanol, merging organic phases, washing 1 time with water, washing 1 time with saturated common salt, recovering n-butanol under reduced pressure to obtain a methyl hesperidin crude product, adding 240mL of ethanol with the mass fraction of 95%, heating to dissolve, filtering while the crude product is hot, cooling filtrate at room temperature for crystallization for 2 days, filtering, and drying.

3. A polysaccharide salt mist mixture for respiratory tract protection comprising a probiotic fermented liquid according to claim 1, characterized in that the sulfated treatment of lentinan is: dissolving lentinan in N, N-dimethylformamide, stirring at room temperature for 30-60min, adding an esterification reagent, stirring in a water bath for 2-3h, adding precooled distilled water to dilute the solution after the reaction is finished, adding sodium hydroxide to neutralize the solution, centrifuging, dialyzing with running water, dialyzing with distilled water, rotationally evaporating and concentrating, adding absolute ethyl alcohol, precipitating with ethanol at 4 ℃, centrifuging, and freeze-drying to obtain white sulfated lentinan.

4. The polysaccharide salt mist mixture for respiratory tract protection comprising a probiotic fermented liquid according to claim 1, wherein the preparation method of linalool oxide is as follows: placing linalool and formic acid in a water bath, controlling the reaction temperature in the water bath to be 60-65 ℃, then dropwise adding hydrogen peroxide water solution for 30min, controlling the temperature in the dropwise adding process to be 65-70 ℃, keeping the temperature and stirring after the dropwise adding, cooling, adding saturated sodium chloride water solution, extracting with benzene, combining extract liquid, washing the extract liquid with 10% sodium carbonate water solution and water to be neutral, drying with anhydrous sodium sulfate, removing benzene residues through evaporation after drying, then distilling under reduced pressure, and collecting fractions with the following boiling ranges of 70-74 ℃ (I) under 799.8 Pa; 75-79 ℃ (II); 80-84 ℃ (III); 85-102 ℃ (IV); and (3) taking fraction (II), namely linalool oxide prepared by the method.

5. A polysaccharide salt mist mixture for respiratory tract protection comprising a probiotic fermented liquid according to claim 3, characterized in that: preparation of esterification reagent: anhydrous pyridine was placed in a 500mL round bottom flask, cooled in an ice water bath, chlorosulfonic acid was slowly added under magnetic stirring, stirring was continued for 30min at room temperature, and after sealing, the flask was refrigerated in a refrigerator.

6. A polysaccharide salt mist mixture for respiratory tract protection comprising a probiotic fermented liquid according to any one of claims 1 to 5, characterized in that: the salt fog mixture is concentrated and soaked into a porous medium to form a chip which is placed in a mask for physiotherapy, or the chip is atomized for physiotherapy.

7. A chip for a mask, characterized by: a salt mist mixture of polysaccharides for respiratory tract protection comprising a probiotic fermented liquid according to any one of claims 1 to 6.

8. A physiotherapy mask comprising the chip of claim 7.