Composite probiotic composition for improving allergic rhinitis as well as preparation method and application of composite probiotic composition

Abstract

The invention provides a compound probiotic composition for improving allergic rhinitis as well as a preparation method and application thereof, and belongs to the technical field of biological preparations. Comprising a composite probiotic liquid, an auxiliary agent, a humectant and a thickening agent, wherein the composite probiotic liquid is composed of a composite bacterium fermentation liquid and probiotic substances, and the composite fermentation liquid is prepared by respectively fermenting saccharomyces boulardii, two bifidobacteria, lactobacillus reuteri, lactobacillus rhamnosus and streptococcus thermophilus. According to the compound probiotic composition, growth of probiotics is promoted through different probiotics, flora balance is adjusted and immunity is improved through mutual cooperation of the different probiotics, rhinitis symptoms are improved, meanwhile, components such as centipeda minima are added to serve as auxiliaries, and the effect of improving rhinitis symptoms is achieved. And the probiotics and the compound probiotics can achieve a synergistic effect to jointly improve the symptoms of the allergic rhinitis.

Description

Technical Field

[0001] The present invention relates to the technical field of biological preparations, and in particular to a composite probiotic composition for improving allergic rhinitis, and a preparation method and application thereof. Background Art

[0002] Allergic rhinitis is usually caused by the body's overreaction to certain substances, such as dust mites, pollen, bacteria and mold. It is an allergic reaction of antigens and antibodies in the nasal cavity. The main symptoms are nasal congestion, runny nose and sneezing, which triggers non-infectious inflammation of the nasal mucosa. It is recurrent and difficult to cure. At the same time, long-term inflammatory reactions can cause swelling and congestion of nasal tissues, affecting the face and the quality of life of patients.

[0003] In the prior art, the method of alleviating allergic rhinitis is usually oral antibiotics (such as cetirizine and budesonide), but long-term use is not good for human health and is prone to drug resistance. Although the use of probiotics can improve or prevent allergic rhinitis, the effect is not ideal, and the treatment effect is only about 70%, and the improvement effect on patients with severe rhinitis is not obvious. Therefore, it is hoped to provide a technical solution that can effectively improve the symptoms of patients with allergic rhinitis. Summary of the invention

[0004] The purpose of the present invention is to provide a product which can effectively improve the symptoms of patients with allergic rhinitis.

[0005] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0006] The present invention provides a composite probiotic composition for improving allergic rhinitis, comprising a composite probiotic liquid and an adjuvant;

[0007] The composite probiotic bacterial liquid is composed of composite bacterial fermentation liquid and probiotic substances, and the composite fermentation liquid is fermented by Saccharomyces boulardii, Bifidobacterium bifidum, Lactobacillus reuteri, Lactobacillus rhamnosus and Streptococcus thermophilus respectively.

[0008] Preferably, the ratio of the composite probiotic solution to the auxiliary agent is: 100 mL: 100-120 mL.

[0009] Preferably, the prebiotics include isomaltulose, red date juice, tangerine juice and honeysuckle water extract.

[0010] Preferably, the composite bacterial fermentation liquid is composed of Saccharomyces boulardii liquid, Bifidobacterium bifidum liquid, Lactobacillus reuteri liquid, Lactobacillus rhamnosus liquid and Streptococcus thermophilus liquid in an effective live bacterial count ratio of 2-4:2-4:1-3:1:2-4;

[0011] The effective viable count of the Saccharomyces boulardii liquid is: (1-2)×107 cfu / mL;

[0012] The effective viable count of the two bifidobacterium bacterial liquids is: (1-2)×10 7 cfu / mL;

[0013] The effective viable count of the Lactobacillus reuteri liquid is: (1-2)×10 7 cfu / mL;

[0014] The effective viable count of the Lactobacillus rhamnosus bacterial solution is: (2-3)×10 7 cfu / mL;

[0015] The effective viable count of the thermophilic streptococcus liquid is: (2-3)×10 7 cfu / mL.

[0016] The present invention provides a method for preparing the composite probiotic composition, comprising the following steps:

[0017] (1) preparing a Saccharomyces boulardii bacterial solution, a Bifidobacterium bisporus bacterial solution, a Lactobacillus reuteri bacterial solution, a Lactobacillus rhamnosus bacterial solution and a Streptococcus thermophilus bacterial solution respectively and mixing the mixture to obtain a composite bacterial fermentation liquid;

[0018] (2) adding 1-5 g of isomaltulose, 10-15 mL of red date juice, 10-15 mL of tangerine juice and 8-12 g of honeysuckle water extract to every 100 mL of the composite bacteria fermentation liquid, mixing and shaking at 30-35° C. for 2-3 h to obtain a composite probiotic bacterial liquid;

[0019] (3) Adding an auxiliary agent to the composite probiotic liquid and stirring to obtain the composite probiotic composition.

[0020] The present invention also provides the use of the composite probiotic composition in preparing a product for improving or preventing allergic rhinitis.

[0021] Preferably, the dosage form of the product includes gel, ointment, and liquid.

[0022] Preferably, when the dosage form of the product is a gel, the auxiliary agent includes an alcohol extract of ophiopogon japonicus, a decoction of dried ginger, lemon essential oil, peppermint essential oil, passion fruit essential oil and a dispersant; the preparation method is: 8 to 12 mL of an alcohol extract of ophiopogon japonicus, 18 to 22 mL of a decoction of dried ginger, 8 to 12 mL of lemon essential oil, 8 to 12 mL of peppermint essential oil, 4 to 8 mL of passion fruit essential oil and 0.1 to 0.5 g of a dispersant polysorbate are added to every 100 mL of 0.6 to 1.0% (w / v) sodium chloride solution.

[0023] Preferably, when the dosage form of the product is a gel, the composite probiotic composition further comprises a moisturizer and a thickener, the moisturizer comprises chitosan and low-molecular hyaluronic acid; the molecular weight of the low-molecular hyaluronic acid is 100 to 500 KDa;

[0024] The thickeners include xanthan gum, gelatin and sodium alginate.

[0025] Preferably, when the dosage form of the product is a liquid, the auxiliary agents include water decoction of goose not eating grass, water decoction of dried ginger, lemon juice, kiwi juice and mint water decoction, and the volume ratio is 1-3:1-3:1-2:1-2:1.

[0026] The composite probiotic composition provided by the present invention is composed of a composite probiotic liquid and an auxiliary agent. In the composite probiotic liquid, Saccharomyces boulardii, Bifidobacterium bifidum, Lactobacillus reuteri, Lactobacillus rhamnosus and Streptococcus thermophilus are rationally compounded to make different floras have a synergistic effect. Adding ingredients such as isomaltulose, red date juice, tangerine juice and honeysuckle water extract has an anti-inflammatory effect, and can promote the growth of probiotics at the same time, thereby ensuring the probiotic content in the product. Goosegrass and dried ginger have a therapeutic effect on allergic rhinitis. In addition, essential oil components such as lemon essential oil and passion fruit essential oil are used in the gel, which have an anti-inflammatory effect, can synergize with composite probiotics, help regulate the balance of nasal flora, improve nasal immunity, and jointly improve allergic rhinitis. After oral administration, the liquid composite probiotic composition can assist in regulating the balance of body flora. After taking it together with ingredients rich in vitamin C such as lemon juice, it can effectively relieve allergic symptoms while improving human immunity and preventing allergies. It can be seen from the experiment that the present invention can significantly improve the symptoms of allergic rhinitis and significantly reduce the IgE antibody level of the allergic rhinitis mouse model, and can effectively alleviate the symptoms of allergic rhinitis. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 The changes in IgE antibody levels in the allergic rhinitis mouse model in Experimental Example 1. DETAILED DESCRIPTION

[0028] In the present invention, Saccharomyces boulardii was purchased from Wuhan Lanabai Pharmaceutical Chemical Co., Ltd. and cultured to an effective viable count of 2×10 7 cfu / mL;

[0029] Two bifidobacteria were purchased from Hubei Mingtuo Biotechnology Co., Ltd. and cultured to 2×10 7 cfu / mL;

[0030] Lactobacillus reuteri was purchased from Hubei Mingtuo Biotechnology Co., Ltd. and cultured to 1×10 7 cfu / mL;

[0031] Lactobacillus rhamnosus was purchased from Hubei Mingtuo Biotechnology Co., Ltd. and cultured to 1×10 7 cfu / mL;

[0032] Streptococcus thermophilus was purchased from Hubei Mingtuo Biotechnology Co., Ltd. and cultured to 2×10 7 cfu / mL;

[0033] LB liquid medium was used as the bacterial culture medium.

[0034] Honeysuckle water extract was purchased from Xi'an Zelin Biotechnology Co., Ltd.

[0035] The technical solutions provided by the present invention are described in detail below in conjunction with the embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0036] Example 1

[0037] A composite probiotic composition for improving allergic rhinitis:

[0038] (1) mixing a Saccharomyces boulardii culture solution, a Bifidobacterium bisporus culture solution, a Lactobacillus reuteri culture solution, a Lactobacillus rhamnosus culture solution and a Streptococcus thermophilus culture solution at an effective viable cell count ratio of 2:4:1:1:4 to obtain a composite bacterial fermentation solution;

[0039] (2) Then, isomaltulose, red date juice, tangerine juice and honeysuckle water extract were added to the composite bacteria fermentation liquid in a ratio of 100 mL: 2 g: 10 mL: 15 mL: 10 g, and the mixture was mixed and shaken at 35° C. for 2 h to obtain a composite probiotic bacterial liquid;

[0040] (3) preparing an auxiliary agent, adding 10 mL of ethanol extract of ophiopogon japonicus, 20 mL of dried ginger decoction, 10 mL of lemon essential oil, 10 mL of peppermint essential oil, 5 mL of passion fruit essential oil and 0.3 g of polysorbate to every 100 mL of 0.9% (w / v) sodium chloride solution to obtain an auxiliary agent;

[0041] 100 mL of auxiliary agent was added to 100 mL of composite probiotic bacterial liquid, and the mixture was stirred. Then, 5 g of chitosan, 3 g of low molecular weight hyaluronic acid (200 KDa), 1 g of xanthan gum, 1 g of gelatin, and 2 g of sodium alginate were added in sequence while stirring to obtain a gel-like composite probiotic composition;

[0042] Among them, the preparation method of the alcohol extract of goose grass is as follows: crush the goose grass into 80 meshes, add 60% (v / v) ethanol, the mass volume ratio is 1:10, extract at 50°C for 2h, filter twice, combine the filtrate to obtain the alcohol extract of goose grass; the preparation method of the dried ginger decoction is as follows: take dried ginger, add water 3 times the volume of dried ginger, boil for 2h, filter the filtrate, and cool to obtain the dried ginger decoction.

[0043] Example 2

[0044] A composite probiotic composition for improving allergic rhinitis:

[0045] (1) mixing a Saccharomyces boulardii culture, a Bifidobacterium bisporus culture, a Lactobacillus reuteri culture, a Lactobacillus rhamnosus culture and a Streptococcus thermophilus culture at an effective viable cell count ratio of 4:2:3:1:2 to obtain a composite bacterial fermentation liquid;

[0046] (2) Then, isomaltulose, red date juice, tangerine juice and honeysuckle water extract were added to the composite bacteria fermentation liquid in a ratio of 100 mL: 1 g: 15 mL: 10 mL: 8 g, and the mixture was mixed and shaken at 30° C. for 3 h to obtain a composite probiotic bacterial liquid;

[0047] (3) preparing an auxiliary agent, adding 12 mL of ethanol extract of ophiopogon japonicus, 18 mL of dried ginger decoction, 8 mL of lemon essential oil, 12 mL of peppermint essential oil, 4 mL of passion fruit essential oil and 0.1 g of dispersant polysorbate to every 100 mL of 0.9% (w / v) sodium chloride solution to obtain an auxiliary agent;

[0048] 110 mL of the above-mentioned auxiliary agent was added to 100 mL of the composite probiotic solution, and the mixture was stirred. Then, 4 g of chitosan, 4 g of low molecular weight hyaluronic acid, 2 g of xanthan gum, 1 g of gelatin, and 2 g of sodium alginate were added in sequence while stirring to obtain a gel-like composite probiotic composition.

[0049] Example 3

[0050] A composite probiotic composition for improving allergic rhinitis:

[0051] (1) mixing a Saccharomyces boulardii culture solution, a Bifidobacterium bisporus culture solution, a Lactobacillus reuteri culture solution, a Lactobacillus rhamnosus culture solution and a Streptococcus thermophilus culture solution at an effective viable cell count ratio of 4:2:2:1:2 to obtain a composite bacterial fermentation solution;

[0052] (2) Then, isomaltulose, red date juice, tangerine juice and honeysuckle water extract were added to the composite bacteria fermentation liquid in a ratio of 100 mL: 3 g: 14 mL: 13 mL: 8 g, and the mixture was mixed and shaken at 35° C. for 2.5 h to obtain a composite probiotic bacterial liquid;

[0053] (3) preparing an auxiliary agent, adding 10 mL of ethanol extract of ophiopogon japonicus, 22 mL of dried ginger decoction, 12 mL of lemon essential oil, 8 mL of peppermint essential oil, 8 mL of passion fruit essential oil and 0.3 g of dispersant polysorbate to every 100 mL of 0.6% (w / v) sodium chloride solution to obtain an auxiliary agent;

[0054] 120 mL of the above-mentioned auxiliary agent was added to 100 mL of the composite probiotic solution, and the mixture was stirred. Then, 5 g of chitosan, 3 g of low molecular weight hyaluronic acid, 1 g of xanthan gum, 1 g of gelatin, and 1 g of sodium alginate were added in sequence while stirring to obtain a gel-like composite probiotic composition.

[0055] Example 4

[0056] Different from Example 1, the method for preparing the auxiliary agent in step (3) is: mixing the decoction of goose grass, the decoction of dried ginger, lemon juice, kiwi juice and mint decoction in a volume ratio of 1:1:1:1:1. Then, 100 mL of the above auxiliary agent is added to every 100 mL of the composite probiotic bacterial liquid to obtain a liquid composite probiotic composition.

[0057] The preparation method of the mint decoction is as follows: take mint, add water twice the volume of the mint, boil for 2 hours, filter the filtrate, and cool to obtain the mint decoction.

[0058] Comparative Example 1

[0059] The difference from Example 1 is that in step (1), the composite bacterial fermentation broth is prepared by mixing Saccharomyces boulardii, Bifidobacterium bisporus and Lactobacillus reuteri in a ratio of 4:4:4 in terms of effective viable count.

[0060] Comparative Example 2

[0061] The difference from Example 1 is that in step (2), oligofructose, inulin and xylan are added to the composite bacteria fermentation broth in a ratio of 100 mL: 5 g: 3 g: 2 g.

[0062] Comparative Example 3

[0063] The difference from Example 1 is that the auxiliary agent in step (3) is 10 mL of ethanol extract of Herba Anemarrhenae, 20 mL of dried ginger decoction, 10 mL of tea tree oil, 10 mL of lavender oil, 5 mL of basil oil and 0.3 g of polysorbate added to every 100 mL of 0.9% (w / v) sodium chloride solution.

[0064] Comparative Example 4

[0065] Different from Example 4, the composite bacteria fermentation liquid is prepared by mixing Saccharomyces boulardii liquid, Bifidobacterium bisporus liquid and Lactobacillus reuteri liquid in a ratio of 4:4:4 in terms of effective live bacteria count.

[0066] Test Example 1

[0067] Experimental animals and groups:

[0068] 80 BALB / c mice aged 6 to 8 weeks were selected, half male and half female, and grouped as shown in Table 1:

[0069] Table 1 Grouping of Test Example 1

[0070] Grouping deal with quantity Blank control group Normal mice, without any treatment 10 Model control group Apply 5g of medical gel to the nose of sensitized mice 10 Example 1 Group 5 g of the composition prepared in Example 1 was applied to the nose of the sensitized mice. 10 Example 2 Group 5 g of the composition prepared in Example 2 was applied to the nose of the sensitized mice. 10 Example 3 Group 5 g of the composition prepared in Example 3 was applied to the nose of the sensitized mice. 10 Comparative Example 1 5 g of the composition prepared in Comparative Example 1 was applied to the nose of the sensitized mice. 10 Comparative Example 2 5 g of the composition prepared in Comparative Example 2 was applied to the nose of the sensitized mice. 10 Comparative Example 3 5 g of the composition prepared in Comparative Example 3 was applied to the nose of the sensitized mice. 10

[0071] Establishment of the allergic rhinitis mouse (sensitized mouse) model: Prepare an OVA suspension containing 25 μg of ovalbumin (OVA), 1 mg of aluminum hydroxide gel, and 0.5 ml of saline. Perform basic sensitization on the mice on the 1st, 5th, 14th, and 21st days, and inject 0.5 mL of OVA suspension intraperitoneally; after basic sensitization, give the mice bilateral nasal instillations of 500 μg OVA plus 20 μl of saline starting on the 28th day, and perform local stimulation for 10 consecutive days. A score greater than 6 points indicates successful modeling.

[0072] After the last intranasal allergen injection, the experimental mice were divided into groups for treatment. The composition of each group was applied to the bilateral nasal walls of the mice, 5 g per nasal cavity, once a day for 10 days.

[0073] 1. Symptoms of rhinitis

[0074] The number of nasal itching, sneezing and runny nose of the experimental mice were counted before administration and within 20 minutes after administration on the 1st, 5th and 10th days. The scoring criteria are shown in Table 2, and the test results are shown in Table 3.

[0075] Table 2 Scoring criteria

[0076] score 0 points 1 point 2 points 3 points Nasal itching frequency Scratching nose 0 times Scratching nose ≤ 2 times Scratching nose ≤ 5 times Scratching nose > 5 times Number of sneezes 0-1 times 2 to 4 times 5-10 times >10 times Runny nose none Flow to the anterior nostril Beyond the anterior nares Tears streaming down one's face

[0077] Table 3 Test Example 1 Results

[0078] Grouping Before administration Day 1 of medication Day 5 of medication Day 10 of medication Blank control 0 0 0 0 Model control group 8 8 8 8 Example 1 Group 8 6 4 2 Example 2 Group 8 7 3 2 Example 3 Group 8 6 3 2 Comparative Example 1 8 7 5 4 Comparative Example 2 8 7 5 3 Comparative Example 3 8 7 5 5

[0079] 2. Changes in serum IgE antibody levels

[0080] Within 3 hours after each administration, peripheral blood of mice was collected and placed in a high-temperature sterilized centrifuge tube. Serum was obtained by centrifugation at 3000 r / min at 4°C for 15 min. The serum IgE levels of mice at 3, 7, and 10 days after administration were detected using an ELISA kit (CUSABIO). The test results were as follows: Figure 1 shown.

[0081] From Table 3 and Figure 1 It can be seen that compared with the model control group, the compositions given in Examples 1 to 3 and Comparative Examples 1 to 3 can improve the symptoms of allergic rhinitis and reduce the IgE antibody level of the allergic rhinitis mouse model, among which the improvement effect of the composite probiotic composition provided by the present invention is the most obvious.

[0082] Test Example 2

[0083] The liquid composite probiotic composition prepared in Example 4 and Comparative Example 4 was subjected to a mouse test. The groups were as follows:

[0084] Table 4 Grouping of Test Example 2

[0085]

[0086]

[0087] The model was established in the same way as in Experiment 1. Each group of mice was gavage-fed 1 hour before each local challenge. The changes in the rhinitis symptoms of mice and the levels of IgE antibodies in the serum of mice within 20 minutes after the last challenge were statistically analyzed. The results are shown in Table 5:

[0088] Table 5 Test Example 2 Results

[0089] Blank control group Model control group Example 4 Group Comparative Example 4 Rating results 0 8 4 6 IgE antibody levels 18.6±3.12 66.24±2.34 32.45±2.64 46.75±3.19

[0090] As shown in Table 5, compared with the model control group and the comparative example 4 group, the liquid composite probiotic composition provided by the present invention can effectively relieve the symptoms of allergic rhinitis after gavage, and reduce the IgE antibody level of the allergic rhinitis mouse model. In addition to the reasonable compounding of different probiotics, the liquid composite probiotic composition provided by the present invention also adds vitamin-rich substances such as lemon juice and kiwi juice, which can not only effectively relieve allergic symptoms, but also adjust the taste.

[0091] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A composite probiotic composition for improving allergic rhinitis, characterized in that: Including compound probiotic liquid and auxiliary agents; The composite probiotic bacterial liquid is composed of composite bacterial fermentation liquid and probiotic substances, and the composite fermentation liquid is fermented by Saccharomyces boulardii, Bifidobacterium bifidum, Lactobacillus reuteri, Lactobacillus rhamnosus and Streptococcus thermophilus respectively.

2. The composite probiotic composition according to claim 1, characterized in that: The ratio of the composite probiotic liquid to the auxiliary agent is: 100mL: 100-120mL.

3. The composite probiotic composition according to claim 2, characterized in that: The beneficial biomass comprises isomaltulose, red date juice, tangerine juice and honeysuckle water extract.

4. The composite probiotic composition according to claim 3, characterized in that: The composite bacterial fermentation liquid is composed of a Saccharomyces boulardii liquid, a Bifidobacterium bisporus liquid, a Lactobacillus reuteri liquid, a Lactobacillus rhamnosus liquid and a Streptococcus thermophilus liquid in an effective live bacterial count ratio of 2-4:2-4:1-3:1:2-4; The effective viable count of the Saccharomyces boulardii liquid is: (1-2)×10 7 cfu / mL; The effective viable count of the two bifidobacterium bacterial liquids is: (1-2)×10 7 cfu / mL; The effective viable count of the Lactobacillus reuteri liquid is: (1-2)×10 7 cfu / mL; The effective viable count of the Lactobacillus rhamnosus bacterial solution is: (2-3)×10 7 cfu / mL; The effective viable count of the thermophilic streptococcus liquid is: (2-3)×10 7 cfu / mL.

5. The method for preparing the composite probiotic composition according to any one of claims 1 to 4, characterized in that: The following steps are involved: (1) preparing a Saccharomyces boulardii bacterial solution, a Bifidobacterium bisporus bacterial solution, a Lactobacillus reuteri bacterial solution, a Lactobacillus rhamnosus bacterial solution and a Streptococcus thermophilus bacterial solution respectively and mixing the mixture to obtain a composite bacterial fermentation liquid; (2) adding 1-5 g of isomaltulose, 10-15 mL of red date juice, 10-15 mL of tangerine juice and 8-12 g of honeysuckle water extract to every 100 mL of the composite bacteria fermentation liquid, mixing and shaking at 30-35° C. for 2-3 h to obtain a composite probiotic bacterial liquid; (3) Adding an auxiliary agent to the composite probiotic liquid and stirring to obtain the composite probiotic composition.

6. Use of the composite probiotic composition according to any one of claims 1 to 4 or the composite probiotic composition prepared by the preparation method according to claim 5 in preparing a product for improving or preventing allergic rhinitis.

7. The use according to claim 6, characterized in that The dosage forms of the product include gel, ointment and liquid.

8. The use according to claim 7, characterized in that When the dosage form of the product is a gel, the auxiliary agent includes an alcohol extract of ophiopogon japonicus, a decoction of dried ginger, lemon essential oil, mint essential oil, passion fruit essential oil and a dispersant; the preparation method is: 8 to 12 mL of an alcohol extract of ophiopogon japonicus, 18 to 22 mL of a decoction of dried ginger, 8 to 12 mL of lemon essential oil, 8 to 12 mL of mint essential oil, 4 to 8 mL of passion fruit essential oil and 0.1 to 0.5 g of a dispersant polysorbate are added to every 100 mL of 0.6 to 1.0% (w / v) sodium chloride solution.

9. The use according to claim 8, characterized in that When the dosage form of the product is a gel, the composite probiotic composition further comprises a moisturizer and a thickener, wherein the moisturizer comprises chitosan and low-molecular hyaluronic acid; the molecular weight of the low-molecular hyaluronic acid is 100 to 500 KDa; The thickeners include xanthan gum, gelatin and sodium alginate.

10. The use according to claim 7, characterized in that: When the dosage form of the product is a liquid, the auxiliary agents include water decoction of goose not eating grass, water decoction of dried ginger, lemon juice, kiwi juice and mint water decoction, and the volume ratio is 1-3:1-3:1-2:1-2:1.

Images